Microorganisms

15 pages, 6031 KiB

Open AccessArticle

Rhizobacteria Increase the Adaptation Potential of Potato Microclones under Aeroponic Conditions

by Oksana V. Tkachenko, Nina V. Evseeva, Kristina Y. Kargapolova, Alena Y. Denisova, Natalia N. Pozdnyakova, Artem A. Kulikov and Gennady L. Burygin

Microorganisms 2023, 11(7), 1866; https://doi.org/10.3390/microorganisms11071866 - 24 Jul 2023

Cited by 1 | Viewed by 1045

Abstract

Adaptation ex vitro is strongly stressful for microplants. Plant-growth-promoting rhizobacteria (PGPR) help to increase the adaptation potential of microplants transplanted from test tubes into the natural environment. We investigated the mechanisms of antioxidant protection of PGPR-inoculated potato microclones adapting to ex vitro growth [...] Read more.

Adaptation ex vitro is strongly stressful for microplants. Plant-growth-promoting rhizobacteria (PGPR) help to increase the adaptation potential of microplants transplanted from test tubes into the natural environment. We investigated the mechanisms of antioxidant protection of PGPR-inoculated potato microclones adapting to ex vitro growth in an aeroponic system. Potato (Solanum tuberosum L. cv. Nevsky) microplants were inoculated in vitro with the bacteria Azospirillum baldaniorum Sp245 and Ochrobactrum cytisi IPA7.2. On days 1 and 7 of plant growth ex vitro, catalase and peroxidase activities in the leaves of inoculated plants were 1.5-fold higher than they were in non-inoculated plants. The activity of ascorbate peroxidase was reduced in both in vitro and ex vitro treatments, and this reduction was accompanied by a decrease in the leaf content of hydrogen peroxide and malondialdehyde. As a result, inoculation contributed to the regulation of the plant pro/antioxidant system, lowering the oxidative stress and leading to better plant survival ex vitro. This was evidenced by the higher values of measured morphological and physiological variables of the inoculated plants, as compared with the values in the control treatment. Thus, we have shown some PGPR-mediated mechanisms of potato plant protection from adverse environmental factors under aeroponic conditions. Full article

(This article belongs to the Special Issue Interaction of Plants and Endophytic Microorganisms: Community, Functions and Applications)

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22 pages, 5594 KiB

Open AccessArticle

Microbial Communities in Ferromanganese Sediments from the Northern Basin of Lake Baikal (Russia)

by Anna Lomakina, Sergei Bukin, Olga Shubenkova, Tatyana Pogodaeva, Vyacheslav Ivanov, Yuri Bukin and Tamara Zemskaya

Microorganisms 2023, 11(7), 1865; https://doi.org/10.3390/microorganisms11071865 - 24 Jul 2023

Cited by 1 | Viewed by 1247

Abstract

We analyzed the amplicons of the 16S rRNA genes and assembled metagenome-assembled genomes (MAGs) of the enrichment culture from the Fe-Mn layer to have an insight into the diversity and metabolic potential of microbial communities from sediments of two sites in the northern [...] Read more.

We analyzed the amplicons of the 16S rRNA genes and assembled metagenome-assembled genomes (MAGs) of the enrichment culture from the Fe-Mn layer to have an insight into the diversity and metabolic potential of microbial communities from sediments of two sites in the northern basin of Lake Baikal. Organotrophic Chloroflexota, Actionobacteriota, and Acidobacteriota, as well as aerobic and anaerobic participants of the methane cycle (Methylococcales and Methylomirabilota, respectively), dominated the communities of the surface layers. With depth, one of the cores showed a decrease in the proportion of the Chloroflexota and Acidobacteriota members and a substantial increase in the sequences of the phylum Firmicutes. The proportion of the Desulfobacteriota and Thermodesulfovibronia (Nitrospirota) increased in another core. The composition of archaeal communities was similar between the investigated sites and differed in depth. Members of ammonia-oxidizing archaea (Nitrososphaeria) predominated in the surface sediments, with an increase in anaerobic methanotrophs (Methanoperedenaceae) and organoheterotrophs (Bathyarchaeia) in deep sediments. Among the 37 MAGs, Gammaproteobacteria, Desulfobacteriota, and Methylomirabilota were the most common in the microbial community. Metagenome sequencing revealed the assembled genomes genes for N, S, and CH₄ metabolism for carbon fixation, and genes encoding Fe and Mn pathways, indicating the likely coexistence of the biogeochemical cycle of various elements and creating certain conditions for the development of taxonomically and functionally diverse microbial communities. Full article

(This article belongs to the Special Issue Microbial Communities in Aquatic Systems: Diversity and Function)

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21 pages, 3479 KiB

Open AccessArticle

A New Proposed Symbiotic Plant–Herbivore Relationship between Burkea africana Trees, Cirina forda Caterpillars and Their Associated Fungi Pleurostomophora richardsiae and Aspergillus nomius

by Lufuno Ethel Nemadodzi and Gerhard Prinsloo

Microorganisms 2023, 11(7), 1864; https://doi.org/10.3390/microorganisms11071864 - 24 Jul 2023

Viewed by 1104

Abstract

Burkea africana is a tree found in savannah and woodland in southern Africa, as well as northwards into tropical African regions as far as Nigeria and Ethiopia. It is used as fuel wood, medicinally to treat various conditions, such as toothache, headache, migraine, [...] Read more.

Burkea africana is a tree found in savannah and woodland in southern Africa, as well as northwards into tropical African regions as far as Nigeria and Ethiopia. It is used as fuel wood, medicinally to treat various conditions, such as toothache, headache, migraine, pain, inflammation, and sexually transmitted diseases, such as gonorrhoea, but also an ornamental tree. The current study investigated the possible symbiotic relationship between B. africana trees and the C. forda caterpillars and the mutual role played in ensuring the survival of B. africana trees/seedlings in harsh natural conditions and low-nutrient soils. Deoxyribonucleic acid isolation and sequencing results revealed that the fungal species Pleurostomophora richardsiae was highly predominant in the leaves of B. africana trees and present in the caterpillars. The second most prominent fungal species in the caterpillars was Aspergillus nomius. The latter is known to be related to a Penicillium sp. which was found to be highly prevalent in the soil where B. africana trees grow and is suggested to play a role in enhancing the effective growth of B. africana trees in their natural habitat. To support this, a phylogenetic analysis was conducted, and a tree was constructed, which shows a high percentage similarity between Aspergillus and Penicillium sp. The findings of the study revealed that B. africana trees not only serve as a source of feed for the C. forda caterpillar but benefit from C. forda caterpillars which, after dropping onto the soil, is proposed to inoculate the soil surrounding the trees with the fungus A. nomius which suggests a symbiotic and/or synergistic relationship between B. africana trees and C. forda caterpillars. Full article

(This article belongs to the Special Issue Plant-Pathogenic Fungi)

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18 pages, 1141 KiB

Open AccessArticle

Growth Response of Non-Conventional Yeasts on Sugar-Rich Media: Part 1: High Production of Lipid by Lipomyces starkeyi and Citric Acid by Yarrowia lipolytica

by Panagiota Diamantopoulou, Dimitris Sarris, Sidoine Sadjeu Tchakouteu, Evangelos Xenopoulos and Seraphim Papanikolaou

Microorganisms 2023, 11(7), 1863; https://doi.org/10.3390/microorganisms11071863 - 24 Jul 2023

Cited by 6 | Viewed by 1199

Abstract

Sugar-rich waste streams, generated in very high quantities worldwide, constitute an important source of environmental pollution. Their eco-friendly conversions into a plethora of added-value compounds through the use of microbial fermentations is currently a very “hot” scientific topic. The aim of this study, [...] Read more.

