Misidentification of Cutibacterium namnetense as Cutibacterium acnes among clinical isolates by MALDI-TOF VitekMS: usefulness of gyrB sequencing and new player in bone infections.

The taxonomy modification of Propionibacterium sp. with the description of new species, especially Cutibacterium namnetense, raises the question of species distribution in routine clinical samples. We performed a retrospective study during 3 years before the implementation of MALDI-TOF. Two hundred sixty-nine isolates were included in the study. MALDI-TOF identification, 16S rRNA, and new developed gyrB partial sequencings were performed. The most representative species was C. acnes in 88% of the cases, regardless of the origin of the clinical sample. Eventually, we identified three C. namnetense strains, representing a 1.1% prevalence over the period of time, including two bone infections. MALDI-TOF databases should be regularly updated to incorporate new species. gyrB sequencing constitutes a both easy and relevant method to identify Cutibacterium sp. especially C. namnetense, a new player in bone infections.

In vitro selection and in vivo confirmation of the antioxidant ability of Lactobacillus brevis MG000874.

AIMS: This study aimed at isolating a probiotic strain from a collection of lactic acid bacteria (LAB) with the high antioxidant property and confirms its potential in d-gal-induced oxidative stress (OS) murine model. METHODS AND RESULTS: The in vitro antioxidant ability of 16 LAB strains was determined in the cell-free supernatant of 3- to 5-day-old culture, intact cells and cell lysates using three different methods for determining the reactive oxygen species scavenging activity and inhibition of lipid peroxidation. The strain, An28, presented the best antioxidant activity and was identified as Lactobacillus brevis MG000874 on the basis of 16 sRNA gene sequencing. The antioxidant potential of L. brevis MG000874 was confirmed in an OS murine model. Albino mice were exposed to d-galactose at a dose of 150 mg kg-1 BW and fed with L. brevis MG000874 (0·2 ml of 1010 CFU per ml cell suspension per animal per day). Antioxidant enzymes were quantified spectrophotometrically in the liver, kidney and serum. Subcutaneous administration of d-gal resulted in decrease in superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) and increase in glutathione-S-transferase (GST) levels in animals. The L. brevis MG000874-treated animals displayed improvement in SOD, CAT and GST in all tissues and GSH in the liver and serum. The safety assessment of L. brevis MG000874 was performed by comparing liver and renal function tests. None of the selected indicators was significantly different from the negative control group. CONCLUSIONS: The antioxidant potential of 16 strains was noticed to be strain specific and in vivo performance of L. brevis MG000874 was found satisfactory in a d-gal murine model. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus brevis MG000874 was identified for its admirable antioxidant property. This strain or/and its metabolites could be further investigated for possible applications in humans and veterinary fields.

The first case of locally acquired tick-borne Babesia microti infection in Canada.

A child with a complicated medical history that included asplenia acquired an infection with Babesia microti in the summer of 2013 and had not travelled outside of Manitoba. Although the clinical findings were subtle, astute laboratory work helped to reach a preliminary identification of Babesia species, while reference laboratory testing confirmed the diagnosis. Blacklegged ticks (Ixodes scapularis) are known to transmit Borrelia burgdorferi and Anaplasma phagocytophilum in the province; however, the present case represents the first known instance of tick-borne B microti, both in Manitoba and in Canada. The expanding territory of the blacklegged tick increases the relevance of this emerging infection. Clinicians, laboratory medical practitioners and public health officials should be aware of B microti as a potential locally acquired infection in Canada.

