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Microorganisms, Volume 9, Issue 9 (September 2021) – 210 articles

Cover Story (view full-size image): We isolated the yeast Saccharomyces cerevisiae mutant with high levels of intracellular proline and found a missense mutation on the PRO1 gene encoding the Gln79His variant of γ-glutamyl kinase Pro1, a key enzyme that controls proline biosynthesis. Biochemical and in silico analyses of the Gln79 variants revealed that the amino acid residue at position 79 is involved in the stabilization of the proline-binding pocket in Pro1 via a hydrogen-bonding network, which plays an important role in feedback inhibition. The proposed mechanism can be applied to construct proline-accumulating yeast strains. View this paper.
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11 pages, 4630 KiB  
Article
Geographical Relationships between Long-Tailed Goral (Naemorhedus caudatus) Populations Based on Gut Microbiome Analysis
by Chang Eon Park, Bum-Joon Cho, Min-Ji Kim, Hee Cheon Park and Jae-Ho Shin
Microorganisms 2021, 9(9), 2002; https://doi.org/10.3390/microorganisms9092002 - 21 Sep 2021
Cited by 3 | Viewed by 3498
Abstract
The long-tailed goral (Naemorhedus caudatus) is an endangered species found in the mountains of eastern and northern Asia. Its populations have declined for various reasons, and this species has been designated as legally protected in South Korea. Although various ecological studies [...] Read more.
The long-tailed goral (Naemorhedus caudatus) is an endangered species found in the mountains of eastern and northern Asia. Its populations have declined for various reasons, and this species has been designated as legally protected in South Korea. Although various ecological studies have been conducted on long-tailed gorals, none have investigated the gut microbiome until now. In the present study, we compared the diversity and composition of the gut microbiome of seven populations of Korean long-tailed gorals. By analyzing the gut microbiome composition for each regional population, it was found that four phyla—Firmicutes, Actinobacteriota, Bacteroidota, and Proteobacteria—were the most dominant in all regions on average. The alpha diversity of the gut microbiome of the goral population in the northern regions was high, while that in the southern regions was low. Through the analysis of beta diversity, the seven long-tailed goral populations have been divided into three groups: the Seoraksan population, the Samcheock population, and the Wangpicheon population. It was possible to confirm the regional migration of the animals using the gut microbiome based on the site-relational network analysis. It was found that the most stable population of long-tailed gorals in Korea was the Seoraksan population, and the closely related groups were the Samcheok and Wangpicheon populations, respectively. Wangpicheon appeared to be a major point of dispersal in the migration route of Korean long-tailed gorals. Full article
(This article belongs to the Special Issue Rumen Microbial Communities)
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16 pages, 5150 KiB  
Article
Preclinical Development of a Bacteriophage Cocktail for Treating Multidrug Resistant Pseudomonas aeruginosa Infections
by Sophie Camens, Sha Liu, Karen Hon, George Spyro Bouras, Alkis James Psaltis, Peter-John Wormald and Sarah Vreugde
Microorganisms 2021, 9(9), 2001; https://doi.org/10.3390/microorganisms9092001 - 21 Sep 2021
Cited by 10 | Viewed by 3403
Abstract
A Pseudomonas aeruginosa (P. aeruginosa) airway infection is one of the predominant causes contributing to the high morbidity and mortality rates in cystic fibrosis (CF) patients. The emergence of antibiotic resistant P. aeruginosa strains has led to an urgent need for [...] Read more.
A Pseudomonas aeruginosa (P. aeruginosa) airway infection is one of the predominant causes contributing to the high morbidity and mortality rates in cystic fibrosis (CF) patients. The emergence of antibiotic resistant P. aeruginosa strains has led to an urgent need for new therapeutic approaches. Bacteriophages (phages) are viruses that can infect and lyse specific bacteria, providing a potential alternative approach in targeting antibiotic-resistant strains. We aim to isolate and characterise novel P. aeruginosa phages for combination in a cocktail to kill P. aeruginosa. One particular phage, PA4, could lyse 14/20 clinical isolates as observed through spot assays. This phage could significantly reduce the growth of bacteria in vitro, as determined through planktonic adsorption and inhibition assays as well as crystal violet- and LIVE/DEAD-stained biofilm assays. A morphological and genomic analysis revealed that PA4 belongs to the Myoviridae family and contained 66,450 bp. The broad infectivity profile, good stability in various pH and temperature conditions, lytic ability and the absence of the absences of antibiotic resistance, toxic and lysogenic genes suggest that PA4 is a good candidate for clinical grade use. Overall, phage therapy represents a promising alternative treatment option to antibiotics when treating a P. aeruginosa infection. Full article
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14 pages, 1343 KiB  
Article
Biofertilizer Activity of Azospirillum sp. B510 on the Rice Productivity in Ghana
by Elsie Sarkodee-Addo, Chihiro Tokiwa, Patrick Bonney, Daniel Asiamah Aboagye, Alex Yeboah, Samuel Oppong Abebrese, Ralph Bam, Eric Kwesi Nartey, Shin Okazaki and Michiko Yasuda
Microorganisms 2021, 9(9), 2000; https://doi.org/10.3390/microorganisms9092000 - 21 Sep 2021
Cited by 1 | Viewed by 3227
Abstract
Rice production in Ghana has become unsustainable due to the extremely nutrient-poor soils. It is caused by inadequate soil fertility management, including the inefficient application of fertilizers. A practical solution could be the biofertilizers, Azospirillum sp. B510. We performed field trials in Ghana [...] Read more.
Rice production in Ghana has become unsustainable due to the extremely nutrient-poor soils. It is caused by inadequate soil fertility management, including the inefficient application of fertilizers. A practical solution could be the biofertilizers, Azospirillum sp. B510. We performed field trials in Ghana and Japan to compare the effects of B510 colonization on selected Ghanaian rice varieties grown. The B510 inoculation significantly enhanced the rice cultivars’ growth and yield. The phenotypic characteristics observed in rice varieties Exbaika, Ex-Boako, AgraRice, and Amankwatia were mainly short length and high tillering capacity. These features are attributed to the host plant (cv. Nipponbare), from which the strain B510 was isolated. Furthermore, Azospirillum species has been identified as the dominant colonizing bacterium of rice rhizosphere across a diverse range of agroecologies in all major rice-growing regions in Ghana. Our results suggest that the utilization of B510 as a bio-fertilizer presents a promising way to improve rice growth, enhance soil fertility, and sustain rice productivity in Ghana. Full article
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15 pages, 1569 KiB  
Article
Re-Emergence and Spread of Haemorrhagic Septicaemia in Germany: The Wolf as a Vector?
by Peter Kutzer, Claudia A. Szentiks, Sabine Bock, Guido Fritsch, Tibor Magyar, Christoph Schulze, Torsten Semmler and Christa Ewers
Microorganisms 2021, 9(9), 1999; https://doi.org/10.3390/microorganisms9091999 - 21 Sep 2021
Cited by 3 | Viewed by 4792
Abstract
Since 2010, outbreaks of haemorrhagic septicaemia (HS) caused by Pasteurella (P.) multocida capsular type B (PmB) emerged in Germany. In 2017, we noticed a close spatiotemporal relationship between HS outbreak sites and wolf (Canis lupus) territories. Thus, [...] Read more.
