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Search Results (149)

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Keywords = 16S and 18S rRNA gene fragments

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23 pages, 2657 KiB  
Article
Enrichment Cultures of Extreme Acidophiles with Biotechnological Potential
by Khussain Valiyev, Aliya Yskak, Elena Latyuk, Alena Artykova, Rakhimbayev Berik, Vadim Chashkov and Aleksandr Bulaev
Mining 2025, 5(3), 49; https://doi.org/10.3390/mining5030049 - 1 Aug 2025
Viewed by 76
Abstract
The purpose of this work was to obtain specialized enrichment cultures from an original extreme acidophilic consortium of extremely acidophilic microorganisms and to study their microbial community composition and biotechnological potential. At temperatures of 25, 35, 40 and 50 °C, distinct enrichments of [...] Read more.
The purpose of this work was to obtain specialized enrichment cultures from an original extreme acidophilic consortium of extremely acidophilic microorganisms and to study their microbial community composition and biotechnological potential. At temperatures of 25, 35, 40 and 50 °C, distinct enrichments of extremely acidophilic microorganisms used in the processes of bioleaching sulfide ores were obtained using nutrient media containing ferrous sulfate, elemental sulfur and a copper sulfide concentrate as nutrient inorganic substrates, with and without the addition of 0.02% yeast extract. The microbial community composition was studied using the sequencing of the V3–V4 hypervariable region of the 16S rRNA genes. The different growth conditions led to changes in the microbial composition and relative abundance of mesophilic and moderately thermophilic, strict autotrophic and mixotrophic microorganisms in members of the genera Acidithiobacillus, Sulfobacillus, Leptospirillum, Acidibacillus, Ferroplasma and Cuniculiplasma. The dynamics of the oxidation of ferrous iron, sulfur, and sulfide minerals (pyrite and chalcopyrite) by the enrichments was also studied in the temperature range of 25 to 50 °C. The study of enrichment cultures using the molecular biological method using the metabarcoding method of variable V3–24 V4 fragments of 16S rRNA genes showed that enrichment cultures obtained under different conditions differed in composition, which can be explained by differences in the physiological properties of the identified microorganisms. Regarding the dynamics of the oxidation of ferrous ions, sulfur, and sulfide minerals (pyrite and chalcopyrite), each enrichment culture was studied at a temperature range of 25 to 50 °C and indicated that all obtained enrichments were capable of oxidizing ferrous iron, sulfur and minerals at different rates. The obtained enrichment cultures may be used in further work to increase bioleaching by using the suitable inoculum for the temperature and process conditions. Full article
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14 pages, 1132 KiB  
Article
Phylogenetic Reclassification of Metarhizium granulomatis and Metarhizium viride Species Complex
by Johanna Würf and Volker Schmidt
Pathogens 2025, 14(8), 745; https://doi.org/10.3390/pathogens14080745 - 29 Jul 2025
Viewed by 239
Abstract
Metarhizium (M.) granulomatis and M. viride have previously been described as pathogens causing hyalohyphomycosis in various species of captive chameleons and bearded dragons (Pogona vitticeps). Previous studies yielded different genotypes of M. granulomatis and M. viride based on sequencing of the [...] Read more.
Metarhizium (M.) granulomatis and M. viride have previously been described as pathogens causing hyalohyphomycosis in various species of captive chameleons and bearded dragons (Pogona vitticeps). Previous studies yielded different genotypes of M. granulomatis and M. viride based on sequencing of the internal transcribed spacer 1-5.8S rDNA (ITS-1-5.8S) and a fragment of the large subunit of the 28S rDNA (LSU). The aim of this study was to clarify the relationships between these genotypes and obtain a more accurate phylogenetic classification by sequencing two different loci of the RNA polymerase II second largest subunit (NRPB2), referred to as RPB1 and RPB2, and the translation elongation factor 1 alpha (EF1α). A total of 23 frozen isolates from 21 lizards, including the first isolates of M. granulomatis and M. viride from Parson’s chameleons (Calumma parsonii), were available for phylogenetic analysis. A total of 13 isolates belonged to the M. granulomatis complex and 10 isolates belonged to the M. viride complex. Following the amplification and sequencing of the protein-coding genes, the resulting nucleotide sequences were analyzed, trimmed and assembled. These were further analyzed with regard to differences in single-nucleotide polymorphisms (SNPs) and amino acid structure. In consideration of the results of the present analyses, a phylogenetic reclassification is recommended. Three different genotypes of M. granulomatis can be distinguished, which can be phylogenetically addressed as subspecies. Six subspecies can be distinguished regarding M. viride. Full article
(This article belongs to the Special Issue Filamentous Fungal Pathogens: 2nd Edition)
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18 pages, 1047 KiB  
Article
Protein Functional Effector (pfe) Noncoding RNAS Are Identical to Fragments from Various Noncoding RNAs
by Roberto Patarca and William A. Haseltine
Int. J. Mol. Sci. 2025, 26(14), 6870; https://doi.org/10.3390/ijms26146870 - 17 Jul 2025
Viewed by 269
Abstract
Protein functional effector (pfe)RNAs were introduced in 2015 as PIWI-interacting-like small noncoding (nc)RNAs and were later categorized as a novel group based on being 2′-O-methylated at their 3′-end, directly binding and affecting protein function, but not levels, and not matching known RNAs. Here, [...] Read more.
