Update of Genetic Diversity of Porcine Circovirus Type 2 in Chile Evidences the Emergence of PCV2d Genotype

Porcine Circovirus 2 (PCV2) can cause multiple clinical conditions known as porcine circovirus-associated diseases (PCVAD). Before the wide availability of PCV2 vaccines, PCVAD resulted in significant losses to the global swine industry. PCV2's rapid evolutionary dynamics are comparable to single-stranded RNA viruses. Thus, shifts in the dominance and distribution of different genotypes may frequently occur, resulting in the emergence and spread of varying PCV2 genotypes and recombinant strains in swine. This study aims at identifying the PCV2 genotypes currently circulating in Chile. Seven hundred thirty-eight samples were obtained from 21 swine farms between 2020 and 2021. The samples were tested using PCR for species detection and genotyping. Sequencing and phylogenetic analyses were conducted in selected samples. PCV2 was detected in 26.9% of the PCR reactions and 67% of the sampled farms. The genotypes were determined in nine farms, PCV2a in one farm, PCV2b in four, and PCV2d in five, with PCV2b and PCV2d co-circulating in one farm. The phylogenetic analysis of twelve ORF2 sequences obtained (PCV2a = 5; PCV2b = 4; PCV2d = 3) showed a PCV2a Chilean strains monophyletic cluster; closely related to Chilean viruses collected in 2012 and 2013. Of the three different PCV2b sequenced viruses, two viruses were close to the root of the PCV2b group, whereas the remaining one grouped with a South Korean virus. PCV2d sequences were closely related to Asian viruses. A previously reported PCV2a/PCV2d recombinant strain was not detected in this study. Our results suggest the emergence and potential shift to PCV2d genotype in Chilean farms.

Case Report: First Report and Phylogenetic Analysis of Porcine Astroviruses in Chile

Porcine Astrovirus (PoAstV) causes mild diarrhea in young pigs and is considered an emerging virus in the swine industry worldwide. PoAstV has high genetic diversity and has been classified into five genetic lineages, PoAstV1–5. In Chile, only human astroviruses have been reported. This study aimed to determine the presence and genetic diversity of PoAstV circulating in intensive pig farms in Chile. Seventeen Chilean intensive swine farms from Valparaíso, Metropolitana, O'Higgins, Ñuble and Araucanía regions were sampled. A selection of oral fluid and fecal material samples from 1–80 days-old pigs were collected and analyzed using next-generation sequencing. The circulation of PoAstV was confirmed in all studied farms. We obtained complete or partial sequences of PoAstV-2 (n = 3), PoAstV-4 (n = 2), and PoAstV-5 (n = 7). In 15 out of 17 farms, we detected more than one lineage co-circulating. Phylogenetic analyses grouped the seven PoAstV-5 strains in a monophyletic cluster, closely related to the United States PoAstV-5 strains. The three PoAstV-2 were located into two separate sub-clusters. PoAstV-4 sequences are also grouped in two different clusters, all related to Japanese strains. Thus, our results indicate that PoAstV circulates in Chile with high frequency and diversity. However, the lack of reference sequences impairs local evolution patterns establishment and regional comparisons. This is the first contribution of PoAstV genomes in Latin America; more studies are needed to understand the diversity and impact of PoAstV on swine health.

Epidemiological and etiological aspects of enterovirus infection in Russia and Vietnam