Un enfant ayant des antécédents médicaux complexes, qui incluaient une asplénie, a contracté une infection à Babesia microti pendant l'été 2013, sans avoir quitté le Manitoba. Même si les résultats cliniques étaient discrets, un travail de laboratoire astucieux a contribué à l'identification préliminaire d'une espèce de Babesia. Le test du laboratoire de référence a confirmé le diagnostic. On sait que les tiques occidentales à pattes noires (Ixodes scapularis) transmettent le Borrelia burgdorferi et l'Anaplasma phagocytophilum dans la province. Le présent cas est toutefois la première occurrence connue de B microti à tique, tant au Manitoba qu'au Canada. L'expansion du territoire de la tique occidentale à pattes noires renforce la pertinence de cette infection émergente. Les cliniciens, les praticiens de laboratoires médicaux et les directeurs de la santé publique devraient savoir que le B microti peut être transmis localement au Canada.

In vivo studies could not confirm in vitro prophylactic synergism between Moringa essential oil and Lactobacillus reuteri (MT180537).

Aerobic vaginitis (AV) is a recently defined vaginal recurring infection, which is treated with antibiotics. However, excessive and prolonged use of antibiotics disrupts healthy vaginal microflora and leads to the emergence of antibiotic resistance among pathogens. This situation has directed researchers to explore alternative antimicrobials. The current study describes in vitro and in vivo antimicrobial efficacy and pharmaceutical interactions between plant essential oils (EOs) and five lactic acid bacteria (LABs), isolated from the healthy vagina, against E. faecalis, one of the major etiological agents of AV. In vitro experiments confirm good antimicrobial activity of both plant EOs and cell free supernatant (CFS) from LABs. Based on high antimicrobial efficacy, Moringa essential oil (MO) was selected to determine its nature of interaction with CFS of five LAB strains. Synergism was recorded between MO and CFS of L. reuteri (MT180537). To validate in vitro findings, prophylactic responses of individual and synergistic application of MO and L. reuteri (MT180537) were evaluated in an E. faecalis (MW051601) induced AV murine model. The prophylactic efficacy was evidenced by a reduction in intensity of clinical symptoms, E. faecalis (MW051601) count per vaginal tissue along with a reduction in AV associated changes in histological markers of infection in animals receiving Moringa essential oil and L. reuteri (MT180537) alone or in combination. However, significant synergism between Moringa essential oil and L. reuteri (MT180537) could not be observed. Our data confirms the importance of in vivo experiments in deducing pharmacological interactions.

Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study.

Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin alongwith marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Antibacterial and bacteriostatic potential of coelomic fluid and body paste of Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) against ampicillin resistant clinical bacterial isolates.

Pheretima posthuma (Vaillant, 1868), a native earthworm of Pakistan and Southeast Asia, has wide utilization in vermicomposting and bioremediation process. In this study, P. posthuma coelomic fluid (PCF) and body paste (PBP) was evaluated as antibacterial agent against ampicillin (AMP) resistant five Gram positive and four Gram negative clinical isolates. The antibacterial effect of different doses (i.e. 25-100 µg/ml) of PCF and PBP along with AMP and azithromycin (AZM) (negative and positive controls, respectively) were observed through disc diffusion and micro-dilution methods. All nine clinical isolates were noticed as AMP resistant and AZM sensitive. Antibacterial effects of PCF and PBP were dose dependent and zone of inhibitions (ZI) against all clinical isolates were between 23.4 ± 0.92 to 0 ± 00 mm. The sensitivity profile of PCF and PBP against clinical isolates was noticed as 44.44 and 55.56%, respectively. Both PCF and PBP showed bacteriostatic (BTS) action against S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Moreover, the cumulative BTS potential of PCF and PBP against all isolates was 66.67 and 55.56%, respectively. The MICs of PCF and PBP were ranged from 50-200 µg/ml against selected isolates. The bacterial growth curves indicated that PCF and PBP inhibited the growth of all isolates at their specific MIC concentrations. However, PBP has better antibacterial potential compared to PCF against selected isolates. Therefore, it is concluded that both PCF and PBP of P. posthuma possess antibacterial and BTS potential against ampicillin resistant clinical isolates. This organism might be considered as a second choice of antibacterial agents and can further be utilized in pharmaceutical industries for novel drug manufacturing by prospecting bioactive potential agents.

Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans.

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. .

