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Microorganisms, Volume 7, Issue 5 (May 2019)

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Open AccessArticle
Parachlamydia acanthamoebae Detected during a Pneumonia Outbreak in Southeastern Finland, in 2017–2018
Microorganisms 2019, 7(5), 141; https://doi.org/10.3390/microorganisms7050141 (registering DOI)
Received: 18 April 2019 / Revised: 9 May 2019 / Accepted: 14 May 2019 / Published: 17 May 2019
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Abstract
Community-acquired pneumonia (CAP) is a common disease responsible for significant morbidity and mortality. However, the definite etiology of CAP often remains unresolved, suggesting that unknown agents of pneumonia remain to be identified. The recently discovered members of the order Chlamydiales, Chlamydia-related bacteria (CRB), [...] Read more.
Community-acquired pneumonia (CAP) is a common disease responsible for significant morbidity and mortality. However, the definite etiology of CAP often remains unresolved, suggesting that unknown agents of pneumonia remain to be identified. The recently discovered members of the order Chlamydiales, Chlamydia-related bacteria (CRB), are considered as possible emerging agents of CAP. Parachlamydia acanthamoebae is the most studied candidate. It survives and replicates inside free-living amoeba, which it might potentially use as a vehicle to infect animals and humans. A Mycoplasma pneumoniae outbreak was observed in Kymenlaakso region in Southeastern Finland during August 2017–January 2018. We determined the occurrence of Chlamydiales bacteria and their natural host, free-living amoeba in respiratory specimens collected during this outbreak with molecular methods. Altogether, 22/278 (7.9%) of the samples contained Chlamydiales DNA. By sequence analysis, majority of the CRBs detected were members of the Parachlamydiaceae family. Amoebal DNA was not detected within the sample material. Our study further proposes that Parachlamydiaceae could be a potential agent causing atypical CAP in children and adolescents. Full article
(This article belongs to the Special Issue Chlamydiae and Chlamydia like Bacteria)
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Open AccessReview
Chlamydia trachomatis and Chlamydia pneumoniae Interaction with the Host: Latest Advances and Future Prospective
Microorganisms 2019, 7(5), 140; https://doi.org/10.3390/microorganisms7050140
Received: 15 April 2019 / Revised: 7 May 2019 / Accepted: 14 May 2019 / Published: 16 May 2019
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Abstract
Research in Chlamydia trachomatis and Chlamydia pneumoniae has gained new traction due to recent advances in molecular biology, namely the widespread use of the metagenomic analysis and the development of a stable genomic transformation system, resulting in a better understanding of Chlamydia pathogenesis. [...] Read more.
Research in Chlamydia trachomatis and Chlamydia pneumoniae has gained new traction due to recent advances in molecular biology, namely the widespread use of the metagenomic analysis and the development of a stable genomic transformation system, resulting in a better understanding of Chlamydia pathogenesis. C. trachomatis, the leading cause of bacterial sexually transmitted diseases, is responsible of cervicitis and urethritis, and C. pneumoniae, a widespread respiratory pathogen, has long been associated with several chronic inflammatory diseases with great impact on public health. The present review summarizes the current evidence regarding the complex interplay between C. trachomatis and host defense factors in the genital micro-environment as well as the key findings in chronic inflammatory diseases associated to C. pneumoniae. Full article
(This article belongs to the Special Issue Chlamydiae and Chlamydia like Bacteria)
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Open AccessReview
Host–Microbe Interactions and Gut Health in Poultry—Focus on Innate Responses
Microorganisms 2019, 7(5), 139; https://doi.org/10.3390/microorganisms7050139
Received: 1 May 2019 / Revised: 14 May 2019 / Accepted: 14 May 2019 / Published: 16 May 2019
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Abstract
Commercial poultry are continually exposed to, frequently pathogenic, microorganisms, usually via mucosal surfaces such as the intestinal mucosa. Thus, understanding host–microbe interactions is vital. Many of these microorganisms may have no or limited contact with the host, while most of those interacting more [...] Read more.
Commercial poultry are continually exposed to, frequently pathogenic, microorganisms, usually via mucosal surfaces such as the intestinal mucosa. Thus, understanding host–microbe interactions is vital. Many of these microorganisms may have no or limited contact with the host, while most of those interacting more meaningfully with the host will be dealt with by the innate immune response. Fundamentally, poultry have evolved to have immune responses that are generally appropriate and adequate for their acquired microbiomes, although this is challenged by commercial production practices. Innate immune cells and their functions, encompassing inflammatory responses, create the context for neutralising the stimulus and initiating resolution. Dysregulated inflammatory responses can be detrimental but, being a highly conserved biological process, inflammation is critical for host defence. Heterogeneity and functional plasticity of innate immune cells is underappreciated and offers the potential for (gut) health interventions, perhaps including exogenous opportunities to influence immune cell metabolism and thus function. New approaches could focus on identifying and enhancing decisive but less harmful immune processes, improving the efficiency of innate immune cells (e.g., targeted, efficient microbial killing) and promoting phenotypes that drive resolution of inflammation. Breeding strategies and suitable exogenous interventions offer potential solutions to enhance poultry gut health, performance and welfare. Full article
(This article belongs to the Special Issue Gut Health in Poultry Production)
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Open AccessArticle
Community- and Hospital-Acquired Klebsiella pneumoniae Urinary Tract Infections in Portugal: Virulence and Antibiotic Resistance
Microorganisms 2019, 7(5), 138; https://doi.org/10.3390/microorganisms7050138
Received: 28 February 2019 / Revised: 30 April 2019 / Accepted: 15 May 2019 / Published: 16 May 2019
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Abstract
Klebsiella pneumoniae is a clinically relevant pathogen and a frequent cause of hospital-acquired (HA) and community-acquired (CA) urinary tract infections (UTI). The increased resistance of this pathogen is leading to limited therapeutic options. To investigate the epidemiology, virulence, and antibiotic resistance profile of [...] Read more.
