16S Genes Research Articles

A rapid and parallel Late Pleistocene/Holocene morphological radiation in a predaceous planktonic water flea: the case of Bythotrephes (Cladocera: Cercopagididae)

Abstract The predatory cladoceran Bythotrephes is one of North America’s most successful and impactive invasive species in freshwater plankton communities. The taxonomic status of the genus Bythotrephes Leydig, 1860 (Crustacea: Cladocera: Cercopagididae) has remained unclear and a subject of intensive debate for over 150 years. We applied an integrative taxonomy approach with multi-gene analysis (mitochondrial COI and 12S genes, and nuclear 18S and 28S genes) on 80 individuals (representing at least four morphospecies) from various regions spanning the genus distribution (North America, Europe, and Asia) to resolve the taxonomic status of species within Bythotrephes. The results of our study strongly support the hypothesis of a single species—Bythotrephes longimanus Leydig, 1860, and ecological morphs should be accepted as junior synonyms. Our work also elucidates the very recent radiation of Bythotrephes, which may have begun rapidly and parallel during the Late Pleistocene, or even after the last glaciation. Finally, we provide a comprehensive biogeographic reconstruction of Bythotrephes dispersal within the Holarctic realm. Europe likely served as a dispersal centre for Bythotrephes from where they spread relatively recently (possibly, less than 10 kyr) and we distinguish five possible dispersal events in its evolutionary history.

DNA sequencing of topotypes helps delineate species distributions in the Ischnocnema verrucosa complex (Anura, Brachycephalidae)

As many new evolutionary lineages are being discovered and formally named, sequencing topotypes when holotypes are not available becomes essential for taxonomy. This study uses a DNA-taxonomy approach to sequence new populations of the Ischnocnema verrucosa species complex (Brazilian Wart Frogs) from different locations, including, for the first time, individuals from the type localities. Phylogenetic analysis of the mitochondrial 16S gene recovered a monophyletic Ischnocnema verrucosa species series composed of three main clades. The most recent common ancestor was estimated to be 33.76 million years ago, and diversification within the three main clades occurred primarily during the Miocene. We delimited eight species-level lineages with high levels of sequence divergence (7% to 16%). Our study highlights the importance of DNA taxonomy and the necessity of protecting and sequencing topotypes in taxonomic studies. Our study also contributes to the conservation and understanding of the genus Ischnocnema and the biodiversity of the Brazilian Atlantic Forest region.

Preliminary study on the potential impact of probiotic combination therapy on Helicobacter pylori infection in children using 16S gene sequencing and untargeted metabolomics approach

ObjectiveThe purpose of this study was to explore the potential mechanism of Helicobacter pylori (Hp) eradication by probiotic therapy through 16S rRNA gene sequencing technology and untargeted metabolomics.MethodsTwenty four Hp-infected children were recruited from the Shanxi Bethune Hospital, and 24 healthy children were recruited as a blank control group. Group A: fecal samples from 24 healthy children. Group B: fecal samples of 24 children with Hp infection. Group B1 (n = 15): fecal samples of group B treated with probiotic therapy for 2 weeks. Group B2 (n = 19): fecal samples of group B treated with probiotic therapy for 4 weeks. The above fecal samples were analyzed by 16S rRNA gene sequencing technology and untargeted metabolomics.ResultsThere was no significant difference in alpha diversity and beta diversity among the four groups, but many bacteria with statistical difference were found in each group at the bacterial genus level and phylum level. LEfSe results showed that in group B, Porphyromonadaceae, Shigella and other microorganisms related to intestinal microecological dysbiosis were enriched. And in group B2, abundant characteristic microorganisms were found, namely Bacillales and Prevotella. KEGG metabolic pathway enrichment analysis showed that groups B1 and B2 were involved in 10 metabolic pathways potentially related to probiotic treatment: purine metabolism, nitrogen metabolism, arginine biosynthesis, alanine, aspartic acid and glutamate metabolism, glyoxylic acid and dicarboxylic acid metabolism, unsaturated fatty acid biosynthesis, fatty acid extension, fatty acid degradation, pyrimidine metabolism, fatty acid biosynthesis.ConclusionProbiotic therapy can inhibit Hp to some extent and can relieve gastrointestinal symptoms, making it a preferred therapy for children with Hp infection and functional abdominal pain. Hp infection can reduce the diversity of intestinal microbes, resulting in the disturbance of intestinal microbiota and changes in the relative abundance of microbiota in children, while probiotic therapy can restore the diversity of intestinal microbes and intestinal microecological balance.

