Saccharopolyspora Rectivirgula Research Articles

Elucidating core microbiota in yellow wine (Huangjiu) through flavor-oriented synthesis and construction of microbial communities

Huangjiu, a traditional Chinese alcoholic beverage with a history spanning thousands of years, holds significant cultural and economic value in China. Despite its importance, the complexity of Huangjiu fermentation and the intricate interactions within its microbial community remain underexplored. This study addresses this gap by identifying the core volatile organic compounds (VOCs) and key microorganisms that define the flavor profile of Huangjiu. We employed HS-SPME-GC–MS along with aroma reconstitution and omission experiments to identify core VOCs, including Isobutanol, Isoamyl alcohol, β-Phenylethanol, and others. Metagenomic sequencing combined with QPCR was used to analyze microbial communities, revealing the temporal and spatial dynamics during fermentation. A synthetic microbial community model was constructed using the core microbes identified: Saccharomyces cerevisiae, Lactobacillus brevis, Saccharopolyspora rectivirgula, Bacillus subtilis, Leuconostoc citreum, Lactobacillus plantarum, Lactobacillus curvatus, Lactobacillus casei, and Aspergillus oryzae. This model successfully replicated Huangjiu’s core VOCs and sensory characteristics, increased alcohol content, and reduced acidity. Our study contributes valuable insights into the microbial influences on Huangjiu quality, paving the way for its enhanced production and providing a foundation for future research in fermented beverages.

Relationship between dynamic changes of microorganisms in Qupi and the quality formation of Fengxiangxing Huairang Daqu.

Fengxiangxing Huairang Daqu (FHD) is one of the major types of Daqu in China. However, the relationship between the microbial community structure at different stages, the changes in the sensory characteristics, fermentation characteristics, volatiles, the most critical process point, and the quality formation of FHD is not clear. Based on microscopic characterization, PacBio SMRT sequencing, and HS-SPME-GC-MS volatile metabolite analysis revealed the relationship between FHD quality formation and the dynamics of Qupi. The results showed that the 12th day of the culture was the most critical process point, highlighting the most significant differences in microbial community structure, sensory characteristics, fermentation characteristics, and flavor substances. Bacillus licheniformis (43.25%), Saccharopolyspora rectivirgula (35.05%), Thermoascus aurantiacus (76.51%), Aspergillus amstelodami (10.81%), and Saccharomycopsis fibuligera (8.88%) were the dominant species in FHD. S. fibuligera, A. amstelodami, and T. aurantiacus were associated with the snow-white color of the FHD epidermis, the yellow color of the interior, and the gray-white color, respectively. The abundance of T. aurantiacus, A. amstelodami, B. licheniformis, and S. rectivirgula was positively associated with the esterifying power and liquefying power of FHD. The abundance of T. aurantiacus and A. amstelodami was positively correlated with the saccharifying power of FHD. The abundance of S. fibuligera was positively related to the fermenting power of FHD. A total of 248 volatiles were detected in Qupi, mainly including alcohols, esters, aldehydes, and ketones. Of them, eleven volatiles had a significant effect on the flavor of Qupi, such as 1-butanol-3-methyl-, hydrazinecarboxamide, ethanol, phenylethyl alcohol, ethyl acetate, 2-octanone, 1-octen-3-ol, formic acid-hexyl ester, (E)-2-octen-1-ol, ethyl hexanoate, and 2(3H)-furanone-dihydro-5-pentyl-. The abundance of B. licheniformis, S. rectivirgula, T. aurantiacus, and S. fibuligera was positively correlated with the alcohols, aromatic compounds, and phenols in FHD. The abundance of S. fibuligera was positively correlated with the acids, esters, and hydrocarbons in FHD. These results indicate important theoretical basis and technical support for controllable adjustment of FHD microbial community structure, stable control of FHD quality, and precise, effective, and large-scale guidance of FHD production.

Environmental factors and particle size shape the community structure of airborne total and pathogenic bacteria in a university campus.