Klebsiella pneumoniae is a clinically relevant pathogen and a frequent cause of hospital-acquired (HA) and community-acquired (CA) urinary tract infections (UTI). The increased resistance of this pathogen is leading to limited therapeutic options. To investigate the epidemiology, virulence, and antibiotic resistance profile of K. pneumoniae in urinary tract infections, we conducted a multicenter retrospective study for a total of 81 isolates (50 CA-UTI and 31 HA-UTI) in Portugal. The detection and characterization of resistance and virulence determinants were performed by molecular methods (PCR, PCR-based replicon typing, and multilocus sequence typing (MLST)). Out of 50 CA-UTI isolates, six (12.0%) carried β-lactamase enzymes, namely blaTEM-156 (n = 2), blaTEM-24 (n = 1), blaSHV-11 (n = 1), blaSHV-33 (n = 1), and blaCTX-M-15 (n = 1). All HA-UTI were extended-spectrum β-lactamase (ESBL) producers and had a multidrug resistant profile as compared to the CA-UTI isolates, which were mainly resistant to ciprofloxacin, levofloxacin, tigecycline, and fosfomycin. In conclusion, in contrast to community-acquired isolates, there is an overlap between virulence and multidrug resistance for hospital-acquired UTI K. pneumoniae pathogens. The study is the first to report different virulence characteristics for hospital and community K. pneumoniae pathogens, despite the production of β-lactamase and even with the presence of CTX-M-15 ESBL, a successful international ST15 clone, which were identified in both settings. This highlights that a focus on genomic surveillance should remain a priority in the hospital environment. Full article
(This article belongs to the Special Issue Multidrug-Resistant Pathogens)
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Open AccessReview
Shiga-Toxin Producing Escherichia Coli in Brazil: A Systematic Review
Microorganisms 2019, 7(5), 137; https://doi.org/10.3390/microorganisms7050137
Received: 29 March 2019 / Revised: 1 May 2019 / Accepted: 4 May 2019 / Published: 16 May 2019
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Abstract
Shiga-toxin producing E. coli (STEC) can cause serious illnesses, including hemorrhagic colitis and hemolytic uremic syndrome. This is the first systematic review of STEC in Brazil, and will report the main serogroups detected in animals, food products and foodborne diseases. Data were obtained [...] Read more.
Shiga-toxin producing E. coli (STEC) can cause serious illnesses, including hemorrhagic colitis and hemolytic uremic syndrome. This is the first systematic review of STEC in Brazil, and will report the main serogroups detected in animals, food products and foodborne diseases. Data were obtained from online databases accessed in January 2019. Papers were selected from each database using the Mesh term entries. Although no human disease outbreaks in Brazil related to STEC has been reported, the presence of several serogroups such as O157 and O111 has been verified in animals, food, and humans. Moreover, other serogroups monitored by international federal agencies and involved in outbreak cases worldwide were detected, and other unusual strains were involved in some isolated individual cases of foodborne disease, such as serotype O118:H16 and serogroup O165. The epidemiological data presented herein indicates the presence of several pathogenic serogroups, including O157:H7, O26, O103, and O111, which have been linked to disease outbreaks worldwide. As available data are concentrated in the Sao Paulo state and almost completely lacking in outlying regions, epidemiological monitoring in Brazil for STEC needs to be expanded and food safety standards for this pathogen should be aligned to that of the food safety standards of international bodies. Full article
(This article belongs to the Special Issue Shiga Toxin-Producing Escherichia coli)
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Open AccessArticle
Presence of Mycobacterium avium Subspecies paratuberculosis Monitored Over Varying Temporal and Spatial Scales in River Catchments: Persistent Routes for Human Exposure
Microorganisms 2019, 7(5), 136; https://doi.org/10.3390/microorganisms7050136
Received: 4 April 2019 / Revised: 2 May 2019 / Accepted: 9 May 2019 / Published: 15 May 2019
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Abstract
Mycobacterium avium subspecies paratuberculosis (Map) was monitored by quantitative PCR over a range of temporal and spatial scales in the River Tywi catchment. This study shows the persistence of Map over a 10-year period with little change, which correlates with the [...] Read more.
Mycobacterium avium subspecies paratuberculosis (Map) was monitored by quantitative PCR over a range of temporal and spatial scales in the River Tywi catchment. This study shows the persistence of Map over a 10-year period with little change, which correlates with the recognised levels of Johne’s disease in British herds over that period (aim 1). Map was quantified within the river at up to 108 cell equivalents L−1 and was shown to be consistently present when monitored over finer timescales (aim 4). Small wastewater treatment plants where the ingress of human-associated Map might be expected had no significant effect (aim 2). Map was found for the first time to be located in natural river foams providing another route for spread via aerosols (aim 5). This study provides evidence for the environmental continuum of Map from the grazing infected animal via rain driven runoff through field drains and streams into main rivers; with detection at a high frequency throughout the year. Should Map need to be monitored in the future, we recommend that weekly or monthly sampling from a fixed location on a river will capture an adequate representation of the flow dynamics of Map in a catchment (aim 3). The human exposure to Map during this process and its impact on human health remains unquantified. Full article
(This article belongs to the Special Issue Non-Tuberculous Mycobacteria: Emerging Diseases and Health Impacts)
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Open AccessArticle
Oral Administration of a Select Mixture of Lactobacillus and Bacillus Alleviates Inflammation and Maintains Mucosal Barrier Integrity in the Ileum of Pigs Challenged with Salmonella Infantis
Microorganisms 2019, 7(5), 135; https://doi.org/10.3390/microorganisms7050135
Received: 25 April 2019 / Revised: 11 May 2019 / Accepted: 12 May 2019 / Published: 15 May 2019
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Abstract
Salmonella is important as both a cause of clinical disease in swine and as a source of food-borne transmission of disease to humans. Lactobacillus and Bacillus are often used as antibiotic substitutes to prevent Salmonella infection. In this study, we evaluated the effects [...] Read more.
Salmonella is important as both a cause of clinical disease in swine and as a source of food-borne transmission of disease to humans. Lactobacillus and Bacillus are often used as antibiotic substitutes to prevent Salmonella infection. In this study, we evaluated the effects of a select mixture of Lactobacillus johnsonii L531, Bacillus licheniformis BL1721 and Bacillus subtilis BS1715 (LBB-mix) in prevention of Salmonella enterica serovar Infantis infection in a pig model. LBB-mix was orally administered to newly weaned piglets for seven days before S. Infantis challenge. LBB-mix pretreatment ameliorated S. Infantis-induced fever, leukocytosis, growth performance loss, and ileal inflammation. Pre-administration of LBB-mix reduced the number of Salmonella in the feces but increased the number of goblet cells in the ileum. S. Infantis infection resulted in an increase in cell death in the ileum, this increase was attenuated by LBB-mix consumption. Claudin 1 and cleaved caspase-1 expression was decreased in the ileum of pigs challenged with S. Infantis, but not in pigs pretreated with LBB-mix. In conclusion, our data indicate that a select LBB-mix has positive effects on controlling S. Infantis infection via alleviating inflammation and maintaining the intestinal mucosal barrier integrity in pigs. Full article
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Open AccessArticle
Combined Effect of Vacuum Packaging, Fennel and Savory Essential Oil Treatment on the Quality of Chicken Thighs
Microorganisms 2019, 7(5), 134; https://doi.org/10.3390/microorganisms7050134
Received: 8 April 2019 / Revised: 8 May 2019 / Accepted: 9 May 2019 / Published: 15 May 2019
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Abstract
The aim of the present work was to evaluate the microbiological quality of chicken thighs after treatment by fennel (Foeniculum vulgare) and savory (Satureja hortensis) essential oil, stored under vacuum packaging (VP) at 4 ± 0.5 °C for a [...] Read more.
