Resumo

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.

Resumo

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.

Resumo

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 [...].

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ABSTRACT Bacteroides fragilis is the strict anaerobic bacteria most commonly found in human infections, and has a high mortality rate. Among other virulence factors, the remarkable ability to acquire resistance to a variety of antimicrobial agents and to tolerate nanomolar concentrations of oxygen explains in part their success in causing infection and colonizing the mucosa. Much attention has been given to genes related to multiple drug resistance derived from plasmids, integrons or transposon, but such genes are also detected in chromosomal systems, like the mar (multiple antibiotic resistance) locus, that confer resistance to a range of drugs. Regulators like MarR, that control expression of the locus mar, also regulate resistance to organic solvents, disinfectants and oxygen reactive species are important players in these events. Strains derived from the parental strain 638R, with mutations in the genes hereby known as marRI (BF638R_3159) and marRII (BF638R_3706) were constructed by gene disruption using a suicide plasmid. Phenotypic response of the mutant strains to hydrogen peroxide, cell survival assay against exposure to oxygen, biofilm formation, resistance to bile salts and resistance to antibiotics was evaluated. The results showed that the mutant strains exhibit statistically significant differences in their response to oxygen stress, but no changes were observed in survival when exposed to bile salts. Biofilm formation was not affected by either gene disruption. Both mutant strains however, became more sensitive to multiple antimicrobial drugs tested. This indicates that as observed in other bacterial species, MarR are an important resistance mechanism in B. fragilis.

Resumo

Enterotoxigenic Bacteroides fragilis (ETBF) is an important part of the human and animal intestinal microbiota and is commonly associated with diarrhea. ETBF strains produce an enterotoxin encoded by the bft gene located in the B. fragilis pathogenicity island (BfPAI). Non-enterotoxigenic B. fragilis (NTBF) strains lack the BfPAI and usually show two different genetic patterns, II and III, based on the absence or presence of a BfPAI-flanking region, respectively. The incidence of ETBF and NTBF strains in fecal samples isolated from children without acute diarrhea or any other intestinal disorders was determined. All 84 fecal samples evaluated were B. fragilis-positive by PCR, four of them harbored the bft gene, 27 contained the NTBF pattern III DNA sequence, and 52 were considered to be NTBF pattern II samples. One sample was positive for both ETBF and NTBF pattern III DNA sequences. All 19 B. fragilis strains isolated by the culture method were bft-negative, 9 belonged to pattern III and 10 to pattern II. We present an updated overview of the ETBF and NTBF incidence in the fecal microbiota of children from Sao Paulo City, Brazil.

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Abstract Colorectal carcinoma is considered the fourth leading cause of cancer deaths worldwide. Several microorganisms have been associated with carcinogenesis, including Enterococcus spp., Helicobacter pylori, enterotoxigenic Bacteroides fragilis, pathogenic E. coli strains and oral Fusobacterium. Here we qualitatively and quantitatively evaluated the presence of oral and intestinal microorganisms in the fecal microbiota of colorectal cancer patients and healthy controls. Seventeen patients (between 49 and 70 years-old) visiting the Cancer Institute of the Sao Paulo State were selected, 7 of whom were diagnosed with colorectal carcinoma. Bacterial detection was performed by qRT-PCR. Although all of the tested bacteria were detected in the majority of the fecal samples, quantitative differences between the Cancer Group and healthy controls were detected only for F. nucleatum and C. difficile. The three tested oral microorganisms were frequently observed, suggesting a need for furthers studies into a potential role for these bacteria during colorectal carcinoma pathogenesis. Despite the small number of patients included in this study, we were able to detect significantly more F. nucleatum and C. difficile in the Cancer Group patients compared to healthy controls, suggesting a possible role of these bacteria in colon carcinogenesis. This finding should be considered when screening for colorectal cancer.

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Bacteroides fragilis colonizes dog guts both as a commensal and as an opportunistic pathogen. This study aims to evaluate virulence factors of 13 B. fragilis strains isolated from dog intestinal tracts and their ability for biofilm formation. Capsules were detected in all the evaluated strains. A total of 61.5% of all strains were biofilm producers. These attributes most likely play an important role in B. fragilis persistent colonization in the gut.

