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Tomatoes (Solanum lycopersicum L.) are nutrient-rich fruits with a high glutamic acid content, making them a suitable source for producing γ-aminobutyric acid (GABA) through the action of lactic acid bacteria (LAB). LAB plays a pivotal role in lactic fermentation, enhancing the taste and nutritional value of food products, often contributing to their health benefits. This study isolated GABA-producing LAB from tomatoes and assessed their potential for producing GABA-enriched fermented tomato juice. The results indicated the isolation of 15 LAB strains from the samples of five different tomatoes. All of these strains demonstrated the capability to produce GABA, with levels ranging from 1.732 to 3.113 mg/mL, as determined using thin-layer chromatography (TLC). Through sequencing, the promising strain TO42, identified as Weissella cibaria, was selected for tomato juice fermentation. Furthermore, GABA-enriched tomato juice was successfully fermented at 15 °Brix for 72 h, resulting in the highest recorded GABA and lactic acid content of 9.185 ± 0.398 mg/mL and 13.05 ± 1.56 g/L, respectively.

O tomate (Solanum lycopersicum L.) é uma fruta rica em nutrientes e com alto teor de ácido glutâmico, tornando-o uma fonte adequada para a produção de ácido γ-aminobutírico (GABA) através da ação de bactérias ácido lácticas (BAL). As BAL desempenham um papel fundamental na fermentação láctica, melhorando o sabor e o valor nutricional dos produtos alimentares, contribuindo frequentemente para os seus benefícios para a saúde. Este estudo teve como objetivo isolar BAL produtoras de GABA de tomates e avaliar seu potencial para a produção de suco de tomate fermentado enriquecido com GABA. Os resultados indicam o isolamento de 15 cepas de BAL a partir de amostras de cinco tomates diferentes. Todas essas cepas demonstraram capacidade de produzir GABA, com níveis variando de 1,732 a 3,113 mg/mL, conforme determinado por cromatografia em camada delgada (CCD). Através de sequenciamento, a cepa promissora TO42, identificada como Weissella cibaria, foi selecionada para fermentação de suco de tomate. Além disso, o suco de tomate enriquecido com GABA foi fermentado com sucesso a 15 °Brix por 72 h, resultando no maior teor registrado de GABA e ácido láctico de 9.185 ± 0.398 mg/mL e 13.05 ± 1.56 g/L, respectivamente.

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To explore the important long noncoding RNA (lncRNA) and its target genes in Tibetan chicken ovarian tissue, whole transcriptome sequencing technology and bioinformatics methods were used to analyze and predict lncRNA, differential expression of mRNA, and lncRNA between the two different breeds using DESeq2 software. The study predicted the Antisense and Cis regulatory target genes of differentially expressed lncRNA and performed functional annotation of GO, and analysis of the KEGG signaling pathway in these target genes. Quantitative fluorescence PCR was conducted in real time to validate the expression of important target genes. The results showed the discovery of 532 differentially expressed lncRNAs, 340 of which upregulated and 292 downregulated, as well as 2314 differentially expressed mRNAs, with 983 being upregulated and 1331 downregulated. The differentially expressed lncRNAs were found to regulate 48 differentially expressed mRNAs in the sense direction, and 14 in the antisense direction. Enrichment analysis on the intersection of target genes of differentially expressed lncRNAs and mRNAs showed enrichment mainly in KEGG signaling pathways such as TGF-ß signaling pathway (ko04350), organic selenium compound metabolism (ko00450), and oocyte meiosis (ko04114). The quantitative fluorescence PCR validation of important target genes such as CYP17A1, PTPN5, ACSL3, Nf2, CYP11A1 showed consistent results with RNA-seq results. This indicates the presence of lncRNA and mRNA with different expression levels and specific expression in the ovarian tissues of Tibetan and Roman chickens, and that genes such as CYP17A1, PTPN5, ACSL3, Nf2, and CYP11A1 may be key factors regulating the laying performance of Tibetan chickens.(AU)

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Abstract Due to extensive application of antibiotics as growth promoters in animal feed, antimicrobial resistance has been increased. To overcome this challenge, rumen microbiologists search for new probiotics to improve the rate of livestock production. The present study was aimed to isolate and evaluate breed-specific lactic acid bacteria (LAB) as potential animal probiotics. The current study was conducted during 10 months from July 2020 to April 2021, in which a total of n=12 strains were isolated from different samples including milk, rumen, and feces of Nilli Ravi Buffaloes. These isolates were evaluated for their antimicrobial potential against common animal pathogens (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). All the isolates were identified using 16S rRNA gene sequencing and the phylogenetic analyses inferred that these strains showed close relations to the species of various genera; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis, and Lactococcus lactis. NMCC-Ru2 has exhibited the enormous potential of antimicrobial activity, 28 mm, for Salmonella typhimurium;23 mm for Listeria monocytogenes 21 mm for E.coil. Highest resistance was seen in NMCC-Ru2 agasint test antbiotic, like 25.5 mm for Tetracycline. Overall results revesl that the probiotic profile of isolates was achieved using standard criteria, particularly with animal probiotic properties

Resumo Devido à extensa aplicação de antibióticos como promotores de crescimento na alimentação animal, a resistência aos antimicrobianos aumentou. Para superar esse desafio, os microbiologistas do rúmen buscam novos probióticos para melhorar a produtividade do gado. O presente estudo teve como objetivo isolar e avaliar bactérias lácticas específicas de raças (BAL) como potenciais probióticos animais. 12 cepas foram isoladas de diferentes amostras, incluindo leite, rúmen e fezes de búfalos Nilli Ravi. Esses isolados foram avaliados quanto ao seu potencial antimicrobiano contra patógenos animais comuns (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). Todos os isolados foram identificados por meio do sequenciamento do gene 16S rRNA e as análises filogenéticas inferiram que essas cepas apresentaram estreita relação com as espécies de vários gêneros; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis, Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis e Lactococcus lactis. O perfil probiótico dos isolados foi obtido usando critérios padrão, particularmente com propriedades probióticas animais.