Since 2010, outbreaks of haemorrhagic septicaemia (HS) caused by Pasteurella (P.) multocida capsular type B (PmB) emerged in Germany. In 2017, we noticed a close spatiotemporal relationship between HS outbreak sites and wolf (Canis lupus) territories. Thus, the main objectives of our study were to investigate the molecular epidemiology of German PmB-HS-isolates and to assess the role of wolves as putative vectors of this pathogen. We collected 83 PmB isolates from HS outbreaks that occurred between 2010 and 2019 and sampled 150 wolves, which were found dead in the years 2017 to 2019, revealing another three PmB isolates. A maximum-likelihood-based phylogeny of the core genomes of 65 PmB-HS-isolates and the three PmB-wolf-isolates showed high relatedness. Furthermore, all belonged to capsular:LPS:MLST genotype B:L2:ST122RIRDC and showed highly similar virulence gene profiles, but clustered separately from 35 global ST122RIRDC strains. Our data revealed that German HS outbreaks were caused by a distinct genomic lineage of PmB-ST122 strains, hinting towards an independent, ongoing epidemiologic event. We demonstrated for the first time, that carnivores, i.e., wolves, might harbour PmB as a part of their oropharyngeal microbiota. Furthermore, the results of our study imply that wolves can carry the pathogen over long distances, indicating a major role of that animal species in the ongoing epidemiological event of HS in Germany. Full article
(This article belongs to the Topic Veterinary Infectious Diseases)
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10 pages, 2677 KiB  
Article
A Mouse Model for Studying Post-Acute Arthritis of Chikungunya
by Aileen Y. Chang, Sarah R. Tritsch, Abigail J. Porzucek, Arnold M. Schwartz, Margaux Seyler-Schmidt, Arielle Glass, Patricia S. Latham, St. Patrick Reid, Gary L. Simon and Christopher N. Mores
Microorganisms 2021, 9(9), 1998; https://doi.org/10.3390/microorganisms9091998 - 21 Sep 2021
Cited by 3 | Viewed by 2512
Abstract
Chikungunya virus (CHIKV) was introduced to the Americas in 2013, causing two million infections across over thirty countries. CHIKV causes a chronic debilitating arthritis in one fourth of infected individuals and currently evidence-based targeted therapies for the treatment of CHIKV arthritis are lacking. [...] Read more.
Chikungunya virus (CHIKV) was introduced to the Americas in 2013, causing two million infections across over thirty countries. CHIKV causes a chronic debilitating arthritis in one fourth of infected individuals and currently evidence-based targeted therapies for the treatment of CHIKV arthritis are lacking. Multiple mouse models of chikungunya have been developed to study acute CHIKV infection. In humans, post-CHIKV arthritis may persist for months to years after viremia from a CHIKV infection has resolved. Therefore, the development of a mouse model of post-acute arthritis of chikungunya may facilitate the study of potential novel therapeutics for this arthritis. In this article we describe the development of a wild-type immunocompetent C57BL/6 mouse model for post-acute arthritis of chikungunya, including a histologic inflammation scoring system, as well as suggestions for how this mouse model may be used to examine the efficacy of novel therapies for CHIKV arthritis. Full article
(This article belongs to the Special Issue Emerging Alphaviruses)
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14 pages, 506 KiB  
Article
Risk of Hemolytic Uremic Syndrome Related to Treatment of Escherichia coli O157 Infection with Different Antimicrobial Classes
by Rajal K. Mody, Robert M. Hoekstra, Magdalena Kendall Scott, John Dunn, Kirk Smith, Melissa Tobin-D’Angelo, Beletshachew Shiferaw, Katie Wymore, Paula Clogher, Amanda Palmer, Nicole Comstock, Kari Burzlaff, Sarah Lathrop, Sharon Hurd and Patricia M. Griffin
Microorganisms 2021, 9(9), 1997; https://doi.org/10.3390/microorganisms9091997 - 21 Sep 2021
Cited by 4 | Viewed by 3328
Abstract
Treatment of Shiga toxin-producing Escherichia coli O157 (O157) diarrhea with antimicrobials might alter the risk of hemolytic uremic syndrome (HUS). However, full characterization of which antimicrobials might affect risk is lacking, particularly among adults. To inform clinical management, we conducted a case-control study [...] Read more.
Treatment of Shiga toxin-producing Escherichia coli O157 (O157) diarrhea with antimicrobials might alter the risk of hemolytic uremic syndrome (HUS). However, full characterization of which antimicrobials might affect risk is lacking, particularly among adults. To inform clinical management, we conducted a case-control study of residents of the FoodNet surveillance areas with O157 diarrhea during a 4-year period to assess antimicrobial class-specific associations with HUS among persons with O157 diarrhea. We collected data from medical records and patient interviews. We measured associations between treatment with agents in specific antimicrobial classes during the first week of diarrhea and development of HUS, adjusting for age and illness severity. We enrolled 1308 patients; 102 (7.8%) developed confirmed HUS. Antimicrobial treatment varied by age: <5 years (12.6%), 5–14 (11.5%), 15–39 (45.4%), ≥40 (53.4%). Persons treated with a β-lactam had higher odds of developing HUS (OR 2.80, CI 1.14–6.89). None of the few persons treated with a macrolide developed HUS, but the protective association was not statistically significant. Exposure to “any antimicrobial” was not associated with increased odds of HUS. Our findings confirm the risk of β-lactams among children with O157 diarrhea and extends it to adults. We observed a high frequency of inappropriate antimicrobial treatment among adults. Our data suggest that antimicrobial classes differ in the magnitude of risk for persons with O157 diarrhea. Full article
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16 pages, 3594 KiB  
Article
Effects of Mixing Volatile Fatty Acids as Carbon Sources on Rhodospirillum rubrum Carbon Metabolism and Redox Balance Mechanisms
by Paloma Cabecas Segura, Quentin De Meur, Audrey Tanghe, Rob Onderwater, Laurent Dewasme, Ruddy Wattiez and Baptiste Leroy
Microorganisms 2021, 9(9), 1996; https://doi.org/10.3390/microorganisms9091996 - 21 Sep 2021
Cited by 7 | Viewed by 2156
Abstract
Rhodospirillum rubrum has a versatile metabolism, and as such can assimilate a broad range of carbon sources, including volatile fatty acids. These carbon sources are gaining increasing interest for biotechnological processes, since they reduce the production costs for numerous value-added compounds and contribute [...] Read more.
Rhodospirillum rubrum has a versatile metabolism, and as such can assimilate a broad range of carbon sources, including volatile fatty acids. These carbon sources are gaining increasing interest for biotechnological processes, since they reduce the production costs for numerous value-added compounds and contribute to the development of a more circular economy. Usually, studies characterizing carbon metabolism are performed by supplying a single carbon source; however, in both environmental and engineered conditions, cells would rather grow on mixtures of volatile fatty acids (VFAs) generated via anaerobic fermentation. In this study, we show that the use of a mixture of VFAs as carbon source appears to have a synergy effect on growth phenotype. In addition, while propionate and butyrate assimilation in Rs. rubrum is known to require an excess of bicarbonate in the culture medium, mixing them reduces the requirement for bicarbonate supplementation. The fixation of CO2 is one of the main electron sinks in purple bacteria; therefore, this observation suggests an adaptation of both metabolic pathways used for the assimilation of these VFAs and redox homeostasis mechanism. Based on proteomic data, modification of the propionate assimilation pathway seems to occur with a switch from a methylmalonyl-CoA intermediate to the methylcitrate cycle. Moreover, it seems that the presence of a mixture of VFAs switches electron sinking from CO2 fixation to H2 and isoleucine production. Full article
(This article belongs to the Special Issue Biotechnological Application of Photosynthetic Bacteria)
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18 pages, 2791 KiB  
Article
The Species-Level Composition of the Fecal Bifidobacterium and Lactobacillus Genera in Indonesian Children Differs from That of Their Mothers
by Mengfan Ding, Bo Yang, Wei Wei Thwe Khine, Yuan-Kun Lee, Endang Sutriswati Rahayu, R. Paul Ross, Catherine Stanton, Jianxin Zhao, Hao Zhang and Wei Chen
Microorganisms 2021, 9(9), 1995; https://doi.org/10.3390/microorganisms9091995 - 21 Sep 2021
Cited by 8 | Viewed by 2568
Abstract
The infant gut microbiota plays a critical role in early life growth and derives mainly from maternal gut and breast milk. This study aimed to analyze the differences in the gut microbiota, namely Bifidobacterium and Lactobacillus communities at species level among breast milk [...] Read more.
