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Sus scrofa papillomatosis (SsP) is a tumour caused by Sus scrofa papillomaviruses (SsPVs). To investigate the presence of SsPVs in China, 354 domestic pig skin samples collected from Guangxi Province were examined for SsPV DNA by PCR. Three SsPV1s (GX12, GX14, and GX18) were identified with a prevalence of 0.847% (3/354). Sequence analysis showed that L1 of SsPV1/GX12 and SsPV1/GX14 had 99.7% and 99.6% nucleotide identify with the reference SsPV1a, respectively. Phylogenetic and evolutionary analyses showed that SsPV1/GX12 and SsPV1/14 clustered into SsPV1a and that SsPV1/GX18 clustered into SsPV1b. Compared with other SsPV L1 and L2 proteins, we found that the SsPV1/GX18 and SsPV1b strains shared the same unique substitutions, and SsPV1/GX12, SsPV1/GX14, and SsPV1a shared almost identical amino acid sequences. This study reports the first detection of SsPV DNA in China based on whole genome information and provides a scientific basis for the development of SsPV pathogenic biology, epidemiology, and prevention, as well as control technology research.
Assuntos
Papillomaviridae , Sus scrofa , Animais , Suínos , Filogenia , Análise de Sequência de DNA , China/epidemiologia , Reação em Cadeia da Polimerase , Papillomaviridae/genéticaRESUMO
De novo synthesis of thiamine (vitamin B1) in plants depends on the action of thiamine thiazole synthase, which synthesizes the thiazole ring, and is encoded by the THI1 gene. Here, we investigated the evolution and diversity of THI1 in Poaceae, where C4 and C3 photosynthetic plants co-evolved. An ancestral duplication of THI1 is observed in Panicoideae that remains in many modern monocots, including sugarcane. In addition to the two sugarcane copies (ScTHI1-1 and ScTHI1-2), we identified ScTHI1-2 alleles showing differences in their sequence, indicating divergence between ScTHI1-2a and ScTHI1-2b. Such variations are observed only in the Saccharum complex, corroborating the phylogeny. At least five THI1 genomic environments were found in Poaceae, two in sugarcane, M. sinensis, and S. bicolor. The THI1 promoter in Poaceae is highly conserved at 300 bp upstream of the start codon ATG and has cis-regulatory elements that putatively bind to transcription factors associated with development, growth, development and biological rhythms. An experiment set to compare gene expression levels in different tissues across the sugarcane R570 life cycle showed that ScTHI1-1 was expressed mainly in leaves regardless of age. Furthermore, ScTHI1 displayed relatively high expression levels in meristem and culm, which varied with the plant age. Finally, yeast complementation studies with THI4-defective strain demonstrate that only ScTHI1-1 and ScTHI1-2b isoforms can partially restore thiamine auxotrophy, albeit at a low frequency. Taken together, the present work supports the existence of multiple origins of THI1 harboring genomic regions in Poaceae with predicted functional redundancy. In addition, it questions the contribution of the levels of the thiazole ring in C4 photosynthetic plant tissues or potentially the relevance of the THI1 protein activity.
Assuntos
Poaceae , Saccharum , Poaceae/metabolismo , Saccharum/genética , Tiamina , Fatores de Transcrição/genética , Folhas de Planta/metabolismoRESUMO
Aim: Two lytic phages were isolated using P. aeruginosa DSM19880 as host and fully characterized. Materials & methods: Phages were characterized physicochemically, biologically and genomically. Results & conclusion: Host range analysis revealed that the phages also infect some multidrug-resistant (MDR) P. aeruginosa clinical isolates. Increasing MOI from 1 to 1000 significantly increased phage efficiency and retarded bacteria regrowth, but phage ph0034 (reduction of 7.5 log CFU/ml) was more effective than phage ph0031 (reduction of 5.1 log CFU/ml) after 24 h. Both phages belong to Myoviridae family. Genome sequencing of phages ph0031 and ph0034 showed that they do not carry toxin, virulence, antibiotic resistance and integrase genes. The results obtained are highly relevant in the actual context of bacterial resistance to antibiotics.