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Abstract Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Resumo Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis (9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

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This study aimed to identify candidate genes regulating ovarian development following ovarian tissue transplantation through transcriptome sequencing (RNA-seq) and weighted gene co-expression network analysis (WGCNA). Ovarian tissues were collected from 10 thirty-day-old donor chickens, and each ovary was divided equally into three parts of similar size, and transplanted in situ into two-day-old recipient chickens. Samples were collected on days 0 (untransplanted), 6, and 12 after transplantation. RNA was extracted from ovarian samples for construction of a transcriptome library, and then sequenced on the IlluminaHiSeqTM2000 sequencing platform. The sequencing results were evaluated to determine differentially expressed genes (DEGs) through GO function and the KEGG pathway analyses, Gene Set Enrichment Analysis (GSEA), WGCNA, and PPI networks. Some candidate genes were further validated through real-time fluorescence quantitative PCR (RT-qPCR). RNA-seq analysis identified 2242 up-regulated and 3095 down-regulated DEGs at 6 days after ovarian transplantation, and 2181 up-regulated and 2129 down-regulated DEGs were identified at 12 days after transplantation. Enrichment analysis showed that the genes were enriched in multiple inflammatory pathways, pathways involved in signal transduction and cellular processes. Through the WGCNA, five modules were constructed, from which 4 and 5 candidate genes were mined at 6 and 12 days after ovarian transplantation, respectively. Transcription factor prediction showed that GTF3A was the most important transcription factor. The results of RT-qPCR verification confirmed that the expression profile of 8 candidate genes was consistent with that of the sequencing results. In summary, this study presents the candidate genes involved in ovarian vascular remodeling and ovarian proliferation after ovarian transplantation.(AU)

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The type series of Leporinus paranensis includes two distinct species, one of which is herein described as new. Leporinus paranensis is redescribed based on its holotype, two paratypes, and additional specimens recently sampled in tributaries of the Grande river, in the north portion of the upper Paraná basin in Brazil. A new species of Leporinus is described based on specimens collected in all major tributaries of the upper Paraná basin in Brazil, and tributaries of the Paraná river in Paraguay. Both species share the presence of three unicuspid teeth on the premaxillary and four on dentary, terminal mouth and three dark midlateral blotches on the body. The new species is distinguished from L. paranensis based on the number of scale series around the caudal peduncle (12 vs. 16). The variation in the body shape of both species were compared through a Principal Component Analysis and showed that they share a similar body shape. Species delimitation analyses using DNA Barcode were applied to compare samples of the new species to congeners and corroborated the uniqueness of the new species. In addition, molecular data revealed that L. bahiensis, L. octofasciatus, and L. taeniatus are possibly closely related to the new species. The conservation status of L. paranensis and the new species are discussed using the IUCN criteria.(AU)

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PICK1 plays a crucial role in mammalian spermatogenesis. Here, we integrated single-molecule long-read and short-read sequencing to comprehensively examine PICK1 expression patterns in adult Baoshan pig (BS) testes. We identified the most important transcript ENSSSCT00000000120 of PICK1, obtaining its full-length coding sequence (CDS) spanning 1254 bp. Gene structure analysis located PICK1 on pig chromosome 5 with 14 exons. Protein structure analysis reflected that PICK1 consisted of 417 amino acids containing two conserved domains, PDZ and BAR_PICK1. Phylogenetic analysis underscored the evolutionary conservation and homology of PICK1 across different mammalian species. Evaluation of protein interaction network, KEGG, and GO pathways implied that interacted with 50 proteins, predominantly involved in glutamatergic synapses, amphetamine addiction, neuroactive ligand-receptor interactions, dopaminergic synapses, and synaptic vesicle recycling, and PICK1 exhibited significant correlation with DLG4 and TBC1D20. Functional annotation identified that PICK1 was involved in 9 GOs, including seven cellular components and two molecular functions. ceRNA network analysis suggested BS PICK1 was regulated by seven miRNA targets. Moreover, qPCR expression analysis across 15 tissues highlighted that PICK1 was highly expressed in the bulbourethral gland and testis. Subcellular localization analysis in ST (Swine Tesits) cells demonstrated that PICK1 significantly localized within the cytoplasm. Overall, our findings shed new light on PICK1's role in BS reproduction, providing a foundation for further functional studies of PICK1.(AU)