Given the dense population on university campuses, indoor and outdoor airborne bacterial contamination may lead to the rapid spread of diseases in a university environment. However, there are few studies of the characteristics of airborne and pathogenic bacterial communities in different sites on a university campus. In this study, we collected particulate matter samples from indoor and outdoor locations at a university in Bengbu City, Anhui Province, China, and analyzed the community characteristics of airborne and pathogenic bacteria using a high-throughput sequencing technique. The results showed that the composition of the dominant airborne and pathogenic bacterial communities was consistent among sites at the phylum and genus levels, with differences in their relative abundance. There were significant differences in the structure of the airborne and pathogenic bacterial communities between indoor and outdoor sites (p < 0.05). An analysis of similarities (ANOSIM) indicated that the structure of airborne bacterial communities in indoor sites was influenced by the room occupancy rate, ventilation conditions, and the extent of indoor furnishing (p < 0.05), while the structure of pathogenic bacterial communities was influenced by the number of individuals and spatial dimensions (p < 0.05). The impact of particle size on the structure of airborne and pathogenic bacterial communities was relatively minor. A total of 194 suspected pathogenic bacterial species were identified, accounting for 0.0001-1.3923% of the total airborne bacteria, all of which were conditional pathogens. Among them, Saccharopolyspora rectivirgula, Acinetobacter johnsonii, and Moraxella osloensis exhibited relatively high relative abundance, accounting for 24.40, 16.22, and 8.66% of the total pathogenic bacteria, respectively. Moreover, 18 emerging or re-emerging pathogenic bacterial species with significant implications for human health were identified, although their relative abundance was relatively low (0.5098%). The relative abundance of pathogenic bacteria in indoor environments was significantly higher than outdoors, with the laboratory and dormitory having the highest levels. The findings of this study provide valuable guidance for the prevention and control of airborne bacterial contamination and the associated health risks in both a campus environment and other public spaces with high occupancy rates.

MMP14high macrophages orchestrate progressive pulmonary fibrosis in SR-Ag-induced hypersensitivity pneumonitis

Fibrotic hypersensitivity pneumonitis (FHP) is a fatal interstitial pulmonary disease with limited treatment options. Lung macrophages are a heterogeneous cell population that exhibit distinct subsets with divergent functions, playing pivotal roles in the progression of pulmonary fibrosis. However, the specific macrophage subpopulations and underlying mechanisms involved in the disease remain largely unexplored. In this study, a decision tree model showed that matrix metalloproteinase-14 (MMP14) had higher scores for important features in the up-regulated genes in macrophages from mice exposed to the Saccharopolyspora rectivirgula antigen (SR-Ag). Using single-cell RNA sequencing (scRNA-seq) analysis of hypersensitivity pneumonitis (HP) mice profiles, we identified MMP14high macrophage subcluster with a predominant M2 phenotype that exhibited higher activity in promoting fibroblast-to myofibroblast transition (FMT). We demonstrated that suppressing toll-like receptor 2 (TLR2) and nuclear factor kappa-B (NF-κB) could attenuate MMP14 expression and exosome secretion in macrophages stimulation with SR-Ag. The exosomes derived from MMP14-overexpressing macrophages were found to be more effective in regulating the transition of fibroblasts through exosomal MMP14. Importantly, it was observed that the transfer of MMP14-overexpressing macrophages into mice promoted lung inflammation and fibrosis induced by SR-Ag. NSC-405020 binding to the hemopexin domain (PEX) of MMP-14 ameliorated lung inflammation and fibrosis induced by SR-Ag in mice. Thus, MMP14-overexpressing macrophages may be an important mechanism contributing to the exacerbation of allergic reactions. Our results indicated that MMP14 in macrophages has the potential to be a therapeutic target for HP.

Bioaerosols in Eastern Canadian Dairy Barns Using Tie- and Free-Stall Housing

Highlights Building characteristics (ventilation and animal density) seem to have an important effect on air quality. Concentration in the air seems to be influenced by amounts in the bedding for certain air quality indicators. Dust concentrations were below OSHA threshold, while some barns exceeded DECOS threshold for endotoxins. This study gives new data into the biological components in the air of dairy barns (bacteria, molds, endotoxins, etc.) Free-stall or tie-stall housing may not be linked with poorer air quality. Abstract. Alternative farming methods make it possible to satisfy public demands for animal welfare while preserving production efficiency, with free housing in dairy farms being an example. The increased movement of cows may have a negative impact on air quality and the presence of etiological agents, increasing the prevalence of lung disease in workers. Free-stall farms, however, are more spacious and modern than tie-stall farms. This study characterizes air quality in free-stall and tie-stall farms (dust, total bacteria, Penicillium/Aspergillus, archaea, and endotoxins). It also focuses on detecting airborne etiological agents and indicators of fecal contamination, as well as assessing the effect of environmental factors on air quality. Five farms of each type (free housing and tie housing) using straw bedding material and equipped with mechanical ventilation were visited. Sampling visits were conducted in winter with no activity (e.g., bedding spreading) in buildings. Dust was evaluated using the DustTrakTM DRX Aerosol Monitor, and bioaerosols were sampled in triplicates for 10 minutes using the SASS®3100 Dry Air Sampler. Finally, soiled bedding was collected throughout the barn. No type of housing seems to be linked with poorer air quality, but some air quality indicators stood out in some outliers. The most recently designed free-housing buildings and the greater air volume may have played a role in the absence of detected differences. Escherichia coli, Enterococcus spp., Clostridium perfringens, Aspergillus fumigatus, Staphylococcus aureus, Saccharopolyspora rectivirgula, Coxiella burnetii, and Klebsiella pneumoniae were detected in high concentrations in both types of buildings. Soiled bedding concentrations, ventilation rates, and animal density seemed to have a significance on air quality in dairy barns. Keywords: Air Quality, Bioaerosols, Housing, Occupational exposure.