Assuntos
Bacteriófagos , Pseudomonas aeruginosa , Bacteriófagos/genética , Especificidade de Hospedeiro , Técnicas In Vitro , Myoviridae/genéticaRESUMO
Limosilactobacillus fermentum (ATCC 23271) was originally isolated from the human intestine and has displayed antimicrobial activity, primarily against Candida species. Complete genome sequencing and comparative analyses were performed to elucidate the genetic basis underlying its probiotic potential. The ATCC 23271 genome was found to contain 2,193,335 bp, with 2123 protein-coding sequences. Phylogenetic analysis revealed that the ATCC 23271 strain shares 941 gene clusters with six other probiotic strains of L. fermentum. Putative genes known to confer probiotic properties have been identified in the genome, including genes related to adhesion, tolerance to acidic pH and bile salts, tolerance to oxidative stress, and metabolism and transport of sugars and other compounds. A search for bacteriocin genes revealed a sequence 48% similar to that of enterolysin A, a protein from Enterococcus faecalis. However, in vitro assays confirmed that the strain has inhibitory activity on the growth of Candida species and also interferes with their adhesion to HeLa cells. In silico analyses demonstrated a high probability of the protein with antimicrobial activity. Our data reveal the genome features of L. fermentum ATCC 23271, which may provide insight into its future use given the functional benefits, especially against Candida infections.
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The objective was to estimate allelic and genotypic frequencies for loci associated with meat quality in a Mexican population of Braunvieh cattle. Information was obtained from 300 animals genotyped with the Genomic Profile Bovine LD chip of 30K and 50K SNPs. After the final edition, including quality control, the data contained information for 12 loci of the CAPN1, CAPN3, CAPN5, CAPN14, DGAT1, DGAT2, TG, ANK1, and MADH3 genes. Allelic and genotypic frequencies and Hardy-Weinberg equilibrium were estimated with the Cervus 3.0.7 software. The studied population markers were in Hardy-Weinberg equilibrium, except for those associated with CAPN1, DGAT1, and MADH3. Frequencies higher than those reported for other breeds were found for genotypes associated with meat softness, higher marbling score, lower quantity of saturated fatty acids, and lower shear force (CAPN1 and DGAT2). There were similarities with frequencies reported for Bos taurus breeds for the CAPN3 and TG genes. For the DGAT1 and ANK1 genes, the frequencies of the desired genotypes were low. A marker for DGAT1 and another for MADH3 were monomorphic. The results of this study are encouraging in terms of the potential of the Braunvieh population studied for breeding programs aiming to increase meat quality. The breed has strengths that could be used either by crossbreeding to generate heterozygous animals or by selection to increase frequencies of valuable alleles.
Assuntos
Carne , Polimorfismo de Nucleotídeo Único , Alelos , Animais , Bovinos/genética , Frequência do Gene , Marcadores Genéticos , GenótipoRESUMO
Lactobacillus delbrueckii subsp. lactis CIDCA 133 (CIDCA 133) has been reported as a potential probiotic strain, presenting immunomodulatory properties. This study investigated the possible genes and molecular mechanism involved with a probiotic profile of CIDCA 133 through a genomic approach associated with in vitro and in vivo analysis. Genomic analysis corroborates the species identification carried out by the classical microbiological method. Phenotypic assays demonstrated that the CIDCA 133 strain could survive acidic, osmotic, and thermic stresses. In addition, this strain shows antibacterial activity against Salmonella Typhimurium and presents immunostimulatory properties capable of upregulating anti-inflammatory cytokines Il10 and Tgfb1 gene expression through inhibition of Nfkb1 gene expression. These reported effects can be associated with secreted, membrane/exposed to the surface and cytoplasmic proteins, and bacteriocins-encoding genes predicted in silico. Furthermore, our results showed the genes and the possible mechanisms used by CIDCA 133 to produce their beneficial host effects and highlight its use as a probiotic microorganism.