Sugar-rich waste streams, generated in very high quantities worldwide, constitute an important source of environmental pollution. Their eco-friendly conversions into a plethora of added-value compounds through the use of microbial fermentations is currently a very “hot” scientific topic. The aim of this study, was to assess the potential of single cell oil (SCO), microbial mass and citric acid (CA) production by non-conventional yeast strains growing on expired (“waste”) glucose. Six yeast strains (viz. Rhodosporidium toruloides DSM 4444, Rhodotorula glutinis NRRL YB-252, R. toruloides NRRL Y-27012, Yarrowia lipolytica LFMB Y-20, Y. lipolytica ACA-DC 50109 and Lipomyces starkeyi DSM 70296) were initially grown in shake flasks with expired glucose used as substrate under nitrogen limitation, in order to “boost” the cellular metabolism towards the synthesis of SCO and CA, and their growth response was quantitatively evaluated. Initial glucose concentration (Glc₀) was adjusted at c. 50 g/L. Besides Y. lipolytica, all other yeast strains produced noticeable SCO quantities [lipid in dry cell weight (DCW) ranging from 25.3% w/w to 55.1% w/w]. Lipids of all yeasts contained significant quantities of oleic acid, being perfect candidates for the synthesis of 2nd generation biodiesel. The highest DCW production (=13.6 g/L) was obtained by L. starkeyi DSM 70296, while both Y. lipolytica strains did not accumulate noticeable lipid quantities, but produced non-negligible CA amounts. The most promising CA-producing strain, namely Y. lipolytica ACA-DC 50109 was further studied in stirred-tank bioreactor systems, while the very promising DCW- and SCO-producing L. starkeyi DSM 70296 was further studied in shake flasks. Both strains were grown on media presenting higher Glc₀ concentrations and the same initial nitrogen quantity as previously. Indeed, L. starkeyi grown at Glc₀ = 85 g/L, produced DCW_max = 34.0 g/L, that contained lipid =34.1% w/w (thus SCO was =11.6 g/L). The strain ACA-DC 50109 in stirred tank bioreactor with Glc₀ ≈ 105 g/L produced CA up to 46 g/L (yield of CA produced on glucose consumed; Y_CA/Glc ≈ 0.45 g/g). Finally, in fed-batch bioreactor experiment, the significant CA quantity of 82.0 g/L (Y_CA/Glc = 0.50 g/g) was recorded. Concluding, “waste” glucose proved to be a suitable substrate for a number of non-conventional yeast strains. Y. lipolytica ACA-DC 50109 produced significant quantities of CA while L. starkeyi DSM 70296 was a very interesting DCW- and SCO-producing candidate. These strains can be used as potential cell factories amenable to convert glucose-based residues into the mentioned metabolic compounds, that present high importance for food, chemical and biofuel facilities. Full article

(This article belongs to the Special Issue Advances in Microbial Metabolites)

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12 pages, 1194 KiB

Open AccessArticle

Prebiotic and Anti-Adipogenic Effects of Radish Green Polysaccharide

by Yu Ra Lee, Hye-Bin Lee, Yoonsook Kim, Kwang-Soon Shin and Ho-Young Park

Microorganisms 2023, 11(7), 1862; https://doi.org/10.3390/microorganisms11071862 - 24 Jul 2023

Cited by 1 | Viewed by 1250

Abstract

Radish (Raphanus sativus L.) greens are consumed as a source of nutrition, and their polysaccharides such as rhamnogalacturonan-I possess certain beneficial properties. This study investigated the prebiotic effects of a radish green polysaccharide (RGP) on gut health and obesity. The prebiotic activity [...] Read more.

Radish (Raphanus sativus L.) greens are consumed as a source of nutrition, and their polysaccharides such as rhamnogalacturonan-I possess certain beneficial properties. This study investigated the prebiotic effects of a radish green polysaccharide (RGP) on gut health and obesity. The prebiotic activity of RGP was evaluated based on the pH changes and short-chain fatty acids (SCFAs) concentration. The results showed that 0.5% RGP had a higher prebiotic activity score than inulin and increased SCFAs production in all five prebiotic strains. Moreover, RGP inhibited fat accumulation in 3T3-L1 adipocytes, indicating its potential to reduce obesity. Overall, these findings suggested that the polysaccharide of radish greens has prebiotic effects and may serve as a beneficial prebiotic for gut health and obesity. Full article

(This article belongs to the Special Issue Functional Foods, Prebiotics and Probiotics Strengthening Intestinal Microbiome Health)

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13 pages, 3782 KiB

Open AccessArticle

Modulation of Cystatin F in Human Macrophages Impacts Cathepsin-Driven Killing of Multidrug-Resistant Mycobacterium tuberculosis

by Manoj Mandal, David Pires, Maria João Catalão, José Miguel Azevedo-Pereira and Elsa Anes

Microorganisms 2023, 11(7), 1861; https://doi.org/10.3390/microorganisms11071861 - 24 Jul 2023

Cited by 1 | Viewed by 1153

Abstract

Tuberculosis (TB) treatment relies primarily on 70-year-old drugs, and prophylaxis suffers from the lack of an effective vaccine. Among the 10 million people exhibiting disease symptoms yearly, 450,000 have multidrug or extensively drug-resistant (MDR or XDR) TB. A greater understanding of host and [...] Read more.

Tuberculosis (TB) treatment relies primarily on 70-year-old drugs, and prophylaxis suffers from the lack of an effective vaccine. Among the 10 million people exhibiting disease symptoms yearly, 450,000 have multidrug or extensively drug-resistant (MDR or XDR) TB. A greater understanding of host and pathogen interactions will lead to new therapeutic interventions for TB eradication. One of the strategies will be to target the host for better immune bactericidal responses against the TB causative agent Mycobacterium tuberculosis (Mtb). Cathepsins are promising targets due to their manipulation of Mtb with consequences such as decreased proteolytic activity and improved pathogen survival in macrophages. We recently demonstrated that we could overcome this enzymatic blockade by manipulating protease inhibitors such as cystatins. Here, we investigate the role of cystatin F, an inhibitor that we showed previously to be strongly upregulated during Mtb infection. Our results indicate that the silencing of cystatin F using siRNA increase the proteolytic activity of cathepsins S, L, and B, significantly impacting pathogen intracellular killing in macrophages. Taken together, these indicate the targeting of cystatin F as a potential adjuvant therapy for TB, including MDR and XDR-TB. Full article

(This article belongs to the Special Issue Prevention, Treatment and Diagnosis of Tuberculosis)

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12 pages, 1429 KiB

Open AccessReview

Analysis of the Intestinal and Faecal Bacterial Microbiota of the Cervidae Family Using 16S Next-Generation Sequencing: A Review

by Irene Pacheco-Torres, David Hernández-Sánchez, Cristina García-De la Peña, Luis A. Tarango-Arámbula, María M. Crosby-Galván and Paulino Sánchez-Santillán

Microorganisms 2023, 11(7), 1860; https://doi.org/10.3390/microorganisms11071860 - 24 Jul 2023

Viewed by 1441

Abstract

The Cervidae family has a wide distribution due to its adaptation to numerous ecological environments, which allows it to develop a diverse microbial community in its digestive tract. Recently, research has focused on the taxonomic composition and functionality of the intestinal and faecal [...] Read more.

The Cervidae family has a wide distribution due to its adaptation to numerous ecological environments, which allows it to develop a diverse microbial community in its digestive tract. Recently, research has focused on the taxonomic composition and functionality of the intestinal and faecal microbiota of different cervid species worldwide, as well as their microbial diversity and variation under different associated factors such as age, sex, diet, distribution, and seasonal variation. In addition, there is special interest in knowing how cervids act as reservoirs of zoonotic pathogenic microorganisms, which represent a threat to public health. This review provides a synthesis of the growing field of microbiota determination in cervids worldwide, focusing on intestinal and faecal samples using 16S next-generation sequencing. It also documents factors influencing microbial diversity and composition, the microorganisms reported as pathogenic/zoonotic, and the perspectives regarding the conservation of these species. Knowing the interactions between bacteria and cervid health can drive management and conservation strategies for these species and help develop an understanding of their evolutionary history and the interaction with emerging disease-causing microorganisms. Full article

(This article belongs to the Special Issue Microbial Genome Analysis and Interpretation Using Computational Approaches)

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17 pages, 2101 KiB

Open AccessArticle

Composite Interventions on Outcomes of Severely and Critically Ill Patients with COVID-19 in Shanghai, China

by Jiasheng Shao, Rong Fan, Chengnan Guo, Xuyuan Huang, Runsheng Guo, Fengdi Zhang, Jianrong Hu, Gang Huang and Liou Cao

Microorganisms 2023, 11(7), 1859; https://doi.org/10.3390/microorganisms11071859 - 23 Jul 2023

Cited by 4 | Viewed by 1351

Abstract

Background: The sixty-day effects of initial composite interventions for the treatment of severely and critically ill patients with COVID-19 are not fully assessed. Methods: Using a Bayesian piecewise exponential model, we analyzed the 60-day mortality, health-related quality of life (HRQoL), and [...] Read more.