A redescription and new generic placement of the New Zealand marine-associated jumping spider Marpissa marina (Goyen, 1892) (Araneae: Salticidae: Euophryini)

ABSTRACT The jumping spider Marpissa marina (Goyen, 1892) is endemic to Aotearoa New Zealand. In this paper, we redescribe the species. Marpissa marina was placed in the Northern Hemisphere genus Marpissa due to its subjective similarity to spiders in that genus. The original description lacked modern morphological diagnoses and useful illustrations. The description was based largely on the female, and therefore lacked the taxonomically important characters seen in the male pedipalp. Previous literature has suggested M. marina may be related to the Australian genus, Maratus Karsch, 1878. We applied an integrative taxonomic approach based on diagnostic morphological characters and molecular analysis for a more detailed species description and assignment into an appropriate genus. Morphological features indicate that M. marina belongs to Maratus. This is supported by phylogenetic analyses using DNA sequences from the nuclear gene Actin 5C and the mitochondrial gene 16S ribosomal RNA. We transfer Marpissa marina into the genus Maratus, as Maratus marinus (Goyen, 1892), new combination.

Never ending diversity: two new species of the genus Allocreadium (Digenea: Allocreadiidae) including new keys to the genus.

Two new species of the genus Allocreadium were isolated from the intestines of the Lake minnow Rhynchocypris percnura caught in the backwater of the Komissarovka River in the South of the Russian Far East. The morphology of A. anastasii n. sp. corresponds to that of Allocreadium sp. from Lake Khar (Mongolia) and Allocreadium sp. Belous, 1952 from the Primorsky region of Russia except for the preacetabular anterior border of the vitelline follicles in A. anastasii n. sp. from the Komissarovka River vs. at anterior half of ventral sucker in Allocreadium sp. Genetic analysis revealed the identity of A. anastasii n. sp. to Allocreadium sp. 1 from the Nezhinka River and Lake Khar. Allocreadium macrolecithum n. sp. was differentiated from Palaearctic Allocreadium spp. by having the following features: respectively large vitelline follicles extending from posterior extremity to anterior margin of the ventral sucker; relatively short caeca reaching the border of middle and posterior thirds of hindbody; and small testes in the middle of hindbody. Interspecific genetic p-distances between Allocreadium spp. were 0.16-7.23% in 28S gene and 18.62-31.54% in Cox1 mtDNA gene. In the phylogenetic tree reconstructed with Maximum parsimony and Bayesian Inference methods, A. anastasii n. sp. and A. macrolecithum n. sp. were nested into different species groups of the genus Allocreadium - sister to A. khankaiense and A. bursense, respectively. Modified dichotomous keys were prepared for 31 Palaearctic species of Allocreadium including A. crassum, A. dogieli, A. papilligerum, A. bursense, A. anastasii n. sp., and A. macrolecithum n. sp.

First detection of D181 genotype of infectious bronchitis in poultry flocks of Morocco