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Fifty one strains of the Bacteroides fragilis group were isolated from 45 fecal samples. Classical phenotypic identification showed that 16 isolates were B. thetaiotaomicron, 12 B. uniformis, 9 B. eggerthii,7 B. vulgatus,3 B. caccae,2 Parabacteroides distasonis with 1 identified B. ovatus and 1 B. fragilis. The 51 strains were tested for susceptibility against 16 antimicrobial agents and the MICs for metronidazole were determined. The tests showed that imipenem, meropenem and chloram-phenicol were the most effective antibiotics (98%, 98% and 92.16% of susceptibility, respectively) followed by ticarcillin/clavulanic acid, piperacillin/tazobactam, rifampin (88.24% susceptibility), moxifloxacin 86.27% and tigecycline 84.31%. Ofloxacin and cefotaxime were the least effective antibiotics with 27.45% and 0% of activity respectively. Only six of the 51 isolated strains were resistant to metronidazole with MICs = 64 mg/L (1 strain) and > 256 mg/L (5 strains).

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The production of Toxic Shock Syndrome Toxin-1 (TSST-1), enterotoxins and bacteriocin-like substances was evaluated in 95 strains of Staphylococcus aureus recovered from raw bovine milk (n=31) and from food samples involved in staphylococcal food poisoning (n=64). Enterotoxigenicity tests with the membrane over agar associated to optimal sensibility plate assays were performed and showed that 96.77% of strains recovered from milk and 95.31% from food samples produced enterotoxins A, B, C, D or TSST-1. Reference strains S. epidermidis, Bacillus cereus, Listeria monocytogenes, Lactobacillus casei, Pseudomonas aeruginosa, S. aureus, Salmonella Typhimurium, Escherichia coli, Enterococcus faecalis and Bacteroides fragilis were used as indicator bacteria in the antagonistic assays, the first five being sensitive to antagonistic substances. Brain heart infusion agar, in pH values ranging from 5.0 to 7.0 in aerobic atmosphere showed to be the optimum condition for antagonistic activity as evaluated with the best producer strains against the most sensitive indicator bacterium, L. monocytogenes. Sensitivity to enzymes confirmed the proteinaceous nature of these substances. Neither bacteriophage activity nor fatty acids were detected and the antagonistic activity was not due to residual chloroform. Results did not establish a positive correlation between the bacteriocinogenic profile and toxigenicity in the tested S. aureus strains.

Avaliou-se a produção de toxina-1 da síndrome do choque tóxico (TSST-1), enterotoxinas e substâncias antagonistas tipo bacteriocina em 95 amostras de Staphylococcus aureus recuperadas de leite bovino in natura (n=31) e de alimentos envolvidos em surto de intoxicação (n=64). Testes de enterotoxigenicidade pelo método da membrana sobre ágar, associado à técnica da sensibilidade ótima em placa, revelaram que 96,77% das amostras do leite e 95,31% daquelas dos alimentos produziram enterotoxinas estafilocócicas tipos A, B, C, D ou TSST-1. Nos ensaios de antagonismo, foram utilizadas como reveladoras amostras de referência de S. epidermidis, Bacillus cereus, Listeria monocytogenes, Lactobacillus casei, Pseudomonas aeruginosa, S. aureus, Salmonella typhimurium, Escherichia coli, Enterococcus faecalis e Bacteroides fragilis, sendo as cinco primeiras sensíveis às substâncias produzidas. As condições ótimas para a atividade antagonista, avaliadas com as melhores produtoras contra a indicadora mais sensível, L. monocytogenes, foram observadas em aerobiose, em ágar infuso de cérebro-coração, nos valores de pH entre 5,0 e 7,0. A sensibilidade a enzimas confirmou a natureza proteica destas substâncias. Não foram detectadas atividades de bacteriófagos nem de ácidos graxos, e a atividade antagonista não foi devido ao clorofórmio residual. Os resultados não mostraram correlação entre o perfil bacteriocinogênico e a toxigenicidade nas amostras de Staphylococcus testadas.