Revealing the developmental characterization of rumen microbiome and its host in newly received cattle during receiving period contributes to formulating precise nutritional strategies

BackgroundMinimizing mortality losses due to multiple stress and obtaining maximum performance are the production goals for newly received cattle. In recent years, vaccination and metaphylaxis treatment significantly decreased the mortality rate of newly received cattle, while the growth block induced by treatment is still obvious. Assessment of blood metabolites and behavior monitoring offer potential for early identification of morbid animals. Moreover, the ruminal microorganisms’ homeostasis is a guarantee of beef steers’ growth and health. The most critical period for newly received cattle is the first-month post-transport. Therefore, analyzing rumen metagenomics, rumen metabolomics, host metabolomics, and their interaction during receiving period (1 day before transport and at days 1/4, 16, and 30 after transport) is key to revealing the mechanism of growth retardation, and then to formulating management and nutritional practices for newly received cattle.ResultsThe levels of serum hormones (COR and ACTH), and pro-inflammatory factors (IL-1β, TNF-α, and IL-6) were highest at day 16, and lowest at day 30 after arrival. Meanwhile, the antioxidant capacity (SOD, GSH-Px, and T-AOC) was significantly decreased at day 16 and increased at day 30 after arrival. Metagenomics analysis revealed that rumen microbes, bacteria, archaea, and eukaryota had different trends among the four different time points. At day 16 post-transport, cattle had a higher abundance of ruminal bacteria and archaea than those before transport, but the eukaryote abundance was highest at day 30 post-transport. Before transport, most bacteria were mainly involved in polysaccharides digestion. At day 4 post-transport, the most significantly enriched KEGG pathways were nucleotide metabolism (pyrimidine metabolism and purine metabolism). At day 16 post-transport, the energy metabolism (glycolysis/gluconeogenesis, pyruvate metabolism) and ruminal contents of MCP and VFAs were significantly increased, but at the same time, energy loss induced by methane yields (Methanobrevibacter) together with pathogenic bacteria (Saccharopolyspora rectivirgula) were also significantly increased. At this time, the most upregulated ruminal L-ornithine produces more catabolite polyamines, which cause oxidative stress to rumen microbes and their host; the most downregulated ruminal 2',3'-cAMP provided favorable growth conditions for pathogenic bacteria, and the downregulated ruminal vitamin B6 metabolism and serum PC/LysoPC disrupt immune function and inflammation reaction. At day 30 post-transport, the ruminal L-ornithine and its catabolites (mainly spermidine and 1,3-propanediamine) were decreased, and the serum PC/LysoPC and 2',3'-cNMPs pools were increased. This is also consistent with the changes in redox, inflammation, and immune status of the host.ConclusionsThis study provides new ideas for regulating the health and performance of newly received cattle during the receiving period. The key point is to manage the newly received cattle about day 16 post-transport, specifically to inhibit the production of methane and polyamines, and the reproduction of harmful bacteria in the rumen, therefore improving the immunity and performance of newly received cattle.6n2QR72-A8x7PGstPh77e2Video

Integrated meta-omics approaches reveal Saccharopolyspora as the core functional genus in huangjiu fermentations

