Operational Taxonomic Units Research Articles

Analysis of bacterial community of Labidochromis caeruleus reared in RAS with various feeds

The electric yellow cichlid (Labidochromis caeruleus), renowned for its exceptional ornamental value, enjoys widespread popularity among aquarium enthusiasts and holds significant economic value in the market. This study employed high-throughput sequencing technology to analyze the microorganisms in the circulating water of the electric yellow cichlid cultivated with six different types of feeds. Statistical results showed that the compound feed group exhibited the highest specific growth and mass gain rates. In contrast, the water flea and brine shrimp groups showed lower rankings with significant differences. 51,084 to 64,626 effective sequences were obtained and clustered into 127 to 848 operational taxonomic units (OTUs). The research findings indicate a remarkable bacterial diversity in the breeding environment of electric yellow cichlids with higher levels in the water flea and brine shrimp groups, and the lowest level in the compound feed group; the dominant bacterial phyla in all six experimental groups include Proteobacteria, Actinobacteria, Parcubacteria, and Cyanobacteria; Comparative analysis using the COGs database revealed significant differences (P<0.05) in the metabolic pathways of 24 functional microbial genes among the experimental groups.

Dissecting the role of soybean rhizosphere-enriched bacterial taxa in modulating nitrogen-cycling functions

Crop roots selectively recruit certain microbial taxa that are essential for supporting their growth. Within the recruited microbes, some taxa are consistently enriched in the rhizosphere across various locations and crop genotypes, while others are unique to specific planting sites or genotypes. Whether these differentially enriched taxa are different in community composition and how they interact with nutrient cycling need further investigation. Here, we sampled bulk soil and the rhizosphere soil of five soybean varieties grown in Shijiazhuang and Xuzhou, categorized the rhizosphere-enriched microbes into shared, site-specific, and variety-specific taxa, and analyzed their correlation with the diazotrophic communities and microbial genes involved in nitrogen (N) cycling. The shared taxa were dominated by Actinobacteria and Thaumarchaeota, the site-specific taxa were dominated by Actinobacteria in Shijiazhuang and by Nitrospirae in Xuzhou, while the variety-specific taxa were more evenly distributed in several phyla and contained many rare operational taxonomic units (OTUs). The rhizosphere-enriched taxa correlated with most diazotroph orders negatively but with eight orders including Rhizobiales positively. Each group within the shared, site-specific, and variety-specific taxa negatively correlated with bacterial amoA and narG in Shijiazhuang and positively correlated with archaeal amoA in Xuzhou. These results revealed that the shared, site-specific, and variety-specific taxa are distinct in community compositions but similar in associations with rhizosphere N-cycling functions. They exhibited potential in regulating the soybean roots’ selection for high-efficiency diazotrophs and the ammonia-oxidizing and denitrification processes. This study provides new insights into soybean rhizosphere-enriched microbes and their association with N cycling.Key points• Soybean rhizosphere affected diazotroph community and enriched nifH, amoA, and nosZ.• Shared and site- and variety-specific taxa were dominated by different phyla.• Rhizosphere-enriched taxa were similarly associated with N-cycle functions.

The Populations of Two Differently Medicine-Used Plants of Hedyotis diffusa and Hedyotis corymbosa Shoot-Assembling Rich Bacterial and Fungal Communities with Varied Compositions but Conserved Structures.

The plant-colonized microbial communities have closely micro-ecological effects on host plant growth and health. There are many medicinal plants in the genus Hedyotis, but it is yet unclear about the shoot-assembled bacterial and fungal communities (SBFC) of Hedyotis plants. Hence, eight plant populations of Hedyotis diffusa (HD) and H. corymbosa (HC) were evaluated with 16S rRNA gene and ITS sequences, for comparing the types, abundance, or/and potential functions of SBFC at plant species- and population levels. In tested HD- and HC-SBFC, 682 fungal operational taxonomic units and 1,329 bacterial zero-radius operational taxonomic units were identified, with rich species compositions and varied alpha diversities. Notably, the SBFC compositions of HD and HC plant populations were exhibited with partly different types and abundances at phylum and genus levels but without significantly different beta diversities at plant species and population levels. Typically, the SBFC of HD and HC plant populations were presented with abundance-different biomarkers, such as Frankiaceae and Bryobacteraceae, and with similar micro-ecological functions of microbial metabolisms of lipids, terpenoids,and xenobiotics. Taken together, HD- and HC-SBFC possessed with varied rich compositions, conservative taxonomic structures, and similar metabolic functions, but with small-scale type and abundance differences at plant species- and population- levels.