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Abstract Maydis leaf blight, caused by Bipolaris maydis, is an important disease of maize crop in Khyber Pakhtunkhwa (KP) Pakistan. Fifteen isolates of the pathogen, collected across KP, were studied for variability based on phenotypic and molecular markers. Significant variability among the isolates was observed when assessed using phenotypic traits such as radial growth, spore concentration, fungicide sensitivity and virulence. The isolates were classified into six culture groups based on colour, texture and margins of the colony. Conidial morphology was also variable. These were either straight or slightly curved and light to dark brown in colour. Fungicide test showed significant variation in the degree of sensitivity against Carbendazim. Isolate Bm8 exhibited maximum radial growth on carbendazim spiked plates. Conversely, isolate Bm15 showed the lowest radial growth. Variations in virulence pattern of the isolates were evident when a susceptible maize variety Azam was inoculated with spores of B. maydis. Genetic variability amongst the isolates was also estimated by RAPD as well as sequencing of ITS region. The RAPD dendrogram grouped all the isolates into two major clusters. Average genetic distance ranged from 0.6% to 100%, indicating a diverse genetic gap among the isolates. Maximum genetic distance was found between isolates Bm9 and Bm10 as well as Bm2 and Bm8. Conversely, isolates Bm13 and Bm15 were at minimum genetic distance. Phylogenetic dendrogram based on sequencing of ITS region grouped all the isolates into a single major cluster. The clusters in both the dendrogram neither correlate to the geographical distribution nor to the morphological characteristics.

Resumo A ferrugem das folhas de maydis, causada por Bipolaris maydis, é uma doença importante da cultura do milho em Khyber Pakhtunkhwa (KP), Paquistão. Quinze isolados do patógeno, coletados em KP, foram estudados quanto à variabilidade com base em marcadores fenotípicos e moleculares. Variabilidade significativa entre os isolados foi observada quando avaliada por meio de características fenotípicas, como crescimento radial, concentração de esporos, sensibilidade a fungicida e virulência. Os isolados foram classificados em seis grupos de cultura com base na cor, textura e margens da colônia. A morfologia dos conídios também foi variável. Estes eram retos ou ligeiramente curvos e de cor marrom-claro a escuro. O teste de fungicida mostrou variação significativa no grau de sensibilidade ao carbendazim. O isolado Bm8 exibiu crescimento radial máximo em placas com adição de carbendazim. Por outro lado, o isolado Bm15 apresentou o menor crescimento radial. As variações no padrão de virulência dos isolados foram evidentes quando uma variedade de milho suscetível Azam foi inoculada com esporos de B. maydis. A variabilidade genética entre os isolados também foi estimada por RAPD, bem como sequenciamento da região ITS. O dendrograma RAPD agrupou todos os isolados em dois grupos principais. A distância genética média variou de 0,6% a 100%, indicando uma lacuna genética diversa entre os isolados. A distância genética máxima foi encontrada entre os isolados Bm9 e Bm10 e também entre Bm2 e Bm8. Por outro lado, os isolados Bm13 e Bm15 estavam a uma distância genética mínima. O dendrograma filogenético baseado no sequenciamento da região ITS agrupou todos os isolados em um único aglomerado principal. Os agrupamentos em ambos os dendrogramas não se correlacionam com a distribuição geográfica nem com as características morfológicas.

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Zinnia sp. and hairy beggartick (Bidens pilosa) plants exhibiting symptoms of possible virus infection were found in the municipality of Santa Bárbara d'Oeste, São Paulo State, Brazil. Flexuous filamentous particles and cytoplasmatic inclusions typical of potyvirus infection were observed by transmission electron microscopy, respectively, in leaf extracts and cells of symptomatic leaves. Infection of both plants with bidens mottle virus (BiMoV) was confirmed by RT-PCR using potyvirus universal primers, followed by nucleotide sequencing of the amplicons. The nearly complete genome sequence of the Brazilian isolate, named BiMoV-BR, is 9700 nucleotides long and shares 95.6 % identity with the corresponding nucleotide sequence of a BiMoV isolate from the United States. BiMoV-BR was mechanically transmitted and caused systemic infection on plants of Zinnia sp., hairy beggarstick, sunflower (Helianthus annuus), and lettuce (Lactuca sativa). Myzus persicae transmitted the virus to Zinnia sp. plants with efficacy of 8 % and 42 %, using one and ten aphids per plant, respectively. This is the first detection of BiMoV in Brazil. Further studies are necessary to evaluate the distribution of this potyvirus in the country.