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Canine brucellosis caused by Brucella canis is a zoonotic disease that causes reproductive alterations in dogs, such as infertility, abortion, and epididymitis. This pathogen is especially prevalent in South America, and due to the lack of official control programs and the growing trend of adopting dogs it constitutes a public health risk that must be addressed. The aim of this study was to determine the prevalence of B. canis infection in kennel, shelter, and household dogs and to characterize the genomic properties of circulating strains, including ure and virB operons and omp25/31 genes. Samples from 771 dogs were obtained, and the infection was detected by blood culture and/or serology in 7.0% of the animals. The complete ure and virB operons and the omp25/31 genes were detected. Interestingly, we found different single-nucleotide polymorphisms (SNPs) in some of the analyzed genes, which could mean a change in the fitness or virulence of these strains. This study provides further evidence about dogs as a source of B. canis strains that can infect people. This also highlights the need to implement official control programs, including the mandatory testing of dogs, especially stray dogs, before adoption.
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Because Canine circovirus (CanineCV) is a new species of the genus Circovirus, several issues related to its epidemiology, pathogenesis and clinical disease remain unknown. Thus, this study aimed to perform the characterization of the first complete genome sequence of CanineCV detected in a dog with diarrhea in Brazil. A stool sample was collected of a ten-month-old female German Shepherd dog which had signs of intermittent hemorrhagic gastroenteritis, vomiting, and a history of eating raw pork. The complete CanineCV genome was sequenced by Next-Generation Sequencing. The sequence had 2,063 nucleotides, showed a typical genomic organization for circovirus, and was grouped with strain 214 described in the United States by phylogenetic analysis. One amino acid change was found in the replicase protein, and because of that it was considered unique to CanineCV. Therefore, the characterization of the complete genome of Brazilian CanineCV can be used in future studies of molecular epidemiology, pathogenesis and development of diagnostic tools for the prevention and control of this disease.(AU)
Como o Canine circovirus (CanineCV) é uma nova espécie do Gênero Circovirus, várias questões relacionadas com a sua epidemiologia, patogenia e doença clínica permanecem desconhecidas. Assim, este estudo objetivou realizar a caracterização da primeira sequência do genoma completo do CanineCV detectado em um cão com diarreia, no Brasil. Uma amostra de fezes foi coletada de um cão da raça Pastor Alemão, fêmea, 10 meses de idade, o qual tinha sinais de gastroenterite hemorrágica intermitente, vômito e uma história de ingestão de carne crua de porco. O genoma completo do CanineCV foi sequenciado pelo Sequenciamento de Nova Geração. A sequência tinha 2.063 nucleotídeos, apresentou uma organização genômica típica para um circovírus e foi agrupado com a cepa 214, descrita nos Estados Unidos pela análise filogenética. Uma mudança de aminoácido foi encontrada na proteína de replicação e por causa disso ela foi considerada única para o CanineCV. Portanto, a caracterização do genoma completo do CanineCV brasileiro pode ser utilizada em futuros estudos de epidemiologia molecular, patogenia e no desenvolvimento de ferramentas de diagnóstico para prevenção e controle desta doença.(AU)
Assuntos
Animais , Cães , Circovirus/genética , Circovirus/isolamento & purificação , Infecções por Circoviridae/epidemiologia , Infecções por Circoviridae/veterinária , Genoma , Gastroenterite/veterináriaRESUMO
ABSTRACT: Because Canine circovirus (CanineCV) is a new species of the genus Circovirus, several issues related to its epidemiology, pathogenesis and clinical disease remain unknown. Thus, this study aimed to perform the characterization of the first complete genome sequence of CanineCV detected in a dog with diarrhea in Brazil. A stool sample was collected of a ten-month-old female German Shepherd dog which had signs of intermittent hemorrhagic gastroenteritis, vomiting, and a history of eating raw pork. The complete CanineCV genome was sequenced by Next-Generation Sequencing. The sequence had 2,063 nucleotides, showed a typical genomic organization for circovirus, and was grouped with strain 214 described in the United States by phylogenetic analysis. One amino acid change was found in the replicase protein, and because of that it was considered unique to CanineCV. Therefore, the characterization of the complete genome of Brazilian CanineCV can be used in future studies of molecular epidemiology, pathogenesis and development of diagnostic tools for the prevention and control of this disease.