Protein functional effector (pfe)RNAs were introduced in 2015 as PIWI-interacting-like small noncoding (nc)RNAs and were later categorized as a novel group based on being 2′-O-methylated at their 3′-end, directly binding and affecting protein function, but not levels, and not matching known RNAs. Here, we document that human pfeRNAs match fragments of GenBank database-annotated human ncRNAs. PDLpfeRNAa matches the 3′-half fragment of a mitochondrial transfer (t)RNA, and PDLpfeRNAb matches a 28S ribosomal (r)RNA fragment. These PDLpfeRNAs are known to bind to tumor programmed death ligand (PD-L)1, enhancing or inhibiting its interaction with lymphocyte PD-1 and consequently tumor immune escape, respectively. In a validated 8-pfeRNA-set classifier for pulmonary nodule presence and benign vs. malignant nature, seven here match one or more of the following: transfer, micro, Y, PIWI, long (lnc)RNAs, and a PDLpfeRNAa fragment. The previously identified chromosomal locations of these pfeRNAs and their matches partially overlap. Another 2-pfeRNA set was previously determined to distinguish between controls, patients with pulmonary tuberculosis, and those with lung cancer. One pfeRNA, previously shown to bind p60-DMAD and affect apoptosis, complements small nucleolar RNA SNORD45C, matching smaller 18S rRNA and lncRNA segments. Thus, pfeRNAs appear to have a common origin with known multifunctional ncRNA fragments. Differential modification may contribute to the multifunctionality of ncRNAs. For instance, for tRNA fragments, stabilizing 3′-end 2′-O-methylation, 3′-aminoacylation, and glycosylation modifications may regulate protein function, translation, and extracellular effects, respectively. One ncRNA gene can encode multiple fragments, multiple genes can encode the same fragment, and differentially modified ncRNA fragments might synergize or antagonize each other. Full article
(This article belongs to the Special Issue Targeting RNA Molecules)
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11 pages, 987 KiB  
Article
Peculiarities of Diagnostic Reliability—Nested PCR Versus SAT in the Identification of Helicobacter pylori
by Barbora Šipková, Michaela Abrahamovská, Janka Klingová, Bianka Prokopová, Jana Krčmáriková, Iveta Cihová and Pavol Sulo
Microorganisms 2025, 13(7), 1498; https://doi.org/10.3390/microorganisms13071498 - 27 Jun 2025
Viewed by 398
Abstract
H. pylori detection via the stool antigen test (SAT) requires 100 times more cells than nested PCR (NPCR) for a 454 bp amplicon, but is significantly more sensitive in identifying positive stool samples. To understand this contradiction, we developed an NPCR assay to [...] Read more.
H. pylori detection via the stool antigen test (SAT) requires 100 times more cells than nested PCR (NPCR) for a 454 bp amplicon, but is significantly more sensitive in identifying positive stool samples. To understand this contradiction, we developed an NPCR assay to amplify a shorter 148 bp segment of the 16S rRNA gene. The assay was extremely sensitive and reliable when adhering to particular rules commonly used in forensic laboratories. The SAT and NPCR for long and short amplicons were compared using stool samples from 208 gastroenterological patients, of which 27.9% were identified as positive according to the SAT and only 6.25% according to the 454 bp NPCR amplicon, but 51.0% in the short 148 bp NPCR. Among 100 asymptomatic volunteers, the prevalence was 35% in the SAT assay and 22% in the long NPCR, but as much as 66.6% of positives were determined in the short 148 bp NPCR. The specificity of the PCR product was determined via DNA sequencing, which confirmed H. pylori’s origin in all NPCR-positive samples. Apparently, the stool contains mostly short fragments of H. pylori DNA, and the most plausible explanation for the SAT/NPCR paradox is the degradation of H. pylori DNA in the digestive system. Full article
(This article belongs to the Section Microbial Biotechnology)
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10 pages, 2975 KiB  
Article
Differential Distribution of Trypanosoma vivax and Trypanosoma theileri in Cattle from Distinct Agroecological Regions of Central Argentina
by Maria Celeste Facelli Fernández, Johann Barolin, Martin Allassia, Javier Hernan Gonzalez, Pablo Martin Beldomenico and Lucas Daniel Monje
Parasitologia 2025, 5(2), 27; https://doi.org/10.3390/parasitologia5020027 - 5 Jun 2025
Viewed by 538
Abstract
Bovine trypanosomiasis, caused by Trypanosoma vivax, affects livestock productivity and is increasingly being reported in South America. This study aimed to detect and characterize Trypanosoma spp. infections, with a focus on T. vivax, in cattle from two distinct agroecological regions of [...] Read more.
