Moraxella haemolytica sp. nov., isolated from a goat with respiratory disease.

An aerobic, haemolytic, Gram-negative and rod-shaped bacterial strain ZY171148T was isolated from the lung of a dead goat with respiratory disease in Southwest China. The strain grew at 24-39 °C, at pH 6.0-9.0 and in the presence of 0.5-2.0% (w/v) NaCl. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain belongs to the genus Moraxella. The nucleotide sequence similarity analysis of the 16S rRNA gene showed that the strain has the highest similarity of 98.1% to Moraxella (M.) caprae ATCC 700019 T. Phylogenomic analysis of 800 single-copy protein sequences indicated that the strain is a member of the genus Moraxella and forms a separated branch on the Moraxella phylogenetic tree. The strain exhibited the highest orthologous average nucleotide identity (OrthoANI) and average amino acid identity (AAI) values of 77.0 and 77.9% to M. nasibovis CCUG 75921T and M. ovis CCUG 354T, respectively. The strain shared the highest digital DNA-DNA hybridization (dDDH) value of 26.2% to M. osloensis CCUG 350T. The genome G + C content of strain ZY171148T was 42.6 mol%. The strain had C18:1 ω9c (41.7%), C18:0 (11.2%), C16:0 (14.1%) and C12:0 3OH (9.7%) as the predominant fatty acids and CoQ-8 as the major respiratory quinone. The strain contained phosphatidylglycerol, phosphatidylethanolamine, cardiolipin, dilysocardiolipin, monolysocardiolipin and phosphatidic acid as the major polar lipids. ß-haemolysis was observed on Columbia blood agar. All results confirmed that strain ZY171148T represents a novel species of the genus Moraxella, for which the name Moraxella haemolytica sp. nov. is proposed, with strain ZY171148T = CCTCC AB 2021471T = CCUG 75920T as the type strain.

Moraxella nasibovis sp. nov., Isolated from a Cow with Respiratory Disease.

Strain ZY190618T, isolated from the nasal cavity of a cow with respiratory disease, was subjected to taxonomic characterization. Cells of the strain were Gram-stain-negative, aerobic and coccus-shaped. Phylogenetic analysis based on 16 S rRNA gene sequences indicated that the strain belonged to the genus Moraxella with the highest similarity of 98.1% to Moraxella nasovis CCUG 75922T. Phylogenomic analysis based on 810 single-copy genes revealed that the strain was a member of the genus Moraxella and formed a deep and separated clade within the genus. The strain showed the highest orthologous average nucleotide identity (OrthoANI) value of 77.1% with Moraxella ovis CCUG 354T and digital DNA-DNA hybridization (dDDH) value of 24.7% with Moraxella equi NCTC 11012T, respectively. The DNA G + C content was 46.5 mol%. The strain optimally grew at 37 °C (temperature range, 24-42 °C), at pH 8.0 (pH range, 6.0-9.0) and with 1.5% (w/v) NaCl (NaCl range, 0.5-3.0%). The strain contained C18:1 ω9c as the sole predominant fatty acid (> 5 %) and CoQ-8 as the major respiratory quinone. The major polar lipids included phosphatidylglycerol, phosphatidylethanolamine, cardiolipin, monolysocardiolipin and hemibismonoacylglycerophosphate. Based on these data, strain ZY190618T clearly represents a novel species in the genus Moraxella, for which the name Moraxella nasibovis sp. nov. (The type strain ZY190618T = CCUG 75921T = CCTCC AB 2021472T) is proposed.

Moraxella nasicaprae sp. nov., Isolated from a Goat with Respiratory Disease.

A novel Gram-stain-negative, aerobic, irregular coccus designated as ZY201224T, was isolated from the nasal cavity of a goat with respiratory disease in a goat farm, located at Jianshui, Yunnan Province, PR China and its taxonomic position was clarified using a polyphasic approach. The strain grew optimally at 37 °C, at pH 8.0 and in the presence of 1% NaCl. Phylogenetic analysis based on 16S rRNA gene sequence and phylogenomic analysis based on 808 single-copy genes revealed that the strain is affiliated to the genus Moraxella and is distinct from the recognized species of the genus. The 16S rRNA gene sequence similarity analysis indicated that the strain is most closely related to Moraxella caviae CCUG 355T with sequence similarity of 98.1%. The genomic OrthoANI and digital DNA-DNA hybridization (dDDH) values between the strain and the type strains of Moraxella species were no higher than 74.7% (Moraxella pluranimalium CCUG 54913T) and 26.0% (Moraxella oblonga NBRC 102422T), respectively. The G + C content of the complete genome sequence was 43.6 mol%. The strain contained CoQ-8 as the major respiratory quinone, and C18:1ω9c, C17:1ω8c, C16:0 and summed feature 3 (C16:1 ω7c and/ or C16:1ω6c) as the predominant fatty acids (> 5%). The major polar lipids comprised phosphatidylglycerol (PG), cardiolipin (CL), monolysocardiolipin (MLCL), phosphatidylethanolamine (PE) and lysophosphatidylglycerol (LPG). Based on these taxonomic characterizations, strain ZY201224T represents a novel species of the genus Moraxella, for which the name Moraxella nasicaprae sp. nov. is proposed. The type strain is ZY201224T (= CCTCC AB 2021474T = NBRC 115473T).

Moraxella nasovis sp. nov., isolated from a sheep with respiratory disease.