Background: The sixty-day effects of initial composite interventions for the treatment of severely and critically ill patients with COVID-19 are not fully assessed. Methods: Using a Bayesian piecewise exponential model, we analyzed the 60-day mortality, health-related quality of life (HRQoL), and disability in 1082 severely and critically ill patients with COVID-19 between 8 December 2022 and 9 February 2023 in Shanghai, China. The final 60-day follow-up was completed on 10 April 2023. Results: Among 1082 patients (mean age, 78.0 years, 421 [38.9%] women), 139 patients (12.9%) died within 60 days. Azvudine had a 99.8% probability of improving 2-month survival (adjusted HR, 0.44 [95% credible interval, 0.24–0.79]), and Paxlovid had a 91.9% probability of improving 2-month survival (adjusted HR, 0.71 [95% credible interval, 0.44–1.14]) compared with the control. IL-6 receptor antagonist, baricitinib and a-thymosin each had a high probability of benefit (99.5%, 99.4%, and 97.5%, respectively) compared to their controls, while the probability of trail-defined statistical futility (HR > 0.83) was high for therapeutic anticoagulation (99.8%; HR, 1.64 [95% CrI, 1.06–2.50]) and glucocorticoid (91.4%; HR, 1.20 [95% CrI, 0.71–2.16]). Paxlovid, Azvudine, and therapeutic anticoagulation showed a significant reduction in disability (p < 0.05) Conclusions: Among severely and critically ill patients with COVID-19 who received 1 or more therapeutic interventions, treatment with Azvudine had a high probability of improved 60-day mortality compared with the control, indicating its potential in a resource-limited scenario. Treatment with an IL-6 receptor antagonist, baricitinib, and a-thymosin also had high probabilities of benefit in improving 2-month survival, among which a-thymosin could improve HRQoL. Treatment with Paxlovid, Azvudine, and therapeutic anticoagulation could significantly reduce disability at day 60. Full article

(This article belongs to the Special Issue Advances in Public Health Microbiology 2023)

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15 pages, 3097 KiB

Open AccessArticle

Gut Protective Effect from Newly Isolated Bacteria as Probiotics against Dextran Sulfate Sodium and Carrageenan-Induced Ulcerative Colitis

by Yuka Ikeda, Ai Tsuji and Satoru Matsuda

Microorganisms 2023, 11(7), 1858; https://doi.org/10.3390/microorganisms11071858 - 23 Jul 2023

Viewed by 1375

Abstract

Gut microbiome dysbiosis might be linked to certain diseases such as inflammatory bowel diseases (IBDs), which are categorized by vigorous inflammation of the gastrointestinal tract. Several studies have shown the favorable anti-inflammatory effect of certain probiotics in IBD therapy. In the present investigation, [...] Read more.

Gut microbiome dysbiosis might be linked to certain diseases such as inflammatory bowel diseases (IBDs), which are categorized by vigorous inflammation of the gastrointestinal tract. Several studies have shown the favorable anti-inflammatory effect of certain probiotics in IBD therapy. In the present investigation, the possible gut protective effects of commensal bacteria were examined in an IBD model mouse that was cost-effectively induced with low molecular weight dextran sulfate sodium (DSS) and kappa carrageenan. Our conclusions show that certain probiotic supplementation could result in the attenuation of the disease condition in the IBD mouse, suggesting a favorable therapeutic capability for considerably improving symptoms of gut inflammation with an impact on the IBD therapy. However, the molecular mechanisms require further investigation. Full article

(This article belongs to the Special Issue Novel Strategies in the Study of the Human Gut Microbiota)

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12 pages, 658 KiB

Open AccessArticle

A Pork Industry in the Backyard: An Analysis of Toxoplasma gondii Infection in Serbia’s Pigs

by Aleksandra Uzelac, Nikola Betić, Nedjeljko Karabasil, Vladimir Ćirković, Olgica Djurković-Djaković and Ivana Klun

Microorganisms 2023, 11(7), 1857; https://doi.org/10.3390/microorganisms11071857 - 23 Jul 2023

Cited by 1 | Viewed by 1143

Abstract

As pork is an important source for Toxoplasma gondii infection, we have analyzed T. gondii genotypes and toxoplasmosis prevalence in pigs in Serbia in the context of production statistics and economics to assess the specific risk to public health. Genotyping was performed using [...] Read more.

As pork is an important source for Toxoplasma gondii infection, we have analyzed T. gondii genotypes and toxoplasmosis prevalence in pigs in Serbia in the context of production statistics and economics to assess the specific risk to public health. Genotyping was performed using MnPCR-RFLP; T. gondii-specific IgG antibodies were detected using a modified agglutination test (MAT); and statistical data were extracted from official records and provided by government authorities. The results indicate that, from 2006 to 2021, the median number of annually slaughtered pigs was 5.6 million, yet only 36.1% were processed by abattoirs. The remainder were “backyard pigs” slaughtered on family farms and homesteads. Toxoplasmosis seroprevalence in market-weight (MW) pigs prior to 2006 was 15.2%, and was 15.1% in 2019. The seroprevalence in owned city cats, likely infected by livestock meat, was 33.2%. ToxoDB#1 was identified in pig tissues. The results indicate that backyard pigs are the backbone of the industry and provide as much as 60% of the pork in Serbia. The seroprevalence in pigs and city cats shows that farms are reservoirs for the parasite. Thus, innovative means of reducing T. gondii infection designed with backyard farmers in mind are needed to reduce the risk to public health. Full article

(This article belongs to the Special Issue Parasitic Diseases in Livestock)

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12 pages, 1871 KiB

Open AccessArticle

Bacterial Flora on Mist Outlet Surfaces in 4D Theaters and Suspended Particle Concentration Characteristics during 4D Movie Screenings

by U Yanagi, Noriko Kaihara, Dai Simazaki, Kanae Bekki, Yoshinori Homma, Chiemi Iba, Atsuto Asai and Motoya Hayashi

Microorganisms 2023, 11(7), 1856; https://doi.org/10.3390/microorganisms11071856 - 22 Jul 2023

Viewed by 1548

Abstract

In this study, we measured suspended particle concentrations during the screening of 4D movies (3 screens and 15 movies) and 2D movies (9 screens and 9 movies) in 3 movie theaters to obtain a more detailed understanding of the situation of suspended particle [...] Read more.

In this study, we measured suspended particle concentrations during the screening of 4D movies (3 screens and 15 movies) and 2D movies (9 screens and 9 movies) in 3 movie theaters to obtain a more detailed understanding of the situation of suspended particle concentrations and adherent bacterial flora in 4D movie theaters, which have been introduced in increasing numbers in recent years. The adherent bacterial flora on the floor and mist outlet surfaces in the 4D movie theaters were collected and analyzed. During the movie showings, the concentrations of suspended particles in 4D movie theaters were significantly higher than those in 2D movie theaters (p < 0.001). A significant increase in suspended particle concentrations due to 4D movie effects was also observed. The results of the α-diversity and β-diversity analyses indicate that the bacterial flora on the surfaces of mist outlets in 4D movie theaters are similar. Moreover, there are many closely related species, and the bacterial flora are rich and contain rare bacterial species. Many of the bacterial genera that are dominant in 4D theaters are suited to aqueous environments, and bacteria in the water supply system may have an impact on the indoor environment. Full article

(This article belongs to the Special Issue Airborne Microbes and Their Potential Influence)

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18 pages, 1789 KiB

Open AccessArticle

Enhancing 1,3-Propanediol Productivity in the Non-Model Chassis Clostridium beijerinckii through Genetic Manipulation

by Jonatã Bortolucci, María-Eugenia Guazzaroni, Teresa Schoch, Peter Dürre and Valeria Reginatto

Microorganisms 2023, 11(7), 1855; https://doi.org/10.3390/microorganisms11071855 - 22 Jul 2023

Cited by 1 | Viewed by 1444

Abstract

Biotechnological processes at biorefineries are considered one of the most attractive alternatives for valorizing biomasses by converting them into bioproducts, biofuels, and bioenergy. For example, biodiesel can be obtained from oils and grease but generates glycerol as a byproduct. Glycerol recycling has been [...] Read more.