Bovine trypanosomiasis, caused by Trypanosoma vivax, affects livestock productivity and is increasingly being reported in South America. This study aimed to detect and characterize Trypanosoma spp. infections, with a focus on T. vivax, in cattle from two distinct agroecological regions of central Argentina: a dairy-producing plain, located in the Espinal ecoregion, and a riparian zone, dedicated to beef production, located in the Delta and Islands of Paraná ecoregion. A total of 220 blood samples were collected from nine cattle farms and analyzed using real-time PCR, melting curve analysis, and the sequencing of 18S rRNA gene fragments. Trypanosoma vivax was detected at low prevalence (2.73%), exclusively in dairy cattle. In contrast, the prevalence of Trypanosoma theileri was much higher (10.91%), and it was found mainly in beef cattle from the riparian region. Phylogenetic analyses confirmed the species identity in all sequenced samples. No trypanosomes were observed by microscopy, and none of the animals showed clinical signs. The results indicate a differential distribution of T. vivax and T. theileri between regions and production systems. Although the study initially focused on T. vivax, the detection of T. theileri highlights the need to consider multiple Trypanosoma species in epidemiological surveys. This study contributes new data on the occurrence of bovine trypanosomes in central Argentina under extensive and semi-intensive management systems. Full article
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11 pages, 1773 KiB  
Article
First Molecular Identification of Zoonotic Babesia odocoilei in Ticks from Romania
by Ioan Cristian Dreghiciu, Diana Hoffman, Simona Dumitru, Ion Oprescu, Mirela Imre, Tiana Florea, Anamaria Plesko, Vlad Iorgoni, Sorin Morariu, Gheorghe Dărăbuș and Marius Stelian Ilie
Microorganisms 2025, 13(6), 1182; https://doi.org/10.3390/microorganisms13061182 - 22 May 2025
Viewed by 637
Abstract
Babesia odocoilei is an emerging zoonotic protozoan parasite primarily associated with cervids, with growing recognition among non-cervid hosts and in terms of potential public health implications. While this species has been documented in North America and parts of Europe, data on its presence [...] Read more.
Babesia odocoilei is an emerging zoonotic protozoan parasite primarily associated with cervids, with growing recognition among non-cervid hosts and in terms of potential public health implications. While this species has been documented in North America and parts of Europe, data on its presence in Romania remain scarce. This study aimed to investigate the presence of Babesia spp. in ticks collected from Romania, providing new information on the existing species and their distribution, as well as their potential epidemiologic significance. A total of 41 Ixodidae ticks were collected from 184 wild boars across six counties from Western and Central Romania. Ticks were identified using morphological assessments, and DNA was extracted from the samples using a standardized protocol. The presence of Babesia spp. was assessed using real-time PCR with primers and a Taq Man probe targeting 116 bp fragments of 18S rRNA genes. Molecular analysis was used to detect Babesia spp. DNA from a single tick sample (1/41, 2.43%), identified as Dermacentor marginatus, from Timiș County. The resulting amplicons were sequenced and compared with reference sequences in GenBank for species confirmation. This finding represents the first molecular identification of B. odocoilei in questing ticks from Romania. The expanding host range and geographic distribution of B. odocoilei raise concerns regarding its zoonotic potential. The presence of this pathogen in Dermacentor marginatus ticks suggests a broader vector competence than previously recognized, and future research should focus on host reservoirs, vector competence, and potential zoonotic transmission, with an emphasis on public health implications, including potential implications for veterinary diagnostics, vector control policies, and public health awareness regarding emerging tick-borne pathogens. Full article
(This article belongs to the Special Issue Ticks and Threats: Insights on Tick-Borne Diseases)
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22 pages, 4602 KiB  
Article
Dynamics of Phyto- and Bacterioplankton in Southern Baikal and Irkutsk Reservoir During the Open Water Period of 2023 According to Metabarcoding Data
by Yuri Galachyants, Darya Petrova, Artem Marchenkov, Maria Nalimova and Yelena Likhoshway
Diversity 2025, 17(6), 369; https://doi.org/10.3390/d17060369 - 22 May 2025
Viewed by 500
Abstract
Artificial reservoirs formed by hydroelectric dams are young ecosystems requiring water quality monitoring, as they often serve local populations. Traditionally, this is performed through hydrochemical and sanitary assessments, alongside phytoplankton composition analysis. This study aimed to assess the seasonal dynamics of microbial communities—both [...] Read more.