In vivo studies could not confirm in vitro prophylactic synergism between Moringa essential oil and Lactobacillus reuteri (MT180537)

Abstract Aerobic vaginitis (AV) is a recently defined vaginal recurring infection, which is treated with antibiotics. However, excessive and prolonged use of antibiotics disrupts healthy vaginal microflora and leads to the emergence of antibiotic resistance among pathogens. This situation has directed researchers to explore alternative antimicrobials. The current study describes in vitro and in vivo antimicrobial efficacy and pharmaceutical interactions between plant essential oils (EOs) and five lactic acid bacteria (LABs), isolated from the healthy vagina, against E. faecalis, one of the major etiological agents of AV. In vitro experiments confirm good antimicrobial activity of both plant EOs and cell free supernatant (CFS) from LABs. Based on high antimicrobial efficacy, Moringa essential oil (MO) was selected to determine its nature of interaction with CFS of five LAB strains. Synergism was recorded between MO and CFS of L. reuteri (MT180537). To validate in vitro findings, prophylactic responses of individual and synergistic application of MO and L. reuteri (MT180537) were evaluated in an E. faecalis (MW051601) induced AV murine model. The prophylactic efficacy was evidenced by a reduction in intensity of clinical symptoms, E. faecalis (MW051601) count per vaginal tissue along with a reduction in AV associated changes in histological markers of infection in animals receiving Moringa essential oil and L. reuteri (MT180537) alone or in combination. However, significant synergism between Moringa essential oil and L. reuteri (MT180537) could not be observed. Our data confirms the importance of in vivo experiments in deducing pharmacological interactions.

Resumo Vaginite aeróbica (VA) é uma infecção vaginal recorrente definida recentemente, que é tratada com antibióticos. No entanto, o uso excessivo e prolongado de antibióticos perturba a microflora vaginal saudável e leva ao surgimento de resistência aos antibióticos entre os patógenos. Esta situação levou os pesquisadores a explorar antimicrobianos alternativos. O presente estudo descreve a eficácia antimicrobiana in vitro e in vivo e as interações farmacêuticas entre óleos essenciais vegetais (OE) e cinco bactérias lácticas (BAL), isoladas de vagina sã, contra E. faecalis, um dos principais agentes etiológicos da AV. Os experimentos in vitro confirmam a boa atividade antimicrobiana de ambos os EOs de plantas e sobrenadante livre de células (CFS) de LABs. Com base na alta eficácia antimicrobiana, o óleo essencial de Moringa (MO) foi selecionado para determinar sua natureza de interação com o sobrenadante livre de células (CFS) de cinco cepas de LAB. Sinergismo foi registrado entre MO e CFS de L. reuteri (MT180537). Para validar os resultados in vitro, as respostas profiláticas da aplicação individual e sinérgica de MO e L. reuteri (MT180537) foram avaliadas em um modelo murino AV induzido por E. faecalis (MW051601). A eficácia profilática foi evidenciada por uma redução na intensidade dos sintomas clínicos, contagem de E. faecalis (MW051601) por tecido vaginal, juntamente com uma redução nas alterações associadas a AV nos marcadores histológicos de infecção em animais que receberam óleo essencial de Moringa e L. reuteri (MT180537) sozinho ou em combinação. No entanto, não foi possível observar sinergismo significativo entre o óleo essencial de Moringa e L. reuteri (MT180537). Nossos dados confirmam a importância dos experimentos in vivo na dedução de interações farmacológicas.