The aim of the present work was to evaluate the microbiological quality of chicken thighs after treatment by fennel (Foeniculum vulgare) and savory (Satureja hortensis) essential oil, stored under vacuum packaging (VP) at 4 ± 0.5 °C for a period of 16 days. The following treatments of chicken thighs were used: Air-packaging control samples (APCS), vacuum-packaging control samples (VPC), vacuum-packaging (VP) control samples with rapeseed oil (VPRO), VP (vacuum-packaging) with fennel essential oil at concentrations 0.2% v/w (VP + F), and VP with savory essential oil at concentration 0.2% v/w (VP + S). The quality assessment of APCS, VPC, VPRO, VP + F and VP + S products was established by microbiological analysis. The microbiological parameters as the total viable counts of bacteria of the Enterobacteriaceae family, lactic acid bacteria (LAB), and Pseudomonas spp. were detected. Bacterial species were identified with the MALDI-TOF MS Biotyper. The combination of essential oils and vacuum packaging had a significant effect (p < 0.05) on the reduction of total viable counts (TVC) compared with control group without vacuum packaging and the untreated control group. Though 15 genera and 46 species were isolated with scores higher than 2.3 from the chicken samples. Full article
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Open AccessReview
Arthritogenic Alphaviruses: A Worldwide Emerging Threat?
Microorganisms 2019, 7(5), 133; https://doi.org/10.3390/microorganisms7050133
Received: 24 April 2019 / Revised: 9 May 2019 / Accepted: 10 May 2019 / Published: 14 May 2019
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Abstract
Arthritogenic alphaviruses are responsible for a dengue-like syndrome associated with severe debilitating polyarthralgia that can persist for months or years and impact life quality. Chikungunya virus is the most well-known member of this family since it was responsible for two worldwide epidemics with [...] Read more.
Arthritogenic alphaviruses are responsible for a dengue-like syndrome associated with severe debilitating polyarthralgia that can persist for months or years and impact life quality. Chikungunya virus is the most well-known member of this family since it was responsible for two worldwide epidemics with millions of cases in the last 15 years. However, other arthritogenic alphaviruses that are as of yet restrained to specific territories are the cause of neglected tropical diseases: O’nyong’nyong virus in Sub-Saharan Africa, Mayaro virus in Latin America, and Ross River virus in Australia and the Pacific island countries and territories. This review evaluates their emerging potential in light of the current knowledge for each of them and in comparison to chikungunya virus. Full article
(This article belongs to the Special Issue Emerging Vector Borne Infections: A Novel Threat for Global Health)
Open AccessArticle
In Vitro Activation of Seed-Transmitted Cultivation-Recalcitrant Endophytic Bacteria in Tomato and Host–Endophyte Mutualism
Microorganisms 2019, 7(5), 132; https://doi.org/10.3390/microorganisms7050132
Received: 19 April 2019 / Revised: 9 May 2019 / Accepted: 10 May 2019 / Published: 14 May 2019
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Abstract
This study was aimed at exploring seed transmission of endophytic bacteria in tomato utilizing aseptic in vitro conditions. Cultivation-based studies were undertaken on two tomato cultivars “Arka Vikas” and “Arka Abha” employing surface sterilized seeds, aseptically germinated seeds and in vitro grown seedlings [...] Read more.
This study was aimed at exploring seed transmission of endophytic bacteria in tomato utilizing aseptic in vitro conditions. Cultivation-based studies were undertaken on two tomato cultivars “Arka Vikas” and “Arka Abha” employing surface sterilized seeds, aseptically germinated seeds and in vitro grown seedlings at different stages. Bacillus sp. appeared primarily as seed externally-associated bacteria. Tissue homogenate from extensively surface-sterilized seeds, day-3 germinating seeds, or 10-day in vitro seedlings did not show any cultivable bacteria on two bacteriological media. Indexing of 4-week old healthy seedlings with seed-coat removal following seed germination showed bacterial association in 50–75% seedlings yielding 106–107 cfu g−1 tissues. Four endophytic bacteria appeared common to both cultivars (Kosakonia, Ralstonia, Sphingomonas, Sphingobium spp.) with three additional species in “Arka Abha”. The bacterial strains showed a manifold increase in growth with host-tissue-extract supplementation. Seed inoculations with single-isolates stimulated germination or enhanced the seedling growth coupled with the activation of additional endophytic bacteria. In vitro seedlings upon recurrent medium-indexing over eight weeks showed gradual emergence of endophytic bacteria. The study reveals the seed internal colonization by different bacterial endophytes in a cultivation-recalcitrant form, their activation to cultivable state during seedling growth and transmission to seedlings with mutualistic effects. Full article
(This article belongs to the Special Issue Plant Control of Symbiotic Microbe Behavior and Reproduction)
Open AccessArticle
The Transcription Factor Sfp1 Regulates the Oxidative Stress Response in Candida albicans
Microorganisms 2019, 7(5), 131; https://doi.org/10.3390/microorganisms7050131
Received: 31 March 2019 / Revised: 7 May 2019 / Accepted: 9 May 2019 / Published: 14 May 2019
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Abstract
Candida albicans is a commensal that inhabits the skin and mucous membranes of humans. Because of the increasing immunocompromised population and the limited classes of antifungal drugs available, C. albicans has emerged as an important opportunistic pathogen with high mortality rates. During infection [...] Read more.
Candida albicans is a commensal that inhabits the skin and mucous membranes of humans. Because of the increasing immunocompromised population and the limited classes of antifungal drugs available, C. albicans has emerged as an important opportunistic pathogen with high mortality rates. During infection and therapy, C. albicans frequently encounters immune cells and antifungal drugs, many of which exert their antimicrobial activity by inducing the production of reactive oxygen species (ROS). Therefore, antioxidative capacity is important for the survival and pathogenesis of C. albicans. In this study, we characterized the roles of the zinc finger transcription factor Sfp1 in the oxidative stress response against C. albicans. A sfp1-deleted mutant was more resistant to oxidants and macrophage killing than wild-type C. albicans and processed an active oxidative stress response with the phosphorylation of the mitogen-activated protein kinase (MAPK) Hog1 and high CAP1 expression. Moreover, the sfp1-deleted mutant exhibited high expression levels of antioxidant genes in response to oxidative stress, resulting in a higher total antioxidant capacity, glutathione content, and glutathione peroxidase and superoxide dismutase enzyme activity than the wild-type C. albicans. Finally, the sfp1-deleted mutant was resistant to macrophage killing and ROS-generating antifungal drugs. Together, our findings provide a new understanding of the complex regulatory machinery in the C. albicans oxidative stress response. Full article
(This article belongs to the Special Issue Molecular Mechanisms of Fungal Virulence and Commensalism)
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Open AccessReview
Advances in Chemical and Biological Methods to Identify Microorganisms—From Past to Present
Microorganisms 2019, 7(5), 130; https://doi.org/10.3390/microorganisms7050130
Received: 12 April 2019 / Revised: 30 April 2019 / Accepted: 8 May 2019 / Published: 13 May 2019
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Abstract
Fast detection and identification of microorganisms is a challenging and significant feature from industry to medicine. Standard approaches are known to be very time-consuming and labor-intensive (e.g., culture media and biochemical tests). Conversely, screening techniques demand a quick and low-cost grouping of bacterial/fungal [...] Read more.