The epidemic situation and etiological factors of enterovirus infection in Russia and Vietnam were analysed and compared. The identified strains of enteroviruses of 47 types, which circulated in Russia in 2018–2019, belonged to different species: Enterovirus species A (CV-A2, CV-A4, CV-A6, CV-A8, CV-A10, CV-A16, EV-A71), as well as Enterovirus species B and Enterovirus species C. The strains isolated from 87 children from southern Vietnam hospitalised in 2018–2019 into infectious hospitals while having enterovirus infection with exanthema were also studied. All identified strains were represented by Enterovirus species A: EV-A71 — 59 strains, CV-A10 — 20 strains, CV-A16 — 5 strains, CV-A6 — two strains and CV-A2 — one strain. Out of 59 viruses EV-A71, 53 strains belonged to genotype C4 and 6 strains belonged to genotype B5. The sequences of EV-A71 strains of genotype C4 from South Vietnam formed a monophyletic cluster with the sequences of EV-A71 viruses which circulated during 2016–2018 in different provinces of China, and they were very close to EV-A71 strains of the same genotype from the Yunnan Province. These strains were genetically different from Russian viruses and Vietnamese viruses identified in the years 2003–2005 and 2011– 2012. Most of the cases of enterovirus infection from southern Vietnam (78%) caused by EV-A71 virus of genotype C4 were reported in three provinces located in southern Vietnam in the Mekong Delta. The epidemic process and the etiology of enterovirus infection in Russia and Vietnam have common features. At the same time, the epidemic situation in these countries is not the same. The incidence of enterovirus infection is influenced by geographic, climatic, economic and demographic factors that differ in two countries. In the majority of territories of Russia, the climate is temperate or cold, seasonal rises in the incidence rates of enterovirus infection usually occur in the summer, when people go on vacation, spend a lot of time outdoors and swim in open reservoirs. In Vietnam, a constant high-level temperature, a high population density and a large proportion of children determine the higher incidence of enterovirus infection, especially in the southern provinces of Vietnam, compared to Russia. The fact that more than 20% of the Vietnamese population lives in the Mekong Delta, which is the largest river in Indochina, has a significant impact on the epidemic process of enterovirus infection in South Vietnam. The Mekong River which flows through China, Laos, Thailand, Cambodia and Vietnam, and carries huge streams of water, including rainstorm waters and sewages, from all of these countries into the southern provinces of Vietnam, which have the highest incidence rates of enterovirus infection in the country. The results of the research underline the importance of active epidemiological and virological surveillance of enterovirus infection, which plays the key role in informing the public health authorities about the changes in the epidemic situation in order to take appropriate measures and develop the prevention strategies. The goal of anti-epidemic and preventive measures is to reduce the incidence of enterovirus infection and the economic burden of this infection for Russia and Vietnam.

Nocardioides baculatus sp. nov., a novel actinomycete isolated from the rhizosphere of Tagetes patula

An aerobic, Gram-stain-positive, non-motile, dull-yellow, short rod-shaped actinomycete strain, designated G10T, was isolated from Tagetes patula (marigold) roots collected from Goyang in the Republic of Korea. The isolate showed best growth on Reasoner's 2A agar at 25 °C, pH 6.5.0 and with 0% NaCl (w/v). The strain was negative for oxidase activity and positive for catalase activity. On the basis of 16S RNA gene sequence similarity, strain G10T was affiliated to the genus Nocardioides and the closest species were Nocardioides glacieisoli HLT3-15T (98.8 %), Nocardioides zhouii HLT2-9T (98.8 %), Nocardioides ganghwensis JC2055T (98.7 %), Nocardioides cavernae YIM A1136T (98.6 %), Nocardioides flavus Y4T (98.5 %), Nocardioides oleivorans DSM 16090T (98.3 %), Nocardioides alpinus Cr7-14T (98.2 %), Nocardioides exalbidus DSM 22017T (98.1 %) and Nocardioides hwasunensis KCTC 19197T (98.1 %). Strain G10T formed a monophyletic cluster with N. glacieisoli HLT3-15T, N. zhouii HLT2-9T and N. hwasunensis KCTC 19197T in all phylogenetic trees. The cell-wall peptidoglycan of strain G10T contained ll-diaminopimelic acid as the diagnostic amino acid. The predominant fatty acids were iso-C16 : 0 and C17 : 1 ω8c. MK-8(H4) was the major isoprenoid quinone. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol. Average nucleotide identity and digital DNA–DNA hybridization values were 78.6–88.7 % and 21.5–36.2 %, respectively, with the type strains of related species of the genus Nocardioides , suggesting that strain G10T represents a novel species. The genome of strain G10T is 4 231 000 bp long with a DNA G+C content of 71.5 mol% and encodes 4071 predicted proteins, six rRNAs and 46 tRNAs. The genome of strain G10T comprises the biosynthetic gene cluster for T3PKS, terpene, NRPS-like fragment and RRE-containing element as secondary metabolites. The results of taxonomic, phylogenetic, biochemical, chemotaxonomic and genomic analysis clearly supported that strain G10T represent a novel species within the genus Nocardioides , for which the name Nocardioides baculatus sp. nov is proposed and the type strain is G10T (=KCTC 49626T=NBRC 114801T).