Biotechnological processes at biorefineries are considered one of the most attractive alternatives for valorizing biomasses by converting them into bioproducts, biofuels, and bioenergy. For example, biodiesel can be obtained from oils and grease but generates glycerol as a byproduct. Glycerol recycling has been studied in several bioprocesses, with one of them being its conversion to 1,3-propanediol (1,3-PDO) by Clostridium. Clostridium beijerinckii is particularly interesting because it can produce a range of industrially relevant chemicals, including solvents and organic acids, and it is non-pathogenic. However, while Clostridium species have many potential advantages as chassis for synthetic biology applications, there are significant limitations when considering their use, such as their limited genetic tools, slow growth rate, and oxygen sensitivity. In this work, we carried out the overexpression of the genes involved in the synthesis of 1,3-PDO in C. beijerinckii Br21, which allowed us to increase the 1,3-PDO productivity in this strain. Thus, this study contributed to a better understanding of the metabolic pathways of glycerol conversion to 1,3-PDO by a C. beijerinckii isolate. Also, it made it possible to establish a transformation method of a modular vector in this strain, therefore expanding the limited genetic tools available for this bacterium, which is highly relevant in biotechnological applications. Full article

(This article belongs to the Special Issue Synthetic Biology Applied to Environmental Microorganisms)

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16 pages, 4837 KiB

Open AccessArticle

A Novel Deoxyribonuclease Low-Molecular-Weight Bacteriocin, Carocin S4, from Pectobacterium carotovorum subsp. carotovorum

by Huang-Pin Wu, Reymund C. Derilo, Shih-Hao Hsu, Jia-Ming Hu and Duen-Yau Chuang

Microorganisms 2023, 11(7), 1854; https://doi.org/10.3390/microorganisms11071854 - 22 Jul 2023

Viewed by 1021

Abstract

Pectobacterium carotovorum subsp. carotovorum (Pcc) is known to produce different types of bacteriocins, active protein substances that inhibit or kill related strains and are known to be induced by several factors. In this paper, we report the discovery, isolation, characterization, [...] Read more.

Pectobacterium carotovorum subsp. carotovorum (Pcc) is known to produce different types of bacteriocins, active protein substances that inhibit or kill related strains and are known to be induced by several factors. In this paper, we report the discovery, isolation, characterization, and functional analysis of Carocin S4, a novel low-molecular-weight bacteriocin (LMWB) from Pcc. A 2750 bp gene fragment was isolated from the chromosomal DNA of Pcc mutant strain rif-TO6, a rifampicin-resistant strain of TO6. The gene contains caroS4K and caroS4I within two open reading frames, which encode CaroS4K and CaroS4I, with molecular weights of about 90 kD and 10 kD, respectively. The unique characteristics of Carocin S4 were revealed after homology analysis with the previously discovered bacteriocins from Pcc. CaroS4K, which shares 23% and 85% homology with CaroS1K and CaroS3K, respectively, is also a deoxyribonuclease. However, unlike the two which can only hydrolyze genomic DNA, CaroS4K hydrolyzes both genomic and plasmid DNA. On the other hand, CaroS4K was found to be 90% homologous with CaroS2K but works differently in killing the target cell, as the latter is a ribonuclease. The optimal reaction temperature for CaroS4K to hydrolyze dsDNA is approximately 50 °C and requires the divalent metal ions Mg²⁺, Ca²⁺, and Zn²⁺ to catalyze its DNase activity. This study reveals another nuclease type of bacteriocin in Pcc, with CaroS4K and CaroS4I functioning as killer and immunity proteins, respectively. Full article

(This article belongs to the Special Issue The Potential of Antimicrobial Activity and Antibiofilm Activity of Bacteriocins)

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9 pages, 311 KiB

Open AccessArticle

Strong Evidence of the Role of Donkeys in the Epidemiology of Leptospira spp. in Semiarid Conditions

by Davidianne de Andrade Morais, Bruno Cesar Nunes, Rafael Rodrigues Soares, Murilo Duarte de Oliveira, Diego Figueiredo da Costa, Hosaneide Gomes de Araújo, João Pessoa Araújo Júnior, Camila Dantas Malossi, Maria Luana Cristiny Rodrigues Silva, Sérgio Santos de Azevedo and Clebert José Alves

Microorganisms 2023, 11(7), 1853; https://doi.org/10.3390/microorganisms11071853 - 22 Jul 2023

Viewed by 947

Abstract

Donkeys (Equus asinus) are historically known for their close relationship to humanity, which raises the need to study zoonotic diseases that affect them. In this perspective, leptospirosis stands out as a disease with an economic and public health impact, and its [...] Read more.

Donkeys (Equus asinus) are historically known for their close relationship to humanity, which raises the need to study zoonotic diseases that affect them. In this perspective, leptospirosis stands out as a disease with an economic and public health impact, and its occurrence is facilitated in times of higher rainfall indexes, especially in large urban centers. In view of the scarcity of information about leptospirosis in donkeys, the objective of this study was to detect the presence of Leptospira spp. and anti-leptospiral antibodies in donkeys rescued by a zoonosis center located in the Caatiga biome, Brazilian semiarid region. Overall, 30 donkeys of both sexes, aged between 4 months and 15 years, were used, from which 64 serum samples were collected and submitted to the microscopic agglutination test (MAT). In addition, 64 samples of urine, vaginal and preputial fluid, in duplicates, were subjected to the polymerase chain reaction (PCR) and microbiological. Sixteen (53.3%) animals tested positive in at least one diagnostic test, 12 (40%) of which were positive at MAT and seven (23.3%) in the molecular and bacteriological detection (urine, vaginal, and preputial fluid samples). This is the first report identifying donkeys infected with Leptospira spp. by molecular and bacteriological diagnosis in Brazil, and the first in the world to detect this agent in their genital fluids. The study also shows that donkeys are commonly exposed to leptospires in the Caatinga biome, and this constitutes a One Health-based concern, demonstrating the importance of broad studies where large numbers of humans and animals coexist when investigating zoonotic infections and when planning and implementing control measures for donkeys-associated leptospirosis. Full article

(This article belongs to the Special Issue Epidemiology of Leptospira sp. Infection: Current Status, Insights and Future Prospects)

17 pages, 2293 KiB

Open AccessArticle

Purification and Characterization of a DegP-Type Protease from the Marine Bacterium Cobetia amphilecti KMM 296

by Yulia Noskova, Oksana Son, Liudmila Tekutyeva and Larissa Balabanova

Microorganisms 2023, 11(7), 1852; https://doi.org/10.3390/microorganisms11071852 - 21 Jul 2023

Cited by 1 | Viewed by 1146

Abstract

A new member of the DegP-type periplasmic serine endoproteases of the S1C family from the marine bacterium Cobetia amphilecti KMM 296 (CamSP) was expressed in Escherichia coli cells. The calculated molecular weight, number of amino acids, and isoelectric point (pI) of the mature [...] Read more.