Fast detection and identification of microorganisms is a challenging and significant feature from industry to medicine. Standard approaches are known to be very time-consuming and labor-intensive (e.g., culture media and biochemical tests). Conversely, screening techniques demand a quick and low-cost grouping of bacterial/fungal isolates and current analysis call for broad reports of microorganisms, involving the application of molecular techniques (e.g., 16S ribosomal RNA gene sequencing based on polymerase chain reaction). The goal of this review is to present the past and the present methods of detection and identification of microorganisms, and to discuss their advantages and their limitations. Full article
(This article belongs to the Special Issue Identification of Microorganisms: Old, New and Future Methods)
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Open AccessArticle
Response of Microbial Communities and Their Metabolic Functions to Drying–Rewetting Stress in a Temperate Forest Soil
Microorganisms 2019, 7(5), 129; https://doi.org/10.3390/microorganisms7050129
Received: 25 March 2019 / Revised: 5 May 2019 / Accepted: 6 May 2019 / Published: 13 May 2019
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Abstract
Global climate change is predicted to alter drought–precipitation patterns, which will likely affect soil microbial communities and their functions, ultimately shifting microbially-mediated biogeochemical cycles. The present study aims to investigate the simultaneous variation of microbial community compositions and functions in response to drought [...] Read more.
Global climate change is predicted to alter drought–precipitation patterns, which will likely affect soil microbial communities and their functions, ultimately shifting microbially-mediated biogeochemical cycles. The present study aims to investigate the simultaneous variation of microbial community compositions and functions in response to drought and following rewetting events, using a soil metaproteomics approach. For this, an established field experiment located in an Austrian forest with two levels (moderate and severe stress) of precipitation manipulation was evaluated. The results showed that fungi were more strongly influenced by drying and rewetting (DRW) than bacteria, and that there was a drastic shift in the fungal community towards a more Ascomycota-dominated community. In terms of functional responses, a larger number of proteins and a higher functional diversity were observed in both moderate and severe DRW treatments compared to the control. Furthermore, in both DRW treatments a rise in proteins assigned to “translation, ribosomal structure, and biogenesis” and “protein synthesis” suggests a boost in microbial cell growth after rewetting. We also found that the changes within intracellular functions were associated to specific phyla, indicating that responses of microbial communities to DRW primarily shifted microbial functions. Microbial communities seem to respond to different levels of DRW stress by changing their functional potential, which may feed back to biogeochemical cycles. Full article
(This article belongs to the Special Issue Towards Integrated Multi-omics Analyses of Environmental Microbiota)
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Open AccessReview
Propionibacterium acnes and Acne Vulgaris: New Insights from the Integration of Population Genetic, Multi-Omic, Biochemical and Host-Microbe Studies
Microorganisms 2019, 7(5), 128; https://doi.org/10.3390/microorganisms7050128
Received: 1 April 2019 / Revised: 8 May 2019 / Accepted: 9 May 2019 / Published: 13 May 2019
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Abstract
The anaerobic bacterium Propionibacterium acnes is believed to play an important role in the pathophysiology of the common skin disease acne vulgaris. Over the last 10 years our understanding of the taxonomic and intraspecies diversity of this bacterium has increased tremendously, and with [...] Read more.
The anaerobic bacterium Propionibacterium acnes is believed to play an important role in the pathophysiology of the common skin disease acne vulgaris. Over the last 10 years our understanding of the taxonomic and intraspecies diversity of this bacterium has increased tremendously, and with it the realisation that particular strains are associated with skin health while others appear related to disease. This extensive review will cover our current knowledge regarding the association of P. acnes phylogroups, clonal complexes and sequence types with acne vulgaris based on multilocus sequence typing of isolates, and direct ribotyping of the P. acnes strain population in skin microbiome samples based on 16S rDNA metagenomic data. We will also consider how multi-omic and biochemical studies have facilitated our understanding of P. acnes pathogenicity and interactions with the host, thus providing insights into why certain lineages appear to have a heightened capacity to contribute to acne vulgaris development, while others are positively associated with skin health. We conclude with a discussion of new therapeutic strategies that are currently under investigation for acne vulgaris, including vaccination, and consider the potential of these treatments to also perturb beneficial lineages of P. acnes on the skin. Full article
(This article belongs to the Special Issue The Functions of the Microbiome in Skin Health and Disease)
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Open AccessArticle
Biocidal Effectiveness of Selected Disinfectants Solutions Based on Water and Ozonated Water against Listeria monocytogenes Strains
Microorganisms 2019, 7(5), 127; https://doi.org/10.3390/microorganisms7050127
Received: 30 March 2019 / Revised: 7 May 2019 / Accepted: 9 May 2019 / Published: 10 May 2019
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Abstract
The aim of this study was to compare the biocidal effectiveness of disinfectants solutions prepared with ozonated and non-ozonated water against Listeria monocytogenes. Six L. monocytogenes strains were the research material (four isolates from food: meat (LMO-M), dairy products (LMO-N), vegetables (LMO-W), [...] Read more.