Identification of Growth-Promoting Bacterial Resources by Investigating the Microbial Community Composition of Polyporus umbellatus Sclerotia

The sclerotium of the edible mushroom Polyporus umbellatus (Zhuling) exhibits various medicinal properties. However, given its long growth cycle and overexploitation, wild resources are facing depletion. Macrofungal growth depends on diverse microbial communities; however, the impact of soil bacteria on P. umbellatus development is unknown. Here, we combined high-throughput sequencing and pure culturing to characterize the diversity and potential function of bacteria and fungi inhabiting the P. umbellatus sclerotium and tested the bioactivities of their isolates. Fungal operational taxonomic units (OTUs) were clustered and classified, revealing 1275 genera. Bacterial OTUs yielded 891 genera. Additionally, 81 bacterial and 15 fungal strains were isolated from P. umbellatus sclerotia. Antagonism assays revealed three bacterial strains (FN2, FL19, and CL15) promoting mycelial growth by producing indole-3-acetic acid, solubilizing phosphate, and producing siderophores, suggesting their role in regulating growth, development, and production of active compounds in P. umbellatus. FN2-CL15 combined with bacterial liquid promoted growth and increased the polysaccharide content of P. umbellatus mycelia. This study reports new bioactive microbial resources for fertilizers or pesticides to enhance the growth and polysaccharide accumulation of P. umbellatus mycelia and offers guidance for exploring the correlation between medicinal macrofungi and associated microbial communities.

Interspecific selection in a diverse mycorrhizal symbiosis.

Coevolution describes evolutionary change in which two or more interacting species reciprocally drive each other's evolution, potentially resulting in trait diversification and ecological speciation. Much progress has been made in analysis of its dynamics and consequences, but relatively little is understood about how coevolution works in multispecies interactions, i.e., those with diverse suites of species on one or both sides of an interaction. Interactions among plant hosts and their mutualistic ectomycorrhizal fungi (ECM) may provide an ecologically unique arena to examine the nature of selection in multispecies interactions. Using native genotypes of Monterey pine (Pinus radiata), we performed a common garden experiment at a field site that contains native stands to investigate selection from ECM fungi on pine traits. We planted seedlings from all five native populations, as well as inter-population crosses to represent intermediate phenotypes/genotypes, and measured seedling traits and ECM fungal traits to evaluate the potential for evolution in the symbiosis. We then combined field estimates of selection gradients with estimates of heritability and genetic variance-covariance matrices for multiple traits of the mutualism to determine which fungal traits drive plant fitness variation. We found evidence that certain fungal operational taxonomic units, families and species-level morphological traits by which ECM fungi acquire and transport nutrients exert selection on plant traits related to growth and allocation patterns. This work represents the first field-based, community-level study measuring multispecific coevolutionary selection in nutritional symbioses.

Smoking induced salivary microbiome dysbiosis and is correlated with lipid biomarkers