Artificial reservoirs formed by hydroelectric dams are young ecosystems requiring water quality monitoring, as they often serve local populations. Traditionally, this is performed through hydrochemical and sanitary assessments, alongside phytoplankton composition analysis. This study aimed to assess the seasonal dynamics of microbial communities—both bacterioplankton and microeukaryotes including phytoplankton—in the Irkutsk Reservoir (IR), which is fed by the cold oligotrophic waters of Southern Baikal (SB). Using parallel metabarcoding of 16S and 18S rRNA gene fragments, we analyzed community composition during the open-water season and evaluated the ecological connectivity between these two freshwater systems. We demonstrated that seasonal changes in microeukaryotic communities were closely linked between SB and IR, with the greatest divergence observed in early summer and progressive convergence by autumn. Metabarcoding confirmed microscopy-based observations while providing higher taxonomic resolution and detecting otherwise overlooked groups. Bacterioplankton communities also exhibited seasonal variation and were shaped by environmental gradients and reservoir characteristics. Cyanobacteria peaked in SB in late summer but did not dominate communities, unlike in mesotrophic lowland reservoirs. These findings demonstrate the value of metabarcoding for freshwater monitoring and provide new insights into microbial community dynamics in river–reservoir systems influenced by oligotrophic lake inflow. Full article
(This article belongs to the Special Issue DNA Barcodes for Evolution and Biodiversity—2nd Edition)
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23 pages, 3021 KiB  
Article
The Combination of Start-Codon-Targeted (SCoT) and Falling Stone (FaSt) Transposon-Specific Primers Provides an Efficient Marker Strategy for Prunus Species
by Beti Ivanovska, Thanyarat Onlamun, Júlia Halász and Attila Hegedűs
Int. J. Mol. Sci. 2025, 26(9), 3972; https://doi.org/10.3390/ijms26093972 - 23 Apr 2025
Viewed by 587
Abstract
A novel primer (FaSt-R) targeting the Prunus-specific Falling Stone (FaSt) non-autonomous transposon was combined with start-codon-targeted (SCoT) primers to assess genetic diversity in 12 cultivars from six Prunus species and 28 cultivars of European plum. Compared to SCoT-only analyses, the [...] Read more.
A novel primer (FaSt-R) targeting the Prunus-specific Falling Stone (FaSt) non-autonomous transposon was combined with start-codon-targeted (SCoT) primers to assess genetic diversity in 12 cultivars from six Prunus species and 28 cultivars of European plum. Compared to SCoT-only analyses, the SCoT–FaSt combination produced fewer total bands but a higher percentage of polymorphic bands, while maintaining comparable values for polymorphism information content, resolving power, gene diversity, and Shannon’s index. SCoT–FaSt markers generated bands across a broader size range, which made gel patterns less dense, enabling the more accurate detection of differentially amplified fragments. Neighbor-joining and principal component analyses confirmed that SCoT–FaSt markers provided sufficient phylogenetic resolution at both interspecific and intraspecific levels. The sequencing of 32 SCoT–FaSt amplicons revealed FaSt elements in 26 fragments, with SCoT primers preferentially annealing to GC-rich exonic and intergenic regions. Seventeen protein-coding and one RNA-coding gene were partially identified, with FaSt elements localized in UTRs and introns of genes with key physiological functions. Comparative analysis indicated a biased distribution of FaSt elements between the Cerasus and Prunus subgenera. In silico findings suggest that FaSt elements are more fragmented in cherry species, potentially contributing to subgeneric divergence. Overall, the SCoT–FaSt marker system is effective for evaluating Prunus genetic diversity, reconstructing phylogenetic relationships, and elucidating the genomic impact of an active Mutator-like transposon. Full article
(This article belongs to the Section Molecular Plant Sciences)
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22 pages, 7129 KiB  
Article
First Report of a Psyllid Vector of ‘Candidatus Phytoplasma pruni’ (Strain 16SrIII-J)
by Tomás Llantén, Sebastián Cabrera, Javiera Fuentes, Camila Gamboa, Constanza González, Alan Zamorano, Tomislav Curkovic, Daniel Burckhardt and Nicola Fiore
Plants 2025, 14(9), 1279; https://doi.org/10.3390/plants14091279 - 23 Apr 2025
Viewed by 712
Abstract
In Graneros, O’Higgins Region, Chile, the mallow psyllid (Russelliana solanicola Tuthill, 1959) from Malva nicaeensis L. was identified as a potential vector of ‘Candidatus Phytoplasma pruni’. Over an 8-month period, 2089 specimens of a species of Psylloidea, including immatures and adults, [...] Read more.
