Resumo Nas últimas décadas, tem havido um interesse crescente no uso de probióticos como uma alternativa aos antibióticos. Pesquisadores conduziram estudos para investigar cepas bacterianas quanto ao seu potencial probiótico. Como outros animais, os peixes também têm várias cepas bacterianas em seus intestinos que possuem propriedades probióticas, embora isso seja limitado a espécies de Bacillus. Portanto, o objetivo deste estudo foi isolar e caracterizar espécies probióticas de Bacillus do intestino de peixe Masheer (Tor Puititora). Quatro isolados bacterianos puros foram selecionados como potenciais cepas probióticas com base em critérios de seleção, incluindo taxa de sobrevivência em ácido e sal biliar. Os isolados exibiram atividade antimicrobiana significativa contra bactérias patogênicas, incluindo Escherichia coli (ATCC8739), Pseudomonas aeruginosa (ATCC9027) e Staphylococcus aureus (ATCC6538). MF2 e MF3 demonstraram zonas claras, incluindo atividade antimicrobiana contra todos os três patógenos indicadores. MF1 e MF4 exibiram atividade antimicrobiana contra E. coli (ATCC8739) e P. aeruginosa (ATCC9027). Além disso, as sequências do gene 16S rRNA de todos os isolados exibiram uma associação próxima com Bacillus tequilensis (KCTC13622), com similaridade de nucleotídeos de 98,63%, 98,25%, 98,80% e 98,35%, respectivamente. Nossos resultados demonstram que esses isolados bacterianos mostram-se promissores como uma alternativa aos antibióticos no sistema alimentar da pesca. Neste estudo, todos os isolados identificados no intestino dos peixes eram subespécies de B. tequilensis. décadas probiótico animais Portanto Tor Puititora. Puititora . Puititora) seleção biliar patogênicas ATCC8739, ATCC8739 ATCC , ATCC9027 (ATCC9027 ATCC6538. ATCC6538 (ATCC6538) MF claras indicadores E (ATCC8739 P ATCC9027. disso S KCTC13622, KCTC13622 KCTC (KCTC13622) 9863 98 63 98,63% 9825 25 98,25% 9880 80 98,80 9835 35 98,35% respectivamente mostramse mostram se pesca B ATCC873 ATCC902 (ATCC902 ATCC653 (ATCC6538 (ATCC873 KCTC1362 (KCTC13622 986 9 6 98,63 982 2 98,25 988 8 98,8 983 3 98,35 ATCC87 ATCC90 (ATCC90 ATCC65 (ATCC653 (ATCC87 KCTC136 (KCTC1362 98,6 98,2 98, 98,3 ATCC8 ATCC9 (ATCC9 ATCC6 (ATCC65 (ATCC8 KCTC13 (KCTC136 (ATCC (ATCC6 KCTC1 (KCTC13 (KCTC1 (KCTC

Abstract Over the past few decades, there has been increasing interest in using probiotics as an alternative to antibiotics. Researchers have conducted studies to investigate bacterial strains for their probiotic potential. Like other animals, fish also have several bacterial strains in their gut that possess probiotic properties, although this is limited to Bacillus species. Therefore, this study aimed to isolate and characterize probiotic Bacillus species from the gut of Masheer fish (Tor Puititora). Four pure bacterial isolates were selected as potential probiotic strains based on selection criteria, including survival rate in acid and bile salt. The isolates exhibited significant antimicrobial activity against pathogenic bacteria, including Escherichia coli (ATCC8739), Pseudomonas aeruginosa (ATCC9027), and Staphylococcus aureus (ATCC6538). MF2 and MF3 demonstrated clear zones, including antimicrobial activity against all three indicator pathogens. MF1 and MF4 exhibited antimicrobial activity against E. coli (ATCC8739) and P. aeruginosa (ATCC9027). Furthermore, the 16S rRNA gene sequences of all isolates exhibited a close association with Bacillus tequilensis (KCTC13622), with nucleotide similarity of 98.63%, 98.25%, 98.80%, and 98.35%, respectively. Our results demonstrate that these bacterial isolates show promise as an alternative to antibiotics in the fisheries food system. In this study, all isolates identified in the fish gut were subspecies of B. tequilensis. decades animals properties Therefore Tor Puititora. Puititora . Puititora) criteria salt bacteria ATCC8739, ATCC8739 ATCC , ATCC9027, ATCC9027 (ATCC9027) ATCC6538. ATCC6538 (ATCC6538) MF zones pathogens E (ATCC8739 P ATCC9027. Furthermore S KCTC13622, KCTC13622 KCTC (KCTC13622) 9863 98 63 98.63% 9825 25 98.25% 9880 80 98.80% 9835 35 98.35% respectively system B ATCC873 ATCC902 (ATCC9027 ATCC653 (ATCC6538 (ATCC873 KCTC1362 (KCTC13622 986 9 6 98.63 982 2 98.25 988 8 98.80 983 3 98.35 ATCC87 ATCC90 (ATCC902 ATCC65 (ATCC653 (ATCC87 KCTC136 (KCTC1362 98.6 98.2 98.8 98.3 ATCC8 ATCC9 (ATCC90 ATCC6 (ATCC65 (ATCC8 KCTC13 (KCTC136 98. (ATCC9 (ATCC6 (ATCC KCTC1 (KCTC13 (KCTC1 (KCTC