The infant gut microbiota plays a critical role in early life growth and derives mainly from maternal gut and breast milk. This study aimed to analyze the differences in the gut microbiota, namely Bifidobacterium and Lactobacillus communities at species level among breast milk as well as maternal and infant feces at different time points after delivery. Fifty-one mother–infant pairs from Indonesia were recruited, and the breast milk and maternal and infant feces were collected and analyzed by high throughput sequencing (16S rRNA, Bifidobacterium groEL and Lactobacillus groEL genes). PCoA results showed bacterial composition was different among breast milk and maternal and infant feces within the first two years. The abundance of Bifidobacterium and Bacteroides were significantly higher in infant feces compared to their maternal feces from birth to two years of age, and maternal breast milk within six months after birth (p < 0.05), whereas the abundance of Blautia, Prevotella, and Faecalibacterium was higher in maternal feces compared to that in breast milk within six months and infant feces within one year after birth, respectively (p < 0.05). The relative abundances of Bacteroides and Lactobacillus was higher and lower in infant feces compared to that in maternal feces only between one and two years of age, respectively (p < 0.05). For Bifidobacterium community at species level, B. adolescentis, B. ruminantium, B. longum subsp. infantis, B. bifidum, and B. pseudolongum were identified in all samples. However, the profile of Bifidobacterium was different between maternal and infant feces at different ages. The relative abundances of B. adolescentis and B. ruminantium were higher in maternal feces compared to those in infant feces from birth to one year of age (p < 0.05), while the relative abundances of B. longum subsp. infantis and B. bifidum were higher in infant feces compared to those in maternal feces beyond three months, and the relative abundance of B. pseudolongum was only higher in infant feces between three and six months (p < 0.05). For Lactobacillus community, L. paragasseri showed higher relative abundance in infant feces when the infant was younger than one year of age (p < 0.05). This study showed bacterial composition at the genus level and Bifidobacterium and Lactobacillus communities at the species level were stage specific in maternal breast milk as well as and maternal and infant feces. Full article
(This article belongs to the Special Issue Gut Microbiota and Metabolism in Different Stages of Life and Health)
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11 pages, 20823 KiB  
Article
Composition and Corrosivity of Extracellular Polymeric Substances from the Hydrocarbon-Degrading Sulfate-Reducing Bacterium Desulfoglaeba alkanexedens ALDC
by Irene A. Davidova, Tiffany R. Lenhart, Mark A. Nanny and Joseph M. Suflita
Microorganisms 2021, 9(9), 1994; https://doi.org/10.3390/microorganisms9091994 - 21 Sep 2021
Cited by 3 | Viewed by 1820
Abstract
Sulfate-reducing bacteria (SRB) often exist as cell aggregates and in biofilms surrounded by a matrix of extracellular polymeric substances (EPSs). The chemical composition of EPSs may facilitate hydrophobic substrate biodegradation and promote microbial influenced corrosion (MIC). Although EPSs from non-hydrocarbon-degrading SRB have been [...] Read more.
Sulfate-reducing bacteria (SRB) often exist as cell aggregates and in biofilms surrounded by a matrix of extracellular polymeric substances (EPSs). The chemical composition of EPSs may facilitate hydrophobic substrate biodegradation and promote microbial influenced corrosion (MIC). Although EPSs from non-hydrocarbon-degrading SRB have been studied; the chemical composition of EPSs from hydrocarbon-degrading SRBs has not been reported. The isolated EPSs from the sulfate-reducing alkane-degrading bacterium Desulfoglaeba alkanexedens ALDC was characterized with scanning and fluorescent microscopy, nuclear magnetic resonance spectroscopy (NMR), and by colorimetric chemical assays. Specific fluorescent staining and 1H NMR spectroscopy revealed that the fundamental chemical structure of the EPS produced by D. alkanexedens is composed of pyranose polysaccharide and cyclopentanone in a 2:1 ratio. NMR analyses indicated that the pyranose ring structure is bonded by 1,4 connections with the cyclopentanone directly bonded to one pyranose ring. The presence of cyclopentanone presumably increases the hydrophobicity of the EPS that may facilitate the accessibility of hydrocarbon substrates to aggregating cells or cells in a biofilm. Weight loss and iron dissolution experiments demonstrated that the EPS did not contribute to the corrosivity of D. alkanexedens cells. Full article
(This article belongs to the Special Issue Petroleum Microbiology)
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34 pages, 1955 KiB  
Review
A Selection of Platforms to Evaluate Surface Adhesion and Biofilm Formation in Controlled Hydrodynamic Conditions
by Luciana C. Gomes and Filipe J. M. Mergulhão
Microorganisms 2021, 9(9), 1993; https://doi.org/10.3390/microorganisms9091993 - 21 Sep 2021
Cited by 20 | Viewed by 3595
Abstract
The early colonization of surfaces and subsequent biofilm development have severe impacts in environmental, industrial, and biomedical settings since they entail high costs and health risks. To develop more effective biofilm control strategies, there is a need to obtain laboratory biofilms that resemble [...] Read more.
The early colonization of surfaces and subsequent biofilm development have severe impacts in environmental, industrial, and biomedical settings since they entail high costs and health risks. To develop more effective biofilm control strategies, there is a need to obtain laboratory biofilms that resemble those found in natural or man-made settings. Since microbial adhesion and biofilm formation are strongly affected by hydrodynamics, the knowledge of flow characteristics in different marine, food processing, and medical device locations is essential. Once the hydrodynamic conditions are known, platforms for cell adhesion and biofilm formation should be selected and operated, in order to obtain reproducible biofilms that mimic those found in target scenarios. This review focuses on the most widely used platforms that enable the study of initial microbial adhesion and biofilm formation under controlled hydrodynamic conditions—modified Robbins devices, flow chambers, rotating biofilm devices, microplates, and microfluidic devices—and where numerical simulations have been used to define relevant flow characteristics, namely the shear stress and shear rate. Full article
(This article belongs to the Special Issue Microbial Films-the Interplay of Physics and Biology)
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23 pages, 17695 KiB  
Article
Comparative Genomics and Specific Functional Characteristics Analysis of Lactobacillus acidophilus
by Zheng Huang, Xingya Zhou, Catherine Stanton, Reynolds Paul Ross, Jianxin Zhao, Hao Zhang, Bo Yang and Wei Chen
Microorganisms 2021, 9(9), 1992; https://doi.org/10.3390/microorganisms9091992 - 20 Sep 2021
Cited by 21 | Viewed by 4542
Abstract
Lactobacillus acidophilus is a common kind of lactic acid bacteria usually found in the human gastrointestinal tract, oral cavity, vagina, and various fermented foods. At present, many studies have focused on the probiotic function and industrial application of L. acidophilus. Additionally, dozens [...] Read more.
Lactobacillus acidophilus is a common kind of lactic acid bacteria usually found in the human gastrointestinal tract, oral cavity, vagina, and various fermented foods. At present, many studies have focused on the probiotic function and industrial application of L. acidophilus. Additionally, dozens of L. acidophilus strains have been genome sequenced, but there has been no research to compare them at the genomic level. In this study, 46 strains of L. acidophilus were performed comparative analyses to explore their genetic diversity. The results showed that all the L. acidophilus strains were divided into two clusters based on ANI values, phylogenetic analysis and whole genome comparison, due to the difference of their predicted gene composition of bacteriocin operon, CRISPR-Cas systems and prophages mainly. Additionally, L. acidophilus was a pan-genome open species with a difference in carbohydrates utilization, antibiotic resistance, EPS operon, surface layer protein operon and other functional gene composition. This work provides a better understanding of L. acidophilus from a genetic perspective, and offers a frame for the biotechnological potentiality of this species. Full article
(This article belongs to the Section Systems Microbiology)
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12 pages, 1750 KiB  
Article
Chagas Disease-Related Mortality in Spain, 1997 to 2018
by Jose-Manuel Ramos-Rincon, Jara Llenas-García, Hector Pinargote-Celorio, Veronica Sánchez-García, Philip Wikman-Jorgensen, Miriam Navarro, Concepción Gil-Anguita, Violeta Ramos-Sesma and Diego Torrus-Tendero
Microorganisms 2021, 9(9), 1991; https://doi.org/10.3390/microorganisms9091991 - 20 Sep 2021
Cited by 4 | Viewed by 1766
Abstract
Background. Chagas disease (CD) is associated with excess mortality in infected people in endemic countries, but little information is available in non-endemic countries. The aim of the study was to analyze mortality in patients admitted to the hospital with CD in Spain. Methods. [...] Read more.