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This work describes the phytochemical study of the extracts from aerial parts of Tibouchina candolleana as well as the evaluation of the antimicrobial activity of extracts, isolated compounds, and semi-synthetic derivatives of ursolic acid against endodontic bacteria. HRGC analysis of the n-hexane extract of T. candolleana allowed identification of b-amyrin, a-amyrin, and b-sitosterol as major constituents. The triterpenes ursolic acid and oleanolic acid were isolated from the methylene chloride extract and identified. In addition, the flavonoids luteolin and genistein were isolated from the ethanol extract and identified. The antimicrobial activity was investigated via determination of the minimum inhibitory concentration (MIC) using the broth microdilution method. Amongst the isolated compounds, ursolic acid was the most effective against the selected endodontic bacteria. As for the semi-synthetic ursolic acid derivatives, only the methyl ester derivative potentiated the activity against Bacteroides fragilis.

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Despite the importance of gastrointestinal diseases and their global distribution, affecting millions of individuals around the world, the role and antimicrobial susceptibility patterns of anaerobic bacteria such as those in the Bacteroides fragilis group (BFG) are still unclear in young children. This study investigated the occurrence and distribution of species in the BFG and enterotoxigenic strains in the fecal microbiota of children and their antimicrobial susceptibility patterns. Diarrheic (n=110) and non-diarrheic (n=65) fecal samples from children aged 0-5 years old were evaluated. BFG strains were isolated and identified by conventional biochemical, physiological and molecular approaches. Alternatively, bacteria and enterotoxigenic strains were detected directly from feces by molecular biology. Antimicrobial drug susceptibility patterns were determined by the agar dilution method according to the guidelines for isolated bacteria. BFG was detected in 64.3 percent of the fecal samples (55 percent diarrheic and 80.4 percent non-diarrheic), and 4.6 percent were enterotoxigenic. Antimicrobial resistance was observed against ampicillin, ampicillin/sulbactam, piperacillin/tazobactam, meropenem, ceftriaxone, clindamycin and chloramphenicol. The data show that these bacteria are prevalent in fecal microbiota at higher levels in healthy children. The molecular methodology was more effective in identifying the B. fragilis group when compared to the biochemical and physiological techniques. The observation of high resistance levels stimulates thoughts about the indiscriminate use of antimicrobial drugs in early infancy. Further quantitative studies are needed to gain a better understanding of the role of these bacteria in acute diarrhea in children.

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The goals of this study were to evaluate if hemoglobin played a role as adjuvant in experimental peritonitis in horses and could cause clinical and haematological alterations that could be used for diagnosis and prognosis of cases of peritonitis. Fifteen adult horses were randomly divided into 5 equal groups, which were injected intraperitoneally with the following suspension: GI: 1x109 colony-forming units (CFU) of E. coli diluted in 500 mL of 0.9% saline solution plus 5 g of hemoglobin; GII: 1x109 CFU of B. fragilis diluted in 500 mL of 0.9% saline plus 5 g of hemoglobin; GIII: 1x109 CFU of E. coli in combination with 1x109  CFU of B. fragilis diluted in 500 ml of 0.9% of saline plus 5 g of hemoglobin; GIV: 500 mL of 0.9% saline plus 5 g of hemoglobin and GV: 500 mL of 0.9% saline. Leukopenia with neutropenia was observed in GI and GIII  and a significant increase in plasma fibrinogen concentration occurred in horses of GI. There was a significant increase in total nucleated cell count in peritoneal fluid in horses of GI, GII, GIII and GIV. Fever, tachycardia, tachypnea, abdominal wall sensibility and tension, diarrhoea, colic, and decreased borborygmi sounds were the most frequent clinical signs observed in horses of GI, GII, GIII and GIV. In conclusion, hemoglobin was able to cause chemical peritonitis in horses, it had an adjuvant effect when associated t

Resumo

Sixteen adult horses were randomly divided into 4 equal groups (GI, GII, GIII and GIV) of 4 animals and each group was injected intraperitoneally with one of the following suspension: GI (100 x 10(7) colony-forming units (CFU) of Escherichia coli diluted in 500ml of 0.9% saline); GII (100 x 10(7) CFU of Bacteroides fragilis in 500ml of 0.9% saline); GIII (100 x 10(7) CFU of E. coli in combination with 100 x 10(7) CFU of B. fragilis in 500ml of 0.9% saline); GIV (500ml of 0.9% saline). Abdominal wall sensitivity to external pressure and tension, diarrhea, decreased intestinal sounds and increased of heart rate were the clinical signs more frequently observed in inoculated horses. Horses inoculated with pure cultures of either E. coli or B. fragilis demonstrated mild and self-limiting peritonitis, while those inoculated with the combination of both bacteria demonstrated clinical signs of higher intensity and duration.