Resumo O estudo teve como objetivo avaliar o impacto da dieta rica em gordura (HFD) e da microbiota intestinal humana sintética (GM) combinada com HFD e dieta alimentar (CD) na indução de diabetes tipo 2 (T2D) usando modelo de camundongos. Para nosso conhecimento, este é o primeiro estudo usando transplante de GM humano selecionado através do método baseado em cultura acoplada à modulação dietética em camundongos para o estabelecimento in vivo de inflamação que leva a T2D e disbiose intestinal. Vinte bactérias (T2D1-T2D20) de amostras de fezes de indivíduos T2D confirmados verificaram ser morfologicamente diferentes e foram submetidas à purificação em meios diferentes aerobicamente e anaerobicamente, o que revelou sete bactérias mais comuns entre 20 isolados com base na caracterização bioquímica. Com base no sequenciamento do gene 16S rRNA, esses sete isolados foram identificados como Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenides (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). Esses sete isolados foram, posteriormente, usados como microbioma intestinal sintético (GM) por seu papel na indução de T2D em camundongos. Linhagens consanguíneas de camundongos albinos foram divididas em quatro grupos e foram alimentadas com CD, HFD, GM + HFD e GM + CD. Camundongos que receberam a dieta modificada com HFD e GM + (CD / HFD) mostraram um aumento altamente significativo (P < 0,05) no peso e na concentração de glicose no sangue, bem como um nível elevado de citocinas inflamatórias (TNF-α, IL-6 e MCP-1) em comparação com os ratos que receberam apenas CD. O sequenciamento do gene 16S rRNA de 11 bactérias fecais obtidas de três animais selecionados aleatoriamente de cada grupo revelou disbiose intestinal em animais que receberam GM. Cepas bacterianas, incluindo Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) e Lactobacillus Gasseri (MT152635D), foram tratadas com dieta modificada / CD) em comparação com as linhagens Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629), que foram isoladas de camundongos recebendo CD / HFD. Em conclusão, esses resultados sugerem que a constituição de GM e dieta desempenham papel significativo na inflamação levando ao início ou/e possivelmente à progressão de T2D.

Abstract The study was aimed to assess impact of high fat diet (HFD) and synthetic human gut microbiota (GM) combined with HFD and chow diet (CD) in inducing type-2 diabetes (T2D) using mice model. To our knowledge, this is the first study using selected human GM transplantation via culture based method coupled dietary modulation in mice for in vivo establishment of inflammation leading to T2D and gut dysbiosis. Twenty bacteria (T2D1-T2D20) from stool samples of confirmed T2D subjects were found to be morphologically different and subjected to purification on different media both aerobically and anerobically, which revealed seven bacteria more common among 20 isolates on the basis of biochemical characterization. On the basis of 16S rRNA gene sequencing, these seven isolates were identified as Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenes (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). The seven isolates were subsequently used as synthetic gut microbiome (GM) for their role in inducing T2D in mice. Inbred strains of albino mice were divided into four groups and were fed with CD, HFD, GM+HFD and GM+CD. Mice receiving HFD and GM+modified diet (CD/HFD) showed highly significant (P<0.05) increase in weight and blood glucose concentration as well as elevated level of inflammatory cytokines (TNF-α, IL-6, and MCP-1) compared to mice receiving CD only. The 16S rRNA gene sequencing of 11 fecal bacteria obtained from three randomly selected animals from each group revealed gut dysbiosis in animals receiving GM. Bacterial strains including Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) and Lactobacillus gasseri (MT152635) were isolated from mice treated with GM+modified diet (HFD/CD) compared to strains Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629) which were isolated from mice receiving CD/HFD. In conclusion, these findings suggest that constitution of GM and diet plays significant role in inflammation leading to onset or/and possibly progression of T2D. .

The incidence of colorectal cancer (CRC) is increasing daily worldwide. Although different aspects of CRC have been studied in other parts of the world, relatively little or almost no information is available in Pakistan about different aspects of this disease at the molecular level. The present study was aimed at determining the frequency and prevalence of K ras gene mutations in Pakistani CRC patients. Tissue and blood samples of 150 CRC patients (64% male and 36% female) were used for PCR amplification of K ras and detection of mutations by denaturing gradient gel electrophoresis, restriction fragment length polymorphism analysis, and nucleotide sequencing. The K ras mutation frequency was found to be 13%, and the most prevalent mutations were found at codons 12 and 13. A novel mutation was also found at codon 31. The dominant mutation observed was a G to A transition. Female patients were more susceptible to K ras mutations, and these mutations were predominant in patients with a nonmetastatic stage of CRC. No significant differences in the prevalence of K ras mutations were observed for patient age, gender, or tumor type. It can be inferred from this study that Pakistani CRC patients have a lower frequency of K ras mutations compared to those observed in other parts of the world, and that K ras mutations seemed to be significantly associated with female patients.