BackgroundThe oral microbiome plays an essential role in maintaining oral homeostasis and health; smoking significantly affects it, leading to microbial dysbiosis. The study aims to investigate changes in the oral microbiome composition of smokers in the Qatari population and establish a correlation with lipid biomarkers.MethodsThe oral microbiota was profiled from saliva samples of 200 smokers and 100 non-smokers in the Qatari population, and 16s rRNA V3-V4 region were sequenced using the Illumina MiSeq platform. The operational taxonomic units (OTUs) were clustered using QIIME and the statistical analysis was performed by R.ResultsNon-smokers exhibited a more diverse microbiome, with significant alpha and beta diversity differences between the non-smoker and smoker groups. Smokers had a higher abundance of Firmicutes, Bacteroidota, Actinobacteriota, Patescibacteria, and Proteobacteria at the phylum level and of Streptococcus, Prevotella, Veillonella, TM7x, and Porphyromonas at the genus level. In contrast, non-smokers had more Bacteroidota, Firmicutes, Proteobacteria, Fusobacteriota, and Patescibacteria at the phylum level, and Prevotella, Streptococcus, Veillonella, Porphromonas, and Neisseria at the genus level. Notably, Streptococcus was significantly positively correlated with LDL and negatively correlated with HDL. Additionally, Streptococcus salivarius, within the genus Streptococcus, was substantially more abundant in smokers.ConclusionThis study highlights the significant influence of smoking on the composition of the oral microbiome by enriching anaerobic microbes and depleting aerobic microbes. Moreover, the observed correlation between Streptococcus abundance and the lipid biomarkers suggests a potential link between smokers-induced salivary microbiome dysbiosis and lipid metabolism. Understanding the impact of smoking on altering the oral microbiome composition and its correlation with chemistry tests is essential for developing targeted interventions and strategies to improve oral health and reduce the risk of diseases.

Integrative taxonomic revision of the African thicket rats (Murinae: Grammomys): how genomics decreases the number of currently recognized species

Abstract Thicket rats of the genus Grammomys are partly arboreal rodents distributed across a large part of sub-Saharan Africa. Previous work showed that their highest diversity is in forests and woodlands of Eastern Africa, that the evolutionary history of the genus roughly mirrors the evolution of African forests, and that the currently delimited species (names) do not correspond to major genetic clades. We used thousands of single nucleotide polymorphisms to perform phylogenomic analysis and to delimit genomic pools (i.e. candidate biological species). The resulting molecular operational taxonomic units were then compared with the most comprehensive sets of other available data (mitochondrial DNA variability, ecological requirements, morphology, karyotypes, sequences from the type material, etc.) and used for an integrative solution to the taxonomy of the genus. Our revision delimited seven species, which is less than the 12 recognized in the most recent literature on African rodents. We conclude that the African thicket rats represent a classic example of challenging problems in current integrative taxonomy. It is now relatively easy to obtain genomic data and to delimit candidate species, but it is much more complicated to give them species names using the valid rules of zoological nomenclature.

Spatial variations impact the soil fungal communities of Larix gmelinii forests in Northeast China

Soil fungi play a critical role in the biogeochemical cycles of forest ecosystems. Larix gmelinii is a strong and important timber tree species, which forms close associations with a wide range of soil fungi. However, the temporal-spatial disparity effects on the assembly of soil fungal communities in L. gmelinii forests are poorly understood. To address these questions, a total of 120 samples, including 60 bulk soil and 60 root samples, were collected from Aershan and Genhe in July (summer) and October (autumn)2021. We obtained 7,788 operational taxonomic units (OTUs) after merging, filtering, and rarefying using high-throughput sequencing. The dominant phyla are Basidiomycota, Ascomycota, Mortierellomycota, and Mucoromycota. There were 13 dominant families, among which the families with average relative abundance more than 5% included Thelephoraceae, Mortierellaceae, Archaeorhizomycoaceae, and Inocybaceae. In the functional guilds, symbiotrophic fungi had a relative advantage in the identified functions, and the relative abundances of pathotrophic and saprotrophic fungi varied significantly between sites. There were 12 families differentially expressed across compartments, 10 families differentially expressed between seasons, and 69 families were differentially expressed between sites. The variation in alpha diversity in the bulk soil was greater than that in the rhizosphere soil. Among the three parts (compartment, season, and site), the site had a crucial effect on the beta diversity of the fungal community. Deterministic processes dominated fungal community assembly in Genhe, whereas stochastic processes dominated in Aershan. Soil physicochemical properties and climatic factors significantly affected fungal community structure, among which soil total nitrogen and pH had the greatest effect. This study highlights that spatial variations play a vital role in the structure and assembly of soil fungal communities in L. gmelinii forests, which is of great significance for us in maintaining the health of the forests.