BackgroundThis paper reports the first pathological and molecular characterization of the novel variant of infectious bronchitis virus (IBV) D181 in poultry flocks in Morocco and Africa.MethodsThe study includes six poultry farms, involving three flocks of layers aged between 28 and 67 weeks and three broiler flocks aged 27, 39 and 42 days from different regions of Morocco. In all affected layer flocks, a severe drop in egg production with poor eggshell quality was reported. Necropsy of dead birds was carried out, and samples of trachea, lungs, oviduct, ovaries, and kidneys were fixed in 10% neutral buffered formalin for histopathologic examinations, while other portions were stored at -20 °C for molecular analysis. Real time RT-qPCR for IBV gene group was performed, and IBV variants were identified. Partial S1 gene sequences were amplified by conventional RT-PCR, sequenced, and aligned for phylogenetic and amino acid similarity analysis.ResultsNecropsy of dead birds revealed misshapen and hemorrhagic ovarian follicles with an edematous oviduct and severe reaction in the cecal tonsils. A caseous material accumulation in the sinus was noted in few birds. In contrast, the broiler flocks exhibited respiratory clinical signs such as difficulty in breathing, sneezing, tracheal rales, watery eyes and lethargy, associated with a decrease in feed consumption. Mortality in broiler ranged from 2 to 15%. Histopathological analysis of samples showed a lympho-plasmocytic inflammation in the oviduct, trachea, and lungs. Individual necrosis of epithelial cells, with sloughing of the bronchial epithelium and accumulation of desquamated cells with mucus in the airways, was observed in some birds. Partial S1 gene sequencing and phylogenetic analyses showed that the Moroccan strains were very closely related to D181 strains isolated in Dutch layers and breeders in 2018. Nucleotide sequence identities reached 90.9–95% with the Dutch isolates (strain CK/NL/D181/2018).ConclusionOur sequencing results demonstrate for the first time that the D181 IBV genotype is circulating in Moroccan poultry. These findings justify permanent monitoring of circulating strains in order to appropriately adjust vaccination strategies to align with the evolving field situation.

Environment, plant genetics, and their interaction shape important aspects of sunflower rhizosphere microbial communities.

Associations with soil microorganisms are crucial for plants' overall health and functioning. While much work has been done to understand drivers of rhizosphere microbiome structure and function, the relative importance of geography, climate, soil properties, and plant genetics remains unclear, as results have been mixed and comprehensive studies across many sites and genotypes are limited. Rhizosphere microbiomes are crucial for crop resistance to pathogens, stress tolerance, nutrient availability, and ultimately yield. Here, we quantify the relative roles of plant genotype, environment, and their interaction in shaping soil rhizosphere communities, using 16S and ITS gene sequencing of rhizosphere soils from 10 genotypes of cultivated sunflower (Helianthus annuus) at 15 sites across the Great Plains of the United States. While site generally outweighed genotype overall in terms of effects on archaeal, bacterial, and fungal richness, community composition, and taxa relative abundances, there was also a significant interaction such that genotype exerted a significant influence on archaeal, bacterial, and fungal microbiomes in certain sites. Site effects were attributed to a combination of spatial distance and differences in climate and soil properties. Microbial taxa that were previously associated with resistance to the fungal necrotrophic pathogen Sclerotinia were present in most sites but differed significantly in relative abundance across sites. Our results have implications for plant breeding and agronomic microbiome manipulations for agricultural improvement across different geographic regions.IMPORTANCEDespite the importance of plant breeding in agriculture, we still have a limited understanding of how plant genetic variation shapes soil microbiome composition across broad geographic regions. Using 15 sites across the Great Plains of North America, we show that cultivated sunflower rhizosphere archaeal, bacterial, and fungal communities are driven primarily by site soil and climatic differences, but genotype can interact with site to influence the composition, especially in warmer and drier sites with lower overall microbial richness. We also show that all taxa that were previously found to be associated with resistance to the fungal pathogen Sclerotinia sclerotiorum were widespread but significantly affected by site, while a subset was also significantly affected by genotype. Our results contribute to a broader understanding of rhizosphere archaeal, bacterial, and fungal community assembly and provide foundational knowledge for plant breeding efforts and potential future microbiome manipulations in agriculture.