Dezesseis eqüinos adultos foram distribuídos aleatoriamente em 4 grupos (GI, GII, GIII e GIV), constituídos por quatro animais, recebendo cada grupo o seguinte inóculo por via intraperitoneal: GI (100 X 10(7) unidades formadoras de colônia (UFC) de Escherichia coli diluídos em 500ml de salina 0,9%); GII (100 X 10(7) UFC de Bacteroides fragilis diluídos em 500ml de salina 0,9%); GIII (100 X 10(7) UFC de E. coli associados a 100 X 10(7) UFC de B. fragilis diluídos em 500ml de salina 0,9%); GIV (testemunho - 500ml de salina 0,9%). Aumento da sensibilidade e tensão da parede abdominal, diarréia, diminuição dos sons intestinais e aumento da freqüência cardíaca foram os sinais mais freqüentemente observados nos eqüinos inoculados com cepas bacterianas. Eqüinos inoculados com culturas puras de E. coli ou B. fragilis apresentaram peritonites brandas e autolimitantes, enquanto que os inoculados com a associação dessas bactérias apresentaram sinais de maior intensidade e duração.

Resumo

Sixteen adult horses were randomly divided into 4 equal groups (GI, GII, GIII and GIV) of 4 animals and each group was injected intraperitoneally with one of the following suspension: Group I, 100 x 107 colony-forming units (CFU) of E. coli diluted in 500 ml of 0.9percent saline; Group II, 100 x 107 CFU of Bacteroides fragilis in 500 ml of 0.9 percent saline; Group III, 100 x 107 CFU of E. coli in combination with 100 x 107 CFU of B. fragilis in 500 ml of 0.9 percent saline; Group IV, 500 ml of 0.9 percent saline. Leukopenia appeared in all animals inoculated with bacteria within the first six hours of the experiment. After this period, leukocytosis was observed in some inoculated horses. Horses inoculated with pure cultures of either E. coli or B. fragilis demonstrated mild and self-limiting peritonitis, whereas those inoculated with a combination of both bacteria demonstrated laboratory findings of higher intensity and duration

Resumo

Dezesseis eqüinos adultos foram aleatoriamente divididos em quatro grupos de quatro animais que receberam inoculaçäo intraperitoneal das seguintes suspensöes: grupo I, 100 x 10 elevado a sétima potência unidades formadoras de colônias (CFU) de E. coli diluídas em 500ml de soluçäo salina a 0,9 por cento: grupo II, 100 x 10 elevado a sétima potência CFU de Bacteroides fragilis em 500ml de soluçäo salina a 0,9 por cento; grupo III, 100 x 10 elevado a sétima potência CFU de E. coli combinados com 100 x 10 elevado a sétima potência CFU de B. fragilis em 500ml de soluçäo salina a 0,9 por cento; grupo IV, 500ml de soluçäo salina a 0,9 por cento. Observou-se aumento significativo do número de leucócitos no líquido peritoneal quatro horas após as inoculaçöes dos animais dos grupos I e II, e oito horas após as inoculaçöes dos animais do grupo III. A contagem mais elevada foi de 516 x 10 elevado a terceira potência leucócitos/mm elevado ao cubo. Aumentos significativos nas concentraçöes de fibrogênio (1 g/dl)e proteína total (9,1 por cento) foram também observados. Eqüinos inoculados com culturas puras, tanto de E. coli quanto de B. fragilis, apresentaram peritonites mais brandas e autolimitantes, enquanto que eqüinos inoculados com associaçäo das duas bactérias apresentaram alteraçöes laboratoriais com maior intensidade e duraçäo