Identification of the core functional microorganisms in food fermentations is necessary to understand the ecological and functional processes for making those foods. Wheat qu, which provides liquefaction and saccharifying power, and affects the flavor quality, is a key ingredient in ancient alcoholic huangjiu fermentation, while core microbiota of them still remains indistinct. In this study, metagenomics, metabolomics, microbial isolation and co-fermentation were used to investigate huangjiu. Although Aspergillus is usually regarded as core microorganism in wheat qu to initiate huangjiu fermentations, our metagenomic analysis showed that bacteria Saccharopolyspora are predominant in wheat qu and responsible for breakdown of starch and cellulose. Metabolic network and correlation analysis showed that Saccharopolyspora rectivirgula, Saccharopolyspora erythraea, and Saccharopolyspora hirsuta made the greatest contributions to huangjiu’s metabolites, consisting of alcohols (phenylethanol, isoamylol and isobutanol), esters, amino acids (Pro, Arg, Glu and Ala) and organic acids (lactate, tartrate, acetate and citrate). S. hirsuta J2 isolated from wheat qu had the highest amylase, glucoamylase and protease activities. Co-fermentations of S. hirsuta J2 with S. cerevisiae HJ resulted in a higher fermentation rate and alcohol content, and huangjiu flavors were more similar to that of traditional huangjiu compared to co-fermentations of Aspergillus or Lactiplantibacillus with S. cerevisiae HJ. Genome of S. hirsuta J2 contained genes encoding biogenic amine degradation enzymes. By S. hirsuta J2 inoculation, biogenic amine content was reduced by 45%, 43% and 62% in huangjiu, sausage and soy sauce, respectively. These findings show the utility of Saccharopolyspora as a key functional organism in fermented food products.

Muc5b plays a role in the development of inflammation and fibrosis in hypersensitivity pneumonitis induced by Saccharopolyspora rectivirgula.

Previously we have shown that a gain-of-function MUC5B promoter variant (rs35705950) is the strongest risk factor for the development of idiopathic pulmonary fibrosis. We have also found that Muc5b overexpression reduces mucociliary clearance in mice, potentially leading to recurrent injury to the bronchoalveolar epithelia. Hypersensitivity pneumonitis (HP) is induced by inhalation of numerous causative antigens that may be affected by mucociliary clearance. We conducted this study to determine the role of Muc5b in a mouse model of HP induced by Saccharopolyspora rectivirgula (SR) antigen. We used Muc5b-deficient and wild-type (WT) mice to determine whether Muc5b plays a role in inflammation and fibrosis at 3 and 6 wk in an SR model of HP. We measured cell concentrations and MUC5B expression in whole lung lavage (WLL) and quantified fibrosis using hydroxyproline assay and second harmonic generation. Muc5b expression in WLL fluid was significantly increased in SR-exposed WT mice compared with saline controls. WT mice challenged with SR developed more inflammation and lung fibrosis at 6 wk compared with 3 wk postexposure. Moreover, we found that 6 wk following challenge with SR, Muc5b-deficient mice had less lung inflammation and less lung fibrosis than Muc5b WT mice. Furthermore, Muc5b-deficient mice had significantly lower concentrations of TGF-β1 in the WLL compared with Muc5b WT mice at 6 wk of exposure. Muc5b appears to play a role in fibrosis in the animal model of HP and this may have implications for HP in humans.

Improved IgG antibody diagnostics of hypersensitivity pneumonitis and pulmonary mycoses by means of newly evaluated serum antibody ranges and frequencies using IgG ImmunoCAP™

Summary Background The ranges of most human IgG antibodies against avian, microbial, and chemical antigens between their cut off values and their maximum values detected by IgG ImmunoCAP™ (Thermo Fisher Diagnostics, Freiburg, Germany) are not well known in pulmonary immune-mediated disorders. In addition, for many antigens it is not yet known how frequently their IgG antibodies appear in patients with these lung diseases. Therefore, we evaluated ranges and frequencies of these IgG antibodies. Methods The sera of 47,200 patients with suspected hypersensitivity pneumonitis (HP, extrinsic allergic alveolitis) or bronchopulmonary mycoses (mainly allergic bronchopulmonary aspergillosis [ABPA]), which were examined for 32 various IgG antibodies against birds, bacteria, molds, yeasts, and chemicals using the IgG ImmunoCAP™ assay, were evaluated retrospectively. Results We found a large spreading of the specific IgG ranges with maximum values from 26 mgA/l for Rhizopus nigricans up to 4640 mgA/l for pigeon. When the maximum values in the literature are also taken into account, the ranges of avian antibodies reach values up to 7280 mgA/l, the ranges of molds and yeasts up to 1707 mgA/l, of bacterial thermoactinomycetes up to 206 mgA/l, and of chemicals up to 139 mgA/l. The evaluated antibody ranges of the individual antigens can be used to decide whether a detected IgG antibody value is weakly, moderately or strongly positive. According to consistent evidence from numerous studies, a strongly positive antibody indicates HP or pulmonary mycosis more likely than a weakly positive antibody. It was found that the antigens of the highest maximum antibody levels—pigeon, budgerigar, parrot, Aspergillus fumigatus—are the causative agents of the most common immune-mediated lung diseases: bird breeder’s lung and pulmonary aspergillosis. Evaluation of the frequencies of eight major IgG antibodies of HP revealed the following rates: pigeon 28%, Aspergillus fumigatus 25%, budgerigar 23%, Penicillium chrysogenum 11%, Saccharopolyspora rectivirgula 7%, Acremonium kiliense 6%, Aureobasidium pullulans 5%, and Thermoactinomyces vulgaris 2%. Conclusion This study is the first to evaluate the ranges not only of avian and Aspergillus fumigatus antigens, as has been done up to now, but also of antibodies against 24 other environmental antigens. Quantification of IgG antibodies regarding their specific ranges can help to improve the serodiagnostics of immune-mediated lung diseases. In the lower ranges IgG antibodies are mainly physiological, while higher ranges correlate with the mentioned diseases as HP and ABPA/allergic bronchopulmonary mycoses (ABPM). The determined frequencies of the eight HP antibodies can be helpful in establishing HP screening tests.