In vivo studies could not confirm in vitro prophylactic synergism between Moringa essential oil and Lactobacillus reuteri (MT180537) / Estudos in vivo não puderam confirmar o sinergismo profilático in vitro entre o óleo essencial de Moringa e Lactobacillus reuteri (MT180537)

Aerobic vaginitis (AV) is a recently defined vaginal recurring infection, which is treated with antibiotics. However, excessive and prolonged use of antibiotics disrupts healthy vaginal microflora and leads to the emergence of antibiotic resistance among pathogens. This situation has directed researchers to explore alternative antimicrobials. The current study describes in vitro and in vivo antimicrobial efficacy and pharmaceutical interactions between plant essential oils (EOs) and five lactic acid bacteria (LABs), isolated from the healthy vagina, against E. faecalis, one of the major etiological agents of AV. In vitro experiments confirm good antimicrobial activity of both plant EOs and cell free supernatant (CFS) from LABs. Based on high antimicrobial efficacy, Moringa essential oil (MO) was selected to determine its nature of interaction with CFS of five LAB strains. Synergism was recorded between MO and CFS of L. reuteri (MT180537). To validate in vitro findings, prophylactic responses of individual and synergistic application of MO and L. reuteri (MT180537) were evaluated in an E. faecalis (MW051601) induced AV murine model. The prophylactic efficacy was evidenced by a reduction in intensity of clinical symptoms, E. faecalis (MW051601) count per vaginal tissue along with a reduction in AV associated changes in histological markers of infection in animals receiving Moringa essential oil and L. reuteri (MT180537) alone or in combination. However, significant synergism between Moringa essential oil and L. reuteri (MT180537) could not be observed. Our data confirms the importance of in vivo experiments in deducing pharmacological interactions.

Vaginite aeróbica (VA) é uma infecção vaginal recorrente definida recentemente, que é tratada com antibióticos. No entanto, o uso excessivo e prolongado de antibióticos perturba a microflora vaginal saudável e leva ao surgimento de resistência aos antibióticos entre os patógenos. Esta situação levou os pesquisadores a explorar antimicrobianos alternativos. O presente estudo descreve a eficácia antimicrobiana in vitro e in vivo e as interações farmacêuticas entre óleos essenciais vegetais (OE) e cinco bactérias lácticas (BAL), isoladas de vagina sã, contra E. faecalis, um dos principais agentes etiológicos da AV. Os experimentos in vitro confirmam a boa atividade antimicrobiana de ambos os EOs de plantas e sobrenadante livre de células (CFS) de LABs. Com base na alta eficácia antimicrobiana, o óleo essencial de Moringa (MO) foi selecionado para determinar sua natureza de interação com o sobrenadante livre de células (CFS) de cinco cepas de LAB. Sinergismo foi registrado entre MO e CFS de L. reuteri (MT180537). Para validar os resultados in vitro, as respostas profiláticas da aplicação individual e sinérgica de MO e L. reuteri (MT180537) foram avaliadas em um modelo murino AV induzido por E. faecalis (MW051601). A eficácia profilática foi evidenciada por uma redução na intensidade dos sintomas clínicos, contagem de E. faecalis (MW051601) por tecido vaginal, juntamente com uma redução nas alterações associadas a AV nos marcadores histológicos de infecção em animais que receberam óleo essencial de Moringa e L. reuteri (MT180537) sozinho ou em combinação. No entanto, não foi possível observar sinergismo significativo entre o óleo essencial de Moringa e L. reuteri (MT180537). Nossos dados confirmam a importância dos experimentos in vivo na dedução de interações farmacológicas.

Peganum harmala can minimize Escherichia coli infection in poultry, but long-term feeding may induce side effects.