In Graneros, O’Higgins Region, Chile, the mallow psyllid (Russelliana solanicola Tuthill, 1959) from Malva nicaeensis L. was identified as a potential vector of ‘Candidatus Phytoplasma pruni’. Over an 8-month period, 2089 specimens of a species of Psylloidea, including immatures and adults, were captured. We only selected the adults used for transmission trials in Catharanthus roseus (L.) G. Don (periwinkle) plants. By nested PCR, using primer pairs for phytoplasma detection in 16S rRNA and IdpA genes, 7 out of 113 (6.2%) periwinkle plants used in transmission trials were found to be infected by phytoplasmas. Insects that fed on these plants also tested positive for the same phytoplasmas. Periwinkle plants never showed virescence and phyllody, as commonly occurs with phytoplasma 16SrIII-J infection due to the effector SAP54. In this case, using primer pairs for the SAP54 gene, an amplification product was never obtained. Virtual restriction fragment length polymorphism (RFLP) analysis of F2nR2 fragments indicated that the phytoplasma, found in both periwinkle plants and insects used in transmission trials, belongs to the 16SrIII-J ribosomal subgroup. The COI gene of the psyllids samples was amplified and sequenced, showing a similarity ranging from 84.84% to 85.02% with R. solanicola from Solanum tuberosum L. The mitochondrial genome of the psyllid was also sequenced, revealing a 14,835 bp circular DNA molecule with 37 genes. The mallow psyllid transmitted the phytoplasma 16SrIII-J to periwinkle plants. The molecular identification of the insect does not match the morphological one, indicating that the mallow psyllid may constitute a cryptic species within the polyphagous R. solanicola species. This is the first report of a psyllid as a vector of the phytoplasma 16SrIII-J. Full article
(This article belongs to the Collection Plant Disease Diagnostics and Surveillance in Plant Protection)
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15 pages, 7280 KiB  
Article
Assembly and Comparative Analysis of the Complete Mitochondrial Genomes of Smilax glabra and Smilax zeylanica
by Guojian Liao, Wenjing Liang, Haixia Yu, Kun Zhang, Linxuan Li, Shixin Feng, Lisha Song, Cuihong Yang, Lingyun Wan, Dongqiang Zeng, Zhanjiang Zhang and Shugen Wei
Genes 2025, 16(4), 450; https://doi.org/10.3390/genes16040450 - 14 Apr 2025
Viewed by 624
Abstract
Background: Smilax glabra (S. glabra) and Smilax zeylanica (S. zeylanica), two medicinally important species within the genus Smilax, have been widely used in Traditional Chinese Medicine (TCM) for the treatment of rheumatism, traumatic injuries, and related ailments. Despite their medicinal [...] Read more.