Lights on tardigrade biodiversity: integrative redescription of Eremobiotus alicatai (Eutardigrada, Isohypsibiidae) with new insights on its morphology, phylogeny, and biogeography

AbstractCurrent knowledge on the genus Eremobiotus still remains limited. Only three species are known, mainly recorded in the Palaearctic region, with Eremobiotus alicatai representing the most common species. In the present study, an integrative re-description of E. alicatai based on the examination of a topotypic population from Sicily (Italy) is given, the morphological characters of which perfectly correspond to those of the type series collected in 1969. A second population of E. alicatai, from Tuscany (Italy), was also investigated from the morphological and the molecular point of view, allowing examining the intraspecific variability of the species. A re-analysis of the morphology of specimens of the type series, along with a morphological analysis of specimens of the two investigated populations of E. alicatai, allowed for an update and correction of certain morphological traits in the species. The topotypic and the Tuscanian populations were both investigated through a molecular approach: COI, ITS2, 18S, and 28S gene sequences were obtained, allowing to update the Isohypsibioidea phylogeny and to discuss the correct placement of the genus Eremobiotus. Moreover, the definition of the morphology of the claws of Eremobiotus-type and new observations regarding the buccal opening are discussed.

Molecular detection of Potential Novel Mansonella species (Filarioidea: Onchocercidae) in equine and Culicoides enderleini from Africa

A new genotype of a filarial worm from the Mansonella genus was determined in three donkeys from Algeria, as well as from four horses and two Culicoides enderleini biting midges from Senegal. Phylogenetic analysis based on the ITS1 and rRNA 5S genes shows similar results clustering this filarial worm within the genus Mansonella with low identity with all known Mansonella species. Further research should be performed to characterise this Mansonella sp. and assess its prevalence in African countries.

Strain-specific differences in reovirus infection of murine macrophages segregate with polymorphisms in viral outer-capsid protein σ3.

Mammalian orthoreovirus (reovirus) strains type 1 Lang (T1L) and type 3 Dearing-RV (T3D-RV) infect the intestine in mice but differ in the induction of inflammatory responses. T1L infection is associated with the blockade of oral immunological tolerance to newly introduced dietary antigens, whereas T3D-RV is not. T1L infection leads to an increase in infiltrating phagocytes, including macrophages, in gut-associated lymphoid tissues that are not observed in T3D-RV infection. However, the function of macrophages in reovirus intestinal infection is unknown. Using cells sorted from infected intestinal tissue and primary cultures of bone-marrow-derived macrophages (BMDMs), we discovered that T1L infects macrophages more efficiently than T3D-RV. Analysis of T1L × T3D-RV reassortant viruses revealed that the viral S4 gene segment, which encodes outer-capsid protein σ3, is responsible for strain-specific differences in infection of BMDMs. Differences in the binding of T1L and T3D-RV to BMDMs also segregated with the σ3-encoding S4 gene. Paired immunoglobulin-like receptor B (PirB), which serves as a receptor for reovirus, is expressed on macrophages and engages σ3. We found that PirB-specific antibody blocks T1L binding to BMDMs and that T1L binding to PirB-/- BMDMs is significantly diminished. Collectively, our data suggest that reovirus T1L infection of macrophages is dependent on engagement of PirB by viral outer-capsid protein σ3. These findings raise the possibility that macrophages function in the innate immune response to reovirus infection that blocks immunological tolerance to new food antigens.IMPORTANCEMammalian orthoreovirus (reovirus) infects humans throughout their lifespan and has been linked to celiac disease (CeD). CeD is caused by a loss of oral immunological tolerance (LOT) to dietary gluten and leads to intestinal inflammation following gluten ingestion, which worsens with prolonged exposure and can cause malnutrition. There are limited treatment options for CeD. While there are genetic risk factors associated with the illness, triggers for disease onset are not completely understood. Enteric viruses, including reovirus, have been linked to CeD induction. We found that a reovirus strain associated with oral immunological tolerance blockade infects macrophages by virtue of its capacity to bind macrophage receptor PirB. These data contribute to an understanding of the innate immune response elicited by reovirus, which may shed light on how viruses trigger LOT and inform the development of CeD vaccines and therapeutic agents.