Contribution of Protein Kinase D1 on Acute Pulmonary Inflammation and Hypersensitivity Pneumonitis Induced by Saccharopolyspora rectivirgula.

Protein kinase D1 (PKD1), a ubiquitously expressed serine/threonine kinase, regulates diverse cellular processes such as oxidative stress, gene expression, cell survival, vesicle trafficking, Ag receptor signaling, and pattern recognition receptor signaling. We found previously that exposure to hypersensitivity pneumonitis (HP) inciting Ag Saccharopolyspora rectivirgula leads to the activation of PKD1 in a MyD88-dependent manner in various types of murine cells in vitro and in the mouse lung in vivo. However, it is currently unknown whether PKD1 plays a role in the S. rectivirgula-induced HP. In this study, we investigated contributions of PKD1 on the S. rectivirgula-induced HP using conditional PKD1-insufficient mice. Compared to control PKD1-sufficient mice, PKD1-insufficient mice showed substantially suppressed activation of MAPKs and NF-κB, expression of cytokines and chemokines, and neutrophilic alveolitis after single intranasal exposure to S. rectivirgula The significantly reduced levels of alveolitis, MHC class II surface expression on neutrophils and macrophages, and IL-17A and CXCL9 expression in lung tissue were observed in the PKD1-insufficient mice repeatedly exposed to S. rectivirgula for 5 wk. PKD1-insuficient mice exposed to S. rectivirgula for 5 wk also showed reduced granuloma formation. Our results demonstrate that PKD1 plays an essential role in the initial proinflammatory responses and neutrophil influx in the lung after exposure to S. rectivirgula and substantially contribute to the development of HP caused by repeated exposure to S. rectivirgula Our findings suggest that PKD1 can be an attractive new molecular target for therapy of S. rectivirgula-induced HP.

ITK independent development of Th17 responses during hypersensitivity pneumonitis driven lung inflammation

T helper 17 (Th17) cells develop in response to T cell receptor signals (TCR) in the presence of specific environments, and produce the inflammatory cytokine IL17A. These cells have been implicated in a number of inflammatory diseases and represent a potential target for ameliorating such diseases. The kinase ITK, a critical regulator of TCR signals, has been shown to be required for the development of Th17 cells. However, we show here that lung inflammation induced by Saccharopolyspora rectivirgula (SR) induced Hypersensitivity pneumonitis (SR-HP) results in a neutrophil independent, and ITK independent Th17 responses, although ITK signals are required for γδ T cell production of IL17A. Transcriptomic analysis of resultant ITK independent Th17 cells suggest that the SR-HP-induced extrinsic inflammatory signals may override intrinsic T cell signals downstream of ITK to rescue Th17 responses in the absence of ITK. These findings suggest that the ability to pharmaceutically target ITK to suppress Th17 responses may be dependent on the type of inflammation.