A new member of the DegP-type periplasmic serine endoproteases of the S1C family from the marine bacterium Cobetia amphilecti KMM 296 (CamSP) was expressed in Escherichia coli cells. The calculated molecular weight, number of amino acids, and isoelectric point (pI) of the mature protein CamSP are 69.957 kDa, 666, and 4.84, respectively. The proteolytic activity of the purified recombinant protease CamSP was 2369.4 and 1550.9 U/mg with the use of 1% bovine serum albumin (BSA) and casein as the substrates, respectively. The enzyme CamSP exhibited maximum activity at pH 6.0–6.2, while it was stable over a wide pH range from 5.8 to 8.5. The optimal temperature for the CamSP protease activity was 50 °C. The enzyme required NaCl or KCl at concentrations of 0.3 and 0.5 M, respectively, for its maximum activity. The Michaelis constant (K_m) and V_max for BSA were determined to be 41.7 µg/mL and 0.036 µg/mL min⁻¹, respectively. The metal ions Zn²⁺, Cu²⁺, Mn²⁺, Li²⁺, Mg²⁺, and Ca²⁺ slightly activated CamSP, while the addition of CoCl₂ to the incubation mixture resulted in a twofold increase in its protease activity. Ethanol, isopropanol, glycerol, and Triton-X-100 increased the activity of CamSP from two- to four-times. The protease CamSP effectively degraded the wheat flour proteins but had no proteolytic activity towards soybean, corn, and the synthetic substrates, α-benzoyl-Arg-p-nitroanilide (BAPNA) and N-Succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanine 4-nitroanilide (SAPNA). Full article

(This article belongs to the Special Issue Marine Microbes: Biotechnological Potential for Processes and Molecules)

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20 pages, 3114 KiB

Open AccessReview

Multifaceted Nature of Lipid Droplets in Viral Interactions and Pathogenesis

by Luis Herrera-Moro Huitron, Luis Adrián De Jesús-González, Macario Martínez-Castillo, José Manuel Ulloa-Aguilar, Carlos Cabello-Gutierrez, Cecilia Helguera-Repetto, Julio Garcia-Cordero and Moisés León Juárez

Microorganisms 2023, 11(7), 1851; https://doi.org/10.3390/microorganisms11071851 - 21 Jul 2023

Cited by 2 | Viewed by 2653

Abstract

Once regarded as inert organelles with limited and ill-defined roles, lipid droplets (LDs) have emerged as dynamic entities with multifaceted functions within the cell. Recent research has illuminated their pivotal role as primary energy reservoirs in the form of lipids, capable of being [...] Read more.

Once regarded as inert organelles with limited and ill-defined roles, lipid droplets (LDs) have emerged as dynamic entities with multifaceted functions within the cell. Recent research has illuminated their pivotal role as primary energy reservoirs in the form of lipids, capable of being metabolized to meet cellular energy demands. Their high dynamism is underscored by their ability to interact with numerous cellular organelles, notably the endoplasmic reticulum (the site of LD genesis) and mitochondria, which utilize small LDs for energy production. Beyond their contribution to cellular bioenergetics, LDs have been associated with viral infections. Evidence suggests that viruses can co-opt LDs to facilitate their infection cycle. Furthermore, recent discoveries highlight the role of LDs in modulating the host’s immune response. Observations of altered LD levels during viral infections suggest their involvement in disease pathophysiology, potentially through production of proinflammatory mediators using LD lipids as precursors. This review explores these intriguing aspects of LDs, shedding light on their multifaceted nature and implications in viral interactions and disease development. Full article

(This article belongs to the Special Issue Latest Review Papers in Virology 2023)

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14 pages, 18729 KiB

Open AccessArticle

A Recombinant Oncolytic Pseudorabies Virus Expressing Interleukin-18, Interferon-Gamma and PH20 Genes Promotes Systemic Antitumor Immunity

by Xiaohui Han, Jingshuai Sun, Xiaocheng Lv, Xiaoyu Tang, Yubin Zheng, Jinyun Ma and Yuan Sun

Microorganisms 2023, 11(7), 1850; https://doi.org/10.3390/microorganisms11071850 - 21 Jul 2023

Viewed by 1289

Abstract

Pseudorabies virus (PRV) is considered to be a promising oncolytic virus that has potential as a cancer gene therapy drug. In this study, PRV-DCD-1-70 was used as a vector to carry exogenous genes IL-18, IFN-γ and PH20 to construct novel recombinant PRV, [...] Read more.

Pseudorabies virus (PRV) is considered to be a promising oncolytic virus that has potential as a cancer gene therapy drug. In this study, PRV-DCD-1-70 was used as a vector to carry exogenous genes IL-18, IFN-γ and PH20 to construct novel recombinant PRV, rPRV-PH20 and rPRV-IL-18-γ-PH20, and their tumorolytic effects were evaluated in vitro and in vivo. Our study showed that recombinant PRV lysed all four tumor cell lines, Pan02, EMT-6, CT26 and H446, and rPRV-IL-18-γ-PH20 showed the best tumor lysis effect. Further studies in mice bearing Pan02 tumors showed that recombinant PRV, especially rPRV-IL-18-γ-PH20, were able to inhibit tumor growth. Moreover, an immunohistochemical analysis indicated that the recombinant PRV effectively increased the infiltration of CD4⁺T and CD8⁺T cells and enhanced the anti-tumor immune response of the organism in vivo. Overall, PRV carrying PH20 and IL-18-γ exogenous genes demonstrated anti-tumor effects, providing a foundation for the further development and application of PRV as a novel tumor oncolytic virus vector. Full article

(This article belongs to the Section Virology)

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19 pages, 838 KiB

Open AccessReview

The Role of Gut Dysbiosis in the Loss of Intestinal Immune Cell Functions and Viral Pathogenesis

by Farzaneh Fakharian, Siva Thirugnanam, David A. Welsh, Woong-Ki Kim, Jay Rappaport, Kyle Bittinger and Namita Rout

Microorganisms 2023, 11(7), 1849; https://doi.org/10.3390/microorganisms11071849 - 21 Jul 2023

Cited by 8 | Viewed by 4340

Abstract

The gut microbiome plays a critical role in maintaining overall health and immune function. However, dysbiosis, an imbalance in microbiome composition, can have profound effects on various aspects of human health, including susceptibility to viral infections. Despite numerous studies investigating the influence of [...] Read more.

The gut microbiome plays a critical role in maintaining overall health and immune function. However, dysbiosis, an imbalance in microbiome composition, can have profound effects on various aspects of human health, including susceptibility to viral infections. Despite numerous studies investigating the influence of viral infections on gut microbiome, the impact of gut dysbiosis on viral infection and pathogenesis remains relatively understudied. The clinical variability observed in SARS-CoV-2 and seasonal influenza infections, and the presence of natural HIV suppressors, suggests that host-intrinsic factors, including the gut microbiome, may contribute to viral pathogenesis. The gut microbiome has been shown to influence the host immune system by regulating intestinal homeostasis through interactions with immune cells. This review aims to enhance our understanding of how viral infections perturb the gut microbiome and mucosal immune cells, affecting host susceptibility and response to viral infections. Specifically, we focus on exploring the interactions between gamma delta (γδ) T cells and gut microbes in the context of inflammatory viral pathogenesis and examine studies highlighting the role of the gut microbiome in viral disease outcomes. Furthermore, we discuss emerging evidence and potential future directions for microbiome modulation therapy in the context of viral pathogenesis. Full article

(This article belongs to the Special Issue Microbiome in Infectious Diseases)

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18 pages, 3154 KiB

Open AccessArticle

Flavonoid Synthesis by Deinococcus sp. 43 Isolated from the Ginkgo Rhizosphere

by Jin Zhou, Kai Zou, Shaodong Fu, Zhenchun Duan, Guoqing Zhang, Xinhong Wu, Jingwen Huang, Shihui Li, Xueduan Liu, Shuangfei Zhang and Yili Liang

Microorganisms 2023, 11(7), 1848; https://doi.org/10.3390/microorganisms11071848 - 21 Jul 2023

Cited by 1 | Viewed by 1646

Abstract

Flavonoids are crucial in physiological and pharmaceutical processes, especially the treatment of cancer and the prevention of cardiovascular and cerebrovascular diseases. Flavonoid-producing plants and fungi have been extensively reported, but bacteria have been much less investigated as a source of flavonoid production. Deinococcus [...] Read more.