The aim of this study was to compare the biocidal effectiveness of disinfectants solutions prepared with ozonated and non-ozonated water against Listeria monocytogenes. Six L. monocytogenes strains were the research material (four isolates from food: meat (LMO-M), dairy products (LMO-N), vegetables (LMO-W), and fish (LMO-R); one clinical strain (LMO-K) and reference strain ATCC 19111). The evaluation of the biocidal effectiveness of disinfectant solutions (QAC—quaternary ammonium compounds; OA—oxidizing agents; ChC—chlorine compounds; IC—iodine compounds; NANO—nanoparticles) was carried out, marking the MBC values. Based on the obtained results, the effectiveness coefficient (A) were calculated. The smaller the A value, the greater the efficiency of disinfection solutions prepared on the basis of ozonated versus non-ozonated water. Ozonated water showed biocidal efficacy against L. monocytogenes. Among tested disinfectentants, independent on type of water used for preparation, the most effective against L. monocytogenes were: QAC 1 (benzyl-C12-18-alkydimethyl ammonium chlorides) (1.00 × 105–1.00 × 104 g/mL) in quaternary ammonium compounds, OA 3 (peracetic acid, hydrogen peroxide, bis (sulphate) bis (peroxymonosulfate)) (3.08 × 104 –3.70 × 103 g/mL) in oxidizing agents, ChC 1 (chlorine dioxide) (5.00 × 108 –7.00 × 107 g/mL) in chlorine compounds, IC 1 (iodine) (1.05–2.15 g/mL) in iodine compounds, and NANO 1 (nanocopper) (1.08 × 10-4 – 1.47 × 104 g/mL) in nanoparticles. The values of the activity coefficient for QAC ranged from 0.10 to 0.40, for OA—0.15–0.84, for ChC—0.25–0.83, for IC—0.45–0.60, and for NANO—0.70–0.84. The preparation of disinfectants solution on the basis of ozonated water, improved the microbicidal efficiency of the tested disinfectant, especially the quaternary ammonium compounds. An innovative element of our work is the use of ozonated water for the preparation of working solutions of the disinfection agents. Use ozonated water can help to reduce the use of disinfectant concentrations and limit the increasing of microbial resistance to disinfectants. This paper provides many new information to optimize hygiene plans in food processing plants and limit the spread of microorganisms such as L. monocytogenes. Full article
Open AccessReview
Members of the Lactobacillus Genus Complex (LGC) as Opportunistic Pathogens: A Review
Microorganisms 2019, 7(5), 126; https://doi.org/10.3390/microorganisms7050126
Received: 15 April 2019 / Revised: 4 May 2019 / Accepted: 8 May 2019 / Published: 10 May 2019
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Abstract
Microorganisms belonging to the Lactobacillus genus complex (LGC) are naturally associated or deliberately added to fermented food products and are widely used as probiotic food supplements. Moreover, these bacteria normally colonize the mouth, gastrointestinal (GI) tract, and female genitourinary tract of humans. They [...] Read more.
Microorganisms belonging to the Lactobacillus genus complex (LGC) are naturally associated or deliberately added to fermented food products and are widely used as probiotic food supplements. Moreover, these bacteria normally colonize the mouth, gastrointestinal (GI) tract, and female genitourinary tract of humans. They exert multiple beneficial effects and are regarded as safe microorganisms. However, infections caused by lactobacilli, mainly endocarditis, bacteremia, and pleuropneumonia, occasionally occur. The relevance of Lactobacillus spp. and other members of the LGC as opportunistic pathogens in humans and related risk factors and predisposing conditions are illustrated in this review article with more emphasis on the species L. rhamnosus that has been more often involved in infection cases. The methods used to identify this species in clinical samples, to distinguish strains and to evaluate traits that can be associated to pathogenicity, as well as future perspectives for improving the identification of potentially pathogenic strains, are outlined. Full article
(This article belongs to the Special Issue Safety Aspects of Lactic Acid Bacteria)
Open AccessArticle
Impact of Binge Alcohol Intoxication on the Humoral Immune Response during Burkholderia spp. Infections
Microorganisms 2019, 7(5), 125; https://doi.org/10.3390/microorganisms7050125
Received: 20 March 2019 / Revised: 2 May 2019 / Accepted: 5 May 2019 / Published: 9 May 2019
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Abstract
Burkholderia pseudomallei, the causative agent of melioidosis can occur in healthy humans, yet binge alcohol use is progressively being recognized as a major risk factor. Currently, no experimental studies have investigated the effects of binge alcohol on the adaptive immune system during [...] Read more.
Burkholderia pseudomallei, the causative agent of melioidosis can occur in healthy humans, yet binge alcohol use is progressively being recognized as a major risk factor. Currently, no experimental studies have investigated the effects of binge alcohol on the adaptive immune system during an active infection. In this study, we used B. thailandensis and B. vietnamiensis, to investigate the impact of a single binge alcohol episode on the humoral response during infection. Eight-week-old female C57BL/6 mice were administered alcohol comparable to human binge drinking (4.4 g/kg) or PBS intraperitoneally 30 min before intranasal infection. Mice infected with B. thailandensis had a 100% survival rate, while those infected with B. vietnamiensis had a 33% survivability rate when a binge alcohol dose was administered. B. thailandensis was detected in blood of mice administered alcohol at only 7 days post infection (PI), while those infected with B. vietnamiensis and receiving alcohol were found throughout the 28-day infection as well as in tissues at day 28 PI. Binge alcohol elevated IgM and delayed IgG specific to the whole cell lysate (WCL) of B. vietnamiensis but not B. thailandensis infections. Differences in immunogenicity of B. pseudomallei near-neighbors provide a framework for novel insights into the effects of binge alcohol’s suppression of the humoral immune response that can cause opportunistic infections in otherwise healthy hosts. Full article
(This article belongs to the Section Medical Microbiology)
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Open AccessReview
Biosynthesis of Polyketides in Streptomyces
Microorganisms 2019, 7(5), 124; https://doi.org/10.3390/microorganisms7050124
Received: 18 March 2019 / Revised: 24 April 2019 / Accepted: 27 April 2019 / Published: 6 May 2019
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Abstract
Polyketides are a large group of secondary metabolites that have notable variety in their structure and function. Polyketides exhibit a wide range of bioactivities such as antibacterial, antifungal, anticancer, antiviral, immune-suppressing, anti-cholesterol, and anti-inflammatory activity. Naturally, they are found in bacteria, fungi, plants, [...] Read more.
Polyketides are a large group of secondary metabolites that have notable variety in their structure and function. Polyketides exhibit a wide range of bioactivities such as antibacterial, antifungal, anticancer, antiviral, immune-suppressing, anti-cholesterol, and anti-inflammatory activity. Naturally, they are found in bacteria, fungi, plants, protists, insects, mollusks, and sponges. Streptomyces is a genus of Gram-positive bacteria that has a filamentous form like fungi. This genus is best known as one of the polyketides producers. Some examples of polyketides produced by Streptomyces are rapamycin, oleandomycin, actinorhodin, daunorubicin, and caprazamycin. Biosynthesis of polyketides involves a group of enzyme activities called polyketide synthases (PKSs). There are three types of PKSs (type I, type II, and type III) in Streptomyces responsible for producing polyketides. This paper focuses on the biosynthesis of polyketides in Streptomyces with three structurally-different types of PKSs. Full article
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Open AccessCommunication
Microbial and Functional Profile of the Ceca from Laying Hens Affected by Feeding Prebiotics, Probiotics, and Synbiotics
Microorganisms 2019, 7(5), 123; https://doi.org/10.3390/microorganisms7050123
Received: 3 April 2019 / Revised: 1 May 2019 / Accepted: 4 May 2019 / Published: 6 May 2019
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Abstract
Diet has an essential influence in the establishment of the cecum microbial communities in poultry, so its supplementation with safe additives, such as probiotics, prebiotics, and synbiotics might improve animal health and performance. This study showed the ceca microbiome modulations of laying hens, [...] Read more.