Endophytic bacterial diversity in the latex-bearing caulosphere of Hevea brasiliensis Müll. Arg.

Rubber trees are a commercial cash crop, and the milky latex or polyisoprene they produce is the natural source of rubber. Little is known about the bacterial populations found in active zone of latex-bearing caulosphere. We employed a tailored cloud microbial bioinformatic approach for the identification and potential hypothetical ecological roles of an uncultured endophytic hidden bacterial community in the active zone of the latex-bearing caulosphere of Hevea brasiliensis. Small pieces of slivers were collected from healthy plant from the village: Belonia, South Tripura, rubber plantation in Northeastern India. These uncultured bacteria were identified using the V3-V4 hypervariable amplicon region of the 16S rDNA gene. A total of 209,586 contigs have been generated. EasyMAP Version 1.0, a cloud-based microbial bioinformatics tool with an integrated QIIME2 pipeline, was used to analyze contigs. We detected 15 phyla and 91 OTUs (operational taxonomic units). Proteobacteria (73.5%) was the most enriched phylum, followed by Firmicutes (13.8%), Bacteroidetes (5.2%), and Actinobacteria (3.2%). Ammonia oxidizers, sulfate reducers, dehalogenation, chitin degradation, nitrite reducers, and aromatic hydrocarbon degraders were the most prevalent functional categories in the active zones of caulosphere. Furthermore, Gammaproteobacteria (49.2%) and Erwinia (29.19%) were the most abundant classes and genera of endophytic bacterial communities. Thus, the presence of a substantial amount of phosphate-solubilizing Gammaproteobacteria (PSB) may stimulate growth, increase plant resilience, suppress disease, and aid in the rubber and sugar breakdown. This is the first report of microbial endophytes associated with Hevea caulosphere.

Effects of Lake Erie dredged material on microbiomes in a farm soil of Northwestern Ohio.

Dredged materials are often considered as candidates for replenishing lost topsoils in the watersheds of rivers and lakes. This study aimed to investigate the impacts of Lake Erie dredged material on the microbial community in a farm soil of Northwestern Ohio. Dredged material from the Toledo Harbor, OH was mixed with a local farm soil at ratios of 0:100, 10:90, 20:80, and 100:0 for soybean growth in a greenhouse for 123 days and was subject to 16S rRNA gene sequencing. α-Diversity analysis revealed that although the original dredged material hosted a highly diverse microbiome, soils blended with the dredged material had similar levels of bacterial diversity to 100% farm soil throughout the experiment. β-Diversity analysis demonstrated that, given the same plant status, that is, with or without soybean, blended soils had similar bacterial communities to 100% farm soil during the experiment. Furthermore, by the end of the experiment, all soils with soybeans merged into one cluster distinctive from those without the plants, indicating that the growth of plants played a dominating role in defining the structure of soil microbiomes. The majority (73.8%) of the operational taxonomy units that were unique to the original dredged material were not detected by the end of the experiment. This study demonstrates that up to 20% of the dredged material can be safely blended into the farm soil without distorting the microbial communities of the latter, implying a potential beneficial use of the dredged material for topsoil restoration.

A comparison of oral bacteriome isolated from periodontal pockets of participants with or without diabetes mellitus in Uganda: a case control study.