Flavonoids are crucial in physiological and pharmaceutical processes, especially the treatment of cancer and the prevention of cardiovascular and cerebrovascular diseases. Flavonoid-producing plants and fungi have been extensively reported, but bacteria have been much less investigated as a source of flavonoid production. Deinococcus sp. 43, a spherical flavonoid-producing bacteria from the Ginkgo rhizosphere, was reported in this study. First, the whole genome of Deinococcus sp. 43 was sequenced and a series of flavonoid anabolic genes were annotated. Simultaneously, High Performance Liquid Chromatography (HPLC) results showed that Deinococcus sp. 43 was capable of producing flavonoids, with a maximum quercetin output of 2.9 mg/L. Moreover, the relative expression of key genes involved in flavonoid synthesis was determined to test the completeness of the flavonoid anabolic pathway. The results of LC-MS analysis demonstrated that the flavonoids produced by Deinococcus sp. 43 were significantly different between intracellular and extracellular environments. The concentration of multiple glycosylated flavonoids was substantially higher in extracellular than intracellular environments, while the majority of flavonoids obtained in intracellular environments were hydroxylated multiple times. Lastly, the flavonoid biosynthetic pathway of Deinococcus sp. 43 was constructed based on the genomic analysis and the detected flavonoids. In conclusion, this study represents the first comprehensive characterization of the flavonoid-producing pathway of Deinococcus. The findings demonstrate that the strain has excellent potential as a genetically engineered strain for the industrial production of flavonoids. Full article

(This article belongs to the Special Issue Advances in Soil Microbiome 2.0)

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16 pages, 8371 KiB

Open AccessArticle

Optimizing Crop Production with Bacterial Inputs: Insights into Chemical Dialogue between Sphingomonas sediminicola and Pisum sativum

by Candice Mazoyon, Stéphane Firmin, Lamine Bensaddek, Audrey Pecourt, Amélie Chabot, Michel-Pierre Faucon, Vivien Sarazin, Fréderic Dubois and Jérôme Duclercq

Microorganisms 2023, 11(7), 1847; https://doi.org/10.3390/microorganisms11071847 - 21 Jul 2023

Cited by 4 | Viewed by 1031

Abstract

The use of biological inputs is an interesting approach to optimize crop production and reduce the use of chemical inputs. Understanding the chemical communication between bacteria and plants is critical to optimizing this approach. Recently, we have shown that Sphingomonas (S.) [...] Read more.

The use of biological inputs is an interesting approach to optimize crop production and reduce the use of chemical inputs. Understanding the chemical communication between bacteria and plants is critical to optimizing this approach. Recently, we have shown that Sphingomonas (S.) sediminicola can improve both nitrogen supply and yield in pea. Here, we used biochemical methods and untargeted metabolomics to investigate the chemical dialog between S. sediminicola and pea. We also evaluated the metabolic capacities of S. sediminicola by metabolic profiling. Our results showed that peas release a wide range of hexoses, organic acids, and amino acids during their development, which can generally recruit and select fast-growing organisms. In the presence of S. sediminicola, a more specific pattern of these molecules took place, gradually adapting to the metabolic capabilities of the bacterium, especially for pentoses and flavonoids. In turn, S. sediminicola is able to produce several compounds involved in cell differentiation, biofilm formation, and quorum sensing to shape its environment, as well as several molecules that stimulate pea growth and plant defense mechanisms. Full article

(This article belongs to the Special Issue Research on Plant—Bacteria Interactions)

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21 pages, 5670 KiB

Open AccessArticle

Cytokine Responses during Mycobacterium tuberculosis H37Rv and Ascaris lumbricoides Costimulation Using Human THP-1 and Jurkat Cells, and a Pilot Human Tuberculosis and Helminth Coinfection Study

by Khethiwe N. Bhengu, Ravesh Singh, Pragalathan Naidoo, Miranda N. Mpaka-Mbatha, Nomzamo Nembe-Mafa and Zilungile L. Mkhize-Kwitshana

Microorganisms 2023, 11(7), 1846; https://doi.org/10.3390/microorganisms11071846 - 21 Jul 2023

Viewed by 1356

Abstract

Background: Helminth infections are widespread in tuberculosis-endemic areas and are associated with an increased risk of active tuberculosis. In contrast to the pro-inflammatory Th1 responses elicited by Mycobacterium tuberculosis (Mtb) infection, helminth infections induce anti-inflammatory Th2/Treg responses. A robust Th2 response has been [...] Read more.

Background: Helminth infections are widespread in tuberculosis-endemic areas and are associated with an increased risk of active tuberculosis. In contrast to the pro-inflammatory Th1 responses elicited by Mycobacterium tuberculosis (Mtb) infection, helminth infections induce anti-inflammatory Th2/Treg responses. A robust Th2 response has been linked to reduced tuberculosis protection. Several studies show the effect of helminth infection on BCG vaccination and TB, but the mechanisms remain unclear. Aim: To determine the cytokine response profiles during tuberculosis and intestinal helminth coinfection. Methods: For the in vitro study, lymphocytic Jurkat and monocytic THP-1 cell lines were stimulated with Mtb H37Rv and Ascaris lumbricoides (A. lumbricoides) excretory-secretory protein extracts for 24 and 48 h. The pilot human ex vivo study consisted of participants infected with Mtb, helminths, or coinfected with both Mtb and helminths. Thereafter, the gene transcription levels of IFN-γ, TNF-α, granzyme B, perforin, IL-2, IL-17, NFATC2, Eomesodermin, IL-4, IL-5, IL-10, TGF-β and FoxP3 in the unstimulated/uninfected controls, singly stimulated/infected and costimulated/coinfected groups were determined using RT-qPCR. Results: TB-stimulated Jurkat cells had significantly higher levels of IFN-γ, TNF-α, granzyme B, and perforin compared to unstimulated controls, LPS- and A. lumbricoides-stimulated cells, and A. lumbricoides plus TB-costimulated cells (p < 0.0001). IL-2, IL-17, Eomes, and NFATC2 levels were also higher in TB-stimulated Jurkat cells (p < 0.0001). Jurkat and THP-1 cells singly stimulated with TB had lower IL-5 and IL-4 levels compared to those singly stimulated with A. lumbricoides and those costimulated with TB plus A. lumbricoides (p < 0.0001). A. lumbricoides-singly stimulated cells had higher IL-4 levels compared to TB plus A. lumbricoides-costimulated Jurkat and THP-1 cells (p < 0.0001). TGF-β levels were also lower in TB-singly stimulated cells compared to TB plus A. lumbricoides-costimulated cells (p < 0.0001). IL-10 levels were lower in TB-stimulated Jurkat and THP-1 cells compared to TB plus A. lumbricoides-costimulated cells (p < 0.0001). Similar results were noted for the human ex vivo study, albeit with a smaller sample size. Conclusions: Data suggest that helminths induce a predominant Th2/Treg response which may downregulate critical Th1 responses that are crucial for tuberculosis protection. Full article

(This article belongs to the Special Issue Parasitic Infection and Host Immunity 2.0)

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25 pages, 1961 KiB

Open AccessReview

The Potential Value of Probiotics after Dental Implant Placement

by Jia Xu, Chenfeng Chen, Shuaiqi Gan, Yihan Liao, Ruijie Fu, Chuping Hou, Shuhan Yang, Zheng Zheng and Wenchuan Chen

Microorganisms 2023, 11(7), 1845; https://doi.org/10.3390/microorganisms11071845 - 20 Jul 2023

Cited by 4 | Viewed by 2027

Abstract

Dental implantation is currently the optimal solution for tooth loss. However, the health and stability of dental implants have emerged as global public health concerns. Dental implant placement, healing of the surgical site, osseointegration, stability of bone tissues, and prevention of peri-implant diseases [...] Read more.

Dental implantation is currently the optimal solution for tooth loss. However, the health and stability of dental implants have emerged as global public health concerns. Dental implant placement, healing of the surgical site, osseointegration, stability of bone tissues, and prevention of peri-implant diseases are challenges faced in achieving the long-term health and stability of implants. These have been ongoing concerns in the field of oral implantation. Probiotics, as beneficial microorganisms, play a significant role in the body by inhibiting pathogens, promoting bone tissue homeostasis, and facilitating tissue regeneration, modulating immune-inflammatory levels. This review explores the potential of probiotics in addressing post-implantation challenges. We summarize the existing research regarding the importance of probiotics in managing dental implant health and advocate for further research into their potential applications. Full article

(This article belongs to the Special Issue Gut Microbiota: Health, Clinical & Beyonds)

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11 pages, 69924 KiB

Open AccessBrief Report

Evaluation of the Inactivation Efficacy of Four Disinfectants for Feline Parvovirus Derived from Giant Panda

by Qianling Peng, Zhisong Yang, Lin Wu, Peilun Yu, Qiang Li, Jingchao Lan, Li Luo, Shan Zhao and Qigui Yan

Microorganisms 2023, 11(7), 1844; https://doi.org/10.3390/microorganisms11071844 - 20 Jul 2023

Viewed by 1270

Abstract

Feline panleukopenia (FPL) is a highly contagious acute infectious disease caused by feline parvovirus (FPV). FPV has also been found in giant pandas with clinical signs of vomiting and mild diarrhea, posing a threat to this vulnerable species. Cleaning and disinfection may be [...] Read more.

