MOLECULAR VIROLOGICAL ANALYSIS OF THE TRANSMISSION CLUSTERS AND RESISTANCE MUTATIONS OF HIV-1 SUBTYPE B IN BULGARIA (2012-2020)

HIV-1 infection in Bulgaria is known for its high level of genetic diversity. Previous studies have indicated that subtype B is the most common strain in Bulgaria, particularly among men who have sex with men, who are at a high risk of transmission. The primary objective of this study was to identify any transmission clusters and transmission resistance in individuals newly diagnosed with HIV-1 who have not yet received antiretroviral therapy (ART). To this end, we sequenced the HIV-1 pol gene in the samples from the study participants using either the Viroseq HIV-1 Genotyping Test (Abbott) and the Applied Biosystems 3130xl genetic analyzer or the TruGene DNA Sequencing System (Siemens Healthcare) and an OpenGene DNA sequencing system. We then subtyped the HIV-1 pol sequences, and further analyzed those that met the criteria for subtype B. The study included a total of 595 HIV-1 subtype B sequences. Our analysis revealed that the majority of those diagnosed with HIV-1 subtype B were male and lived in Sofia region. The most common transmission mode was through sexual intercourse among men who have sex with men, followed by heterosexual transmission. We also observed the presence of multiple transmission clusters , and a low percentage of transmitted drug resistance mutations (TDRMs). Overall, our study confirms that HIV-1 subtype B remains the most dominant strain in Bulgaria

Phylogenetic, Genomic and Morphological Investigations of Three Lance Nematode Species (\u3ci\u3eHoplolaimus\u3c/i\u3e spp.)

Lance nematodes (Hoplolaimus spp.) are migratory ecto-endo plant-parasitic. They have been found from a wide range of the world that feed on the roots of a diversity of monocotyledonous and dicotyledonous plants, and have caused a great agricultural damage. Since more taxonomic knowledge and molecular references are demanded for the lance nematode phylogeny and population study, four chapters of lance nematode researches on three species were presented here: (1) A new species, Hoplolaimus smokyensis n. sp., was discovered from a mixed forest sample of maple (Acer sp.), hemlock (Tsuga sp.) and silverbell (Halesia carolina) from the Great Smoky Mountains National Park. It is characterized by possession of a lateral field with four incisures, an excretory pore posterior to the hemizonid, esophageal glands with three nuclei, phasmids anterior and posterior to the vulva, and the epiptygma absent. Phylogenetic analyses based on ribosomal and mitochondrial gene sequences also suggest H. smokyensis n. sp. to be an independent lineage distinct from all other reported Hoplolaimus species. (2) Additional morphological characteristics of Hoplolaimus columbus were described. Photos of its esophageal gland cell nuclei, a H. columbus male and abnormal female tails were presented. (3) The first complete de novo assembly of mitochondrial genome of Hoplolaimus columbus using Whole Genome Amplification and Illumina MiSeq technique was reported as a circularized DNA of 25228bp. The annotation results using two genetic codes were diagnosed and compared. Including H. columbus, phylogenetic relationships, gene content and gene order arrangement of 92 taxa nematodes were analyzed. (4) The phylogenetic informativeness of mitochondrial genes in Nematoda phylum is analyzed with two quantitative methods using mitochondrial genomes of 93 nematode species, including H. columbus and H. galeatus. Results from both methods agree with each other, indicate that the nad5 and nad4 contain higher informativeness than other candidates. Traditional markers like the cox1 and cytb genes contain medium informativeness. The nad4l and nad3 contain the lowest informativeness comparing with other protein-coding genes. Results also indicate that the phylogenetic informativeness is independent of the molecular sequence length of a phylogenetic marker. Concatenated-genes marker could present better phylogenetic informativeness if selected genes are higher informative

Zoonotic Cryptosporidium spp. in Wild Rodents and Shrews

There has been a significant increase in the number of reported human cryptosporidiosis cases in recent years. The aim of this study is to estimate the prevalence of Cryptosporidium spp. in wild rodents and shrews, and investigate the species and genotype distribution to assess zoonotic risk. Partial 18S rRNA gene nested-PCR reveals that 36.8, 53.9 and 41.9% of mice, voles and shrews are infected with Cryptosporidium species. The highest prevalence occurred in the Microtus agrestis (field vole) and Myodes glareolus (bank vole). Interestingly, bank voles caught in fields were significantly more often Cryptosporidium-positive compared to those caught in forests. The proportion of infected animals increases from over-wintered (spring and summer) to juveniles (autumn) suggesting acquired immunity in older animals. Based on Sanger sequencing and phylogenetic analyses, Apodemus flavicollis (yellow-necked mouse) is commonly infected with zoonotic C. ditrichi. Voles carry multiple different Cryptosporidium sp. and genotypes, some of which are novel. C. andersoni, another zoonotic species, is identified in the Craseomys rufocanus (grey-sided vole). Shrews carry novel shrew genotypes. In conclusion, this study indicates that Cryptosporidium protozoan are present in mouse, vole and shrew populations around Finland and the highest zoonotic risk is associated with C. ditrichi in Apodemus flavicollis and C. andersoni in Craseomys rufocanus. C. parvum, the most common zoonotic species in human infections, was not detected

First documented occurrences of Cladonia krogiana and C. rangiformis in north America