Peganum harmala seed extracts have been frequently reported to possess antibacterial potential through in vitro studies, but in vivo studies have acquired less attention. The present study was therefore designed to investigate its efficacy on the course of colibacillosis and effects of long-term feeding on selected parameters of general health in chickens. Two experiments were conducted in this regard. Experiment 1 (a pilot study) was performed to determine the dose of a field strain of Escherichia coli (O1:K1) required to induce clinical symptoms in 4- and 15-d-old specific-pathogen-free chickens. A successful induction of colibacillosis, in terms of clinical signs, mortality, and pathological lesions in addition to reisolation of the pathogen was observed by inoculating 4- and 15-d-old chicks with 4.3 log(10) and 6.4 log(10) cfu of E. coli, respectively, by intraperitoneal injection. Using these doses experiment 2 (main study) consisting of a single experiment with 3 parts was performed. Parts A and B generated the information regarding efficacy of the extract against infection in 4- and 15-d-old chickens applying different treatment schemes, whereas the effects of continuous feeding of the extract were assessed in part C. Whereas no protective effect of the extract could be recorded in young chickens, significant differences (P < 0.05) with regard to BW, clinical score, gross lesion score, and total granulocyte counts were observed in 15-d-old birds. Bacterial recovery per gram of tissue and reisolation frequency were lower in treated birds. The continuous feeding of the extract for 6 wk resulted in an augmentation in relative liver weight and depletion in alkaline phosphatase, protein, albumin, and globulin. It can be concluded that the crude extract of Peganum harmala possesses limited antimicrobial activity against E. coli in vivo and long-term continuous feeding may induce undesired effects. Furthermore, the study underlines the value of in vivo experiments and the diverse picture that herbal products, in this case Peganum harmala, may deliver by testing them against specific pathogens.

Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study / Avaliação antimicrobiana e toxicológica de Origanum vulgare: um estudo in vivo

Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin along with marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study

Abstract Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin alongwith marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Resumo Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study / Avaliação antimicrobiana e toxicológica de Origanum vulgare: um estudo in vivo

Abstract Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin alongwith marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Resumo Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Antibacterial and bacteriostatic potential of coelomic fluid and body paste of Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) against ampicillin resistant clinical bacterial isolates / Potencial antibacteriano e bacteriostático do fluido celômico e pasta corporal de Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) contra isolados bacterianos clínicos resistentes à ampicilina

Abstract Pheretima posthuma (Vaillant, 1868), a native earthworm of Pakistan and Southeast Asia, has wide utilization in vermicomposting and bioremediation process. In this study, P. posthuma coelomic fluid (PCF) and body paste (PBP) was evaluated as antibacterial agent against ampicillin (AMP) resistant five Gram positive and four Gram negative clinical isolates. The antibacterial effect of different doses (i.e. 25-100 µg/ml) of PCF and PBP along with AMP and azithromycin (AZM) (negative and positive controls, respectively) were observed through disc diffusion and micro-dilution methods. All nine clinical isolates were noticed as AMP resistant and AZM sensitive. Antibacterial effects of PCF and PBP were dose dependent and zone of inhibitions (ZI) against all clinical isolates were between 23.4 ± 0.92 to 0 ± 00 mm. The sensitivity profile of PCF and PBP against clinical isolates was noticed as 44.44 and 55.56%, respectively. Both PCF and PBP showed bacteriostatic (BTS) action against S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Moreover, the cumulative BTS potential of PCF and PBP against all isolates was 66.67 and 55.56%, respectively. The MICs of PCF and PBP were ranged from 50-200 µg/ml against selected isolates. The bacterial growth curves indicated that PCF and PBP inhibited the growth of all isolates at their specific MIC concentrations. However, PBP has better antibacterial potential compared to PCF against selected isolates. Therefore, it is concluded that both PCF and PBP of P. posthuma possess antibacterial and BTS potential against ampicillin resistant clinical isolates. This organism might be considered as a second choice of antibacterial agents and can further be utilized in pharmaceutical industries for novel drug manufacturing by prospecting bioactive potential agents.