Background: Smilax glabra (S. glabra) and Smilax zeylanica (S. zeylanica), two medicinally important species within the genus Smilax, have been widely used in Traditional Chinese Medicine (TCM) for the treatment of rheumatism, traumatic injuries, and related ailments. Despite their medicinal significance, research on the mitochondrial DNA (mtDNA) of Smilax species remains limited. Methods: We utilized NovaSeq 6000 and PromethION sequencing platforms to assemble the complete mitochondrial genomes of Smilax glabra and Smilax zeylanica, and conducted in-depth comparative genomic and evolutionary analyses. Results: The complete mitochondrial genomes of S. glabra and S. zeylanica were assembled and annotated, with total lengths of 535,215 bp and 471,049 bp, respectively. Both genomes encode 40 unique protein-coding genes (PCGs), composed of 24 core and 16 non-core genes, alongside multiple tRNA and rRNA genes. Repetitive element analysis identified 158 and 403 dispersed repeats in S. glabra and S. zeylanica, respectively, as well as 123 and 139 simple sequence repeats (SSRs). RNA editing site predictions revealed C-to-U conversions in both species. Additionally, chloroplast-to-mitochondrial DNA migration analysis detected 34 homologous fragments in S. glabra and 28 homologous fragments in S. zeylanica. Phylogenetically, S. glabra and S. zeylanica cluster within the Liliales order and Smilacaceae family, closely related to Lilium species. Collinearity analysis indicated numerous syntenic blocks between Smilax and three other Liliopsida species, though gene order was not conserved. Conclusions: This study presents high-quality mitochondrial genome assemblies for S. glabra and S. zeylanica, providing valuable insights into molecular identification and conservation efforts of these traditional medicinal plants. Full article
(This article belongs to the Section Plant Genetics and Genomics)
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16 pages, 1024 KiB  
Article
Global Archaeal Diversity Revealed Through Massive Data Integration: Uncovering Just Tip of Iceberg
by Antonios Kioukis, Antonio Pedro Camargo, Pavlos Pavlidis, Ioannis Iliopoulos, Nikos C Kyrpides and Ilias Lagkouvardos
Microorganisms 2025, 13(3), 598; https://doi.org/10.3390/microorganisms13030598 - 5 Mar 2025
Viewed by 1084
Abstract
The domain of Archaea has gathered significant interest for its ecological and biotechnological potential and its role in helping us to understand the evolutionary history of Eukaryotes. In comparison to the bacterial domain, the number of adequately described members in Archaea is [...] Read more.
The domain of Archaea has gathered significant interest for its ecological and biotechnological potential and its role in helping us to understand the evolutionary history of Eukaryotes. In comparison to the bacterial domain, the number of adequately described members in Archaea is relatively low, with less than 1000 species described. It is not clear whether this is solely due to the cultivation difficulty of its members or, indeed, the domain is characterized by evolutionary constraints that keep the number of species relatively low. Based on molecular evidence that bypasses the difficulties of formal cultivation and characterization, several novel clades have been proposed, enabling insights into their metabolism and physiology. Given the extent of global sampling and sequencing efforts, it is now possible and meaningful to question the magnitude of global archaeal diversity based on molecular evidence. To do so, we extracted all sequences classified as Archaea from 500 thousand amplicon samples available in public repositories. After processing through our highly conservative pipeline, we named this comprehensive resource the ‘Global Archaea Diversity’ (GAD), which encompassed nearly 3 million molecular species clusters at 97% similarity, and organized it into over 500 thousand genera and nearly 100 thousand families. Saline environments have contributed the most to the novel taxa of this previously unseen diversity. The majority of those 16S rRNA gene sequence fragments were verified by matches in metagenomic datasets from IMG/M. These findings reveal a vast and previously overlooked diversity within the Archaea, offering insights into their ecological roles and evolutionary importance while establishing a foundation for the future study and characterization of this intriguing domain of life. Full article
(This article belongs to the Special Issue Earth Systems: Shaped by Microbial Life)
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11 pages, 14331 KiB  
Article
Fatal Toxoplasmosis in Red Kangaroos (Macropus rufus) in East China
by Haiyan Gong, Quan Wang, Yinghong Jin, Suoping Qiu, Zhaoguo Chen, Xiangan Han, Zongyan Chen and Wei Jiang
Pathogens 2025, 14(2), 202; https://doi.org/10.3390/pathogens14020202 - 19 Feb 2025
Viewed by 1105
Abstract
Background: As a ubiquitous apicomplexan parasite, Toxoplasma gondii causes huge economic losses and poses a great threat to the health of animals, including humans, worldwide. In some kangaroo species, T. gondii can be fatal. To date, little information is available on T. gondii [...] Read more.