The taxonomy of two uncultivated fungal mammalian pathogens is revealed through phylogeny and population genetic analyses

Ever since the uncultivated South American fungal pathogen Lacazia loboi was first described 90 years ago, its etiology and evolutionary traits have been at the center of endless controversies. This pathogen infects the skin of humans and as long believed, dolphin skin. However, recent DNA analyses of infected dolphins placed its DNA sequences within Paracoccidioides species. This came as a surprise and suggested the human and dolphin pathogens may be different species. In this study, population genetic analyses of DNA from four infected dolphins grouped this pathogen in a monophyletic cluster sister to P. americana and to the other Paracoccidioides species. Based on the results we have emended the taxonomy of the dolphin pathogen as Paracoccidioides cetii and P. loboi the one infecting human. Our data warn that phylogenetic analysis of available taxa without the inclusion of unusual members may provide incomplete information for the accurate classification of anomalous species.

Genomic Characterization of Strains From a Cluster of Infant Botulism Type A in a Small Town in Colorado, United States

Three cases of infant botulism were reported in a small Colorado town between 1981 and 1984. The first two cases occurred in 1981, 6 months apart, and the third case occurred in 1984. Clostridium botulinum type A was isolated from stool of all three case patients and from environmental samples of the patient’s homes. An epidemiological investigation and follow-up study were conducted from 1981 to 1986 and concluded the cases were likely related. In this study, we sought to determine whether the C. botulinum type A clinical isolates were related to each other and to isolates obtained from environmental samples. We performed whole genome sequencing (WGS) for 17 isolates associated with this potential cluster of infant botulism. Fifteen isolates were confirmed to be C. botulinum type A(B) and contained botulinum toxin gene subtypes A1 and B5 by WGS; these strains formed a monophyletic cluster in a phylogeny and were considered closely related to each other (0–18 high-quality single-nucleotide polymorphisms), but distinct from other C. botulinum type A(B) in Colorado and elsewhere in the United States. Results of our study suggest that the three infant botulism cases could have represented a cluster due to a C. botulinum type A(B) strain present in the environment.

Sequence Read Depth Analysis of a Monophyletic Cluster of Y Chromosomes Characterized by Structural Rearrangements in the AZFc Region Resulting in DYS448 Deletion and DYF387S1 Duplication

The azoospermia factor c region (AZFc), located in the long arm of the human Y chromosome, is frequently involved in chromosome rearrangements, mainly due to non-allelic homologous recombination events that occur between the nearly identical sequences (amplicon) that comprises it. These rearrangements may have major phenotypic effects like spermatogenic failure or other pathologies linked to male infertility. Moreover, they may also be relevant in forensic genetics, since some of the Y chromosome short tandem repeats (Y-STRs) commonly used in forensic analysis are located in amplicons or in inter-amplicon sequences of the AZFc. In a previous study, we identified four phylogenetically related samples with a null allele at DYS448 and a tetrallelic pattern at DYF387S1, two Y-STRs located in the AZFc. Through NGS read depth analysis, we found that the unusual Y-STR pattern may be due to a 1.6 Mb deletion arising concurrently or after a 3.5 Mb duplication event. The observed large genomic rearrangement results in copy number reduction for the RBMY gene family as well as duplication of other AZFc genes. Based on the diversity of 16 additional Y-STRs, we estimated that the duplication/deletion event occurred at least twenty generations ago, suggesting that it has not been affected by negative selection.