Resumo Pheretima posthuma (Vaillant, 1868), uma minhoca nativa do Paquistão e sudeste da Ásia, tem ampla utilização em processos de vermicompostagem e biorremediação. Neste estudo, o fluido celômico de P. posthuma (PCF) e a pasta corporal (PBP) foram avaliados como agente antibacteriano contra cinco isolados clínicos Gram-positivos e quatro Gram-negativos resistentes à ampicilina (AMP). O efeito antibacteriano de diferentes doses (ou seja, 25-100 µg / ml) de PCF e PBP juntamente com AMP e azitromicina (AZM) (controles negativo e positivo, respectivamente) foi observado por meio de métodos de difusão em disco e microdiluição. Todos os nove isolados clínicos foram notados como resistentes a AMP e sensíveis a AZM. Os efeitos antibacterianos de PCF e PBP foram dependentes da dose e a zona de inibição (ZI) contra todos os isolados clínicos foi entre 23,4 ± 0,92 a 0 ± 00 mm. O perfil de sensibilidade do PCF e PBP contra isolados clínicos foi observado como 44,44% e 55,56%, respectivamente. Tanto o PCF quanto o PBP mostraram ação bacteriostática (BTS) contra S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Além disso, o potencial BTS cumulativo de PCF e PBP contra todos os isolados foi de 66,67% e 55,56%, respectivamente. Os MICs de PCF e PBP variaram de 50-200 µg / ml contra isolados selecionados. As curvas de crescimento bacteriano indicaram que o PCF e o PBP inibiram o crescimento de todos os isolados em suas concentrações específicas de MIC. No entanto, PBP tem melhor potencial antibacteriano em comparação com PCF contra isolados selecionados. Portanto, conclui-se que tanto o PCF quanto o PBP de P. posthuma possuem potencial antibacteriano e BTS contra isolados clínicos resistentes à ampicilina. Esse organismo pode ser considerado como uma segunda escolha de agentes antibacterianos e pode ainda ser utilizado nas indústrias farmacêuticas para a fabricação de novos medicamentos por meio da prospecção de agentes com potencial bioativo.

Antibacterial and bacteriostatic potential of coelomic fluid and body paste of Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) against ampicillin resistant clinical bacterial isolates / Potencial antibacteriano e bacteriostático do fluido celômico e pasta corporal de Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) contra isolados bacterianos clínicos resistentes à ampicilina

Pheretima posthuma (Vaillant, 1868), a native earthworm of Pakistan and Southeast Asia, has wide utilization in vermicomposting and bioremediation process. In this study, P. posthuma coelomic fluid (PCF) and body paste (PBP) was evaluated as antibacterial agent against ampicillin (AMP) resistant five Gram positive and four Gram negative clinical isolates. The antibacterial effect of different doses (i.e. 25-100 µg/ml) of PCF and PBP along with AMP and azithromycin (AZM) (negative and positive controls, respectively) were observed through disc diffusion and micro-dilution methods. All nine clinical isolates were noticed as AMP resistant and AZM sensitive. Antibacterial effects of PCF and PBP were dose dependent and zone of inhibitions (ZI) against all clinical isolates were between 23.4 ± 0.92 to 0 ± 00 mm. The sensitivity profile of PCF and PBP against clinical isolates was noticed as 44.44 and 55.56%, respectively. Both PCF and PBP showed bacteriostatic (BTS) action against S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Moreover, the cumulative BTS potential of PCF and PBP against all isolates was 66.67 and 55.56%, respectively. The MICs of PCF and PBP were ranged from 50-200 µg/ml against selected isolates. The bacterial growth curves indicated that PCF and PBP inhibited the growth of all isolates at their specific MIC concentrations. However, PBP has better antibacterial potential compared to PCF against selected isolates. Therefore, it is concluded that both PCF and PBP of P. posthuma possess antibacterial and BTS potential against ampicillin resistant clinical isolates. This organism might be considered as a second choice of antibacterial agents and can further be utilized in pharmaceutical industries for novel drug manufacturing by prospecting bioactive potential agents.