Background: As a ubiquitous apicomplexan parasite, Toxoplasma gondii causes huge economic losses and poses a great threat to the health of animals, including humans, worldwide. In some kangaroo species, T. gondii can be fatal. To date, little information is available on T. gondii infection in the red kangaroos in east China. At a zoo in east China, thirteen red kangaroos consecutively developed clinical signs from July to November 2016, resulting in the deaths of seven, three of which were analyzed in this study. Methods: In the present study, ascitic fluid, blood and samples from the brain, heart, liver, spleen, lung, kidney, and mesenteric lymph nodes of three dead red kangaroos were collected. The pathogen was explored through microscopic observation, nested PCR, immunofluorescence antibody test (IFAT), hematoxylin–eosin (HE) staining, and immunohistochemistry (IHC) staining, respectively. Meanwhile, the potential source of the infection was also investigated by testing the blood of stray cats in the zoo for T. gondii using nested PCR. Results: Three dead red kangaroos were subjected to a necropsy, and organisms resembling T. gondii were detected in their ascitic fluids under microscope. This infection was further confirmed by a nested PCR assay, which resulted in a successful amplification and sequencing of the 433 bp fragment of the T. gondii 5.8S rRNA gene in all the dissected tissues, including heart, liver, spleen, lung, kidney, lymph nodes, cecum, and brain, as well as in body fluids (blood and ascitic fluid). Furthermore, the tachyzoites were observed in the heart, liver, spleen, lymph nodes, cecum, and brain through IFAT and HE staining. Administration of classic drugs (sulfadiazine and pyrimethamine) against T. gondii significantly alleviated the clinical signs of the sick kangaroos. The possible source of this infection was traced to a native stray cat, as T. gondii DNA was detected in its blood. Conclusions: In the present study, lethal T. gondii infection in red kangaroos has been described for the first time in east China, highlighting the necessity and urgency for close and long-term surveillance of this parasite infection in captive animals. The same strain of T. gondii detected in kangaroos as that found in stray cats wandering in the same area emphasizes the importance of controlling stray cat populations to mitigate the risk of Toxoplasma transmission to other animals. Full article
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18 pages, 4593 KiB  
Article
Diversity of Cytauxzoon spp. (Piroplasmida: Theileriidae) in Wild Felids from Brazil and Argentina
by Ana Cláudia Calchi, Joares A. May-Júnior, Vinícius Baggio-Souza, Laura Berger, Renata Fagundes-Moreira, Rafaela Mallmann-Bohn, Laíza de Queiroz Viana Braga, Murillo Daparé Kirnew, Matheus Folgearini Silveira, Roberto Andres Navarrete Ampuero, Charlotte O. Moore, Ricardo Bassini-Silva, Heitor Miraglia Herrera, Edward Bealmear Breitschwerdt, Ricardo G. Maggi, Eduardo Eizirik, Rosangela Zacarias Machado, Fabiana Lopes Rocha, João Fabio Soares and Marcos Rogério André
Pathogens 2025, 14(2), 148; https://doi.org/10.3390/pathogens14020148 - 4 Feb 2025
Cited by 2 | Viewed by 1650
Abstract
Domestic and wild felids are frequently parasitized by apicomplexan protozoa in the genus Cytauxzoon. Expanding species diversity has recently been described within this genus, with potential implications for epidemiology and pathogenesis. In light of these findings, this study assessed the genetic diversity [...] Read more.
Domestic and wild felids are frequently parasitized by apicomplexan protozoa in the genus Cytauxzoon. Expanding species diversity has recently been described within this genus, with potential implications for epidemiology and pathogenesis. In light of these findings, this study assessed the genetic diversity of Cytauxzoon spp. in wild felids (n = 66) from different eco-regions of Brazil and Argentina. Of the 66 blood samples analyzed, 53 (80.3%) were 18S rRNA gene PCR-positive for Cytauxzoon spp., including 43 jaguars (Panthera onca) and 10 ocelots (Leopardus pardalis). Panthera onca specimens (100%, 43/43) were most frequently infected, followed by Leopardus pardalis (76.9%; 10/13). Cytauxzoon spp. were not detected in Leopardus braccatus (n = 1) or Puma concolor (n = 9). Phylogenetic analyses of fragments of the 18S rRNA, cytB, and cox-1 gene sequences from jaguars were closely related to Cytauxzoon felis. In contrast, sequences from ocelots were more closely associated with Cytauxzoon brasiliensis. Distance and haplotype analysis further confirmed the circulation of at least two distinct genovariants of C. felis among jaguars, as evidenced by their close positioning and low genetic divergence (0–0.14% for 18S rRNA, 0.37–0.56% for cytB, and 0.08–0.74% for cox-1). Additionally, sequence data from ocelots suggested that multiple genovariants of C. brasiliensis are circulating among these cats in different Brazilian eco-regions. Our study provides evidence of two distinct Cytauxzoon organisms parasitizing free-ranging and captive jaguars and ocelots, respectively, in Brazil and Argentina. Full article
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10 pages, 1378 KiB  
Brief Report
The Occurrence of Borrelia burgdorferi sensu lato in Ixodes ricinus Ticks Collected from Nature-Educational and Tourist Trails in the Poprad Landscape Park
by Sylwia Koczanowicz, Magdalena Nowak-Chmura, Anna Kocoń, Grzegorz Rączka and Marek Asman
Pathogens 2025, 14(2), 117; https://doi.org/10.3390/pathogens14020117 - 26 Jan 2025
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Abstract
Throughout Europe, including Poland, Ixodes ricinus ticks are the main vector of numerous pathogenic agents that pose a serious threat to public health. Southern Poland attracts many tourists with its scenic landscapes and abundant recreational opportunities. These areas are ideal habitats for wild [...] Read more.