Background. Chagas disease (CD) is associated with excess mortality in infected people in endemic countries, but little information is available in non-endemic countries. The aim of the study was to analyze mortality in patients admitted to the hospital with CD in Spain. Methods. A retrospective, observational study using the Spanish National Hospital Discharge Database. We used the CD diagnostic codes of the 9th and 10th International Classification of Diseases to retrieve CD cases from the national public registry from 1997 to 2018. Results. Of the 5022 hospital admissions in people with CD, there were 56 deaths (case fatality rate (CFR) 1.1%, 95% confidence interval (CI) 0.8%, 1.4%), 20 (35.7%) of which were considered directly related to CD. The median age was higher in those who died (54.5 vs. 38 years; p < 0.001). The CFR increased with age, peaking in the 70–79-year (7.9%, odds ratio (OR) 6.27, 95% CI 1.27, 30.90) and 80–89-year (16.7%, OR 14.7, 95% CI 2.70, 79.90) age groups. Men comprised a higher proportion of those who died compared to survivors (50% vs. 22.6%; p < 0.001). Non-survivors were more likely to have neoplasms (19.6% vs. 3.4%; p < 0.001), heart failure (17.9% vs. 7.2%; p = 0.002), diabetes (12.5% vs. 3.7%; p = 0.001), chronic kidney failure (8.9% vs. 1.6%; p < 0.001), and HIV (8.9% vs. 0.8%; p < 0.001). In the multivariable analysis, the variables associated with mortality were age (adjusted OR (aOR) 1.05; 95% CI: 1.03, 1.07), male sex (aOR 1.79, 95% CI 1.03, 3.14), cancer (aOR: 4.84, 95% CI 2.13, 11.22), and HIV infection (aOR 14.10 95% CI 4.88, 40.73). Conclusions. The case fatality rate of CD hospitalization was about 1%. The mortality risk increased with age, male sex, cancer, and HIV infection. Full article
(This article belongs to the Special Issue Chagas Disease (American Trypanosomiasis))
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16 pages, 3863 KiB  
Article
Enhancing Teak (Tectona grandis) Seedling Growth by Rhizosphere Microbes: A Sustainable Way to Optimize Agroforestry
by Leardwiriyakool Chaiya, Paiboolya Gavinlertvatana, Neung Teaumroong, Wasu Pathom-aree, Amornrat Chaiyasen, Rungroch Sungthong and Saisamorn Lumyong
Microorganisms 2021, 9(9), 1990; https://doi.org/10.3390/microorganisms9091990 - 19 Sep 2021
Cited by 7 | Viewed by 2920
Abstract
With its premium wood quality and resistance to pests, teak is a valuable tree species remarkably required for timber trading and agroforestry. The nursery stage of teak plantation needs critical care to warrant its long-term productivity. This study aimed to search for beneficial [...] Read more.
With its premium wood quality and resistance to pests, teak is a valuable tree species remarkably required for timber trading and agroforestry. The nursery stage of teak plantation needs critical care to warrant its long-term productivity. This study aimed to search for beneficial teak rhizosphere microbes and assess their teak-growth-promoting potentials during nursery stock preparation. Three teak rhizosphere/root-associated microbes, including two teak rhizobacteria (a nitrogen-fixing teak root endophyte-Agrobacterium sp. CGC-5 and a teak rhizosphere actinobacterium-Kitasatospora sp. TCM1-050) and an arbuscular mycorrhizal fungus (Claroideoglomus sp. PBT03), were isolated and used in this study. Both teak rhizobacteria could produce in vitro phytohormones (auxins) and catalase. With the pot-scale assessments, applying these rhizosphere microbes in the form of consortia offered better teak-growth-promoting activities than the individual applications, supported by significantly increased teak seedling biomass. Moreover, teak-growth-promoting roles of the arbuscular mycorrhizal fungus were highly dependent upon the support by other teak rhizobacteria. Based on our findings, establishing the synergistic interactions between beneficial rhizosphere microbes and teak roots was a promising sustainable strategy to enhance teak growth and development at the nursery stage and reduce chemical inputs in agroforestry. Full article
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14 pages, 3904 KiB  
Article
Application of Fungus Enzymes in Spent Mushroom Composts from Edible Mushroom Cultivation for Phthalate Removal
by Bea-Ven Chang, Chiao-Po Yang and Chu-Wen Yang
Microorganisms 2021, 9(9), 1989; https://doi.org/10.3390/microorganisms9091989 - 19 Sep 2021
Cited by 12 | Viewed by 3348
Abstract
Spent mushroom composts (SMCs) are waste products of mushroom cultivation. The handling of large amounts of SMCs has become an important environmental issue. Phthalates are plasticizers which are widely distributed in the environment and urban wastewater, and cannot be effectively removed by conventional [...] Read more.
Spent mushroom composts (SMCs) are waste products of mushroom cultivation. The handling of large amounts of SMCs has become an important environmental issue. Phthalates are plasticizers which are widely distributed in the environment and urban wastewater, and cannot be effectively removed by conventional wastewater treatment methods. In this study, SMCs are tested for their ability to remove phthalates, including benzyl butyl phthalate (BBP), di-n-butyl phthalate (DBP), and diethyl phthalate (DEP). Batch experiments reveal that BBP, DBP, and DEP can be degraded by the SMC enzyme extracts of four edible mushrooms: Pleurotus eryngii, Pleurotus djamor, Pleurotus ostreatus, and Auricularia polytricha. Potential fungus enzymes associated with BBP, DBP, and DEP degradation in SMCs (i.e., esterases, oxygenases, and oxidases/dehydrogenases) are uncovered by metaproteomic analysis using mass spectrometry. Bioreactor experiments indicate that the direct application of SMCs can remove BBP, DBP, and DEP from wastewater, through adsorption and biodegradation. The results of this study extend the application of white-rot fungi without laccases (e.g., Auricularia sp.) for the removal of organic pollutants which are not degraded by laccases. The application of SMCs for phthalate removal can be developed into a mycoremediation-based green and sustainable technology. Full article
(This article belongs to the Special Issue Fungal Biodiversity for Bioremediation)
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18 pages, 882 KiB  
Review
Pathogen Biocontrol Using Plant Growth-Promoting Bacteria (PGPR): Role of Bacterial Diversity
by Hao Wang, Runjin Liu, Ming Pei You, Martin J. Barbetti and Yinglong Chen
Microorganisms 2021, 9(9), 1988; https://doi.org/10.3390/microorganisms9091988 - 18 Sep 2021
Cited by 71 | Viewed by 8809
Abstract
A vast microbial community inhabits in the rhizosphere, among which, specialized bacteria known as Plant Growth-Promoting Rhizobacteria (PGPR) confer benefits to host plants including growth promotion and disease suppression. PGPR taxa vary in the ways whereby they curtail the negative effects of invading [...] Read more.
A vast microbial community inhabits in the rhizosphere, among which, specialized bacteria known as Plant Growth-Promoting Rhizobacteria (PGPR) confer benefits to host plants including growth promotion and disease suppression. PGPR taxa vary in the ways whereby they curtail the negative effects of invading plant pathogens. However, a cumulative or synergistic effect does not always ensue when a bacterial consortium is used. In this review, we reassess the disease-suppressive mechanisms of PGPR and present explanations and illustrations for functional diversity and/or stability among PGPR taxa regarding these mechanisms. We also provide evidence of benefits when PGPR mixtures, rather than individuals, are used for protecting crops from various diseases, and underscore the critical determinant factors for successful use of PGPR mixtures. Then, we evaluate the challenges of and limitations to achieving the desired outcomes from strain/species-rich bacterial assemblages, particularly in relation to their role for plant disease management. In addition, towards locating additive or synergistic outcomes, we highlight why and how the benefits conferred need to be categorized and quantified when different strains/species of PGPR are used in combinations. Finally, we highlight the critical approaches needed for developing PGPR mixtures with improved efficacy and stability as biocontrols for utilization in agricultural fields. Full article
(This article belongs to the Special Issue Microbial Interactions in Soil)
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18 pages, 1799 KiB  
Article
Wastewater from the Edible Oil Industry as a Potential Source of Lipase- and Surfactant-Producing Actinobacteria
by Pamela Welz, Gustav Swanepoel, Shandré Weels and Marilize Le Roes-Hill
Microorganisms 2021, 9(9), 1987; https://doi.org/10.3390/microorganisms9091987 - 18 Sep 2021
Cited by 10 | Viewed by 2399
Abstract
Wastewaters generated from various stages of edible oil production in a canola processing facility were collected with the aim of determining the presence of lipase-producing actinobacteria of potential industrial significance. The high chemical oxygen demand (COD) readings (up to 86,700 mg L−1 [...] Read more.