Diabetes mellitus predisposes patients to increased incidence and severe forms of periodontal disease. Currently, information on the bacterial diversity of patients with diabetes mellitus and periodontitis in Uganda is scanty. This study set out to describe the bacteria associated with periodontitis in patients with diabetes mellitus in Uganda, as part of a larger study describing the association between periodontal disease and diabetes mellitus. This was a case control involving 45 samples of gingival crevicular fluid collected from participants with periodontitis, the cases being 26 participants with diabetes mellitus and controls 19 participants without diabetes mellitus. Sequencing using the 16s Oxford nanopore long read protocol was followed by a bioinformatics analysis pipeline for alpha and beta diversity indices in the two groups. Multivariate tests were done to determine the differences in the bacterial composition in the two groups. Of the 739 Operational Taxonomic Units and 500 phyla identified, 37.9% (280/739) were from participants with diabetes mellitus. Analysis of beta diversity revealed a dissimilarity between the two study groups (CAP score = 0) with a significant association noted between periodontitis and the subgingival bacteria (P = 0.001). Diabetes mellitus reduced the quantity and altered the composition of the subgingival microbiome in the study participants.

Time-series metaproteogenomics of a high-CO2 aquifer reveals active viruses with fluctuating abundances and broad host ranges

Abstract Ecosystems subject to mantle degassing are of particular interest for understanding global biogeochemistry, as their microbiomes are shaped by prolonged exposure to high CO2 and have recently been suggested to be highly active. While the genetic diversity of bacteria and archaea in these deep biosphere systems have been studied extensively, little is known about how viruses impact these microbial communities. Here, we show that the viral community in a high-CO2 cold-water geyser (Wallender Born, Germany) undergoes substantial fluctuations over a period of 12 days, although the corresponding prokaryotic community remains stable, indicating a newly observed “infect to keep in check” strategy that maintains prokaryotic community structure. We characterized the viral community using metagenomics and metaproteomics, revealing 8,654 viral operational taxonomic units (vOTUs). CRISPR spacer-to-protospacer matching linked 278 vOTUs to 32 hosts, with many vOTUs sharing hosts from different families. High levels of viral structural proteins present in the metaproteome (several structurally annotated based on AlphaFold models) indicate active virion production at the time of sampling. Viral genomes expressed many proteins involved in DNA metabolism and manipulation, and encoded for auxiliary metabolic genes, which likely bolster phosphate and sulfur metabolism of their hosts. The active viral community encodes genes to facilitate acquisition and transformation of host nutrients, and appears to consist of many nutrient-demanding members, based on abundant virion proteins. These findings indicate viruses are inextricably linked to the biogeochemical cycling in this high-CO2 environment and substantially contribute to prokaryotic community stability in the deep biosphere hotspots.

Characterizations of gut bacteriome, mycobiome, and virome of healthy individuals living in sea-level and high-altitude areas.

The contribution of gut microbiota to human high-altitude adaptation remains inadequatelyunderstood. Here a comparative analysis of gut microbiota was conducted between healthy individuals living at sealevel and high altitude using deep whole-metagenome shotgun sequencing, to investigate the adaptivemechanisms of gut microbiota in plateau inhabitants. The results showed the gut bacteriomes in high-altitude individuals exhibited greater within-samplediversity and significant alterations in both bacterial compositional and functional profiles when compared to thoseof sea-level individuals, indicating the potential selection of unique bacteria associated with high-altitudeenvironments. The strain-level investigation revealed enrichment of Collinsella aerofaciens and Akkermansiamuciniphila in high-altitude populations. The characteristics of gut virome and gut mycobiome were alsoinvestigated. Compared to sea-level subjects, high-altitude subjects exhibited a greater diversity in their gut virome,with an increased number of viral operational taxonomic units (vOTUs) and unique annotated genes. Finally,correlation analyses revealed 819 significant correlations between 42 bacterial species and 375 vOTUs, while nosignificant correlations were observed between bacteria and fungi or between fungi and viruses. The findings have significantly contributed to an enhanced comprehension of the mechanismsunderlying the high-altitude geographic adaptation of the human gut microbiota.