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The current study describes the presence of Bacillus cereus (B. cereus) in contaminated foods of animal source and ready for human consumption with highlighting on their virulence contributing factors by detection of its virulence genes in addition to identification of their sequencing. Three hundred sixty food samples categorized as (228) meat products and (132) milk products were examined for B. cereus isolation and all of these isolates were confirmed by biochemical tests. Eighteen strains obtained from different food samples were examined for the attendance of a number of virulence genes (nheA, cytK, entFM, bceT and hblC genes) using uniplex PCR method. Furthermore, the B. cereus strains were valued for the sequencing of described genes. Generally 24.44% (88/360) food samples classified as 11.11% (40/360) meat products and 13.33% (48/360) milk products carried B. cereus according to cultural and biochemical properties, with geometric mean (1.5×107±0.15 CFU/g or mL) . The highest counts (above 105 CFU/g or mL) were originated from milk products (with geometric mean 2.2×107±0.22 CFU/g or mL) more than meat products (with geometric mean 1×107±0.19 CFU/g or mL). The results revealed that all of our isolates had one or more virulence (enterotoxin) genes. In our research, the most predominant genes were nheA (100%), followed by cytK (61.11%), entFM (33.33%), bceT (11.11%) then hblC (5.56%). Molecular method detected that overall, 5 strains (27.78%) harbored only 1 gene (nheA), 7 strains (38.88%) harbored 2 genes which classified as 5 strains (27.78%) (nheA and cytK), 2 strains (11.11%) have (nheA and entFM). Moreover, 5 strains (27.78%) have 3 genes classified as 3 strains (16.67%) harbored (nheA, cytK and entFM), 1 strain (5.56%) had (nheA, cytK and hblC), and 1 strain (5.56%) had (nheA, cytK and bceT). Only 1 strain (5.56%) carried 4 tested virulence genes (nheA, cytK, entFM and bceT) genes. The most prevalent gene in meat and dairy foods was nheA (100%). The nucleotide sequences of (bceT, cytK, entFM, hblC and nheA genes) of B. cereus strains were deposited in GenBank under accession no. (MW911824, MW911825, MW911826, MW911827 and MW911828), respectively. Our study was established to indicate the presence of virulent B. cereus in meat and milk products ready for human consumption as a result of deficient hygienic actions. So, a plain for good hygienic measures should be modified to avoid causing serious health problems to human due to ingestion of such products.

O presente estudo descreve a presença de Bacillus cereus em alimentos contaminados de origem animal e prontos para consumo humano, com destaque para seus fatores de contribuição de virulência por meio da detecção de seus genes de virulência, além da identificação de seu sequenciamento. Trezentas e sessenta amostras de alimentos categorizados como produtos cárneos (228) e produtos lácteos (132) foram examinadas para isolamento de B. cereus, e todos esses isolados foram confirmados por testes bioquímicos. Dezoito cepas obtidas de diferentes amostras de alimentos foram examinadas para a presença de uma série de genes de virulência (genes nheA, cytK, entFM, bceT e hblC) usando o método de PCR uniplex. Além disso, as cepas de B. cereus foram avaliadas para o sequenciamento dos genes descritos. De forma geral, 24,44% (88/360) das amostras de alimentos classificados como produtos cárneos (11,11%; 40/360) e produtos lácteos (13,33%; 48/360) transportavam B. cereus, de acordo com as propriedades culturais e bioquímicas, com média geométrica de 1,5 × 10 7 ± 0,15 CFU/g ou mL. Os resultados revelaram que todos os nossos isolados tinham um ou mais genes de virulência (enterotoxina). Em nossa pesquisa, os genes mais predominantes foram nheA (100%), seguidos de cytK (61,11%), entFM (33,33%), bceT (11,11%) e hblC (5,56%). O método molecular detectou que, no geral, 5 cepas (27,78%) apresentavam apenas 1 gene (nheA) e 7 cepas (38,88%) continham 2 genes que foram classificados como 5 cepas (27,78%) (nheA e cytK), 2 cepas (11,11%) possuíam (nheA e entFM). Além disso, 5 cepas (27,78%) continham 3 genes classificados como 3 cepas (16,67%) hospedados (nheA, cytK e entFM), 1 cepa (5,56%) tinha (nheA, cytK e hblC) e 1 cepa (5,56%) teve (nheA, cytK e bceT). Apenas 1 cepa (5,56%) carregava 4 genes de virulência testados (nheA, cytK, entFM e bceT). As sequências de nucleotídeos (genes bceT, cytK, entFM, hblC e nheA) de cepas de B. cereus foram depositadas no GenBank sob o número de acesso (MW911824, MW911825, MW911826, MW911827 e MW911828), respectivamente. Nosso estudo foi estabelecido para indicar a virulência de B. cereus em carnes e produtos lácteos prontos para consumo humano como resultado de ações higiênicas deficientes. Portanto, deve ser estabelecido um plano com boas medidas de higiene para evitar sérios problemas de saúde humana por causa da ingestão de tais produtos.

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Dental abscess in ruminants is an acute polymicrobial infection, usually resulting from periodontal disease or endodontic infection, with consequences for animal health and welfare. The present study aimed to describe the bacterial microbiota of dental abscesses in Blastocerus dichotomus. Biological material from mandibular dental abscesses, punctured with a sterile syringe and needle during routine veterinary curative procedures or necropsies, was collected from three ex-situ marsh deer. Bacteria were identified using high-throughput sequencing of the 16S ribosomal RNA gene. The three specimens had the presence of facial bulging, and two died because of severe emaciation with a history of progressive weight loss. Bacteroides (38.6%), Fusobacterium (36.65%), and Porphyromonas (7.49%) represented the most abundant genera and Fusobacterium necrophorum (35.69%), Porphyromonas levii (3.12%) and Porphyromonas gulae (1.78%) were among the ten most represented species in the microbiota of mandibular abscess in Blastocerus dichotomus. These molecular findings demonstrate a broader diversity of species in the polymicrobial nature of dental abscesses in B. dichotomus than was previously reported when culture-dependent methods were used in the diagnosis.