Diet has an essential influence in the establishment of the cecum microbial communities in poultry, so its supplementation with safe additives, such as probiotics, prebiotics, and synbiotics might improve animal health and performance. This study showed the ceca microbiome modulations of laying hens, after feeding with dry whey powder as prebiotics, Pediococcus acidilactici as probiotics, and the combination of both as synbiotics. A clear grouping of the samples induced per diet was observed (p < 0.05). Operational taxonomic units (OTUs) identified as Olsenella spp., and Lactobacillus crispatus increased their abundance in prebiotic and synbiotic treatments. A core of the main functions was shared between all metagenomes (45.5%), although the genes encoding for the metabolism of butanoate, propanoate, inositol phosphate, and galactose were more abundant in the prebiotic diet. The results indicated that dietary induced-changes in microbial composition did not imply a disturbance in the principal biological roles, while the specific functions were affected. Full article
(This article belongs to the Special Issue Gut Health in Poultry Production)
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Open AccessArticle
Bioconversion of Flavonoid Glycosides from Hippophae rhamnoides Leaves into Flavonoid Aglycones by Eurotium amstelodami
Microorganisms 2019, 7(5), 122; https://doi.org/10.3390/microorganisms7050122
Received: 29 March 2019 / Revised: 30 April 2019 / Accepted: 30 April 2019 / Published: 5 May 2019
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Abstract
The flowering process has been reported to play crucial roles in improving the flavor and efficacy of fermented tea. Hippophae rhamnoides leaves containing many beneficial ingredients are a suitable plant source for tea processing. In this study, we isolated a β-glucosidase-producing fungus Eurotium [...] Read more.
The flowering process has been reported to play crucial roles in improving the flavor and efficacy of fermented tea. Hippophae rhamnoides leaves containing many beneficial ingredients are a suitable plant source for tea processing. In this study, we isolated a β-glucosidase-producing fungus Eurotium amstelodami BSX001 from the fermented tea and used Hippophae rhamnoides leaves (HRL) as a substrate to explore the detailed process of bioconversion of some important functional factors. The results show that the contents of total phenolic compounds and flavonoids increased significantly after seven days, especially flavonoid aglycones (e.g., quercetin, kaempferol, and isorhamnetin). Such compounds greatly enhance the antioxidative activity of fermented products. Metabolic analysis of the standard compounds (rutin, quercetin-3-glucoside, kaempferol-3-glucoside, quercetin, isorhamnetin-3-glucoside, isorhamnetin, and kaempferol) further confirmed the effective biotransformation by E. amstelodami. Mechanisms of the bioconversion could be involved in deglycosylation, dihydroxylation, and O-methylation. Our findings expand the understanding of tea fermentation process and provide further guidance for the fermented tea industry. Full article
(This article belongs to the Special Issue The Mechanisms and Applications of Microbial Electrocatalysis)
Open AccessReview
Role of Gut Microbiota in Hepatocarcinogenesis
Microorganisms 2019, 7(5), 121; https://doi.org/10.3390/microorganisms7050121
Received: 20 March 2019 / Revised: 23 April 2019 / Accepted: 3 May 2019 / Published: 5 May 2019
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Abstract
Hepatocellular carcinoma (HCC), one of the leading causes of death worldwide, has a causal nexus with liver injury, inflammation, and regeneration that accumulates over decades. Observations from recent studies have accounted for the involvement of the gut–liver axis in the pathophysiological mechanism responsible [...] Read more.
Hepatocellular carcinoma (HCC), one of the leading causes of death worldwide, has a causal nexus with liver injury, inflammation, and regeneration that accumulates over decades. Observations from recent studies have accounted for the involvement of the gut–liver axis in the pathophysiological mechanism responsible for HCC. The human intestine nurtures a diversified colony of microorganisms residing in the host ecosystem. The intestinal barrier is critical for conserving the normal physiology of the gut microbiome. Therefore, a rupture of this barrier or dysbiosis can cause the intestinal microbiome to serve as the main source of portal-vein endotoxins, such as lipopolysaccharide, in the progression of hepatic diseases. Indeed, increased bacterial translocation is a key sign of HCC. Considering the limited number of clinical studies on HCC with respect to the microbiome, we focus on clinical as well as animal studies involving the gut microbiota, with the current understandings of the mechanism by which the intestinal dysbiosis promotes hepatocarcinogenesis. Future research might offer mechanistic insights into the specific phyla targeting the leaky gut, as well as microbial dysbiosis, and their metabolites, which represent key pathways that drive HCC-promoting microbiome-mediated liver inflammation and fibrosis, thereby restoring the gut barrier function. Full article
(This article belongs to the Special Issue Host-Gut Microbiota Interactions)
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Open AccessArticle
Feeding Essential Oils to Neonatal Holstein Dairy Calves Results in Increased Ruminal Prevotellaceae Abundance and Propionate Concentrations
Microorganisms 2019, 7(5), 120; https://doi.org/10.3390/microorganisms7050120
Received: 3 April 2019 / Revised: 26 April 2019 / Accepted: 29 April 2019 / Published: 1 May 2019
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Abstract
Since antibiotic use in animal production has become a public health concern, great efforts are being dedicated to find effective and viable alternatives. While essential oils (EO) have become attractive candidates for use in the livestock industry, their mode of action and microbial [...] Read more.