Integrative taxonomic revision of the grasshopper genera Parapetasia Bolívar, 1884, and Loveridgacris Rehn, 1954 (Orthoptera, Pyrgomorphidae), with description of a new species of Loveridgacris

The taxonomic status of the Pyrgomorphid genera Parapetasia Bolívar, 1884, and Loveridgacris Rehn, 1954 is complex and challenging. Here, we use a combination of morphological, distributional, and genetic data to revise the two genera and provide new information on their diversity. We describe a new species, Loveridgacris tectiferussp. nov., from Tanzania and formally resurrect the status of Parapetasia rammei as a valid species within Parapetasia, resulting in two species in Parapetasia (P. femorata and P. rammei) and two in Loveridgacris (L. impotens and L. tectiferussp. nov.). We also sequenced the COI and 16S genes of 10 Pyrgomorphidae species and provided the first phylogeny of the group. Our data show that all species are clearly distinct and represent molecular operational taxonomic units (mOTUs), with the exceptions of L. impotens and L. tectiferussp. nov., which are morphologically clearly distinct but for which the concatenated sequence alignments of the two individual gene datasets (COI and 16S) do not provide sufficient information. In addition, high interspecific distances were found between Parapetasia and Loveridgacris. Moreover, the complete mitogenomes of L. impotens and L. tectiferussp. nov. were sequenced using next-generation sequencing technology. The total lengths of the assembled mitogenomes were 15,592 bp and 15,737 bp, representing 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and one D-loop region, respectively. To aid in identification, we present a key for the two genera, including a key to species. This study provides insights into the morphology, distribution, and phylogeny of Pyrgomorphidae in Africa.

Circular DNA enhanced amplification-free CRISPR/Cas12a assays for end-user friendly ultra-sensitive Porphyromonas gingivalis diagnosis

Periodontitis not only leads to tooth loss but is also associated with various systemic symptoms. Porphyromonas gingivalis (Pg) is the microorganism most closely linked to periodontitis. A rapid and accurate Pg detection method holds significant potential health benefits for patients. While several nucleic acid amplification-based diagnostic methods have been developed, there is a current lack of rapid diagnostic tools for Pg, particularly at chair-side level. Here, two detection methods for the Pg 16S gene based on the CRISPR/Cas12a system have been developed. To achieve ultra-high sensitivity in the developed CRISPR/Cas12a assays without pre-amplification, we designed a circular DNA molecule (termed as Cir-amplifier) which amplifies the signal output of the Cas12a reactions in a cascade response manner. The Cir-amplifier enhanced Fluorescence-Cas12a reaction achieved a minimum detectable limit of 0.39 CFU of Pg within 80-minute under room-temperature. In addition, the Cir-amplifier enhanced Lateral Flow Detection (LFD)-Cas12a reaction has demonstrated the ability to detect a minimum of 3.9 CFU of Pg in 90 min, with results observable by the naked eye. The potential of the two methods for Pg detection in clinical samples has been demonstrated, showing 100 % sensitivity compared to the traditional real-time PCR method. In summary, we have established two efficient detection methods for Pg that eliminate the need for pre-amplification, hence no amplicon contaminations, making them promising candidates for chair-side Pg diagnosis. Furthermore, the Cir-amplifier has been demonstrated to be adaptable to CRISPR/Cas systems producing various signal formats. The Cir-amplifier is expected to become a universal CRISPR/Cas enhancer.

The terrestrial flatworm Microplana scharffi (Geoplanidae, Microplaninae): mitochondrial genome, phylogenetic proximity to the Bipaliinae and genes related to regeneration

A genome skimming approach of sequencing was undertaken on a subfamily of terrestrial flatworms that had been neglected in genomic studies until now, namely the Microplaninae as represented here by Microplana scharffi. A single run of short-read sequencing enabled retrieval of the complete mitogenome, the two paralogous versions of the 18S gene, the elongation factor gene EF1α, plus two genes involved in the regeneration process, namely those coding for ß-CATENIN-1 and adenomatous polyposis coli (APC). The 15,297 bp mitogenome lacks a functional tRNA-Ala and has a mandatory alternative TTG start codon in its cox1 gene. The multiprotein phylogeny, inferred from mitogenome proteins, positions M. scharffi as sister-group to the Bipaliinae with maximum support, although the organisation of the mitogenomes shows features previously never observed among Bipaliinae, such as the conserved 32 bp overlap between ND4 and ND4L. Similarly to what has been observed in recent publications on other species of Geoplanidae, the two types of 18S genes display strongly different coverages and are only 90.57% identical. Additionally, alien DNA was identified in the pool of contigs in the form of the complete mitochondrial genome of Lumbricus rubellus, confirming previous observations on the feeding habits of M. scharffi.