Throughout Europe, including Poland, Ixodes ricinus ticks are the main vector of numerous pathogenic agents that pose a serious threat to public health. Southern Poland attracts many tourists with its scenic landscapes and abundant recreational opportunities. These areas are ideal habitats for wild fauna, which serve as the main reservoirs and hosts for these pathogens and ticks. The large population and biodiversity of these hosts facilitate the proliferation of ticks. The aim of this study was to determine the potential exposure of humans to ticks and tick-borne pathogens such as Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, and Babesia spp., along the nature-educational and tourist trails of the Poprad Landscape Park. From 2020 to 2021, ticks were collected using the flagging method on three tourist trails and nature-educational paths within the Poprad Landscape Park. DNA was isolated from 213 I. ricinus ticks using the ammonia method. To detect pathogens in ticks, PCR and nested PCR methods were used. To detect B. burgdorferi s.l. and A. phagocytophilum, two pairs of primers specific to the flaB gene fragment and 16S rRNA gene fragment were used, respectively. For Babesia spp. detection, primers specific to the 18S rRNA gene were used. The amplification products were separated electrophoretically and visualized under ultraviolet light. In total, among the 213 examined ticks, B. burgdorferi s.l. was detected in 31% of the samples. Neither A. phagocytophilum nor Babesia spp. were detected in the studied material. These results indicate a potentially high risk of ticks and tick-borne B. burgdorferi s.l. infections for residents and tourists in the recreational areas of the Poprad Landscape Park. Full article
(This article belongs to the Special Issue Ticks and Tick-Borne Pathogens in a Changing World)
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16 pages, 5330 KiB  
Article
Mitochondrial Genome Characteristics Reveal Evolution of Danxiaorchis yangii and Phylogenetic Relationships
by Xuedie Liu, Huolin Luo, Zhong-Jian Liu and Bo-Yun Yang
Int. J. Mol. Sci. 2025, 26(2), 562; https://doi.org/10.3390/ijms26020562 - 10 Jan 2025
Cited by 1 | Viewed by 1088
Abstract
Danxiaorchis yangii is a fully mycoheterotrophic orchid that lacks both leaves and roots, belonging to the genus Danxiaorchis in the subtribe Calypsoinae. In this study, we assembled and annotated its mitochondrial genome (397,867 bp, GC content: 42.70%), identifying 55 genes, including 37 protein-coding [...] Read more.
Danxiaorchis yangii is a fully mycoheterotrophic orchid that lacks both leaves and roots, belonging to the genus Danxiaorchis in the subtribe Calypsoinae. In this study, we assembled and annotated its mitochondrial genome (397,867 bp, GC content: 42.70%), identifying 55 genes, including 37 protein-coding genes (PCGs), 16 tRNAs, and 2 rRNAs, and conducted analyses of relative synonymous codon usage (RSCU), repeat sequences, horizontal gene transfers (HGTs), and gene selective pressure (dN/dS). Additionally, we sequenced and assembled its plastome, which has a reduced size of 110,364 bp (GC content: 36.60%), comprising 48 PCGs, 26 tRNAs, and 4 rRNAs. We identified 64 potential chloroplast DNA fragments transferred to the mitogenome. Phylogenomic analysis focusing on 33 mitogenomes, with Vitis vinifera as the outgroup, indicated that D. yangii is grouped as follows: D. yangii + ((Dendrobium wilsonii + Dendrobium wilsonii henanense) + Phalaenopsis aphrodite). Phylogenetic analysis based on 83 plastid PCGs from these species showed that D. yangii is grouped as follows: (D. yangii + Pha. aphrodite) + (Den. wilsonii + Den. henanense). Gene selective pressure analysis revealed that most mitochondrial and plastid genes in D. yangii are under purifying selection, ensuring functional stability, and certain genes may have undergone positive selection or adaptive evolution, reflecting the species’ adaptation to specific ecological environments. Our study provides valuable data on the plastomes and mitogenomes of D. yangii and lays the groundwork for future research on genetic variation, evolutionary relationships, and the breeding of orchids. Full article
(This article belongs to the Special Issue Advances in Plant Genomics and Genetics: 2nd Edition)
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