Metagenomics-based insights into the microbial community profiling and flavor development potentiality of baijiu Daqu and huangjiu wheat Qu

Daqu and wheat Qu are saccharification and fermenting agents in Chinese huangjiu and baijiu production. This study aimed to investigate the difference between Daqu and wheat Qu in physicochemical indices, microbial communities, functional genes, and the metabolic network of key microbes responsible for flavor synthesis by whole-metagenome sequencing and metabolite analysis. Herein, physicochemical indices indicated that compared with wheat Qu, Daqu exhibited higher protease and cellulase activity and acidity, and lower glucoamylase and amylase enzyme activity. Metagenomic sequencing reveals that although Daqu and wheat Qu community composition have significant differences at species level, they have similar functional genes. Daqu were enriched in Pediococcus pentosaceus, Weissella paramesenteroides, Rasamsonia emersonii and Byssochlamys spectabilis (22.48% of the total abundance), while wheat Qu harbored greater abundances of Saccharopolyspora (54.78%, Saccharopolyspora rectivirgula, Saccharopolyspora shandongensis, Saccharopolyspora hirsuta, Saccharopolyspora spinose, and Saccharopolyspora erythraea). From a functional perspective, the important functions of Daqu and wheat Qu are both amino acid metabolism and carbohydrate metabolism. Meanwhile, a combined analysis among microbiota, functional genes, and dominant flavors indicated S. shandongensis, S. rectivirgula, and S. spinose might be the main contributor to the synthesis of flavor compounds in wheat Qu, while R. emersonii, W. paramesenteroides, Leuconostoc citreum, Leuconostoc mesenteroides, Weissella cibaria and P. pentosaceus may make the greatest contribution to flavor compounds synthesis in Daqu. This study reveals the microbial and functional dissimilarities of Daqu and wheat Qu, and helps elucidating different metabolic roles of microbes during flavor formation.

The Role of Tyrosine Kinase Itk in T helper 17 and T regulatory cells in Hypersensitivity Pneumonitis

Abstract The balance of inflammatory and suppressive cytokines is critical in controlling inflammatory responses, and the pro- and anti-inflammatory cytokines IL17A and IL10 has been implicated in numerous pulmonary inflammatory diseases. The tyrosine kinase, Itk, plays a critical role in T cell activation. Itk is required for the development of Th17 cells and their production of IL17A in allergic lung inflammation. Furthermore, Type I regulatory and Foxp3+ T regulatory (Tregs) cells, producers of IL10, are positively and negatively regulated by Itk respectively. Farmer’s lung, a subset of hypersensitivity pneumonitis, develops due to repeated exposure to the bacteria Saccharopolyspora rectivirgula (SR) and is dependent on IL17A and regulated by IL10. Surprisingly, exposure to SR drives robust CD4+ T cell IL17A response even in the absence of Itk, with pulmonary inflammation. Transcriptomic analysis of sort purified WT and Itk−/− IL17A producing CD4+ T cells from SR-exposed mice revealed an enrichment of Notch signaling pathway in the absence of Itk. SR also induced the Itk independent development of a population of IL17A producing Foxp3+ Tregs cells, and a significant decrease in IL10 producing Tr1 cells. These data suggest that Itk regulates the expression of IL10, and pathogenic Th17 cells via Notch signaling. These studies suggest that TCR signaling through Itk differentially regulates the development of inflammatory Th17 cells and suppressive Tregs and Tr1 cells in response to SR exposure. Understanding how Itk modulates the development of Th17/Treg cytokine responses will allow us to better understand the precise role of Itk in the regulating the balance of pro- and anti-inflammatory cytokine production during airway inflammation.

Methods for Bioaerosol Characterization: Limits and Perspectives for Human Health Risk Assessment in Organic Waste Treatment

Bioaerosol characterization represents a major challenge for the risk assessment and management of exposed people. One of the most important bioaerosol sources is the organic waste collection and treatment. This work analyzed and discussed the literature with the purpose of investigating the main techniques used nowadays for bioaerosol monitoring during organic waste treatment. The discussion includes an overview on the most efficient sampling, DNA extraction, and analysis methods, including both the cultural and the bio-molecular approach. Generally, an exhaustive biological risk assessment is not applied due to the organic waste heterogeneity, treatment complexity, and unknown aerosolized emission rate. However, the application of bio-molecular methods allows a better bioaerosol characterization, and it is desirable to be associated with standardized cultural methods. Risk assessment for organic waste workers generally includes the evaluation of the potential exposition to pathogens and opportunistic pathogens or to other microorganisms as biomarkers. In most cases, Saccharopolyspora rectivirgula, Legionella spp., Aspergillus spp., and Mycobacterium spp. are included. Future perspectives are focused on identifying common composting biomarkers, on investigating the causality process between chronic bioaerosol exposure and disease onset, and finally, on defining common exposure limits.