Since antibiotic use in animal production has become a public health concern, great efforts are being dedicated to find effective and viable alternatives. While essential oils (EO) have become attractive candidates for use in the livestock industry, their mode of action and microbial targets in food animals remain largely uncharacterized. To gain further insight, we investigated the rumen environment of neonatal calves fed calf starter pellets and milk replacer supplemented with a commercial blend of EO. Propionate concentrations were not only found to be higher in EO-fed calves compared to controls (P < 0.05), but ruminal bacterial communities also differed greatly. For instance, the abundance of Firmicutes was significantly lower in samples from EO-fed calves than in controls, which appeared to be mostly due to lower Lachnospiraceae levels (P < 0.05). In contrast, Bacteriodetes were more abundant in EO-fed calves compared to controls, which was primarily the result of higher Prevotellaceae (P < 0.05). Notably, two bacterial operational taxonomic units (OTUs) were significantly more abundant in EO-fed calves; SD_Bt-00966 was found to be a close relative of Prevotella ruminicola (97%), while SD_Bt-00978 likely corresponded to an uncharacterized species of Gammaproteobacteria. In addition, Pearson correlation and canonical correspondence analyses revealed potential associations between other ruminal bacterial OTUs and either short chain fatty acids (SCFA) parameters or metrics for calf growth. Together, these results support that EO supplementation in growing dairy calves can modulate rumen function through SCFA production and growth of specific rumen bacterial groups. Full article
(This article belongs to the Section Gut Microbiota)
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Open AccessArticle
Efficient Epstein-Barr Virus Progeny Production Mediated by Cancer-Derived LMP1 and Virally-Encoded microRNAs
Microorganisms 2019, 7(5), 119; https://doi.org/10.3390/microorganisms7050119
Received: 26 March 2019 / Revised: 25 April 2019 / Accepted: 28 April 2019 / Published: 30 April 2019
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Abstract
Epstein-Barr virus (EBV) genomes, particularly their latent genes, are heterogeneous among strains. The heterogeneity of EBV-encoded latent membrane protein 1 (LMP1) raises the question of whether there are functional differences between LMP1 expressed by cancer-associated EBV and that by non-cancerous strains. Here, we [...] Read more.
Epstein-Barr virus (EBV) genomes, particularly their latent genes, are heterogeneous among strains. The heterogeneity of EBV-encoded latent membrane protein 1 (LMP1) raises the question of whether there are functional differences between LMP1 expressed by cancer-associated EBV and that by non-cancerous strains. Here, we used bacterial artificial chromosome (BAC)-cloned EBV genomes retaining all virally encoded microRNA (miRNA) genes to investigate the functions of cancer-derived LMP1 in the context of the EBV genome. HEK293 cells were stably transfected with EBV-BAC clone DNAs encoding either nasopharyngeal carcinoma (NPC)-derived CAO-LMP1 (LMP1CAO) or LMP1 from a prototype B95-8 strain of EBV (LMP1B95-8). When an EBV-BAC clone DNA encoding LMP1CAO was stably transfected into HEK293 cells, it generated many more stable transformants than the control clone encoding LMP1B95-8. Furthermore, stably transfected HEK293 cells exhibited highly efficient production of progeny virus. Importantly, deletion of the clustered viral miRNA genes compromised the ability to produce progeny viruses. These results indicate that cancer-derived LMP1 and viral miRNAs together are necessary for efficient production of progeny virus, and that the resulting increase in efficiency contributes to EBV-mediated epithelial carcinogenesis. Full article
(This article belongs to the Special Issue Recent Advances in Understanding Epstein-Barr Virus)
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Open AccessCase Report
A Bird’s-Eye View of Chronic Unilateral Conjunctivitis: Remember about Chlamydia psittaci
Microorganisms 2019, 7(5), 118; https://doi.org/10.3390/microorganisms7050118
Received: 31 March 2019 / Revised: 17 April 2019 / Accepted: 26 April 2019 / Published: 30 April 2019
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Abstract
Chlamydia psittaci causes psittacosis in humans, mainly in people in contact with birds in either the setting of occupational or companion bird exposure. Infection is associated with a range of clinical manifestations from asymptomatic infection to severe atypical pneumonia and systemic disease. C. [...] Read more.
Chlamydia psittaci causes psittacosis in humans, mainly in people in contact with birds in either the setting of occupational or companion bird exposure. Infection is associated with a range of clinical manifestations from asymptomatic infection to severe atypical pneumonia and systemic disease. C. psittaci has also been associated with ocular adnexal lymphoma in human patients. The current paper describes successful doxycycline treatment of a male patient suffering from C. psittaci chronic unilateral conjunctivitis, most probably linked to the visit of a South African wildlife reserve. Increased awareness among general and occupational physicians, ophthalmologists, clinicians, and the public on the potential of C. psittaci to cause ocular infection is needed. Full article
(This article belongs to the Special Issue Chlamydiae and Chlamydia like Bacteria)
Open AccessArticle
Occurrence and Function of the Na+-Translocating NADH:Quinone Oxidoreductase in Prevotella spp.
Microorganisms 2019, 7(5), 117; https://doi.org/10.3390/microorganisms7050117
Received: 5 February 2019 / Revised: 8 April 2019 / Accepted: 25 April 2019 / Published: 27 April 2019
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Abstract
Strictly anaerobic Prevotella spp. are characterized by their vast metabolic potential. As members of the Prevotellaceae family, they represent the most abundant organisms in the rumen and are typically found in monogastrics such as pigs and humans. Within their largely anoxic habitats, these [...] Read more.
Strictly anaerobic Prevotella spp. are characterized by their vast metabolic potential. As members of the Prevotellaceae family, they represent the most abundant organisms in the rumen and are typically found in monogastrics such as pigs and humans. Within their largely anoxic habitats, these bacteria are considered to rely primarily on fermentation for energy conservation. A recent study of the rumen microbiome identified multiple subunits of the Na+-translocating NADH:quinone oxidoreductase (NQR) belonging to different Prevotella spp. Commonly, the NQR is associated with biochemical energy generation by respiration. The existence of this Na+ pump in Prevotella spp. may indicate an important role for electrochemical Na+ gradients in their anaerobic metabolism. However, detailed information about the potential activity of the NQR in Prevotella spp. is not available. Here, the presence of a functioning NQR in the strictly anaerobic model organism P. bryantii B14 was verified by conducting mass spectrometric, biochemical, and kinetic experiments. Our findings propose that P. bryantii B14 and other Prevotella spp. retrieved from the rumen operate a respiratory NQR together with a fumarate reductase which suggests that these ruminal bacteria utilize a sodium motive force generated during respiratory NADH:fumarate oxidoreduction. Full article
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Open AccessArticle
Advanced Strategies for Food-Grade Protein Production: A New E. coli/Lactic Acid Bacteria Shuttle Vector for Improved Cloning and Food-Grade Expression
Microorganisms 2019, 7(5), 116; https://doi.org/10.3390/microorganisms7050116
Received: 30 March 2019 / Revised: 19 April 2019 / Accepted: 24 April 2019 / Published: 27 April 2019
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Abstract
Food-grade production of recombinant proteins in Gram-positive bacteria, especially in LAB (i.e., Lactococcus, Lactobacillus, and Streptococcus), is of great interest in the areas of recombinant enzyme production, industrial food fermentation, gene and metabolic engineering, as well as antigen delivery for oral [...] Read more.