Pheretima posthuma (Vaillant, 1868), uma minhoca nativa do Paquistão e sudeste da Ásia, tem ampla utilização em processos de vermicompostagem e biorremediação. Neste estudo, o fluido celômico de P. posthuma (PCF) e a pasta corporal (PBP) foram avaliados como agente antibacteriano contra cinco isolados clínicos Gram-positivos e quatro Gram-negativos resistentes à ampicilina (AMP). O efeito antibacteriano de diferentes doses (ou seja, 25-100 µg / ml) de PCF e PBP juntamente com AMP e azitromicina (AZM) (controles negativo e positivo, respectivamente) foi observado por meio de métodos de difusão em disco e microdiluição. Todos os nove isolados clínicos foram notados como resistentes a AMP e sensíveis a AZM. Os efeitos antibacterianos de PCF e PBP foram dependentes da dose e a zona de inibição (ZI) contra todos os isolados clínicos foi entre 23,4 ± 0,92 a 0 ± 00 mm. O perfil de sensibilidade do PCF e PBP contra isolados clínicos foi observado como 44,44% e 55,56%, respectivamente. Tanto o PCF quanto o PBP mostraram ação bacteriostática (BTS) contra S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Além disso, o potencial BTS cumulativo de PCF e PBP contra todos os isolados foi de 66,67% e 55,56%, respectivamente. Os MICs de PCF e PBP variaram de 50-200 µg / ml contra isolados selecionados. As curvas de crescimento bacteriano indicaram que o PCF e o PBP inibiram o crescimento de todos os isolados em suas concentrações específicas de MIC. No entanto, PBP tem melhor potencial antibacteriano em comparação com PCF contra isolados selecionados. Portanto, conclui-se que tanto o PCF quanto o PBP de P. posthuma possuem potencial antibacteriano e BTS contra isolados clínicos resistentes à ampicilina. Esse organismo pode ser considerado como uma segunda escolha de agentes antibacterianos e pode ainda ser utilizado nas indústrias farmacêuticas para a fabricação de novos medicamentos por meio da prospecção de agentes com potencial bioativo.

Antibacterial and bacteriostatic potential of coelomic fluid and body paste of Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) against ampicillin resistant clinical bacterial isolates

Abstract Pheretima posthuma (Vaillant, 1868), a native earthworm of Pakistan and Southeast Asia, has wide utilization in vermicomposting and bioremediation process. In this study, P. posthuma coelomic fluid (PCF) and body paste (PBP) was evaluated as antibacterial agent against ampicillin (AMP) resistant five Gram positive and four Gram negative clinical isolates. The antibacterial effect of different doses (i.e. 25-100 µg/ml) of PCF and PBP along with AMP and azithromycin (AZM) (negative and positive controls, respectively) were observed through disc diffusion and micro-dilution methods. All nine clinical isolates were noticed as AMP resistant and AZM sensitive. Antibacterial effects of PCF and PBP were dose dependent and zone of inhibitions (ZI) against all clinical isolates were between 23.4 ± 0.92 to 0 ± 00 mm. The sensitivity profile of PCF and PBP against clinical isolates was noticed as 44.44 and 55.56%, respectively. Both PCF and PBP showed bacteriostatic (BTS) action against S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Moreover, the cumulative BTS potential of PCF and PBP against all isolates was 66.67 and 55.56%, respectively. The MICs of PCF and PBP were ranged from 50-200 µg/ml against selected isolates. The bacterial growth curves indicated that PCF and PBP inhibited the growth of all isolates at their specific MIC concentrations. However, PBP has better antibacterial potential compared to PCF against selected isolates. Therefore, it is concluded that both PCF and PBP of P. posthuma possess antibacterial and BTS potential against ampicillin resistant clinical isolates. This organism might be considered as a second choice of antibacterial agents and can further be utilized in pharmaceutical industries for novel drug manufacturing by prospecting bioactive potential agents.