Wastewaters generated from various stages of edible oil production in a canola processing facility were collected with the aim of determining the presence of lipase-producing actinobacteria of potential industrial significance. The high chemical oxygen demand (COD) readings (up to 86,700 mg L−1 in some samples) indicated that the wastewater exhibited the nutritional potential to support bacterial growth. A novel approach was developed for the isolation of metagenomic DNA from the oil-rich wastewater samples. Microbiota analysis of the buffer tank and refinery condensate tank wastewater samples showed a dominance of Cutibacterium acnes subsp. defendens, followed by a limited number of other actinobacterial genera, indicating the presence of a highly specialized actinobacterial population. Cultured isolates with typical actinobacterial morphology were analyzed for their ability to produce lipases and biosurfactants. Two strains, designated as BT3 and BT4, exhibited the highest lipase production levels when grown in the presence of tributyrin and olive oil (1.39 U mg−1 crude protein and 0.8 U mg−1 crude protein, respectively) and were subsequently definitively identified by genome sequencing to be related to Streptomyces albidoflavus. Cultivation of the strains in media containing different types of oils did not markedly increase the level of enzyme production, with the exception of strain BT4 (1.0 U mg−1 crude protein in the presence of peanut oil). Genome sequencing of the two strains, BT3 and BT4, revealed the presence of a range of lipase and esterase genes that may be involved in the production of the enzymes detected in this study. The presence of gene clusters involved in the production of biosurfactants were also detected, notably moreso in strain BT3 than BT4. Full article
(This article belongs to the Section Microbial Biotechnology)
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16 pages, 2918 KiB  
Article
Anaerobic Fungal Mevalonate Pathway Genomic Biases Lead to Heterologous Toxicity Underpredicted by Codon Adaptation Indices
by Ethan T. Hillman, Elizabeth M. Frazier, Evan K. Shank, Adrian N. Ortiz-Velez, Jacob A. Englaender and Kevin V. Solomon
Microorganisms 2021, 9(9), 1986; https://doi.org/10.3390/microorganisms9091986 - 18 Sep 2021
Cited by 4 | Viewed by 2225
Abstract
Anaerobic fungi are emerging biotechnology platforms with genomes rich in biosynthetic potential. Yet, the heterologous expression of their biosynthetic pathways has had limited success in model hosts like E. coli. We find one reason for this is that the genome composition of [...] Read more.
Anaerobic fungi are emerging biotechnology platforms with genomes rich in biosynthetic potential. Yet, the heterologous expression of their biosynthetic pathways has had limited success in model hosts like E. coli. We find one reason for this is that the genome composition of anaerobic fungi like P. indianae are extremely AT-biased with a particular preference for rare and semi-rare AT-rich tRNAs in E coli, which are not explicitly predicted by standard codon adaptation indices (CAI). Native P. indianae genes with these extreme biases create drastic growth defects in E. coli (up to 69% reduction in growth), which is not seen in genes from other organisms with similar CAIs. However, codon optimization rescues growth, allowing for gene evaluation. In this manner, we demonstrate that anaerobic fungal homologs such as PI.atoB are more active than S. cerevisiae homologs in a hybrid pathway, increasing the production of mevalonate up to 2.5 g/L (more than two-fold) and reducing waste carbon to acetate by ~90% under the conditions tested. This work demonstrates the bioproduction potential of anaerobic fungal enzyme homologs and how the analysis of codon utilization enables the study of otherwise difficult to express genes that have applications in biocatalysis and natural product discovery. Full article
(This article belongs to the Special Issue Unleashing the Hidden Potential of Anaerobic Fungi)
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13 pages, 959 KiB  
Article
Whole-Genome Sequencing (WGS) of Carbapenem-Resistant K. pneumoniae Isolated in Long-Term Care Facilities in the Northern Italian Region
by Alessandra Piccirilli, Sabrina Cherubini, Anna Maria Azzini, Evelina Tacconelli, Giuliana Lo Cascio, Laura Maccacaro, Alda Bazaj, Laura Naso, Gianfranco Amicosante, LTCF-Veneto Working Group and Mariagrazia Perilli
Microorganisms 2021, 9(9), 1985; https://doi.org/10.3390/microorganisms9091985 - 17 Sep 2021
Cited by 16 | Viewed by 2628
Abstract
K. pneumoniae (KPN) is one of the widest spread bacteria in which combined resistance to several antimicrobial groups is frequent. The most common β-lactamases found in K. pneumoniae are class A carbapenemases, both chromosomal-encoded (i.e., NMCA, IMI-1) and plasmid-encoded (i.e., GES-enzymes, IMI-2), VIM, [...] Read more.
K. pneumoniae (KPN) is one of the widest spread bacteria in which combined resistance to several antimicrobial groups is frequent. The most common β-lactamases found in K. pneumoniae are class A carbapenemases, both chromosomal-encoded (i.e., NMCA, IMI-1) and plasmid-encoded (i.e., GES-enzymes, IMI-2), VIM, IMP, NDM, OXA-48, and extended-spectrum β-lactamases (ESBLs) such as CTX-M enzymes. In the present study, a total of 68 carbapenem-resistant KPN were collected from twelve long-term care facilities (LTCFs) in the Northern Italian region. The whole-genome sequencing (WGS) of each KPN strain was determined using a MiSeq Illumina sequencing platform and analysed by a bacterial analysis pipeline (BAP) tool. The WGS analysis showed the prevalence of ST307, ST512, and ST37 as major lineages diffused among the twelve LTCFs. The other lineages found were: ST11, ST16, ST35, ST253, ST273, ST321, ST416, ST1519, ST2623, and ST3227. The blaKPC-2, blaKPC-3, blaKPC-9, blaSHV-11, blaSHV-28, blaCTX-M-15, blaOXA-1, blaOXA-9, blaOXA-23, qnrS1, qnrB19, qnrB66, aac(6′)-Ib-cr, and fosA were the resistance genes widespread in most LTCFs. In this study, we demonstrated the spreading of thirteen KPN lineages among the LTCFs. Additionally, KPC carbapenemases are the most widespread β-lactamase. Full article
(This article belongs to the Special Issue ß-Lactamases)
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19 pages, 2230 KiB  
Review
Sugarcane Mosaic Disease: Characteristics, Identification and Control
by Guilong Lu, Zhoutao Wang, Fu Xu, Yong-Bao Pan, Michael P. Grisham and Liping Xu
Microorganisms 2021, 9(9), 1984; https://doi.org/10.3390/microorganisms9091984 - 17 Sep 2021
Cited by 22 | Viewed by 10898
Abstract
Mosaic is one of the most important sugarcane diseases, caused by single or compound infection of Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), and/or Sugarcane streak mosaic virus (SCSMV). The compound infection of mosaic has become increasingly serious in the last few [...] Read more.