Funding Information: We thank Scott LaGreca for providing a photograph of the specimen of C. rangiformis at BM that was reportedly collected on Bermuda; Bruce Allen and the late Ronald Pursell for allowing us to cite their collections of Homalothecium sericeum from Newfoundland; Zdeněk Palice for enabling us to compare his collection of C. krogiana from the Czech Republic with material from Canada and Norway; Nathalie Djan-Chékar (Provincial Museum of Newfoundland and Labrador) for co-organizing the 2007 Tuckerman Workshop; and Irwin Brodo, James Lendemer, and an anonymous reviewer for their helpful comments on the manuscript. Permission to collect lichens in Fundy National Park and New River Beach Provincial Park (NRBPP) was approved by Renee Wissink (Parks Canada) and Martin MacMullin (NB Department of Tourism, Heritage, and Culture). Fieldwork in NRBPP and follow-up studies were supported in part by the New Brunswick Wildlife Trust Fund and the New Brunswick Environmental Trust Fund. Publisher Copyright: © 2021, New York Botanical Garden. All rights reserved.– Cladonia krogiana, previously known only from Norway and the Czech Republic, is reported here for North America from two localities near the Bay of Fundy, New Brunswick, Canada. It occurs there on open, rocky banks of clear, free-flowing rivers, habitats similar to those in which it has been found in Norway. We also document the occurrence of C. rangiformis in North America, based on collections from two localities on the southwest coast of Conception Bay, on the Avalon Peninsula of the island of Newfoundland, Canada. It is possibly an accidental, but naturalized, introduction in this area, where European settlement began in the early 1600s. A molecular phylogenetic analysis confirmed the identity of one of the Newfoundland specimens. The IGS rDNA haplotype to which it belongs is the same as the most widely distributed haplotype of C. rangiformis in Europe and Macaronesia. Previous reports of C. rangiformis for continental North America are based on misidentifications. A 19th century collection reportedly made on the island of Bermuda, while correctly identified, is of uncertain provenance.Peer reviewe

Can plastid genome sequencing be used for species identification in the Lauraceae?

Using DNA barcoding for species identification remains challenging for many plant groups. New sequencing approaches such as complete plastid genome sequencing may provide some increased power and practical benefits for species identification beyond standard plant DNA barcodes. We undertook a case study comparing standard DNA barcoding to plastid genome sequencing for species discrimination in the ecologically and economically important family Lauraceae, using 191 plastid genomes for 131 species from 25 genera, representing the largest plastome data set for Lauraceae to date. We found that the plastome sequences were useful in correcting some identification errors and for finding new and cryptic species. However, plastome data overall were only able to discriminate c. 60% of the species in our sample, with this representing a modest improvement from 40 to 50% discrimination success with the standard plant DNA barcodes. Beyond species discrimination, the plastid genome sequences revealed complex relationships in the family, with 12/25 genera being non-monophyletic and with extensive incongruence relative to nuclear ribosomal DNA. These results highlight that although useful for improving phylogenetic resolution in the family and providing some species-level insights, plastome sequences only partially improve species discrimination, and this reinforces the need for large-scale nuclear data to improve discrimination among closely related species

A multidisciplinary approach in tackling invasive species: barcoding, morphology, and metataxonomy of the leafhopper Arboridia adanae

The leafhopper genus Arboridia includes several species that feed on Vitis vinifera and cause leaf chlorosis. We report the first alien Arboridia infestation in Italy in 2021 in an Apulian vineyard. To confirm the taxonomic status of the species responsible for crop damage, and reconstruct its demographic history, we barcoded individuals from Apulia together with Arboridia spp. from Crete (Greece), A. adanae from Central Turkey and other specimens of the presumed sister species, A. dalmatina from Dalmatia (Croatia). Molecular phylogenies and barcoding gap analysis identified clades not associated with sampling locations. This result is incongruent with classical specimen assignment and is further supported by morphological analyses, which did not reveal significant differences among the populations. Therefore, we propose A. dalmatina as a junior synonym of A. adanae, which would become the only grapevine-related Arboridia species in the eastern Mediterranean. To further characterise A. adanae evolution, we performed a molecular clock analysis that suggested a radiation during the Pleistocene glaciations. Finally, to assess whether the Apulian individuals carried microorganisms of agricultural relevance, we sequenced their bacterial microbiota using 16S rRNA amplicon sequencing identifying three phytopathogens not generally associated with Arboridia activities as well as Wolbachia in one Apulian haplogroup. We discuss the agricultural implications of this infestatio

House dust mites possess a polymorphic, single domain putative peptidoglycan d,l endopeptidase belonging to the NlpC/P60 Superfamily

AbstractA 14kDa protein homologous to the γ-d-glutamyl-l-diamino acid endopeptidase members of the NlpC/P60 Superfamily has been described in Dermatophagoides pteronyssinus and Dermatophagoides farinae but it is not clear whether other species produce homologues. Bioinformatics revealed homologous genes in other Sarcopteformes mite species (Psoroptes ovis and Blomia tropicalis) but not in Tetranychus urticae and Metaseiulus occidentalis. The degrees of identity (similarity) between the D. pteronyssinus mature protein and those from D. farinae, P. ovis and B. tropicalis were 82% (96%), 77% (93%) and 61% (82%), respectively. Phylogenetic studies showed the mite proteins were monophyletic and shared a common ancestor with both actinomycetes and ascomycetes. The gene encoding the D. pteronyssinus protein was polymorphic and intronless in contrast to that reported for D. farinae. Homology studies suggest that the mite, ascomycete and actinomycete proteins are involved in the catalysis of stem peptide attached to peptidoglycan. The finding of a gene encoding a P60 family member in the D. pteronyssinus genome together with the presence of a bacterial promotor suggests an evolutionary link to one or more prokaryotic endosymbionts

Description, Distribution, and Relevance of Viruses of the Forest Pathogen Gremmeniella abietina

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