Hemotropic mycoplasmas in bats from forest fragments, state of Paraná, southern Brazil

The order Chiroptera is the second largest group of mammals with bats being identified as reservoir of several viral zoonoses, although, little is known about their role in other groups of pathogens, including hemotropic Mycoplasma spp. To date, hemoplasma species have been found infecting several species of bats with high genetic diversity between 16S rRNA gene sequences. On this study, we aimed to identify the occurrence and characterize 16S and 23S rRNA genes of hemoplasma species in four bats species (Artibeus lituratus, Carollia perspicillata, Sturnira lilium and Sturnira tildae) from forest fragments in Paraná State, southern Brazil, using PCR-based assays. Spleen tissue samples were collected, DNA extracted and further screened by a pan‑hemoplasma PCR assay. All samples consistently amplified the mammal endogenous gapdh gene. One out of 15 (6.66%; 95% CI: 0.2-31%) bats tested positive for hemotropic Mycoplasma sp. by the PCR assay targeting the 16S rRNA gene. Sequencing of the 16S rRNA gene fragment from the hemoplasma-positive bat showed 99.14% identity with hemotropic Mycoplasma sp. detected in Sturnira parvidens from Belize. Sequencing of the 23S rRNA gene fragment from the hemoplasma-positive bat showed 86.17% identity with ‘Candidatus Mycoplasma haemosphiggurus’ detected in orange-spined hairy dwarf porcupines (Sphiggurus villosus) from Southern Brazil.

Phylogenetic position of Odorrana macrotympana (Yang, 2008) (Anura, Ranidae) and extension of its geographical distribution

Based on a 16S rRNA gene fragment, a molecular phylogeny for the genus Odorrana Fei, Ye & Huang, 1990 was reconstructed, the validity of the poorly-known ranid species O. macrotympana (Yang, 2008) was confirmed and its phylogenetic position was evaluated. In addition, we report the first country record of O. macrotympana from Myanmar, based on our new records from Htamanthi Wildlife Sanctuary, Sagaing Division and present a supplementary description of this species. This report also constitutes the first record of O. macrotympana from outside of China.

Assessment of Plasmodium falciparum anti-malarial drug resistance markers in pfcrt and pfmdr1 genes in isolates from Honduras and Nicaragua, 2018–2021

Background Central America and the island of Hispaniola have set out to eliminate malaria by 2030. However, since 2014 a notable upturn in the number of cases has been reported in the Mosquitia region shared by Nicaragua and Honduras. In addition, the proportion of Plasmodium falciparum malaria cases has increased significantly relative to vivax malaria. Chloroquine continues to be the first-line drug to treat uncomplicated malaria in the region. The objective of this study was to evaluate the emergence of chloroquine resistant strains of P. falciparum using a genetic approach. Plasmodium vivax populations are not analysed in this study. Methods 205 blood samples from patients infected with P. falciparum between 2018 and 2021 were analysed. The pfcrt gene fragment encompassing codons 72–76 was analysed. Likewise, three fragments of the pfmdr1 gene were analysed in 51 samples by nested PCR and sequencing. Results All samples revealed the CVMNK wild phenotype for the pfcrt gene and the N86, Y184F, S1034C, N1042D, D1246 phenotype for the pfmdr1 gene. Conclusions The increase in falciparum malaria cases in Nicaragua and Honduras cannot be attributed to the emergence of chloroquine-resistant mutants. Other possibilities should be investigated further. This is the first study to report the genotype of pfmdr1 for five loci of interest in Central America.

Toxorhynchites (Lynchiella) caatingensis sp. nov. (Diptera: Culicidae) from the semi-arid region of Brazil

This publication describes a new species of mosquito of the genus Toxorhynchites Theobald, 1901, subgenus Lynchiella Lahille, 1904. The species was found during an exploratory survey of the Culicidae in the Caatinga biome, which is unique to Brazil and characterized by a semi-arid climate. Because of the specific characteristics of the biome, it has potential for endemism of species that have adapted to its restrictive conditions, particularly those imposed by severe drought. The new species, Toxorhynchites (Lynchiella) caatingensis, is a phytotelmic species that inhabits bromeliads and has an unknown biology. It belongs to a species complex that is hereafter referred to as the Violaceus Complex, which comprises three species that are found in the same type of habitat and have similar morphology: Tx. caatingensis n. sp., Tx. mariae (Bourroul, 1904) and Tx. violaceus (Wiedemann, 1820). We describe the larva, pupa, adult female and male genitalia of specimens collected as larvae from the tank bromeliad Aechmea aquilega (Salisb.) Griseb. The main distinctive characteristics of the species are the presence of three lateral tufts on the terminal abdominal segments (one pale and the others dark); a blue midline on the abdominal sterna; in the pupa, the pentagonal shape of the paddle and the length of seta 6-V. PCR reactions were carried out for the mitochondrial CO1 gene fragment, which placed the new species in the subgenus Lynchiella. No match was found with any species recorded in GenBank. The subgenus now includes 17 species.