Comparative analysis of Seabuckthorn and its significant component in enhancing fish mucosal immunity

Seabuckthorn and its significant component, gallic acid, were found to alleviate foodborne enteritis in zebrafish, a model species. To investigate the protective effects on mucosal immunity in cultured fish, seabuckthorn or gallic acid was added to soybean meal feed in a study conducted on juvenile grass carp. After a 5-week feeding trial, histopathology, enteritis-related gene expression, transcriptomics, and 16S rRNA gene sequencing were performed on the hindgut and liver tissues. The results showed that dietary inclusion of seabuckthorn (9.5 g/kg) or gallic acid (1.2 g/kg) prevented enteritis, evidenced by an increase in intestinal villi length and a decrease in goblet cell numbers. The gallic acid group showed benefits in preserving villi length, while seabuckthorn was advantageous in controlling goblet cells. On the molecular level, downregulation of pro-inflammatory genes and upregulation of anti-inflammatory genes were observed. Specifically, seabuckthorn was more effective in controlling cytokine expression, while gallic acid better protected the epithelial barrier function. Omics analysis revealed an upregulated hepatic “PPAR signaling pathway” in the seabuckthorn group and downregulated intestinal “platelet activation” in the gallic acid group, suggesting reduced inflammation and improved immune homeostasis. Regarding intestinal microbiota, both groups showed reduced pathogenic Campilobacterota and Aeromonas, and increased Bacteroides and Cetobacteria, potentially enhancing mucosal immunity. For anti-inflammatory processes, hepatic “fatty acid degradation”, “tryptophan metabolism”, and “FoxO signaling pathway” were elevated only in the seabuckthorn group. Gallic acid was advantaged with more “mitophagy” and “autophagy”. Fish-fed seabuckthorn or gallic acid were challenged with Aeromonas hydrophila to assess impacts on other mucosal surfaces. In the gill, compared to the soybean meal group, significantly higher survival rates, reduced CD4 inflammatory signals, and increased filament space were observed. Comparing seabuckthorn with its active component, seabuckthorn performed better in lipid metabolism, resistance to A. hydrophila, anti-inflammation, and probiotic abundance, while gallic acid mainly showed benefits for mucosal barrier protection, such as increased gill filament space. In summary, seabuckthorn and gallic acid improved mucosal immunity in fish, particularly in the gut and gills, enhancing resistance to bacterial infection and foodborne inflammation. Thus, gallic acid and seabuckthorn may be optimal phytotherapy choices in aquaculture.

Characterization of newborn gut microbiota according to the pre-gestational maternal nutritional status and delivery mode.

The aim of this study is to characterize the composition of the newborn gut microbiota based on the maternal pre-pregnancy nutritional status and the delivery mode. A biological sample was collected from the anal mucosa of the newborns between 24 and 48h after delivery, as it was not possible to collect a meconium sample at that time. A general data collection questionnaire was administered. The microbiome of the samples was analyzed by next-generation sequencing of the hypervariable regions v3-v4 of the 16S gene. Alpha diversity analyses were performed using the Observed Richness and Shannon diversity index metrics and Beta diversity analyses were conducted using Nonmetric multidimensional scaling with Weighted Unifrac, Differential abundance analysis was performed using a Negative Binomial Wald Test with maximum likelihood estimation for coefficients of Generalized Linear Models. Newborns of obese mothers exhibited lower alpha diversity compared to newborns of mothers with adequate BMI (body mass index). We observed variation in the composition of the microbial community in newborn stool samples, both from mothers with overweight/obesity and those with adequate pre-pregnancy BMI. We observed a visible correlation between the mode of delivery and the newborn's microbiota. We found variation in the overall composition of the microbial community in the stools of newborns, regardless of the delivery mode. The results of our study demonstrate differences in the microbiota of neonates born via cesarean section compared to those born vaginally as well as differences in newborns of mothers with overweight/obesity compared to those with an adequate pre-pregnancy BMI.