O abcesso dentário em ruminantes é uma infeção polimicrobiana aguda, geralmente resultante de doença periodontal ou infeção endodôntica, com consequências para a saúde e bem-estar animal. O presente estudo teve como objetivo descrever a microbiota bacteriana de abscessos dentários em Blastocerus dichotomus. Material biológico de abscessos dentários mandibulares, puncionados com seringa e agulha estéril durante procedimentos curativos veterinários de rotina ou necropsias, foi coletado de três cervos do pantanal criados ex situ. As bactérias foram identificadas usando sequenciamento de alto rendimento do gene 16S ribossomal RNA. Os três espécimes apresentavam abaulamento facial e dois faleceram por emagrecimento severo com histórico de emagrecimento progressivo. Bacteroides (38,6%), Fusobacterium (36,65%) e Porphyromonas (7,49%) representaram os gêneros mais abundantes e Fusobacterium necrophorum (35,69%), Porphyromonas levii (3,12%) e Porphyromonas gulae (1,78%) estavam entre as dez espécies mais representadas na microbiota do abscesso mandibular em Blastocerus dichotomus. Esses achados moleculares demonstram uma diversidade mais ampla de espécies na natureza polimicrobiana dos abscessos dentários em B. dichotomus, do que o relatado anteriormente quando métodos dependentes de cultura foram utilizados no diagnóstico.

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Heart and blood vessel disorders, such as coronary heart disease, brain vessel disease, rheumatic heart disease, and others, are together referred to as cardiovascular disease (CVD). In this study, we sought to determine how mitochondrial Leucine Transfer RNA genes and CVDs are related (MT-L1 and MT-L2). From CVD patients in Peshawar, a total of 27 saliva samples were taken. Leu-tRNA genes expressed by mitochondria were amplified using polymerase chain reaction after DNA was removed. Ten samples were sent for sequencing after PCR and gene cleaning. We obtained all of the sequenced results, which were subsequently aligned and evaluated against the mitochondrial revised Cambridge Reference Sequence (rCRS). However, in our sequenced samples, Leu-tRNA MT-L1 and MT-L2 genes were determined to be unaltered. Thus, it is suggested that a large population be taken into account while screening for mutations in the mitochondrial encoded Leu-tRNA MT-L1 and MT-L2 genes of cardiac patients in areas of Pakistan. Additionally, it is recommended that patients with cardiac problems should also have other mitochondrial encoded genes checked for potential mutations. This could result in the identification of genetic markers that could be used for early CVD screening in Pakistan.

Distúrbios do coração e dos vasos sanguíneos, como doença cardíaca coronária, doença dos vasos cerebrais, doença cardíaca reumática entre outros, são referidos juntos como doença cardiovascular (DCV). Neste estudo, procuramos determinar como os genes mitocondriais do RNA de transferência de leucina e as DCVs estão relacionados (MT-L1 e MT-L2). Foi coletado um total de 27 amostras de saliva de pacientes com DCV em Peshawar. Genes de Leu-tRNA expressos por mitocôndrias foram amplificados usando reação em cadeia da polimerase (PCR) após a remoção do DNA. Dez amostras foram enviadas para sequenciamento após PCR e limpeza gênica. Obtivemos todos os resultados sequenciados, que foram posteriormente alinhados e avaliados em comparação com a Sequência de Referência de Cambridge revisada (rCRS). No entanto, em nossas amostras sequenciadas, os genes Leu-tRNA MT-L1 e MT-L2 foram determinados como inalterados. Assim, sugere-se que uma grande população seja levada em consideração durante a triagem de mutações nos genes Leu-tRNA MT-L1 e MT-L2 mitocondriais codificados de pacientes cardíacos em áreas do Paquistão. Além disso, recomenda-se que outros genes mitocondriais codificados de pacientes com problemas cardíacos também sejam verificados quanto a possíveis mutações. Isso pode resultar na identificação de marcadores genéticos que podem ser usados para triagem precoce de DCV no Paquistão.

Resumo

Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis ​​(9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

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The present study explored the potential of leaf litter as a source of fungi able to produce ligninolytic enzymes for the biodegradation of anthraquinone dyes. Within the colonies isolated from the leaf litter, only three colonies of two species Trametes were selected based on the detection of oxidation and decolorization halos in Petri dishes with PDA (potato-dextrose-agar) + Guaicol and PDA + RBBR (Remazol Brilliant Blue R). The identification of the colonies was done through sequencing of the ITS region. The enzymatic activity of Lac (lacase), MnP (manganês peroxidase) and LiP (lignina peroxidase) was analyzed by spectrophotometry during fermentation in PD+RBBR imedium. Isolates A1SSI01 and A1SSI02 were identified as Trametes flavida, while A5SS01 was identified as Trametes sp. Laccase showed the highest enzymatic activity, reaching 452.13 IU.L-1 (A1SSI01, 0.05% RBBR) after 96h. Isolate A1SSI02 reached the highest percentage of decolorization, achieving 89.28% in seven days. The results imply that these Trametes isolates can be highly effective in waste treatment systems containing toxic anthraquinone dyes. Keywords: laccase, peroxidases, basidiomycete, litter and biodecolorization.