The genome of the diatom Chaetoceros tenuissimus carries an ancient integrated fragment of an extant virus

Diatoms are one of the most prominent oceanic primary producers and are now recognized to be distributed throughout the world. They maintain their population despite predators, infections, and unfavourable environmental conditions. One of the smallest diatoms, Chaetoceros tenuissimus, can coexist with infectious viruses during blooms. To further understand this relationship, we sequenced the C. tenuissimus strain NIES-3715 genome. A gene fragment of a replication-associated gene from the infectious ssDNA virus (designated endogenous virus-like fragment, EVLF) was found to be integrated into each 41 Mb of haploid assembly. In addition, the EVLF was transcriptionally active and conserved in nine other C. tenuissimus strains from different geographical areas, although the primary structures of their proteins varied. The phylogenetic tree further suggested that the EVLF was acquired by the ancestor of C. tenuissimus. Additionally, retrotransposon genes possessing a reverse transcriptase function were more abundant in C. tenuissimus than in Thalassiosira pseudonana and Phaeodactylum tricornutum. Moreover, a target site duplication, a hallmark for long interspersed nuclear element retrotransposons, flanked the EVLF. Therefore, the EVLF was likely integrated by a retrotransposon during viral infection. The present study provides further insights into the diatom-virus evolutionary relationship.

Assessment of Plasmodium Falciparum Antimalarial Drug Resistance Markers in pfcrt and pfmdr1 Genes in Isolates from Honduras and Nicaragua, 2018-2021

Background: Central America and the island of Hispaniola have set out to eliminate malaria by 2030. However, since 2014 a notable upturn in the number of cases has been reported in La Mosquitia region shared by Nicaragua and Honduras. In addition, the proportion of Plasmodium falciparum malaria cases has increased significantly relative to vivax malaria. Chloroquine continues to be the first line drug to treat uncomplicated malaria in the region. The objective of this study was to evaluate the emergence of chloroquine resistant strains using a genetic approach. Methods: 205 blood samples from patients infected with P. falciparum between 2018 and 2021 were analyzed. The pfcrt gene fragment encompassing codons 72-76 was analyzed. Likewise, three fragments of the pfmdr1 gene were analyzed in 51 samples by nested PCR and sequencing. Results: All samples revealed the CVMNK wild phenotype for the pfcrt gene and the N86, Y184F, S1034C, N1042D, D1246 phenotype for the pfmdr1 gene. Conclusions: The increase in falciparum malaria cases in Nicaragua and Honduras cannot be attributed to the emergence of chloroquine-resistant mutants. Other possibilities should be investigated further. This is the first study to report the genotype of pfmdr1 for five loci of interest in Central America.

The version of microevolutionary relationships between biotypes close to Elymus caninus (Poaceae) according to the sequencing of the nuclear gene GBSS1 (waxy)

A comparative study of the sequences of the GBSS1 gene fragment in accessions of species close to Elymus caninus:E. prokudinii, E. viridiglumis, E. goloskokovii, as well as a number of morphologically deviating forms (MDF) from the territoryof Russia and Kazakhstan was carried out. The StH-genomic constitution was established or confirmed in all studied taxa andMDF, and the microevolutionary relationships between species were assessed by constructing separate NJ dendrograms based onexons and introns together and exons only. Differences in the location of Y subgenomes in reference StY-genomic species on twotypes of constructed dendrograms were noted. Possible evolutionary reasons for these differences are discussed. A characteristicfeature of all taxa close to E. caninus is the presence of only St2clones of the St subgenome, which are closer in composition tothe North American ancestral line Pseudoroegneria spicata than to the Asiatic line ascending to P. strigosa. The reference Polishaccession E. caninus can_5274 is farthest from the Asian accessions in the St2subgenome, but closer to the diploid carrier ofthe St genome P. spicata. According to the levels of differentiation of the H subgenome, all putative relatives of E. caninus havevariants of the H1subgenome around the Asian diploid host Hordeum jubatum, while being divided into two distinct subgroups.Only the accessions of four reference species gravitate towards the North American species Hordeum californicum – the carrierof the H2gene variant. It should be noted that the gene variants of the Ural endemic E. uralensis are close to the main group ofE. caninus accessions in both subgenomes. Together with the results of sexual hybridization, this fact gives grounds to considerE. uralensis as closely related to the main group of taxa deserving the intraspecific rank of E. caninus s. l.