First complete-genome documentation of HIV-1 intersubtype superinfection with transmissions of diverse recombinants over time to five recipients

Human immunodeficiency virus type 1 (HIV-1) recombinants in the world are believed to be generated through recombination between distinct HIV-1 strains among coinfection or superinfection cases. However, direct evidence to support transmission of HIV-1 recombinants from a coinfected/superinfected donor to putative recipient is lacking. Here, we report on the origin and evolutionary relationship between a set of recombinants from a CRF01_AE/CRF07_BC superinfected putative donor and diverse CRF01_AE/CRF07_BC recombinants from five putative recipients. Interviews on sociodemographic characteristics and sexual behaviors for these six HIV-1-infected men who have sex with men showed that they had similar ways of partner seeking: online dating sites and social circles. Phylogenetic and recombination analyses demonstrated that the near-full-length genome sequences from six patients formed a monophyletic cluster different from known HIV-1 genotypes in maximum likelihood phylogenetic trees, were all composed of CRF01_AE and CRF07_BC fragments with two common breakpoints on env, and shared 4–7 breakpoints with each other. Moreover, 3’ half-genomes of recombinant strains from five recipients had identical/similar recombinant structures with strains at longitudinal samples from the superinfected donor. Recombinants from the donor were paraphyletic, whereas five recipients were monophyletic or polyphyletic in the maximum clade credibility tree. Bayesian analyses confirmed that the estimated time to the most recent common ancestor (tMRCA) of CRF01_AE and CRF07_BC strains of the donor was 2009.2 and 2010.7, respectively, and all were earlier than the emergence of recombinants from five recipients. Our results demonstrated that the closely related unique recombinant forms of HIV-1 might be the descendent of a series of recombinants generated gradually in a superinfected patient. This finding highlights the importance of early initiation of antiretroviral therapy as well as tracing and testing of partners in patients with multiple HIV-1 infection.

Morphological description, DNA barcodes and phylogenetic placement of a new mite species: Dinogamasus saengdaoae sp. nov. (Mesostigmata: Laelapidae) found in the acarinarium of carpenter bees in Thailand

Dinogamasus saengdaoae Attasopa & Ferrari sp. nov. is described based on adult females from the abdominal pouch of females of Xylocopa tenuiscapa (Westwood) in Chiang Mai Province, Northern Thailand. The new species belongs to the D. perkinsi (Oudemans) group (sensu LeVeque) and can be distinguished from its congeners by the combination of the following characters: (1) dorsal shield covering opisthosoma neither laterally nor posteriorly; (2) opisthonotal soft cuticle with a pair of relatively long setae posteriorly; (3) setae pd1, pd2 on genu I and ad3, pd3, pl1, pl2 on both genu and tibia I conical. Maximum likelihood-based analysis of newly-generated DNA barcodes shows that the sequenced specimens of D. saengdaoae sp. nov. form a monophyletic cluster, and parsimony analysis of a previously available morphological dataset indicates that the species comprises a strongly-supported clade with D. perkinsi and D. piperi LeVeque. We provide an additional couplet for Lundqvist’s key for the species of Dinogamasus Kramer to facilitate identification of D. saengdaoae sp. nov..

Origin and Spread of the Dengue Virus Type 1, Genotype V in Senegal, 2015–2019

Dengue virus (DENV) is the most widespread arthropod-borne virus, with the number and severity of outbreaks increasing worldwide in recent decades. Dengue is caused by genetically distinct serotypes, DENV-1–4. Here, we present data on DENV-1, isolated from patients with dengue fever during an outbreak in Senegal and Mali (Western Africa) in 2015–2019, that were analyzed by sequencing the envelope (E) gene. The emergence and the dynamics of DENV-1 in Western Africa were inferred by using maximum likelihood and Bayesian methods. The DENV-1 grouped into a monophyletic cluster that was closely related to those from Southeast Asia. The virus appears to have been introduced directly into Medina Gounass (Suburb of Dakar), Senegal (location probability = 0.301, posterior = 0.76). The introduction of the virus in Senegal occurred around 2014 (95% HPD = 2012.88–2014.84), and subsequently, the virus moved to regions within Senegal (e.g., Louga and Fatick), causing intense outbreaks in the subsequent years. The virus appears to have been introduced in Mali (a neighboring country) after its introduction in Senegal. In conclusion, we present evidence that the outbreak caused by DENV-1 in urban environments in Senegal and Mali after 2015 was caused by a single viral introduction from Asia.