Mosaic is one of the most important sugarcane diseases, caused by single or compound infection of Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), and/or Sugarcane streak mosaic virus (SCSMV). The compound infection of mosaic has become increasingly serious in the last few years. The disease directly affects the photosynthesis and growth of sugarcane, leading to a significant decrease in cane yield and sucrose content, and thus serious economic losses. This review covers four aspects of sugarcane mosaic disease management: first, the current situation of sugarcane mosaic disease and its epidemic characteristics; second, the pathogenicity and genetic diversity of the three viruses; third, the identification methods of mosaic and its pathogen species; and fourth, the prevention and control measures for sugarcane mosaic disease and potential future research focus. The review is expected to provide scientific literature and guidance for the effective prevention and control of mosaic through resistance breeding in sugarcane. Full article
(This article belongs to the Section Plant Microbe Interactions)
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13 pages, 4785 KiB  
Article
Lactobacillus johnsonii L531 Alleviates the Damage Caused by Salmonella Typhimurium via Inhibiting TLR4, NF-κB, and NLRP3 Inflammasome Signaling Pathways
by Shiyan Chen, Yanan Li, Bingxin Chu, Lanxin Yuan, Ning Liu, Yaohong Zhu and Jiufeng Wang
Microorganisms 2021, 9(9), 1983; https://doi.org/10.3390/microorganisms9091983 - 17 Sep 2021
Cited by 18 | Viewed by 2850
Abstract
Salmonella Typhimurium (S. Typhimurium) is an aggressive zoonotic pathogen that causes enteritis and diarrhea. Antibiotic therapy is still the primary method at present. However, the increasing emergence of multi-drug resistant bacteria weakens the therapeutic efficacy of antibiotics. Probiotics have been widely studied [...] Read more.
Salmonella Typhimurium (S. Typhimurium) is an aggressive zoonotic pathogen that causes enteritis and diarrhea. Antibiotic therapy is still the primary method at present. However, the increasing emergence of multi-drug resistant bacteria weakens the therapeutic efficacy of antibiotics. Probiotics have been widely studied as an alternative antibiotic therapy. In this study, we established an IPEC-J2 cell model of S. Typhimurium infection, aiming to determine the protective effect of Lactobacillus johnsonii L531 (L. johnsonii L531) on S. Typhimurium infection. As our data showed, S. Typhimurium infection resulted in a robust inflammatory response demonstrated by promoted protein levels of the inflammatory-related pathway (TLR4, MyD88, p-IκBα, and p-p65), increased cytokine levels of IL-6, IL-1β, IL-18, and TNF-α, and activated the NLRP3 inflammasome via promoting its assembly. However, L. johnsonii L531 pre-incubation inhibited the activation of the above inflammatory signaling pathways and reduced the expression levels of pro-inflammatory cytokines. In addition, L. johnsonii L531 alleviated the damage of S. Typhimurium to tight junctions ZO-1, Occludin, and Claudin-1. In summary, our findings suggested that L. johnsonii L531 alleviated S. Typhimurium-induced tight junction injury by inhibiting the TLR4/NF-κB/NLRP3 inflammasome signaling pathway. Full article
(This article belongs to the Topic Veterinary Infectious Diseases)
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11 pages, 2241 KiB  
Article
A yceI Gene Involves in the Adaptation of Ralstonia solanacearum to Methyl Gallate and Other Stresses
by Kai-Hao Wang, De-Hong Zheng, Gao-Qing Yuan, Wei Lin and Qi-Qin Li
Microorganisms 2021, 9(9), 1982; https://doi.org/10.3390/microorganisms9091982 - 17 Sep 2021
Viewed by 1963
Abstract
Ralstonia solanacearum is a plant-pathogenic bacterium causing plant bacterial wilt, and can be strongly inhibited by methyl gallate (MG). Our previous transcriptome sequencing of MG-treated R. solanacearum showed that the yceI gene AVT05_RS03545 of Rs-T02 was up-regulated significantly under MG stress. In this [...] Read more.
Ralstonia solanacearum is a plant-pathogenic bacterium causing plant bacterial wilt, and can be strongly inhibited by methyl gallate (MG). Our previous transcriptome sequencing of MG-treated R. solanacearum showed that the yceI gene AVT05_RS03545 of Rs-T02 was up-regulated significantly under MG stress. In this study, a deletion mutant (named DM3545) and an over-expression strain (named OE3545) for yceI were constructed to confirm this hypothesis. No significant difference was observed among the growth of wild-type strain, DM3545 and OE3545 strains without MG treatment. Mutant DM3545 showed a lower growth ability than that of the wild type and OE3545 strains under MG treatment, non-optimal temperature, or 1% NaCl. The ability of DM3545 for rhizosphere colonization was lower than that of the wild-type and OE3545 strains. The DM3545 strain showed substantially reduced virulence toward tomato plants than its wild-type and OE3545 counterpart. Moreover, DM3545 was more sensitive to MG in plants than the wild-type and OE3545 strains. These results suggest that YceI is involved in the adaptability of R. solanacearum to the presence of MG and the effect of other tested abiotic stresses. This protein is also possibly engaged in the virulence potential of R. solanacearum. Full article
(This article belongs to the Section Plant Microbe Interactions)
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14 pages, 1726 KiB  
Article
A Pectin-Rich, Baobab Fruit Pulp Powder Exerts Prebiotic Potential on the Human Gut Microbiome In Vitro
by Martin Foltz, Alicia Christin Zahradnik, Pieter Van den Abbeele, Jonas Ghyselinck and Massimo Marzorati
Microorganisms 2021, 9(9), 1981; https://doi.org/10.3390/microorganisms9091981 - 17 Sep 2021
Cited by 10 | Viewed by 4031
Abstract
Increasing insight into the impact of the gut microbiota on human health has sustained the development of novel prebiotic ingredients. This exploratory study evaluated the prebiotic potential of baobab fruit pulp powder, which consists of pectic polysaccharides with unique composition as compared to [...] Read more.
Increasing insight into the impact of the gut microbiota on human health has sustained the development of novel prebiotic ingredients. This exploratory study evaluated the prebiotic potential of baobab fruit pulp powder, which consists of pectic polysaccharides with unique composition as compared to other dietary sources, given that it is rich in low methoxylated homogalacturonan (HG). After applying dialysis procedures to remove simple sugars from the product (simulating their absorption along the upper gastrointestinal tract), 48 h fecal batch incubations were performed. Baobab fruit pulp powder boosted colonic acidification across three simulated human adult donors due to the significant stimulation of health-related metabolites acetate (+18.4 mM at 48 h), propionate (+5.5 mM at 48 h), and to a lesser extent butyrate (0.9 mM at 48 h). Further, there was a trend of increased lactate levels (+2.7 mM at 6h) and reduced branched chain fatty acid (bCFA) levels (−0.4 mM at 48 h). While Bacteroidetes levels increased for all donors, donor-dependent increases in Bifidobacteria, Lactobacilli, and Firmicutes were observed, stressing the potential interindividual differences in microbial composition modulation upon Baobab fruit pulp powder treatment. Overall, Baobab fruit pulp powder fermentation displayed features of selective utilization by host microorganisms and, thus, has promising prebiotic potential (also in comparison with the ‘gold standard’ prebiotic inulin). Further research will be required to better characterize this prebiotic potential, accounting for the interindividual differences, while aiming to unravel the potential resulting health benefits. Full article
(This article belongs to the Special Issue Nutritional Regulation on Gut Microbiota)
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15 pages, 3809 KiB  
Article
Antifungal Streptomyces spp., Plausible Partners for Brood-Caring of the Dung Beetle Copris tripartitus
by Sung Hun Kim, Goeun Park, Jin-Soo Park and Hak Cheol Kwon
Microorganisms 2021, 9(9), 1980; https://doi.org/10.3390/microorganisms9091980 - 17 Sep 2021
Cited by 5 | Viewed by 2162
Abstract
The dung beetle Copris tripartitus Waterhouse (Coleoptera: Scarabaeidae) is a coprophagous insect that lives in and feeds primarily on the feces of mammalian herbivores and is known to protect their offspring from the pathogen-rich environment by performing parental care for brood balls. Brood [...] Read more.