Este estudo investigou o potencial da serapilheira como fonte de fungos capazes de produzir enzimas ligninolíticas para a biodegaradação de corante antraquinônico. Entre as colônias isoladas do material de serapilheira, apenas três colônias de duas espécies de Trametes. foram selecionados com base na detecção de halos de oxidação e descoloração em placas de Petri com PDA (Batata-Dextrose-Ágar) + Guaicol e PDA + RBBR (Azul Brilhante de Remazol R). A identificação das colônias foi através do sequenciamento da região ITS. A atividade enzimática de Lac (lacase), MnP (manganês peroxidase) e LiP (lignina peroxidase) foi avaliada por espectrofotometria durante a fermentação em meio BD+RBBR. Os isolados A1SSI01 e A1SSI02 foram identificados como Trametes flavida, enquanto A5SS01 foi identificado como Trametes sp. A lacase mostrou a maior atividade enzimática, atingindo 452,13 UI.L-1 (A1SSI01, 0,05% RBBR) após 96h. O isolado A1SSI02 alcançou o maior percentual de descoloração, atingindo 89,28% em sete dias. Os resultados sugerem que esses isolados de Trametes podem ser eficazes em sistemas de tratamento de resíduos contendo corantes antraquinônicos tóxicos.

Resumo

Abstract Several species of Cichla successfully colonized lakes and reservoirs of Brazil, since the 1960s, causing serious damage to local wildlife. In this study, 135 peacock bass were collected in a reservoir complex in order to identify if they represented a single dominant species or multiple ones, as several Cichla species have been reported in the basin. Specimens were identified by color pattern, morphometric and meristic data, and using mitochondrial markers COI, 16S rDNA and Control Region (CR). Overlapping morphological data and similar coloration patterns prevented their identification using the taxonomic keys to species identification available in the literature. However, Bayesian and maximum likelihood from sequencing data demonstrated the occurrence of a single species, Cichla kelberi. A single haplotype was observed for the 16S and CR, while three were detected for COI, with a dominant haplotype present in 98.5% of the samples. The extreme low diversity of the transplanted C. kelberi evidenced a limited number of founding maternal lineages. The success of this colonization seems to rely mainly on abiotic factors, such as increased water transparency of lentic environments that favor visual predators that along with the absence of predators, have made C. kelberi a successful invader of these reservoirs.

Resumo Muitas espécies de Cichla colonizaram com sucesso lagos e reservatórios do Brasil desde os anos 1960, causando graves prejuízos à vida selvagem nesses locais. Neste estudo, 135 tucunarés foram coletados em um complexo de reservatórios a fim de identificar se representavam uma espécie dominante ou múltiplas espécies, uma vez que diversas espécies de Cichla foram registradas na bacia. Os espécimes foram identificados com base na coloração, dados morfométricos e merísticos, e por marcadores mitocondriais COI, 16S rDNA e Região Controle (RC). A sobreposição dos dados morfométricos e o padrão similar de coloração impediram a identificação utilizando as chaves de identificação disponíveis na literatura. Entretanto, as análises bayesiana e de máxima verossimilhança de dados moleculares demonstraram a ocorrência de uma única espécie, Cichla kelberi. Um único haplótipo foi observado para o 16S e RC, enquanto três foram detectados para o COI, com um haplótipo dominante presente em 98,5% das amostras. A baixa diversidade nos exemplares introduzidos de C. kelberi evidenciou um número limitado de linhagens maternas fundadoras. O sucesso da invasão parece depender de fatores abióticos, como a maior transparência da água de ambientes lênticos que favorece predadores visuais que, atrelado à ausência de predadores, fez do C. kelberi um invasor bem-sucedido nesses reservatórios.

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Abstract Bacteria were isolated from samples of Fresh Apple juices from shops of three different localities of Lahore. Analysis of samples from Liberty, Anarkali and Yateem khana Markets show different levels of contamination. There were pathogenic and non-pathogenic bacteria in all samples and were identified by the morphological and biochemical tests. Most of the plasmids of pathogenic bacteria were 4kb in their molecular size. Ribotyping of 16S ribosomal RNA gene sequencing was done to confirm Helicobacter pylori strain and Gluconobacter oxydans. The highest sensitivity of 210mm was shown by Enterobacter sp. against Aztheromysine disk (15µg) while Micrococcus sp. was highly resistant against all of the Antibiotics applied. The antibiotic resistance of pathogenic bacteria was also checked against Ricinus communis plant's extracts, all isolated bacterial pathogens were resistant but only, E.coli was inhibited at 300µl of the extracts. Presence of pathogenic bacteria in Apple juice samples was due to contamination of sewage water in drinking water while some of these pathogenic bacteria came from Apple's tree and other from store houses of fruits.

Resumo As bactérias foram isoladas de amostras de suco de maçã fresco de lojas de três diferentes localidades de Lahore. A análise de amostras dos mercados Liberty, Anarkali e Yateem khana mostram diferentes níveis de contaminação. Havia bactérias patogênicas e não patogênicas em todas as amostras e foram identificadas pelos testes morfológicos e bioquímicos. A maioria dos plasmídeos de bactérias patogênicas tinha 4 kb em seu tamanho molecular. A ribotipagem do sequenciamento do gene do RNA ribossômico 16S foi realizada para confirmar a cepa de Helicobacter pylori e Gluconobacter oxydans. A maior sensibilidade de 210 mm foi mostrada por Enterobacter sp. contra disco de azteromisina (15µg) enquanto Micrococcus sp. foi altamente resistente a todos os antibióticos aplicados. A resistência a antibióticos de bactérias patogênicas também foi verificada contra extratos de plantas de Ricinus communis, todos os patógenos bacterianos isolados foram resistentes, mas apenas E. coli foi inibida em 300µl dos extratos. A presença de bactérias patogênicas nas amostras de suco de maçã deveu-se à contaminação da água de esgoto na água potável, enquanto algumas dessas bactérias patogênicas vieram da árvore da maçã e outras de armazéns de frutas.