Feline panleukopenia (FPL) is a highly contagious acute infectious disease caused by feline parvovirus (FPV). FPV has also been found in giant pandas with clinical signs of vomiting and mild diarrhea, posing a threat to this vulnerable species. Cleaning and disinfection may be one of the most efficacious ways to prevent FPV spread in the habitat of giant pandas. This study evaluated the inactivation effect of peracetic acid (PAA), povidone-iodine (PVP-I), glutaral and deciquam solution (JM) and Virkon S. The tissue culture infective dose (TCID₅₀) assay indicated that the virus may be totally inactivated by JM, PAA and Virkon S. Meanwhile, the hemagglutination (HA) assay showed a high inactivation efficiency of PAA and Virkon S. The analysis of Western blot revealed that PAA, Virkon S and JM can inhibit the structural protein synthesis. Taken together, our findings demonstrated that PAA could rapidly and efficiently inactivate FPV, representing an efficacious disinfectant for FPV control. Full article

(This article belongs to the Special Issue Emerging Viral Zoonoses)

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11 pages, 2478 KiB

Open AccessArticle

Seroepidemiology Study of Porcine Epidemic Diarrhea Virus in Mexico by Indirect Enzyme-Linked Immunosorbent Assay Based on a Recombinant Fragment of N-Terminus Domain Spike Protein

by Eduardo García-González, José Luis Cerriteño-Sánchez, Julieta Sandra Cuevas-Romero, José Bryan García-Cambrón, Francisco Jesus Castañeda-Montes and Francisco Villaseñor-Ortega

Microorganisms 2023, 11(7), 1843; https://doi.org/10.3390/microorganisms11071843 - 20 Jul 2023

Cited by 1 | Viewed by 1129

Abstract

Porcine epidemic diarrhea (PED) is an intestinal disease caused by the porcine epidemic diarrhea virus (PEDV) and affects Mexico’s swine industry. Despite the disease initially being described in Mexico in 2013, there has been no research into the virus’s seroepidemiology carried out in [...] Read more.

Porcine epidemic diarrhea (PED) is an intestinal disease caused by the porcine epidemic diarrhea virus (PEDV) and affects Mexico’s swine industry. Despite the disease initially being described in Mexico in 2013, there has been no research into the virus’s seroepidemiology carried out in Mexico. Thus, the goal of this study was to develop an indirect ELISA (iELISA) based on a recombinant N-terminal domain truncated spike (S) protein (rNTD-S) of PEDV to evaluate serum obtained from different pig-producing states in Mexico. A total of 1054 sera were collected from pig farms, slaughterhouses, and backyard production in the states of Aguascalientes, Guanajuato, Hidalgo, Jalisco, Morelos, Queretaro, Sinaloa, and Veracruz between 2019 and 2021. The rNTD-S protein was expressed in E. coli BL21 (DE3) cells. Negative and positive serum samples used in the iELISA were previously tested by Western blot. According to our findings, 61.66% of the serum samples (650/1054) were positive, with Jalisco having the highest percentage of positive samples, at a rate of 21.44% (226/1054). This is the first seroepidemiology study of PEDV carried out in Mexico, revealing that the virus is still circulating since the initial outbreak; furthermore, it provides an overview of PEDV’s spread and high level of persistence across the country’s key swine-producing states. Full article

(This article belongs to the Special Issue The Pathogenic Epidemiology of Important Swine Diseases)

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20 pages, 3305 KiB

Open AccessArticle

Lichenysin-like Polypeptide Production by Bacillus licheniformis B3-15 and Its Antiadhesive and Antibiofilm Properties

by Vincenzo Zammuto, Maria Giovanna Rizzo, Claudia De Pasquale, Guido Ferlazzo, Maria Teresa Caccamo, Salvatore Magazù, Salvatore Pietro Paolo Guglielmino and Concetta Gugliandolo

Microorganisms 2023, 11(7), 1842; https://doi.org/10.3390/microorganisms11071842 - 20 Jul 2023

Cited by 2 | Viewed by 1458

Abstract

We report the ability of the crude biosurfactant (BS B3-15), produced by the marine, thermotolerant Bacillus licheniformis B3-15, to hinder the adhesion and biofilm formation of Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213 to polystyrene and human cells. First, we attempted [...] Read more.

We report the ability of the crude biosurfactant (BS B3-15), produced by the marine, thermotolerant Bacillus licheniformis B3-15, to hinder the adhesion and biofilm formation of Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213 to polystyrene and human cells. First, we attempted to increase the BS yield, optimizing the culture conditions, and evaluated the surface-active properties of cell-free supernatants. Under phosphate deprivation (0.06 mM) and 5% saccharose, the yield of BS (1.5 g/L) increased by 37%, which could be explained by the earlier (12 h) increase in lchAA expression compared to the non-optimized condition (48 h). Without exerting any anti-bacterial activity, BS (300 µg/mL) prevented the adhesion of P. aeruginosa and S. aureus to polystyrene (47% and 36%, respectively) and disrupted the preformed biofilms, being more efficient against S. aureus (47%) than P. aeruginosa (26%). When added to human cells, the BS reduced the adhesion of P. aeruginosa and S. aureus (10× and 100,000× CFU/mL, respectively) without altering the epithelial cells’ viability. As it is not cytotoxic, BS B3-15 could be useful to prevent or remove bacterial biofilms in several medical and non-medical applications. Full article

(This article belongs to the Special Issue New Antimicrobial Strategies for Medical Implantation)

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13 pages, 2312 KiB

Open AccessArticle

Revealing the Bacterial Quorum-Sensing Effect on the Biofilm Formation of Diatom Cylindrotheca sp. Using Multimodal Imaging

by Cuiyun Yang, Guojuan Song, Jiyoung Son, Logan Howard and Xiao-Ying Yu

Microorganisms 2023, 11(7), 1841; https://doi.org/10.3390/microorganisms11071841 - 20 Jul 2023

Cited by 2 | Viewed by 1667

Abstract

Diatoms contribute to carbon fixation in the oceans by photosynthesis and always form biofouling organized by extracellular polymeric substances (EPS) in the marine environment. Bacteria-produced quorum-sensing signal molecules N-acyl homoserine lactones (AHLs) were found to play an important role in the development of [...] Read more.

Diatoms contribute to carbon fixation in the oceans by photosynthesis and always form biofouling organized by extracellular polymeric substances (EPS) in the marine environment. Bacteria-produced quorum-sensing signal molecules N-acyl homoserine lactones (AHLs) were found to play an important role in the development of Cylindrotheca sp. in previous studies, but the EPS composition change was unclear. This study used the technology of alcian blue staining and scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and time-of-flight secondary ion mass spectrometry (ToF-SIMS) to directly observe the biofilm formation process. The results showed that AHLs promote the growth rates of diatoms and the EPS secretion of biofilm components. AHLs facilitated the diatom-biofilm formation by a forming process dependent on the length of carbon chains. AHLs increased the biofilm thickness and the fluorescence intensity and then altered the three-dimensional (3D) structures of the diatom-biofilm. In addition, the enhanced EPS content in the diatom-biofilm testified that AHLs aided biofilm formation. This study provides a collection of new experimental evidence of the interaction between bacteria and microalgae in fouling biofilms. Full article

(This article belongs to the Special Issue Host–Biofilm Interactions 2.0)

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10 pages, 2345 KiB

Open AccessArticle

Contamination of Hotel Water Distribution Systems by Legionella Species: Environmental Surveillance in Campania Region, South Italy

by Valeria Di Onofrio, Mariangela Pagano, Marco Santulli, Annamaria Rossi, Renato Liguori, Mirella Di Dio and Giorgio Liguori

Microorganisms 2023, 11(7), 1840; https://doi.org/10.3390/microorganisms11071840 - 20 Jul 2023

Viewed by 1285

Abstract

The COVID-19 pandemic period was marked by the absence or reduced circulation of some infectious diseases. Legionellosis may have been affected by the prevention measures adopted to counter COVID-19. Legionellosis is a form of pneumonia interstitial that is normally transmitted via aerosol-containing bacteria [...] Read more.