Resumo

Os gêneros Bacteroides e Parabacteroides estão envolvidos em sérias patologias como abscessos em vários órgãos e bacteremias em homens e animais. Essas bactérias são caracterizadas pela resistência aos antimicrobianos e B. fragilis é a principal bactéria anaeróbia isolada do intestino que pode formar biofilme. O objetivo desse estudo foi isolar Bacteroides e Parabacteroides do trato intestinal de cães para avaliar a sensibilidade antimicrobiana e a ação de concentrações subinibitórias de antimicrobianos na formação de biofilme. No período compreendido entre janeiro a junho de 2011, 30 cepas dos gêneros Bacteroides e Parabacteroides foram isolados de 50 cães atendidos na Unidade Hospitalar Veterinária da UECE. As amostras foram coletadas por meio de swab retal e semeadas em meio seletivo agar Bacteroides Bile Esculina e incubadas em anaerobiose a 37 oC por 48h. As espécies dos gêneros Bacteroides e Parabacteroides foram identificadas por meio de provas bioquímicas e cromatografia gasosa. Foi avaliada a sensibilidade a 10 antimicrobianos pela técnica de microdiluição em caldo e as cepas de B. fragilis formadoras de biofilme foram classificadas e as mais fortemente aderentes em número de quatro foram utilizadas no estudo sobre a ação da concentração subinibitória dos antimicrobianos sobre a formação de biofilme. O biofilme formado dessas quatro cepas sob a ação de antimicrobianos foi avaliado por meio de microscopia confocal. As espécies mais frequentemente isoladas foram B. fragilis, P. distasonis e B. vulgatus. As cepas apresentaram alta sensibilidade ao cloranfenicol, imipenem, metronidazol e amoxicilina/ácido clavulânico. As cepas foram menos sensíveis a cefoxitina e a sensibilidade a tetraciclina, clindamicina e penicilina foram baixas. As concentrações inibitórias mínimas das quinolonas testadas para as cepas isoladas foram semelhantes as descritas na literatura. Os antimicrobianos testados reduziram a formação de biofilme nas quatro cepas exceto enrofloxacina, que agiu induzindo a formação de biofilme em metade das cepas testadas.(AU)

Resumo

Os gêneros Bacteroides e Parabacteroides estão envolvidos em sérias patologias como abscessos em vários órgãos e bacteremias em homens e animais. Essas bactérias são caracterizadas pela resistência aos antimicrobianos e B. fragilis é a principal bactéria anaeróbia isolada do intestino que pode formar biofilme. O objetivo desse estudo foi isolar Bacteroides e Parabacteroides do trato intestinal de cães para avaliar a sensibilidade antimicrobiana e a ação de concentrações subinibitórias de antimicrobianos na formação de biofilme. No período compreendido entre janeiro a junho de 2011, 30 cepas dos gêneros Bacteroides e Parabacteroides foram isolados de 50 cães atendidos na Unidade Hospitalar Veterinária da UECE. As amostras foram coletadas por meio de swab retal e semeadas em meio seletivo agar Bacteroides Bile Esculina e incubadas em anaerobiose a 37 oC por 48h. As espécies dos gêneros Bacteroides e Parabacteroides foram identificadas por meio de provas bioquímicas e cromatografia gasosa. Foi avaliada a sensibilidade a 10 antimicrobianos pela técnica de microdiluição em caldo e as cepas de B. fragilis formadoras de biofilme foram classificadas e as mais fortemente aderentes em número de quatro foram utilizadas no estudo sobre a ação da concentração subinibitória dos antimicrobianos sobre a formação de biofilme. O biofilme formado dessas quatro cepas sob a ação de antimicrobianos foi avaliado por meio de microscopia confocal. As espécies mais frequentemente isoladas foram B. fragilis, P. distasonis e B. vulgatus. As cepas apresentaram alta sensibilidade ao cloranfenicol, imipenem, metronidazol e amoxicilina/ácido clavulânico. As cepas foram menos sensíveis a cefoxitina e a sensibilidade a tetraciclina, clindamicina e penicilina foram baixas. As concentrações inibitórias mínimas das quinolonas testadas para as cepas isoladas foram semelhantes as descritas na literatura. Os antimicrobianos testados reduziram a formação de biofilme nas quatro cepas exceto enrofloxacina, que agiu induzindo a formação de biofilme em metade das cepas testadas.