Anastomotic Leakage after Colorectal Surgery is Associated with a Specific Tissue-Microbiome Signature

Abstract Background Anastomotic leakage (AL) occurs in 5-15% of colorectal resections. In addition to well-known risk factors, recent evidence suggests that the gut microbiota might critically affect anastomotic healing. However, studies in humans are limited. Aims To explore differences in pre-operative microbiome of patients with or without AL and identify anastomosis-protective or damaging bacterial species. Methods Clinical records of patients undergoing left colon or rectal resection with anastomosis at our department between 2013 and 2021 were screened. AL cases were matched equally to controls by propensity score, based on common AL risk factors. The microbiome was analyzed by amplification and sequencing of the V4 region of the 16S gene of DNA extracted from formalin-fixed paraffin-embedded mucosa taken at the proximal and distal margin of the resected specimen, mixed in equal amounts. Differential abundance of identified operational taxonomic units (OTUs) in AL-cases and controls was tested by using the R-package DESeq2. Results Among 588 patients screened, 41 cases and 42 matched controls were eligible for microbiome analysis. Patient characteristics were similar except for a higher percentage of rectal resections in the AL group. Out of 5406 OTUs identified, 197 OTUs were differentially abundant between the two groups (adjusted-p value <0.05). Of those, 175 OTUs were more prevalent in the AL-group, in particular Fusobacterium periodonticum (32% vs. 5% of patients) and bacteria from the Hyphomicrobium genera (39-44% vs. 12-21%); 22 were more prevalent in the control group, in particular Akkermansia muciniphila (26 vs. 17%) and Eubacterium sireum (19 vs. 5% of patients). Conclusion Our analysis unravels AL-associated microbiome-signatures and identifies bacterial species highly prevalent in particular in patients with AL. These findings could be used for future research to develop targeted antibiotic therapy that helps prevent AL.

Endophytic Fungal Diversity in Hardwickia binata: Bridging the Gap between Traditional and Modern Techniques

Endophytic fungus is crucial for maintaining plant health and defense mechanisms, acting as protective barriers against pathogens, and producing medicinally beneficial bioactive compounds. Genome sequencing and metagenomics have significantly enhanced the understanding of fungal diversity and metabolic capabilities, enabling the identification of new genes and substances. Traditional culture-dependent methods have been complemented by culture-independent techniques, offering a more comprehensive view of fungal diversity. Using both culture-dependent and culture-independent techniques, the present research investigation explored the diversity of endophytic fungi encountered in the foliage of Hardwickia binata. The study examined the topographical characteristics and nutritional content of soil samples collected from the locality of the selected plant sample, H. binata, to better comprehend the effects on the plant’s growth. The balanced nutrient constituted approximately a pH of 7.2, which suggested an alkaline nature and promoted plant development. The ratio of nitrogen, phosphorous, and potassium remained 3:1:1. A total of 25 fungal isolates, categorized into 17 morphotypes, were obtained using the culture-dependent approach; Curvularia and Nigrospora emerged as the most common genera. Furthermore, the prediction of the ITS2 secondary structure supports the identification of species, highlighting a wide variety of fungal species present in H. binata. The culture-independent approach generated 69,570 high-quality sequences, identifying 269 Operational Taxonomic Units (OTUs). The dominant Ascomycota phylum, along with various genera, indicated a rich fungal community associated with H. binata. This study advances the understanding of the endophytic fungus communities that are associated with H. binata and the nature of soil ecology. The findings emphasize the significance of holistic techniques in the study of microbial dynamics within plant systems as well as their implications for ecosystem management and plant health.

Characterization and evaluation of actinomycete from the Protaetia brevitarsis Larva Frass

Protaetia brevitarsis larvae (PBL) are soil insects important for the soil organic carbon cycle, and PBL frass not only contains a large amount of humic acid but also affects the diversity, novelty, and potential functions of actinomycetes. Here, we characterized and assessed the actinomycete. The operational taxonomic unit (OTU) data showed that 90% of the actinomycetes cannot be annotated to species, and pure culture and genome analysis showed that 35% of the strains had the potential to be new species, indicating the novelty of PBL frass actinomycetes. Additionally, genome annotation showed that many gene clusters related to antifungal, antibacterial and insecticidal compound synthesis were identified, and confrontation culture confirmed the antifungal activities of the actinomycetes against soil-borne plant pathogenic fungi. The incubation experiment results showed that all isolates were able to thrive on media composed of straw powder and alkaline lignin. These results indicated that PBL hindgut-enriched actinomycetes could survive in soil by using the residual lignocellulose organic matter from plant residues, and the antibiotics produced not only give them a competitive advantage among soil microflora but also have a certain inhibitory effect on plant diseases and pests. This study suggests that the application of PBL frass can not only supplement soil humic acid but also potentially affect the soil microbiota of cultivated land, which is beneficial for the healthy growth of crops.