Molecular and Morphological Characteristics of a Novel Cyst Nematode in the Rhizosphere of Artemisia lavandulaefolia DC. in Gansu Province, Northwest China.

Cyst nematodes are obligate parasitic nematodes found in the fields of many cultivated crops. These nematodes, which have great economic importance, pose a threat to food security, though they are frequently ignored or misdiagnosed as pests because of covert parasitism. A cyst nematode population parasitizing on Artemisia lavandulaefolia DC., one of the traditional Chinese medicines was collected in Gansu Province. The species was diagnosed using integrative taxonomy and molecular approaches. The cyst population is spherical or lemon-shaped, light brown or dark brown in color, with a long neck and a protruding vulval cone. The stylet of the second-stage juvenile is strong, and the front end of the ball at the base of the stylet is concave; the median bulb and excretory pore are prominent; the tail is blunt and circular, and the transparent tail is usually shorter than the stylet. A phylogenetic analysis was carried out using the internal transcribed spacer (ITS) and 28S genes of ribosomal DNA, which further confirmed the presence of Cactodera chenopodiae. According to our literature review, this is the first report on C. chenopodiae in Compositae. By following this research, we can better understand the challenges posed by A. lavandulaefolia DC. and develop effective strategies for managing its spread and impacts. This will help to protect vulnerable ecosystems and ensure the sustainability of agricultural and forestry activities in affected areas.

Effects of Sucralose Supplementation on Glycemic Response, Appetite, and Gut Microbiota in Subjects with Overweight or Obesity: A Randomized Crossover Study Protocol.

Sucralose stands as the most common non-nutritive sweetener; however, its metabolic effects have sparked significant controversy over the years. We aim to examine the effects of sucralose daily intake on glycemia, subjective appetite, and gut microbiota (GM) changes in subjects with overweight or obesity. In this randomized, crossover, and controlled trial, 23 participants with a body mass index between 25 kg/m2 and 39.9 kg/m2 will be assigned to one of two interventions to receive either sucralose (2 mg/kg/day equivalent to 40% of the acceptable daily intake) or glucose (control) for 4 weeks, each phase separated by a 4-week washout period. The glycemic response will be determined during a meal tolerance test, subjective appetite will be evaluated using a visual analog scale, and GM changes will be analyzed by next-generation sequencing of the bacterial rRNA 16S gene from fecal samples. All measures will be performed before and after intervention periods. We hypothesize that sucralose supplementation induces changes in glycemic response, subjective appetite, and gut microbiota in overweight and obese participants. This protocol was approved by the Ethics Committee of the UJAT (No. 0721) and was registered in the Australian New Zealand Clinical Trials Registry (ACTRN12621001531808).

A molecular approach to identify parrotfish (Sparisoma) species during early ontogeny.

Sparisoma species (parrotfish) comprise an important functional group contributing to coral-reef resilience. The morphological diagnostic characteristics for species identification are clearly described for adult forms but not for the early stages. Consequently, many taxonomical listings of Sparisoma larvae are restricted to the genus level. The aims of this study are to determine whether the morphological and molecular identification techniques are useful to assign the species taxonomic level to Sparisoma larvae occurring in the Gulf of Mexico and whether there is a set of diagnostic features that could be used to discriminate between species in larvae of different developmental stages. Morphological assignment of Sparisoma was performed based on morphological and meristic features for 30 larvae collected in the Gulf of Mexico from late August to mid-September 2015. To corroborate and complement the morphological assignments, molecular identification was carried out using DNA sequences from regions of two mitochondrial genes, mitochondrial cytochrome oxidase I (mtDNA COI) and mitochondrial 16S rRNA (mtDNA 16S rRNA). COI and 16S gene trees for Sparisoma and related fish taxa were constructed using sequences available in the NCBI (National Center for Biotechnology Information) GenBank and BOLD (Barcode of Life Data) databases. Two morphotypes were identified based on morphology, but no diagnostic characteristics for species discrimination were found. Molecular identification, in contrast, successfully discriminated four early development stages of Sparisoma atomarium, three stages of Sparisoma radians, and two stages of Sparisoma chrysopterum and Sparisoma aurofrenatum, therefore demonstrating the successful and necessary application of molecular taxonomic approaches for species-level identifications of Sparisoma larvae.