The COVID-19 pandemic period was marked by the absence or reduced circulation of some infectious diseases. Legionellosis may have been affected by the prevention measures adopted to counter COVID-19. Legionellosis is a form of pneumonia interstitial that is normally transmitted via aerosol-containing bacteria (genus Legionella), that could be present in contaminated water sources and is often associated with travel and with staying in hotels. In this work, the data of the environmental surveys carried out by ARPA Campania in accommodation facilities since 2019 were analyzed for a better understanding of the dispersion patterns of L. pneumophila associated with the environment and to evaluate the variation of the data during the pandemic period. The aim was to provide a better understanding of Legionella at different geographic scales and to define a predictive epidemiological method. Results: In 2019, the Legionella genus contaminated 37.7% of all tourist facilities evaluated. In 2020, the Legionella genus contaminated 44.4% of all tourist facilities evaluated. In 2021, the Legionella genus contaminated 54.2% of all tourist facilities evaluated. Conclusions: Legionella pneumophila was the most prevalent species in our community, serogroup 1 was the most frequently isolated and the most implicated risk factor of contamination was the temperature of water in circulation. Full article

(This article belongs to the Special Issue Clinical and Environmental Surveillance for the Prevention of Legionellosis)

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19 pages, 1249 KiB

Open AccessArticle

Physiological Conditions Leading to Maternal Subclinical Ketosis in Holstein Dairy Cows Can Impair the Offspring’s Postnatal Growth and Gut Microbiome Development

by Jessica Halfen, Nathaly Ana Carpinelli, Sergio Lasso-Ramirez, Tainara Cristina Michelotti, Emily C. Fowler, Benoit St-Pierre, Erminio Trevisi and Johan S. Osorio

Microorganisms 2023, 11(7), 1839; https://doi.org/10.3390/microorganisms11071839 - 19 Jul 2023

Cited by 4 | Viewed by 1156

Abstract

Maternal metabolic disruptions, such as ketosis, can have adverse effects on fetal development and influence postnatal factors. Twelve Holstein calves were randomly enrolled in this study at birth and monitored until 8 weeks of age. The study was conducted from fall 2018 until [...] Read more.

Maternal metabolic disruptions, such as ketosis, can have adverse effects on fetal development and influence postnatal factors. Twelve Holstein calves were randomly enrolled in this study at birth and monitored until 8 weeks of age. The study was conducted from fall 2018 until spring 2019. After completing the data collection period, calves were classified according to their respective dams ketotic condition after parturition. This classification was based on dam blood β-hydroxybutyrate < 1.4 mmol/L nonketotic (NONKET; n = 6 calves) or ≥1.4 mmol/L subclinical-ketotic (SK; n = 6 calves). SK calves had greater birth body weight (p = 0.05) but exhibited a slower growth rate compared to NONKET calves from 1 to 8 weeks (p = 0.02). At birth, SK calves had lower (p < 0.01) levels of non-esterified fatty acids and bilirubin compared to NONKET calves. Analysis of feces alpha diversity indicates that by 3 weeks, NONKET calves had greater diversity, richness, and evenness. Butyricicoccus pullicaecorum and Gallibacterium anatis were more abundant in SK calves (p < 0.05) at 3 weeks. In contrast, NONKET calves had a greater (p < 0.05) abundance of Sharpae azabuensis at 3 weeks. These findings suggest that subclinical ketosis in cows can impact the in-utero development, postnatal growth, and maturing gut microbiome of their offspring. Full article

(This article belongs to the Special Issue Gut Microbiota Development in Farm Animals 2.0)

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9 pages, 1173 KiB

Open AccessCommunication

M2 Polarization and Inhibition of Host Cell Glycolysis Contributes Intracellular Survival of Salmonella Strains in Chicken Macrophage HD-11 Cells

by Haiqi He, Kenneth J. Genovese, Ryan J. Arsenault, Christina L. Swaggerty, Casey N. Johnson, J. Allen Byrd and Michael H. Kogut

Microorganisms 2023, 11(7), 1838; https://doi.org/10.3390/microorganisms11071838 - 19 Jul 2023

Cited by 1 | Viewed by 1102

Abstract

Salmonella enterica is a group of facultative, gram-negative bacteria. Recently, new evidence indicated that Salmonella could reprogram the host metabolism to increase energy or metabolites available for intracellular replication. In this study, using a chicken-specific kinomic immunometabolism peptide array analysis, we found that [...] Read more.

Salmonella enterica is a group of facultative, gram-negative bacteria. Recently, new evidence indicated that Salmonella could reprogram the host metabolism to increase energy or metabolites available for intracellular replication. In this study, using a chicken-specific kinomic immunometabolism peptide array analysis, we found that infection by S. Enteritidis induced significant phosphorylation changes in many key proteins of the glycolytic pathway in chicken macrophage HD-11 cells, indicating a shift in glycolysis caused by Salmonella infection. Nitric oxide production and changes of glycolysis and mitochondrial oxidative phosphorylation (OXPHOS) represented by extracellular acidification rate (ECAR) and oxygen consumption rate (OCR), respectively, were measured in chicken macrophages infected with three Salmonella strains (S. Enteritidis, S. Heidelberg, and S. Senftenberg). The infection reduced glycolysis and enhanced OXPHOS in chicken macrophages as indicated by changes of ECAR and OCR. Salmonella strains differentially affected macrophage polarization and glycolysis. Among three strains tested, S. Enteritidis was most effective in downregulating glycolysis and promoting M2 polarization as measured by ECAR, ORC, and NO production; while S. Senftenberg did not alter glycolysis and may promote M1 polarization. Our results suggested that downregulation of host cell glycolysis and increase of M2 polarization of macrophages may contribute to increased intracellular survival of S. Enteritidis. Full article

(This article belongs to the Section Food Microbiology)

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13 pages, 2068 KiB

Open AccessArticle

Molecular Characterization of Salmonella Phage Wara Isolated from River Water in Brazil

by Danitza Xiomara Romero-Calle, Francisnei Pedrosa-Silva, Luiz Marcelo Ribeiro Tomé, Vagner Fonseca, Raquel Guimarães Benevides, Leila Thaise Santana de Oliveira Santos, Tulio de Oliveira, Mateus Matiuzzi da Costa, Luiz Carlos Junior Alcantara, Vasco Ariston de Carvalho Azevedo, Bertram Brenig, Thiago M. Venancio, Craig Billington and Aristóteles Góes-Neto

Microorganisms 2023, 11(7), 1837; https://doi.org/10.3390/microorganisms11071837 - 19 Jul 2023

Cited by 1 | Viewed by 1447

Abstract

Antimicrobial resistance is increasing despite new treatments being employed, so novel strategies are required to ensure that bacterial infections remain treatable. Bacteriophages (phages; bacteria viruses) have the potential to be used as natural antimicrobial methods to control bacterial pathogens such as Salmonella spp. [...] Read more.

Antimicrobial resistance is increasing despite new treatments being employed, so novel strategies are required to ensure that bacterial infections remain treatable. Bacteriophages (phages; bacteria viruses) have the potential to be used as natural antimicrobial methods to control bacterial pathogens such as Salmonella spp. A Salmonella phage, Wara, was isolated from environmental water samples at the Subaé River Basin, Salvador de Bahia, Brazil. The basin has environmental impacts in its main watercourses arising from the dumping of domestic and industrial effluents and agricultural and anthropological activities. The phage genome sequence was determined by Oxford Nanopore Technologies (ONT) MinION and Illumina HiSeq sequencing, and assembly was carried out by Racon (MinION) and Unicycler (Illumina, Illumina + MinION). The genome was annotated and compared to other Salmonella phages using various bioinformatics approaches. MinION DNA sequencing combined with Racon assembly gave the best complete genome sequence. Phylogenetic analysis revealed that Wara is a member of the Tequintavirus genus. A lack of lysogeny genes, antimicrobial resistance, and virulence genes indicated that Wara has therapeutic and biocontrol potential against Salmonella species in healthcare and agriculture. Full article

(This article belongs to the Special Issue Biotechnological Applications of Bacteriophages and Enteric Viruses)

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