Effects of Maize/Peanut Intercropping and Nitrogen Fertilizer Application on Soil Fungal Community Structure

Maize/peanut intercropping may improve soil health through reducing nitrogen (N) fertilization. However, the effects of maize/peanut intercropping combined with reduced N fertilization on the soil fungal community structure have not been well reported. Using a long-term localized micro-zone experiment, we investigated the combined effects of intercropping and N fertilizer application on soil fungal community diversity and composition. Three cropping patterns (maize/peanut intercropping, maize monoculture, and peanut monoculture) and three N application levels (0 kg·hm−2, 150 kg·hm−2, and 300 kg·hm−2) were assessed. The results showed that the total numbers of fungal species and unique species (operational taxonomic units, OTUs) in both maize and peanut soils tended to first increase and then decrease with increasing N application. Compared with monoculture, the numbers of total OTUs and unique OTUs in intercropped maize soil decreased by 4.14% and 12.79%, respectively, but the total numbers of OTUs and unique OTUs in peanut soil increased by 1.08% and 3.78%, respectively. With increasing N application, the soil fungal Ace and Chao indices of maize soil first increased and then decreased, while the fungal Shannon, Ace, and Chao indices of peanut soil decreased. Compared with the monoculture system, intercropping significantly reduced the maize soil fungal Ace and Chao indices but increased the peanut soil fungal Shannon, Ace, and Chao indices. Nitrogen application and intercropping significantly altered the fungal community structure of maize soil, while N application had no significant effect on the fungal community structure of peanut soil, though intercropping significantly changed the fungal community structure of peanut soil. At the phylum level, Ascomycota, Basidiomycota, Mortierellomycota, unclassified_k_Fungi, and Chytridiomycota were the dominant taxa. Redundancy analysis (RDA) showed that soil nitrate (NO3−) content was the main environmental factor shaping the soil fungal community. In conclusion, excessive N fertilization (300 kg·hm−2) can reduce soil fungal community diversity; maize/peanut intercropping reversed the negative effect of N application on fungal community of peanut soil, but not that of maize soil. Soil NO3− content is the primary environmental driver of soil fungal communities.

Species boundaries of the whiptail catfish Rineloricaria (Siluriformes: Loricariidae) from the Paraíba do Sul River drainage, southeastern Brazil, with species redescriptions and description of a new species.

Species of the catfish genus Rineloricaria are common in the Paraíba do Sul River basin, in southeastern Brazil; here we present a revision of the taxonomic diversity and geographic distribution of the species of the genus inhabiting the basin, based on novel morphologic and molecular data. Five species delimitation methods based on cytochrome C oxidase subunit 1 nucleotide sequences yielded comparable molecular operational taxonomic units. The automatic barcode gap discovery, assemble species by automatic partitioning, barcode index number, and Bayesian implementation of the Poisson tree process methods supported the recognition of five evolutionary lineages. These taxonomic units were assigned to the previously described Rineloricaria nigricauda, Rineloricaria steindachneri, Rineloricaria zawadzkii, and Rineloricaria nudipectoris, and an additional undescribed species. R. zawadzkii was further divided into two intraspecific geographically structured lineages using the generalized mixed Yule coalescent delimitation method. A maximum likelihood phylogenetic analysis revealed that the five lineages from the Paraíba do Sul have closer relationships to different species from southern and southeastern Brazil (Ribeira de Iguape, Lagoa dos Patos, Uruguay, Paraguay, and Parana river basins) than to each other. Based on the analysis of lectotypes, recently collected material, and specimens from ichthyological collections, the poorly described R. nigricauda and R. steindachneri are redescribed following current descriptive standards. The undescribed species from the middle and upper Paraíba do Sul River basin is formally described. The description of a new species, along with the description of species boundaries in R. nigricauda and R. steindachneri, contributes to the knowledge of the ichthyofauna of the Paraíba do Sul River basin and adjacent coastal drainages of southeastern Brazil. An identification key for the species of Rineloricaria occurring in the Paraíba do Sul River basin is provided.