The dung beetle Copris tripartitus Waterhouse (Coleoptera: Scarabaeidae) is a coprophagous insect that lives in and feeds primarily on the feces of mammalian herbivores and is known to protect their offspring from the pathogen-rich environment by performing parental care for brood balls. Brood balls under continuous management by dung beetle are rarely contaminated by entomopathogenic fungi compared to abandoned brood balls. On the supposition that dung beetles may benefit from mutualistic bacteria that protect their offspring against fungal pathogens, we evaluated the antifungal activities of bacteria isolated from the dung beetle and brood ball. As a result, bacterial isolates, mainly streptomycetes, manifested potent and broad-spectrum antifungal activity against various fungi, including entomopathogens. Of the isolates, Streptomyces sp. AT67 exhibited pronounced antifungal activities. Culture-dependent and independent approaches show that this strain has occurred continuously in dung beetles that were collected over three years. Moreover, metabolic profiling and chemical investigation demonstrated that the strain produced an antifungal polyene macrocyclic lactam, sceliphrolactam, as a major product. Our findings imply that specific symbiotic bacteria of C. tripartitus are likely to contribute brood ball hygiene by inhibiting fungal parasites in the environment. Full article
(This article belongs to the Special Issue Microbiota in Insects)
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22 pages, 27077 KiB  
Article
Fifty Generations of Amitosis: Tracing Asymmetric Allele Segregation in Polyploid Cells with Single-Cell DNA Sequencing
by Valerio Vitali, Rebecca Rothering and Francesco Catania
Microorganisms 2021, 9(9), 1979; https://doi.org/10.3390/microorganisms9091979 - 17 Sep 2021
Cited by 5 | Viewed by 2228
Abstract
Amitosis is a widespread form of unbalanced nuclear division whose biomedical and evolutionary significance remain unclear. Traditionally, insights into the genetics of amitosis have been gleaned by assessing the rate of phenotypic assortment. Though powerful, this experimental approach relies on the availability of [...] Read more.
Amitosis is a widespread form of unbalanced nuclear division whose biomedical and evolutionary significance remain unclear. Traditionally, insights into the genetics of amitosis have been gleaned by assessing the rate of phenotypic assortment. Though powerful, this experimental approach relies on the availability of phenotypic markers. Leveraging Paramecium tetraurelia, a unicellular eukaryote with nuclear dualism and a highly polyploid somatic nucleus, we probe the limits of single-cell whole-genome sequencing to study the consequences of amitosis. To this end, we first evaluate the suitability of single-cell sequencing to study the AT-rich genome of P. tetraurelia, focusing on common sources of genome representation bias. We then asked: can alternative rearrangements of a given locus eventually assort after a number of amitotic divisions? To address this question, we track somatic assortment of developmentally acquired Internal Eliminated Sequences (IESs) up to 50 amitotic divisions post self-fertilization. To further strengthen our observations, we contrast empirical estimates of IES retention levels with in silico predictions obtained through mathematical modeling. In agreement with theoretical expectations, our empirical findings are consistent with a mild increase in variation of IES retention levels across successive amitotic divisions of the macronucleus. The modest levels of somatic assortment in P. tetraurelia suggest that IESs retention levels are largely sculpted at the time of macronuclear development, and remain fairly stable during vegetative growth. In forgoing the requirement for phenotypic assortment, our approach can be applied to a wide variety of amitotic species and could facilitate the identification of environmental and genetic factors affecting amitosis. Full article
(This article belongs to the Special Issue Microbial Genetics and Evolution)
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18 pages, 2035 KiB  
Article
Transovarial Transmission of Borrelia hermsii by Its Tick Vector and Reservoir Host Ornithodoros hermsi
by Tom G. Schwan and Sandra J. Raffel
Microorganisms 2021, 9(9), 1978; https://doi.org/10.3390/microorganisms9091978 - 17 Sep 2021
Cited by 3 | Viewed by 2352
Abstract
Transovarial passage of relapsing fever spirochetes (Borrelia species) by infected female argasid ticks to their progeny is a widespread phenomenon. Yet this form of vertical inheritance has been considered rare for the North American tick Ornithodoros hermsi infected with Borrelia hermsii. [...] Read more.
Transovarial passage of relapsing fever spirochetes (Borrelia species) by infected female argasid ticks to their progeny is a widespread phenomenon. Yet this form of vertical inheritance has been considered rare for the North American tick Ornithodoros hermsi infected with Borrelia hermsii. A laboratory colony of O. hermsi was established from a single infected female and two infected males that produced a population of ticks with a high prevalence of transovarial transmission based on infection assays of single and pooled ticks feeding on mice and immunofluorescence microscopy of eggs and larvae. Thirty-eight of forty-five (84.4%) larval cohorts (groups of larvae originating from the same egg clutch) transmitted B. hermsii to mice over four and a half years, and one hundred and three single and one hundred and fifty-three pooled nymphal and adult ticks transmitted spirochetes during two hundred and fourteen of two hundred and fifty-six (83.6%) feedings on mice over seven and a half years. The perpetuation of B. hermsii for many years by infected ticks only (without acquisition of spirochetes from vertebrate hosts) demonstrates the reservoir competence of O. hermsi. B. hermsii produced the variable tick protein in eggs and unfed larvae infected by transovarial transmission, leading to speculation of the possible steps in the evolution of borreliae from a tick-borne symbiont to a tick-transmitted parasite of vertebrates. Full article
(This article belongs to the Special Issue Borrelia Ecology and Evolution: Ticks and Hosts and the Environment)
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16 pages, 1761 KiB  
Review
Subtelomeric Chromatin in the Fission Yeast S. pombe
by Rajesh K. Yadav, Atsushi Matsuda, Brandon R. Lowe, Yasushi Hiraoka and Janet F. Partridge
Microorganisms 2021, 9(9), 1977; https://doi.org/10.3390/microorganisms9091977 - 17 Sep 2021
Cited by 3 | Viewed by 3083
Abstract
Telomeres play important roles in safeguarding the genome. The specialized repressive chromatin that assembles at telomeres and subtelomeric domains is key to this protective role. However, in many organisms, the repetitive nature of telomeric and subtelomeric sequences has hindered research efforts. The fission [...] Read more.
Telomeres play important roles in safeguarding the genome. The specialized repressive chromatin that assembles at telomeres and subtelomeric domains is key to this protective role. However, in many organisms, the repetitive nature of telomeric and subtelomeric sequences has hindered research efforts. The fission yeast S. pombe has provided an important model system for dissection of chromatin biology due to the relative ease of genetic manipulation and strong conservation of important regulatory proteins with higher eukaryotes. Telomeres and the telomere-binding shelterin complex are highly conserved with mammals, as is the assembly of constitutive heterochromatin at subtelomeres. In this review, we seek to summarize recent work detailing the assembly of distinct chromatin structures within subtelomeric domains in fission yeast. These include the heterochromatic SH subtelomeric domains, the telomere-associated sequences (TAS), and ST chromatin domains that assemble highly condensed chromatin clusters called knobs. Specifically, we review new insights into the sequence of subtelomeric domains, the distinct types of chromatin that assemble on these sequences and how histone H3 K36 modifications influence these chromatin structures. We address the interplay between the subdomains of chromatin structure and how subtelomeric chromatin is influenced by both the telomere-bound shelterin complexes and by euchromatic chromatin regulators internal to the subtelomeric domain. Finally, we demonstrate that telomere clustering, which is mediated via the condensed ST chromatin knob domains, does not depend on knob assembly within these domains but on Set2, which mediates H3K36 methylation. Full article
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15 pages, 1891 KiB  
Article
Mussel Mass Mortality and the Microbiome: Evidence for Shifts in the Bacterial Microbiome of a Declining Freshwater Bivalve
by Jordan C. Richard, Lewis J. Campbell, Eric M. Leis, Rose E. Agbalog, Chris D. Dunn, Diane L. Waller, Susan Knowles, Joel G. Putnam and Tony L. Goldberg
Microorganisms 2021, 9(9), 1976; https://doi.org/10.3390/microorganisms9091976 - 17 Sep 2021
Cited by 16 | Viewed by 3371
Abstract
Freshwater mussels (Unionida) are suffering mass mortality events worldwide, but the causes remain enigmatic. Here, we describe an analysis of bacterial loads, community structure, and inferred metabolic pathways in the hemolymph of pheasantshells (Actinonaias pectorosa) from the Clinch River, USA, during [...] Read more.