Role of the Innate Immune System in the Development of Hypersensitivity Pneumonitis

Hypersensitivity pneumonitis (HP) is an extrinsic allergic alveolitis disease that is constantly affecting the farmers. It is triggered by the continuous exposure of environmental antigens, specifically antigens that grow in hot humid conditions in hay like Saccharopolyspora rectivirgula (SR). While a role of T cells has been established in the development of this diseases, the role of the innate immune system is still largely unknown in the development of lung inflammation leading to HP. To study this disease, we used a mouse model of HP, induced by SR, along with Rag−/− mice which only have an innate immune system and lack an adaptive immune system (and thus have no T cells). We hypothesized that the development of Hypersensitivity pneumonitis requires the presence of T (and B) cells, at the same time, there will be a lower percentage and number of inflammatory cells in the lungs of Rag−/− mice upon exposure to SR. WT and Rag−/− mice were exposed to SR. Lungs, spleen and lymph nodes where collected to analyze their response. We examined the percentage and number of myeloid and Innate Lymphoid Cells (ILC), as well as their ability to produce cytokines. We found that there was no significant difference in the number of Innate Lymphoid cells and myeloid cells between SR‐exposed WT and Rag−/− mice. These results may be due to the low concentration of bacteria used for these experiments. In future experiments, we propose to use a higher concentration of SR to induce inflammation.Support or Funding InformationThis work was supported in part by grants from the NIH‐AI38570, NIH‐AI20701, and NIH‐AI129422. NIH‐NIGMS #2R25GM096955 (RISE). VIP‐Veterinary Investigation Program.

Genetic variability of T cell responses in hypersensitivity pneumonitis identified using the BXD genetic reference panel.

Hypersensitivity pneumonitis (HP) is an interstitial lung disease that may progress to fibrosis and significant risk of death. HP develops following repeated exposures to inhaled environmental antigens; however, only a fraction of the exposed population develops the disease, suggesting that host genetics contribute to disease susceptibility. We used the BXD family of mice with the Saccharopolyspora rectivirgula (SR) model of HP to investigate the role of genetics in susceptibility to HP. The BXD family is derived from a B6 mother and a D2 father and has been used to map susceptibility loci to numerous diseases. B6, D2, and BXD progeny strains were exposed to SR for 3 wk, and the development of HP was monitored. The B6 and D2 strains developed alveolitis; however, the cellular composition was neutrophilic in the D2 strain and more lymphocytic in the B6 strain. Hematoxylin-eosin staining of lung sections revealed lymphoid aggregates in B6 lungs, whereas D2 lungs exhibited a neutrophilic infiltration. Twenty-eight BXD strains of mice were tested, and the results reveal significant heritable variation for numbers of CD4+ or CD8+ T cells in the air spaces. There was significant genetic variability for lymphoid aggregates and alveolar wall thickening. We mapped a significant quantitative trait locus (QTL) on chromosome 18 for CD8+CD69+ T cells that includes cadherin 2 (Cdh2), an excellent candidate gene associated with epithelial-mesenchymal transition, which is upregulated in lungs of strains with HP. These results demonstrate that the BXD family is a valuable and translationally relevant model to identify genes contributing to HP and to devise early and effective interventions.

The role of tyrosine kinase Itk in T helper 17 and T regulatory cells in IL17 mediated hypersensitivity pneumonitis

Abstract The balance of inflammatory and suppressive cytokines is critical in controlling inflammatory responses, and the pro- and anti-inflammatory cytokines IL17A and IL10 has been implicated in numerous pulmonary inflammatory diseases. The tyrosine kinase, Itk, plays a critical role in T cell activation. We have previously shown that Itk−/− mice do not develop allergic airway disease coupled with reduced production of IL17A in the lungs. In vitro, Itk is required for the development of Th17 cells and their production of the pro-inflammatory cytokine IL17A. Furthermore, Type I regulatory and Foxp3+ T regulatory (Tregs) cells, suppressive producers of IL10, are positively and negatively regulated by Itk respectively. Farmer’s lung is a subset of hypersensitivity pneumonitis caused by repeated exposure to the bacteria Saccharopolyspora rectivirgula (SR), and is dependent on IL17A and regulated by IL10. We find that surprisingly, exposure to SR drives robust IL17A response produced by CD4+ T cells even in the absence of Itk, accompanied by the development of pulmonary inflammation. SR also induced Itk independent development of a population of population of Foxp3+ Tregs cells that produce IL17A, accompanied by a significant decrease in Tr1 cells compared to WT mice. These studies suggest that TCR signaling through Itk differentially regulates the development of inflammatory Th17 cells and suppressive Tregs and Tr1 cells. Understanding how Itk modulates the development of Th17/Treg cytokine responses will allow us to better understand the precise role of Itk in the production of IL17A and IL10 in airway inflammation.