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Abstract Naturally occurring mutations in morphogenetic protein 15 (BMP15) are associated with decreased ovulation rate (OR), litter size (LS), and sterility. It is of a great interest to elucidate BMP15 gene in Cholistani sheep breed to uplift socio-economic status and the knowledge of Cholistani sheep breeding in Southern Punjab, Pakistan. In our study, a total of 50 infertile Cholistani sheep aged between 2-6 years and having no blood relation were screened for BMP15 mutations. For this purpose, a high-quality DNA was extracted from the blood of sheep followed by primer designing, Polymerase Chain Reaction (PCR) amplification, DNA sequencing, and in silico analyses. Out of total 50 samples, 9 samples including case 1 (T3), case 2 (T8), case 3 (T17), case 4 (T22), case 5 (T25), case 6 (T33), case 7 (T40), case 8 (T44), and case 9 (T47) were found positive for a variety of already reported and novel BMP15 mutations. Further in silico analyses of the observed mutations have shown the functional impact of these mutations on different characteristics (molecular weight, theoretical PI, estimated half-life, instability index, sub-cellular localization, and 3D confirmation) of the encoded proteins, possibly altering the normal functionality. In a nutshell, findings of this study have confirmed the possible essential role of the BMP15 mutations in the infertility of the Cholistani sheep.

Resumo Mutações de ocorrência natural na proteína morfogenética 15 (BMP15) estão associadas à diminuição da taxa de ovulação (TO), tamanho da ninhada (TN) e esterilidade. Estudar a BMP15 na raça Cholistani para elevar o status socioeconômico e o conhecimento da criação de ovinos Cholistani no sul de Punjab, Paquistão. Em nosso estudo, 50 ovelhas Cholistani inférteis sem parentesco sanguíneo foram rastreadas para mutações BMP15. Para tanto, um DNA de alta qualidade foi extraído do sangue dessas ovelhas, seguido de concepção do primer, amplificação da reação em cadeia da polimerase (PCR), sequenciamento de DNA e análises in silico. Do total de 50 amostras, 9, incluindo caso 1 (T3), caso 2 (T8), caso 3 (T17), caso 4 (T22), caso 5 (T25), caso 6 (T33), caso 7 (T40), caso 8 (T44) e caso 9 (T47), foram consideradas positivas para uma variedade de mutações BMP15 novas e já relatadas. Mais análises in silico das mutações observadas mostraram o impacto funcional dessas mutações em diferentes características (peso molecular, PI teórico, meia-vida estimada, índice de instabilidade, localização subcelular e confirmação 3D) das proteínas codificadas, possivelmente alterando a funcionalidade normal. Nossos achados confirmaram o possível papel essencial das mutações BMP15 na infertilidade de ovelhas Cholistani.

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Abstract Among Bemisia tabaci species, the invasive MEAM1 and MED species are key agricultural pests for many crops. In Brazil, most part of B. tabaci population outbreaks were associated with MEAM1, which, since 1990s quickly spread across the entire country. Later in 2014, the MED was identified in Brazil, initially more restricted to greenhouses, but suddenly reaching new areas in the South and Southeast open regions. Thus, our objective was to investigate the geographical distribution of MEAM1 and MED on open field crops in Brazil. MEAM1 is still the predominant species on open field crops such as soybean, cotton, and tomato. The sequencing of a cytochrome c oxidase subunit I (COI) gene fragment revealed a single haplotype of MEAM1, suggesting the establishment of a single MEAM1 strain in the country. The haplotypes found for MEAM1 and MED are genetically related to the globally dispersed strains, Jap1 and Mch1, respectively. Continuous monitoring of B. tabaci species is crucial because landscape alterations, climatic changes, and pest management methods may shift the B. tabaci species distribution and dominance in Brazilian crop areas.

Resumo Dentre as espécies de Bemisia tabaci, as espécies invasoras MEAM1 e MED se destacam como pragas de grande importância para várias culturas. No Brasil, a maior parte dos surtos populacionais de mosca-branca são associados a presença da espécie MEAM1, que a partir 1990 se espalhou por todo o país. Por outro lado, em 2014 a espécie MED foi identificada no Brasil, inicialmente restrita a casas de vegetação, mas rapidamente se difundindo em novas áreas nas regiões Sul e Sudeste do Brasil. Assim, nosso objetivo foi investigar a distribuição geográfica das espécies MEAM1 e MED em grandes culturas no Brasil. A espécie MEAM1 continua sendo predominante nas monoculturas como algodão, soja e tomate. O sequenciamento de um fragmento do gene citocromo c oxidase subunidade I (COI) revelou a presença de um haplótipo para MEAM1, sugerindo o estabelecimento de apenas uma linhagem no país. Os haplótipos encontrados para MEAM1 e MED são geneticamente relacionados as linhagens globalmente dispersas Jap1 e Mch1, respectivamente. O monitoramento contínuo das espécies de B. tabaci é crucial pois as mudanças na paisagem, mudanças climáticas e métodos de manejo das pragas podem alterar a dominância e a distribuição dessas espécies nas áreas agrícolas do Brasil.