The gills and skin microbiota of five pelagic fish species from the Atlantic Ocean.

The gills and skin microbiota and microbiome of wild fish remain far more under-investigated compared to that of farmed fish species, despite that these animal-microbe interactions hold the same ecophysiological roles in both cases. In this study, the gills and skin bacterial microbiota profiles and their presumptive bacterial metabolisms were investigated in five open-sea fishes: bullet tuna (Auxis sp.), common dolphinfish (Coryphaena hippurus), Atlantic little tunny (Euthynnus alletteratus), Atlantic bonito (Sarda sarda) and Atlantic white marlin (Kajikia albida). Gills and skin tissues were collected from two to three individuals per species, from specimens caught by recreational trolling during summer of 2019, and their bacterial 16S rRNA gene diversity was analysed by high-throughput sequencing. The gills bacterial communities among the five species were clearly different but not the skin bacterial microbiota. The dominant operational taxonomic units belonged to the Moraxellaceae, Pseudomonadaceae, Rhodobacteraceae, Staphylococcaceae and Vibrionaceae families. Despite the differences in taxonomic composition, the presumptive bacterialmetabolisms between the gills and skin of the five fishes investigated here were ≥ 94% similarand were dominated by basic metabolism, most likely reflecting the continuous exposure of these tissues in the surrounding seawater.

A DNA barcode reference library of Croatian mosquitoes (Diptera: Culicidae): implications for identification and delimitation of species, with notes on the distribution of potential vector species

BackgroundMosquitoes pose a risk to human health worldwide, and correct species identification and detection of cryptic species are the most important keys for surveillance and control of mosquito vectors. In addition to traditional identification based on morphology, DNA barcoding has recently been widely used as a complementary tool for reliable identification of mosquito species. The main objective of this study was to create a reference DNA barcode library for the Croatian mosquito fauna, which should contribute to more accurate and faster identification of species, including cryptic species, and recognition of relevant vector species.MethodsSampling was carried out in three biogeographical regions of Croatia over six years (2017–2022). The mosquitoes were morphologically identified; molecular identification was based on the standard barcoding region of the mitochondrial COI gene and the nuclear ITS2 region, the latter to identify species within the Anopheles maculipennis complex. The BIN-RESL algorithm assigned the COI sequences to the corresponding BINs (Barcode Index Number clusters) in BOLD, i.e. to putative MOTUs (Molecular Operational Taxonomic Units). The bPTP and ASAP species delimitation methods were applied to the genus datasets in order to verify/confirm the assignment of specimens to specific MOTUs.ResultsA total of 405 mosquito specimens belonging to six genera and 30 morphospecies were collected and processed. Species delimitation methods assigned the samples to 31 (BIN-RESL), 30 (bPTP) and 28 (ASAP) MOTUs, with most delimited MOTUs matching the morphological identification. Some species of the genera Culex, Aedes and Anopheles were assigned to the same MOTUs, especially species that are difficult to distinguish morphologically and/or represent species complexes. In total, COI barcode sequences for 34 mosquito species and ITS2 sequences for three species of the genus Anopheles were added to the mosquito sequence database for Croatia, including one individual from the Intrudens Group, which represents a new record for the Croatian mosquito fauna.ConclusionWe present the results of the first comprehensive study combining morphological and molecular identification of most mosquito species present in Croatia, including several invasive and vector species. With the exception of some closely related species, this study confirmed that DNA barcoding based on COI provides a reliable basis for the identification of mosquito species in Croatia.Graphical