A novel Gram-stain-negative, aerobic, coccus-shaped bacteria, designated ZY201115T, was isolated from the nasal cavity of a sheep with respiratory disease in Yunnan Province, south-west China, and its taxonomic affiliation was studied by applying a polyphasic approach. The strain grew at 18-41 °C (optimum, 37 °C), at pH 6.0-9.0 (optimum, pH 8.0) and in 0.5-3.0% (w/v) NaCl (optimum, 1.0 % NaCl). Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain is affiliated to the genus Moraxella with highest similarity to Moraxella bovis ATCC 10900T (96.6 %). Phylogenomic analysis based on 811 single-copy genes also indicated that the strain represents a novel species in the genus Moraxella and formed a deep and separated clade with Moraxella caviae NCTC 10293T. The highest genomic orthologous average nucleotide identity and digital DNA-DNA hybridization values between the strain and the type strains in the genus Moraxella were 73.7% (M. caviae NCTC 10293T) and 25.3% (Moraxella osloensis CCUG 350T), respectively. The G+C content of the complete genome sequence was 42.1 mol%. The predominant fatty acids (>5 %) were C18:1 ω9c, C17:1 ω8c, C12:03OH and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c). The major polar lipids were phosphatidylglycerol, cardiolipin, monolysocardiolipin, phosphatidylethanolamine and hemibismonoacylglycerophosphate. The major respiratory quinone was CoQ-8. On the basis of the results of phylogenetic, phenotypic and chemotaxonomic characterizations, strain ZY201115T clearly represents a novel species of the genus Moraxella, for which the name Moraxella nasovis sp. nov. is proposed. The type strain is ZY201115T (=CCTCC AB 2021473T=CCUG 75922T).

Genetic and Pathogenic Characterisation of a Virulent Akabane Virus Isolated from Goats in Yunnan, China.

Introduction: Akabane virus (AKAV) has been detected in a variety of host species in China, but there are only limited records of its occurrence in goats. However, more attention needs to be paid to understanding the diversity of viruses in this species. The aim of the study was to explore the genotype characteristics and variation trend of AKAV and their relationship with virulence in Yunnan, China. Material and Methods: Blood samples were collected from goats during routine surveillance of goat diseases in Yunnan province in 2019. The AKAV CX-01 strain was isolated using BHK-21 cells. To understand pathogenicity, the virus was intraperitoneally (IP) and intracerebrally (IC) inoculated into suckling mice and tissue samples were subsequently analysed histopathologically and immunohistochemically. Results: Akabane virus CX-01 strain induced encephalitis and impairment of the central nervous system with fatal consequences. Phylogenetic analysis based on the ORF sequences of the small segments indicated that the AKAV isolate used was most closely related to the GD18134/2018 Chinese midge and bovine NM BS/1strains, while phylogenetic analysis based on the medium segments showed a close relationship between CX-01 and the Chinese GLXCH01 strain. Conclusion: The CX-01 isolate was related to AKAV genogroup Ia and probably originated from a recombination of different strains.

Substitution 3A protein of foot-and-mouth disease virus of attenuated ZB strain rescued the viral replication and infection in bovine cells.

The non-structural protein 3A of foot-and-mouth disease virus (FMDV) plays an important role in viral replication, virulence and determination of host range. Previously we identified genomic changes in gene encoding 3A protein between the attenuated ZBatt strain and its parental virulent strain during the attenuation process. However, the effects of changes in 3A protein on viral replication and infection of the rabbit-attenuated ZBatt virus during the attenuation process are poorly understood. In this study, a chimeric virus, rZBatt-3A, was constructed by introducing the 3A gene of virulent ZB virus into its attenuated vaccine ZB strain. Subsequently, the biological characteristics between rZBatt-3A and its parental virus (rZBatt) were compared. The relative expression level of four host cell proteins that interact with FMDV 3A were also analyzed. The results showed that the chimeric virus rZBatt-3A exhibited significantly different growth properties and plaque phenotypes from rZBatt in primary fetal bovine kidney (BK) cells. Cytopathic effect (CPE) of the rZBatt-3A was observed in BK cells with smaller plaque size, but CPE from the rZBatt could not be observed. The viral RNA replication was higher in rZBatt-3A-infected BK cells than in rZBatt-infected cells at 24 hpi (P < .05). In addition, the relative mRNA expression level of Ubiquilin 1 (UBQLN1) was significantly increased in rZBatt-3A-infected BK cells than in rZBatt-infected cells (P < .01) suggesting that UBQLN1 may be associated with 3A protein changes. Thus, the substitution of 3A protein altered the replication efficiency of attenuated ZB virus in bovine cells. Our data suggested that changes in 3A protein might be associated with the attenuation of ZB virus, which shed more lights in molecular mechanisms about attenuation of FMDV.

Genome Sequence of Corynebacterium pseudotuberculosis Strain KM01, Isolated from the Abscess of a Goat in Kunming, China.

Caseous lymphadenitis (CLA) is an acute, pyogenic, and contagious disease of goat that imposes considerable economic losses for farmers, and it is caused by Corynebacterium pseudotuberculosis Herein, we introduce the genome sequencing of C. pseudotuberculosis strain KM01, isolated from an abscess of a Saanen goat from Kunming, China. The genome contains 2,198 genes, the total length of the genes was 2,337,666 bp, and the GC content was 52.18%. The number of tandem repeat sequences was 44, the total length of the tandem repeat sequences was 1,970 bp (0.0772% of the genome), the number of minisatellite DNAs was 36, and there were 48 tRNAs and 12 rRNAs.