Food-grade production of recombinant proteins in Gram-positive bacteria, especially in LAB (i.e., Lactococcus, Lactobacillus, and Streptococcus), is of great interest in the areas of recombinant enzyme production, industrial food fermentation, gene and metabolic engineering, as well as antigen delivery for oral vaccination. Food-grade expression relies on hosts generally considered as safe organisms and on clone selection not dependent on antibiotic markers, which limit the overall DNA manipulation workflow, as it can be carried out only in the expression host and not in E. coli. Moreover, many commercial expression vectors lack useful elements for protein purification. We constructed a “shuttle” vector containing a removable selective marker, which allows feasible cloning steps in E. coli and subsequent protein expression in LAB. In fact, the cassette can be easily excised from the selected recombinant plasmid, and the resulting marker-free vector transformed into the final LAB host. Further useful elements, as improved MCS, 6xHis-Tag, and thrombin cleavage site sequences were introduced. The resulting vector allows easy cloning in E. coli, can be quickly converted in a food-grade expression vector and harbors additional elements for improved recombinant protein purification. Overall, such features make the new vector an improved tool for food-grade expression. Full article
(This article belongs to the Special Issue Recombinant Protein Expression in Microorganisms)
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Open AccessArticle
Inhibiting Methanogenesis in Rumen Batch Cultures Did Not Increase the Recovery of Metabolic Hydrogen in Microbial Amino Acids
Microorganisms 2019, 7(5), 115; https://doi.org/10.3390/microorganisms7050115
Received: 29 March 2019 / Revised: 19 April 2019 / Accepted: 23 April 2019 / Published: 27 April 2019
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Abstract
There is an interest in controlling rumen methanogenesis as an opportunity to both decrease the emissions of greenhouse gases and improve the energy efficiency of rumen fermentation. However, the effects of inhibiting rumen methanogenesis on fermentation are incompletely understood even in in vitro [...] Read more.
There is an interest in controlling rumen methanogenesis as an opportunity to both decrease the emissions of greenhouse gases and improve the energy efficiency of rumen fermentation. However, the effects of inhibiting rumen methanogenesis on fermentation are incompletely understood even in in vitro rumen cultures, as the recovery of metabolic hydrogen ([H]) in the main fermentation products consistently decreases with methanogenesis inhibition, evidencing the existence of unaccounted [H] sinks. We hypothesized that inhibiting methanogenesis in rumen batch cultures would redirect [H] towards microbial amino acids (AA) biosynthesis as an alternative [H] sink to methane (CH4). The objective of this experiment was to evaluate the effects of eight inhibitors of methanogenesis on digestion, fermentation and the production of microbial biomass and AA in rumen batch cultures growing on cellulose. Changes in the microbial community composition were also studied using denaturing gradient gel electrophoresis (DGGE). Inhibiting methanogenesis did not cause consistent changes in fermentation or the profile of AA, although the effects caused by the different inhibitors generally associated with the changes in the microbial community that they induced. Under the conditions of this experiment, inhibiting methanogenesis did not increase the importance of microbial AA synthesis as a [H] sink. Full article
(This article belongs to the Section Systems Microbiology)
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Open AccessArticle
The Influence of Fungicide Treatments on Mycobiota of Grapes and Its Evolution During Fermentation Evaluated by Metagenomic and Culture-Dependent Methods
Microorganisms 2019, 7(5), 114; https://doi.org/10.3390/microorganisms7050114
Received: 1 April 2019 / Revised: 20 April 2019 / Accepted: 24 April 2019 / Published: 26 April 2019
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Abstract
The present study evaluated the impact of organic and conventional fungicide treatments compared with untreated samples (no fungicides were used) on the grape berry yeast community of the Montepulciano variety. The yeast dynamics during the spontaneous fermentation using culture-dependent and -independent methods was [...] Read more.
The present study evaluated the impact of organic and conventional fungicide treatments compared with untreated samples (no fungicides were used) on the grape berry yeast community of the Montepulciano variety. The yeast dynamics during the spontaneous fermentation using culture-dependent and -independent methods was also evaluated. Results showed a reduction of yeast biodiversity by conventional treatments determining a negative influence on fermenting yeasts in favor of oxidative yeasts such as Aerobasidium pullulans. Starmerella bacillaris was significantly more present in organic samples (detected by next generation sequencing (NGS)), while Hanseniaspopa uvarum was significantly less present in untreated samples (detected by the culture-dependent method). The fermenting yeasts, developed during the spontaneous fermentation, were differently present depending on the fungicide treatments used. Culture-dependent and -independent methods exhibited the same most abundant yeast species during the spontaneous fermentation but a different relative abundance. Differently, the NGS method was able to detect a greater biodiversity (lower abundant species) in comparison with the culture-dependent method. In this regard, the methodologies used gave a different picture of yeast dynamics during the fermentation process. The results indicated that the fungal treatments can influence the yeast community of grapes leading must fermentation and the final composition of wine. Full article
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Open AccessArticle
Nontuberculous Mycobacteria Persistence in a Cell Model Mimicking Alveolar Macrophages
Microorganisms 2019, 7(5), 113; https://doi.org/10.3390/microorganisms7050113
Received: 25 March 2019 / Revised: 22 April 2019 / Accepted: 24 April 2019 / Published: 26 April 2019
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Abstract
Nontuberculous Mycobacteria (NTM) respiratory infections have been gradually increasing. Here, THP-1 cells were used as a model to evaluate intracellular persistence of three NTM species (reference and clinical strains) in human alveolar macrophages. The contribution of phagosome acidification, nitric oxide (NO) production and [...] Read more.
Nontuberculous Mycobacteria (NTM) respiratory infections have been gradually increasing. Here, THP-1 cells were used as a model to evaluate intracellular persistence of three NTM species (reference and clinical strains) in human alveolar macrophages. The contribution of phagosome acidification, nitric oxide (NO) production and cell dead on NTM intracellular fate was assessed. In addition, strains were characterized regarding their repertoire of virulence factors by whole-genome sequencing. NTM experienced different intracellular fates: M. smegmatis and M. fortuitum ATCC 6841 were cleared within 24h. In contrast, M. avium strains (reference/clinical) and M. fortuitum clinical strain were able to replicate. Despite this fact, unexpectedly high percentages of acidified phagosomes were found harbouring rab7, but not CD63. All NTM were able to survive in vitro at acidic pHs, with the exception of M. smegmatis. Our data further suggested a minor role for NO in intracellular persistence and that apoptosis mediated by caspase 8 and 3/7, but not necrosis, is triggered during NTM infection. Insights regarding the bacteria genomic backbone corroborated the virulence potential of M. avium and M. fortuitum. In conclusion, the phenotypic traits detected contrast with those described for M. tuberculosis, pointing out that NTM adopt distinct strategies to manipulate the host immune defense and persist intracellularly. Full article
(This article belongs to the Special Issue Virulence Studies of Pathogenic Mycobacteria of Humans and Animal)
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