Resumo Pheretima posthuma (Vaillant, 1868), uma minhoca nativa do Paquistão e sudeste da Ásia, tem ampla utilização em processos de vermicompostagem e biorremediação. Neste estudo, o fluido celômico de P. posthuma (PCF) e a pasta corporal (PBP) foram avaliados como agente antibacteriano contra cinco isolados clínicos Gram-positivos e quatro Gram-negativos resistentes à ampicilina (AMP). O efeito antibacteriano de diferentes doses (ou seja, 25-100 µg / ml) de PCF e PBP juntamente com AMP e azitromicina (AZM) (controles negativo e positivo, respectivamente) foi observado por meio de métodos de difusão em disco e microdiluição. Todos os nove isolados clínicos foram notados como resistentes a AMP e sensíveis a AZM. Os efeitos antibacterianos de PCF e PBP foram dependentes da dose e a zona de inibição (ZI) contra todos os isolados clínicos foi entre 23,4 ± 0,92 a 0 ± 00 mm. O perfil de sensibilidade do PCF e PBP contra isolados clínicos foi observado como 44,44% e 55,56%, respectivamente. Tanto o PCF quanto o PBP mostraram ação bacteriostática (BTS) contra S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Além disso, o potencial BTS cumulativo de PCF e PBP contra todos os isolados foi de 66,67% e 55,56%, respectivamente. Os MICs de PCF e PBP variaram de 50-200 µg / ml contra isolados selecionados. As curvas de crescimento bacteriano indicaram que o PCF e o PBP inibiram o crescimento de todos os isolados em suas concentrações específicas de MIC. No entanto, PBP tem melhor potencial antibacteriano em comparação com PCF contra isolados selecionados. Portanto, conclui-se que tanto o PCF quanto o PBP de P. posthuma possuem potencial antibacteriano e BTS contra isolados clínicos resistentes à ampicilina. Esse organismo pode ser considerado como uma segunda escolha de agentes antibacterianos e pode ainda ser utilizado nas indústrias farmacêuticas para a fabricação de novos medicamentos por meio da prospecção de agentes com potencial bioativo.

Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans / Disbiose intestinal, inflamação e diabetes tipo 2 em camundongos usando microbiota intestinal sintética de humanos diabéticos

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 [...].

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 [...].

Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans

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-, 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. .

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-, 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.

Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans / Disbiose intestinal, inflamação e diabetes tipo 2 em camundongos usando microbiota intestinal sintética de humanos diabéticos

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. .

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.

Recurrence and mortality after first venous thromboembolism in a large population-based cohort.

Essentials Reports on recurrence and mortality after a first venous thromboembolism (VTE) vary considerably. We describe rates of recurrence and mortality in patients with a first VTE from the Tromsø study. The overall recurrence rate was 3.9 per 100 person-years, but this varied widely with time. Despite advances in VTE management, the rates of adverse events are still fairly high. SUMMARY: Background Previous reports on recurrence and mortality rates after a first episode of venous thromboembolism (VTE) vary considerably. Advances in the management and treatment of VTE during the last 15 years may have influenced the rates of clinical outcomes. Aim To estimate the rates of recurrence and mortality after a first VTE in patients recruited from a large population-based cohort. Method From the Tromsø study, patients (n = 710) with a first, symptomatic, objectively confirmed VTE were included and followed in the period 1994-2012. Recurrent episodes of VTE were identified from multiple sources and carefully validated by review of medical records. Incidence rates and cumulative incidence rates with 95% confidence intervals (CIs) of VTE recurrence and mortality were calculated. Results The mean age of the patients was 68 years (range 28-102 years), and 166 (23.4%) had cancer at the time of first VTE. There were 114 VTE recurrences and 333 deaths during a median study period of 7.7 years (range 0.04-18.2 years). The risk of recurrence was highest during the first year. The overall 1-year recurrence rate was 7.8 (95% CI 5.8-10.6) per 100 person-years (PY), whereas the recurrence rate in the remaining follow-up period (1-18 years) was 3.0 (95% CI 2.4-3.8) per 100 PY. The overall 1-year all-cause mortality rate was 29.9 (95% CI 25.7-34.8) per 100 PY, and in those without cancer the corresponding rate was 23.6 (95% CI 17.8-31.3) per 100 PY. Conclusion Despite advances in VTE management, the rates of adverse events remained fairly high, particularly in the first year following a first VTE.