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This article reports on a golden-billed saltator Saltator aurantiirostris Vieillot, 1817, kept in captivity outside its natural distribution area, in the proximities of the Itatiaia National Park, as a new host for Isospora similisi Coelho, Berto, Neves, Oliveira, Flausino & Lopes, 2013. Additionally, a supplementary molecular identification is provided through the sequencing of three non-overlapping loci of mitochondrial DNA and one locus of the 18S small subunit ribosomal RNA (18S) gene. All the taxonomic features of the I. similisi oocysts shed by S. aurantiirostris were equivalent to those originally described from Saltator similis d'Orbigny & Lafresnaye, 1837. The new sequenced loci were identical, or showed 99.9% similarity, to the samples of I. similisi from S. similis and S. aurantiirostris, confirming the same species from both hosts. Lastly, I. similisi is estimated as a junior synonym of Isospora formarum McQuistion & Capparella, 1992, due to the morphological similarities and wide distribution of its hosts in the Neotropical region. Therefore, this study encourages future taxonomic inquiries into I. similisi collected from other Saltator spp. in order to establish this synonymization of I. formarum with I. similisi, and hence, its wide distribution and dispersion in the Neotropical region, including across the Andes mountains.(AU)

Este artigo relata um bico-duro Saltator aurantiirostris Vieillot, 1817, mantido em cativeiro fora de sua área de distribuição natural, nas proximidades do Parque Nacional de Itatiaia, como novo hospedeiro de Isospora similisi Coelho, Berto, Neves, Oliveira, Flausino & Lopes, 2013. Adicionalmente, uma identificação molecular suplementar foi fornecida por intermédio do sequenciamento de três loci não sobrepostos de DNA mitocondrial e um locus da subunidade pequena do gene 18S do RNA ribossômico (18S). Todas os caracteres taxonômicos dos oocistos de I. similisi, eliminados por S. aurantiirostris, foram equivalentes àqueles originalmente descritos de Saltator similis d'Orbigny & Lafresnaye, 1837. Os novos loci sequenciados foram idênticos, ou tiveram 99,9% de similaridade, com as amostras de I. similisi de S. similis e S. aurantiirostris, confirmando a mesma espécie de ambos os hospedeiros. Por fim, I. similisi foi estimada como sinonímia júnior de Isospora formarum McQuistion & Capparella, 1992, devido às semelhanças morfológicas e ampla distribuição de seus hospedeiros na região Neotropical. Portanto, este estudo incentiva futuras investigações taxonômicas sobre I. similisi, recuperados de outros Saltator spp., a fim de estabelecer essa sinonimização de I. formarum com I. similisi e, consequentemente, sua ampla distribuição e dispersão na região Neotropical, inclusive Cordilheira dos Andes.(AU)

Resumo

The current study characterizes the genetic distribution of virulence and antimicrobial resistance of Streptococcus lutetiensis and Streptococcus equinus isolated from cows with clinical mastitis using whole genome sequencing (WGS). Although they are not the protagonist species within the genus Streptococcus, recent studies have isolated these species associated with bovine mastitis. In addition, these species are reported and isolated from humans and other animals. A total of four strains of S. lutetiensis and one of S. equinus were isolated from five cows with identified cases of clinical mastitis at a dairy farm near Ithaca, New York. Nineteen genes associated with antimicrobial resistance and 20 genes associated with virulence were identified in the analyzed strains. All strains presented genes associated with resistance: alr, ddl, gdpD, kasA, murA, lsa(E), msr(D), mef(A), gidB, and LiaF. Resistance genes associated with several different classes of antibiotics have also been reported. Sixteen virulence-associated genes were identified in all strains. Based on our findings, we conclude that the studied species have the potential to cause mastitis in cattle, and further studies are important to elucidate their role.

O presente estudo caracteriza a distribuição genética de virulência e resistência antimicrobiana de Streptococcus lutetiensis e Streptococcus equinus isolados de vacas com mastite clínica usando sequenciamento completo do genoma. Apesar de não serem as espécies protagonistas dentro do gênero Streptococcus, estudos recentes têm isolado essas espécies associadas à mastite bovina. Além disso, essas espécies são relatadas e isoladas de humanos e outros animais. Um total de quatro cepas de S. lutetiensis e uma de S. equinus foram isoladas de cinco vacas com casos identificados de mastite clínica em uma fazenda leiteira perto de Ithaca, Nova York. Dezenove genes associados à resistência antimicrobiana e 20 genes associados à virulência foram identificados nas cepas analisadas. Todas as linhagens apresentaram genes associados à resistência: alr, ddl, gdpD, kasA, murA, lsa(E), msr(D), mef(A), gidB e LiaF. Genes de resistência associados a várias classes diferentes de antibióticos também foram relatados. Dezesseis genes associados à virulência foram identificados em todas as cepas. Com base em nossos achados, concluímos que as espécies estudadas têm potencial para causar mastite em bovinos e mais estudos são importantes para elucidar seu papel.