Microbial exposure to dairy farmers’ dwellings and COPD occurrence

ABSTRACTDairy farming is a risk factor for chronic obstructive pulmonary disease (COPD). The aim was to determine predictive markers either in blood samples or in dwelling dust samples by comparing COPD and healthy controls with or without farming activity.Dust was collected and analyzed by real-time quantitative PCR. ELISA and DELFIA® were performed to assay the level of specific IgG and IgE of 10 targeted microorganisms.The dwelling exposure of farmers was higher than in the non-farmers (Especially Eurotium amstelodami and Lichtheimia corymbifera). The IgG response against Wallemia sebi and Saccharopolyspora rectivirgula was more often higher in the farmers than the non-farmers. However, exposure and sensitization to the microorganisms tested cannot explain the occurrence of COPD in the dairy farmers’ population. COPD development is probably caused by multiple factors associated with exposure over a period of several years.

Hypersensitivity pneumonitis onset and severity is regulated by CD103 dendritic cell expression.

BackgroundPulmonary dendritic cells drive lung responses to foreign antigens, including Saccharopolyspora rectivirgula, a causative agent of hypersensitivity pneumonitis. While the airway inflammatory mechanisms involved in hypersensitivity pneumonitis are well described, the mechanisms leading to the break in homeostasis and hypersensitivity pneumonitis onset are not well-described, and could involve CD103+ dendritic cells, which are found at baseline and during inflammatory responses in the lung. However, recent demonstration of the ability of CD103+ dendritic cells to induce inflammatory responses starkly contrasts with their classically described role as regulatory cells. These discrepancies may be attributable to the lack of current information on the importance of CD103 expression and modulation on these cells during inflammatory episodes.MethodsTo verify the importance of CD103 expression in the regulation of hypersensitivity pneumonitis, wild-type and Cd103-/- mice were exposed intranasally to S. rectivirgula and airway inflammation was quantified. Surface expression of CD103 in response to S. rectivirgula exposure was studied and cell transfers were used to determine the relative importance of CD103 expression on dendritic cells and T cells in regulating the inflammation in hypersensitivity pneumonitis.ResultsCd103-/- mice developed an exacerbated inflammatory response as early as 18h following S. rectivirgula exposure. CD103 expression on dendritic cells was downregulated quickly following S. rectivirgula exposure, and cell transfers demonstrated that CD103 expression on dendritic cells specifically (and not T cells) regulates the onset and severity of this response.ConclusionAll in all, we demonstrate that CD103 expression by dendritic cells, but not T cells, is crucial for homeostasis maintenance and the regulation of the TH17 airway inflammatory response in hypersensitivity pneumonitis.

Effect of genetics on the development of Hypersensitivity Pneumonitis (HP) using the BXD recombinant inbred mouse reference population

Abstract HP is an interstitial lung disease that develops following repeated exposure to a variety of inhaled environmental antigens. The disease is characterized by alveolitis, granulomas and in some patients, develops into a chronic form associated with fibrosis. Not all individuals exposed to HP causative antigens develop disease suggesting that genetic factors play a role in susceptibility; however, the host co-factor(s) that play a role in susceptibility are unknown. We are using the innovative systems genetics tool, the BXD panel of recombinant inbred mice with the Saccharopolyspora rectivirgula (SR) model of HP to identify genetic loci and candidate genes associated with disease development. We exposed the parental strains, C57Bl/6 and DBA/2, and sixteen BXD strains to SR for 3 weeks and analyzed the lungs 18h after the last exposure. Bronchoalveolar lavage (BAL) was performed and cells recovered from the BAL fluid were analyzed by flow cytometry to determine the frequency of infiltrating immune cells. The results reveal variability in the cellular composition of the BAL; some BXD strains exhibited a neutrophilic alveolitis whereas other BXD strains exhibited a more lymphocytic alveolitis. BAL recovered from mice of the BXD45 strain was 68% neutrophils and 7% T lymphocytes, whereas BAL from the BXD73 strain was 92% neutrophils and 2% T lymphocytes. In addition, there were significant differences in the level of activated CD4+ and CD8+ T cells across the strains. H&amp;E staining of lung tissue revealed significant differences in the extent of granuloma formation across strains. These data support the contention that genetic differences critically influence SR responsiveness given that environmental factors were held constant.