Changes in Soil Bacterial and Fungal Community Composition and Functional Groups During the Artificial Restoration of Degraded Grassland of "Black-Soil Mountain".

About 35% of grassland in Sanjiangyuan area of China has degenerated into black-soil mountain. Artificial grassland is considered to be an effective measure to alleviate the severely degraded grassland in the alpine region of the three rivers and has been widely used. However, the pattern, potential function, and changes of carbon and nitrogen contents of soil microorganisms in degraded grassland in Heimushan by planting artificial grassland are still unclear. In this study, mixed-sown artificial alpine grassland (AG) was the focus of our study, whereas degraded black-soil mountain grassland (BG) and natural alpine grassland (NG) served as controls. Illumina 16S and ITS gene sequence analyses were used to analyze the community structure of the soil bacteria and fungi. The functional groups of NG, AG, and BG were predicted using the FAPROTAX and FUNGuild databases. In addition, the levels of soil carbon, nitrogen, and soil enzyme activities were evaluated. The results indicated a significant increase in the aboveground biomass of BG due to the planting artificial grassland. Moreover, the contents of total carbon (TC), total nitrogen (TN), ammonium nitrogen ( ), microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), and leucine aminopeptidase (LAP) increased in the soil. Planting artificial grasslands changed the composition of bacterial and fungal communities. Among these, the bacterial community was more sensitive to planting artificial grasslands. The relative abundance of bacterial functional groups involved in carbon and nitrogen cycling changed significantly, suggesting that bacteria may play a role in regulating nutrient cycling during artificial grassland planting. Soil TC, TN, LAP, and affected the microbial community structure related to carbon and nitrogen. and β-1,4-glucosidase were carbon and nitrogen factors, respectively, that affected functional changes in fungi. These results indicate that planting artificial grasslands can effectively enhance the productivity of degraded black-soil mountain and regulate soil microbial communities and soil physical and chemical properties.

A NEW SPECIES OF ACANTHOBOTHRIUM (CESTODA: ONCHOPROTEOCEPHALIDEA), PARASITE OF THE GIANT ELECTRIC RAY NARCINE ENTEMEDOR JORDAN AND STARKS, 1895 (BATOIDEA: TORPEDINIFORMES) FROM MEXICO.

Spiral valves from specimens of the giant electric ray Narcine entemedor Jordan & Starks, 1895 were examined in search of tapeworms at 2 localities of the Mexican tropical Pacific Ocean. Acanthobothrium oceguerai n. sp. is described herein based on material from Ventanilla, Oaxaca and from Acapulco Bay, Guerrero. The new species is a category 6 species, distinguished by being apolytic, retaining proglottids on the strobila until they are gravid, having strobila of 166-322 proglottids, having a small scolex and very long bothridia relative to the scolex, and having abaxial prongs that are short and thin in comparison to the axial prongs, which are longer and more robust. Acanthobothrium oceguerai n. sp. can be differentiated from other members of category 6 by the hooks, which are shorter, more robust, and smaller than those of the other members of this category. The phylogenetic analysis based on the 28S rRNA locus placed Acanthobothrium oceguerai n. sp. as sister to an unidentified species of larval Acanthobothrium from Philadelphia, Pennsylvania. In addition, sequences of the mitochondrial cytochrome oxidase subunit I gene and nuclear 18S rDNA were generated to provide support for future taxon sampling. Acanthobothrium oceguerai n. sp. is the fifth species of cestode reported from N. entemedor in the tropical eastern Pacific.