RESUMO: Como o Canine circovirus (CanineCV) é uma nova espécie do Gênero Circovirus, várias questões relacionadas com a sua epidemiologia, patogenia e doença clínica permanecem desconhecidas. Assim, este estudo objetivou realizar a caracterização da primeira sequência do genoma completo do CanineCV detectado em um cão com diarreia, no Brasil. Uma amostra de fezes foi coletada de um cão da raça Pastor Alemão, fêmea, 10 meses de idade, o qual tinha sinais de gastroenterite hemorrágica intermitente, vômito e uma história de ingestão de carne crua de porco. O genoma completo do CanineCV foi sequenciado pelo Sequenciamento de Nova Geração. A sequência tinha 2.063 nucleotídeos, apresentou uma organização genômica típica para um circovírus e foi agrupado com a cepa 214, descrita nos Estados Unidos pela análise filogenética. Uma mudança de aminoácido foi encontrada na proteína de replicação e por causa disso ela foi considerada única para o CanineCV. Portanto, a caracterização do genoma completo do CanineCV brasileiro pode ser utilizada em futuros estudos de epidemiologia molecular, patogenia e no desenvolvimento de ferramentas de diagnóstico para prevenção e controle desta doença.
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The aim is to describe the molecular epidemiology and perform a genomic characterization of hepatitis B virus (HBV) circulating in Mar del Plata and to identify the origin and diversification patterns of the most prevalent genotype. The S gene and the region encompassing the X gene, basal core promoter (BCP), and precore (preC) was analyzed in 56 samples. They were genotyped as: 80% F1b, 9% A2, 7% D3, and 2% D1. A recombinant F4/D2 genome was detected. The double substitution G1764A/A1762T at the BCP (reduced HBeAg expression) was found in 20% F1b, 2% A2, 2% D1, and 2% D3 samples. A unique D3 presented the G1896A substitution at the preC (HBeAg negative phenotype). A 13% of the samples showed mutations at the HBsAg "a" immunodeterminant (escape from neutralizing antibodies). Mutations at the polymerase (antiviral resistance) were found in 52% of the samples. Coalescent analysis of subgenotype F1b, the most prevalent in the city, showed that viral diversification in Mar del Plata started by year 2000. F1b was the most prevalent genotype detected, being a characteristic of actual HBV infections in Mar del Plata. Local HBV exhibit clinically relevant mutations, but a minority of them was shown to be associated to potential vaccination escape or antiviral resistance. Nevertheless, further studies are needed to determine whether any of these mutants could pose a threat to prevention, diagnosis, or treatment.
Assuntos
Evolução Molecular , Genótipo , Vírus da Hepatite B/classificação , Vírus da Hepatite B/genética , Hepatite B/virologia , Adulto , Idoso , Argentina/epidemiologia , Feminino , Antígenos do Núcleo do Vírus da Hepatite B/genética , Antígenos de Superfície da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Regiões Promotoras Genéticas , Transativadores/genética , Proteínas Virais Reguladoras e Acessórias , Adulto JovemRESUMO
Genus Gammapapillomavirus (Gamma-PV) is the most diverse and largest clade within the Papillomaviridae family. A novel set of degenerate primers targeting the E1 gene was designed and further used in combination with the well-known CUT PCR assay to assess HPV prevalence and genus distribution in a variety of cutaneous samples from 448 immunocompetent individuals. General HPV, Gamma-PV and mixed infections prevalence were significantly higher in actinic keratosis with respect to benign and malignant neoplasms, respectively (pâ¯=â¯0.0047, pâ¯=â¯0.0172, pâ¯=â¯0.00001). Gamma-PVs were significantly more common in actinic keratosis biopsies than Beta- and Alpha-PVs (pâ¯=â¯0.002). The full-length genome sequence of a novel putative Gamma-PV type was amplified by 'hanging droplet' long-range PCR and cloned. The novel virus, designated HPV210, clustered within species Gamma-12. This study provides an additional tool enabling detection of HPV infections in skin and adds new insights about possible early roles of Gamma-PVs in the development of cutaneous malignant lesions.