Freshwater mussels (Unionida) are suffering mass mortality events worldwide, but the causes remain enigmatic. Here, we describe an analysis of bacterial loads, community structure, and inferred metabolic pathways in the hemolymph of pheasantshells (Actinonaias pectorosa) from the Clinch River, USA, during a multi-year mass mortality event. Bacterial loads were approximately 2 logs higher in moribund mussels (cases) than in apparently healthy mussels (controls). Bacterial communities also differed between cases and controls, with fewer sequence variants (SVs) and higher relative abundances of the proteobacteria Yokenella regensburgei and Aeromonas salmonicida in cases than in controls. Inferred bacterial metabolic pathways demonstrated a predominance of degradation, utilization, and assimilation pathways in cases and a predominance of biosynthesis pathways in controls. Only two SVs correlated with Clinch densovirus 1, a virus previously shown to be strongly associated with mortality in this system: Deinococcota and Actinobacteriota, which were associated with densovirus-positive and densovirus-negative mussels, respectively. Overall, our results suggest that bacterial invasion and shifts in the bacterial microbiome during unionid mass mortality events may result from primary insults such as viral infection or environmental stressors. If so, bacterial communities in mussel hemolymph may be sensitive, if generalized, indicators of declining mussel health. Full article
(This article belongs to the Section Environmental Microbiology)
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16 pages, 6698 KiB  
Article
Atomic Force Microscopy to Characterize Antimicrobial Peptide-Induced Defects in Model Supported Lipid Bilayers
by Kathleen W. Swana, Ramanathan Nagarajan and Terri A. Camesano
Microorganisms 2021, 9(9), 1975; https://doi.org/10.3390/microorganisms9091975 - 17 Sep 2021
Cited by 3 | Viewed by 2258
Abstract
Antimicrobial peptides (AMPs) interact with bacterial cell membranes through a variety of mechanisms, causing changes extending from nanopore formation to microscale membrane lysis, eventually leading to cell death. Several AMPs also disrupt mammalian cell membranes, despite their significantly different lipid composition and such [...] Read more.
Antimicrobial peptides (AMPs) interact with bacterial cell membranes through a variety of mechanisms, causing changes extending from nanopore formation to microscale membrane lysis, eventually leading to cell death. Several AMPs also disrupt mammalian cell membranes, despite their significantly different lipid composition and such collateral hemolytic damage hinders the potential therapeutic applicability of the AMP as an anti-microbial. Elucidating the mechanisms underlying the AMP–membrane interactions is challenging due to the variations in the chemical and structural features of the AMPs, the complex compositional variations of cell membranes and the inadequacy of any single experimental technique to comprehensively probe them. (1) Background: Atomic Force Microscopy (AFM) imaging can be used in combination with other techniques to help understand how AMPs alter the orientation and structural organization of the molecules within cell membranes exposed to AMPs. The structure, size, net charge, hydrophobicity and amphipathicity of the AMPs affect how they interact with cell membranes of differing lipid compositions. (2) Methods: Our study examined two different types of AMPs, a 20-amino acid, neutral, α-helical (amphipathic) peptide, alamethicin, and a 13-amino acid, non-α-helical cationic peptide, indolicidin (which intramolecularly folds, creating a hydrophobic core), for their interactions with supported lipid bilayers (SLBs). Robust SLB model membranes on quartz supports, incorporating predominantly anionic lipids representative of bacterial cells, are currently not available and remain to be developed. Therefore, the SLBs of zwitterionic egg phosphatidylcholine (PC), which represents the composition of a mammalian cell membrane, was utilized as the model membrane. This also allows for a comparison with the results obtained from the Quartz Crystal Microbalance with Dissipation (QCM-D) experiments conducted for these peptides interacting with the same zwitterionic SLBs. Further, in the case of alamethicin, because of its neutrality, the lipid charge may be less relevant for understanding its membrane interactions. (3) Results: Using AFM imaging and roughness analysis, we found that alamethicin produced large, unstable defects in the membrane at 5 µM concentrations, and completely removed the bilayer at 10 µM. Indolicidin produced smaller holes in the bilayer at 5 and 10 µM, although they were able to fill in over time. The root-mean-square (RMS) roughness values for the images showed that the surface roughness caused by visible defects peaked after peptide injection and gradually decreased over time. (4) Conclusions: AFM is useful for helping to uncover the dynamic interactions between different AMPs and cell membranes, which can facilitate the selection and design of more efficient AMPs for use in therapeutics and antimicrobial applications. Full article
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11 pages, 1818 KiB  
Article
Microbiota of Cow’s Milk with Udder Pathologies
by Mariya V. Gryaznova, Mikhail Y. Syromyatnikov, Yulia D. Dvoretskaya, Sergey A. Solodskikh, Nikolay T. Klimov, Vitaliy I. Mikhalev, Vitaliy I. Zimnikov, Evgeniy V. Mikhaylov and Vasily N. Popov
Microorganisms 2021, 9(9), 1974; https://doi.org/10.3390/microorganisms9091974 - 17 Sep 2021
Cited by 11 | Viewed by 2489
Abstract
Mastitis is the most common disease for cattle, causing great economic losses for the global dairy industry. Recent studies indicate the multi-agent and microbiome diversity of this disease. To understand the nature of mastitis and investigate the role of the microbiome in the [...] Read more.
Mastitis is the most common disease for cattle, causing great economic losses for the global dairy industry. Recent studies indicate the multi-agent and microbiome diversity of this disease. To understand the nature of mastitis and investigate the role of the microbiome in the development of pathologies in the udder of bovines, we performed NGS sequencing of the 16S rRNA gene of cow’s milk with pathologies of the udder. The obtained data show a significant increase in the Cutibacterium, Blautia, Clostridium sensu stricto 2, Staphylococcus, Streptococcus and Microbacterium genera for groups of cows with udder pathologies. Increasing relative abundance of the Staphylococcus and Streptococcus genera was associated with subclinical mastitis. Our data show that a relative increase in abundance of the Staphylococcus and Microbacterium genera may be an early sign of infection. We have shown, for the first time, an increase in the Colidextribacter, Paeniclostridium and Turicibacter genera in groups of cows with mastitis. These results expand our understanding of the role of the microbiome in the development of bovine mastitis. Full article
(This article belongs to the Section Veterinary Microbiology)
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14 pages, 990 KiB  
Article
Cork Oak Forests Soil Bacteria: Potential for Sustainable Agroforest Production
by Francisca Reis, Ana João Pereira, Rui M. Tavares, Paula Baptista and Teresa Lino-Neto
Microorganisms 2021, 9(9), 1973; https://doi.org/10.3390/microorganisms9091973 - 16 Sep 2021
Cited by 5 | Viewed by 2100
Abstract
Plant growth promoting rhizobacteria (PGPR) are in increasing demand due to their role in promoting sustainable practices, not only in agriculture but also in forestry. Keeping in mind the future application of PGPR for increasing cork oak sustainability, the aim of this study [...] Read more.
Plant growth promoting rhizobacteria (PGPR) are in increasing demand due to their role in promoting sustainable practices, not only in agriculture but also in forestry. Keeping in mind the future application of PGPR for increasing cork oak sustainability, the aim of this study was to find cork oak PGPR isolates with increased nutrient solubilisation traits, able to promote root morphological changes and/or antagonize cork oak bark phytopathogens. Soils from three cork oak forests with distinct bioclimates (humid, semi-humid and semi-arid) were used for isolating bacteria. From the 7634 colony-forming units, 323 bacterial isolates were biochemically assayed for PGPR traits (siderophores production, phosphate solubilizing and organic acids production), and 51 were found to display all these traits. These PGPR were able to induce root morphological changes on Arabidopsis thaliana, like suppression of primary root growth, increase of lateral roots or root hairs formation. However, the most proficient PGPR displayed specific ability in changing a single root morphological trait. This ability was related not only to bacterial genotype, but also with the environment where bacteria thrived and isolation temperature. Bacteria from semi-arid environments (mainly Bacillus megaterium isolates) could hold a promising tool to enhance plant development. Other isolates (Serratia quinivorens or B. cereus) could be further explored for biocontrol purposes. Full article
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