Assuntos
Gammapapillomavirus/genética , Gammapapillomavirus/isolamento & purificação , Ceratose Actínica/virologia , Reação em Cadeia da Polimerase/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Gammapapillomavirus/classificação , Humanos , Masculino , Pessoa de Meia-Idade , Infecções por Papillomavirus/virologia , Adulto JovemRESUMO
Bacillus cereus strains were isolated from dried foods, which included international brands of spices from South East Asia, Mexico and India purchased from several retail stores, samples of powdered infant formula (PIF), medicated fish feed and dietary supplements. The genetic diversity of 64 strains from spices and PIF was determined using a multiplex endpoint PCR assay designed to identify hemolysin BL, nonhemolytic enterotoxin, cytotoxin K, and enterotoxin FM toxin genes. Thirteen different B. cereus toxigenic gene patterns or profiles were identified among the strains. Randomly selected B. cereus strains were sequenced and compared with reference Genomic Groups from National Center Biotechnology Information using bioinformatics tools. A comprehensive multi-loci sequence analysis (MLSA) was designed using alleles from 25 known MLST genes specifically tailored for use with whole genome assemblies. A cohort of representative genomes of strains from a few FDA regulated commodities like dry foods and medicated fish feed was used to demonstrate the utility of the 25-MLSA approach for rapid clustering and identification of Genome Groups. The analysis clustered the strains from medicated fish feed, dry foods, and dietary supplements into phylogenetically-related groups. 25-MLSA also pointed to a greater diversity of B. cereus strains from foods and feed than previously recognized. Our integrated approach of toxin gene PCR, and to our knowledge, whole genome sequencing (WGS) based sequence analysis, may be the first of its kind that demonstrates enterotoxigenic potential and genomic diversity in parallel.
Assuntos
Bacillus cereus/genética , Bacillus cereus/metabolismo , Enterotoxinas/biossíntese , Microbiologia de Alimentos/métodos , Alimentos em Conserva/microbiologia , Fórmulas Infantis/microbiologia , Bacillus cereus/isolamento & purificação , Enterotoxinas/genética , Genes Bacterianos , Genoma Bacteriano/genética , Proteínas Hemolisinas/genética , Humanos , Índia , México , Tipagem de Sequências Multilocus , Reação em Cadeia da Polimerase Multiplex , Filogenia , Prevalência , Sequenciamento Completo do GenomaRESUMO
Tacaiuma virus (TCMV) is antigenically characterized as a member of the Anopheles A complex in the Orthobunyavirus genus, Peribunyaviridae family (Bunyavirales order). Clinically, the TCMV infection is characterized by acute febrile illness with myalgia and arthralgia lasting three to five days. However, the genomic and evolutionary aspect of this virus has not been elucidated. In this study, we described the complete coding sequences of three segments of two TCMV strains isolated in Brazil and three complete coding sequences of the small segment of three TCMV strains. All the strains sequenced in this study showed the typical genomic organization of orthobunyaviruses that infect vertebrates, except for the absence of the open reading frame that encodes the well-described non-structural small protein. This study presents the genomic and evolutionary characterization of TCMV strains and would be helpful for diagnostic purposes and epidemiology.