Resumo

Quinze eqüinos adultos foram aleatoriamente divididos em 5 grupos com três animais cada, os quais foram inoculados intraperitonealmente com as seguintes suspensões: Grupo 1: 1x109unidades formadoras de colônias (UFC) de E. coli diluídas em 500 mL de solução salina a 0,9% adicionadas de 5 gramas de hemoglobina, Grupo 2: 1x109UFC de B. fragilis diluídas em 500 mL de solução salina a 0,9% adicionadas de 5 gramas de hemoglobina, Grupo 3: 1x109UFC de E. coli em combinação com 1x109UFC de B. fragilis diluídas em 500 mL de solução salina a 0,9% adicionadas de 5 gramas de hemoglobina, Grupo 4: 500 mL de solução salina a 0,9% adicionadas de 5 gramas de hemoglobina e Grupo 5: 500 mL de solução salina a 0,9%. Leucopenia com neutrofilia foram observadas nos grupos 1 e 3 dentro das primeiras seis horas do estudo e aumento da concentração plasmática de fibrinogênio nos eqüinos do Grupo 1 foi observado das 48 até as 120 horas após a inoculação (HAI). Houve aumento significativo na contagem total de células nucleadas do líquido peritoneal dos eqüinos dos Grupos 1, 2, 3 e 4 das 10 HAI até as 120 HAI. Os teores de proteína total do líquido peritoneal aumentaram significativamente das 2 HAI até o término das observações, nos eqüinos do Grupo 1; das 10 HAI até as 72 HAI, nos eqüinos do Grupo 2; das 6 HAI até as 72 HAI, nos eqüinos do grupo 3; e das 4 HAI até as 60 HAI nos eqüinos do Grupo 4, enquanto que aumento dos níveis de fibrinogênio no líquido peritoneal ocorreram das 2 as 120 HAI nos eqüinos do Grupo 1. Hipertermia, taquicardia, taquipnéia, aumento da tensão e sensibilidade da parede abdominal, diarréia, mímica de dor e diminuição dos sons intestinais foram os sinais clínicos mais freqüentemente observados nos eqüinos dos grupos 1, 2, 3 e 4... (resumo completo, clicar acesso eletrônico abaixo)

Fifteen adult horses were randomly divided into 5 equal groups which was injected intraperitoneally with the following suspension: Group 1: 1x109 colony-forming units (CFU) of E. coli diluted in 500 mL of 0.9% saline plus 5 gr. of hemoglobin, Group 2: 1x109 CFU of B. fragilis diluted in 500 mL of 0.9% saline plus 5 gr. of hemoglobin, Group 3: 1x109 CFU of E. coli in combination with 1x109 CFU of B. fragilis diluted in 500 mL of 0.9% saline plus 5 gr. of hemoglobin, Group 4: 500 mL of 0.9% saline plus 5 gr. of hemoglobin and Group 5: 500 mL of 0.9% saline. Leukopenia with neutropenia was observed in groups 1 and 3 within the first 6 hours of the study and significant increase in plasma fibrinogen concentration in horses of group 1 was observed from 48 till 120 hours after inoculation (HAI). There was a significant increase in total nucleated cell count in peritoneal fluid in horses of groups 1, 2, 3 and 4 from 10 HAI till 120 HAI. Total protein had a significant increase from 2 HAI till the end of the study in G1, from 10 HAI till 72 HAI in G2, from 6 HAI till 72 HAI in G3 and from 4 HAI till 60 HAI in G4, while there was significant increased fibrinogen levels from 2 to 120 HAI in G1. Fever, tachicardia, tachipnea, abdominal wall sensitivity and tension, diarrhoea, colic, decreased borborigmi sounds were the most frequent clinical signs observed in horses of G1, G2, G3 and G4 similarly to others in natural and experimental peritonitis. Animals of Group 5 showed no physical signs alterations. Horses of Groups 2 and 4 demonstrated mild and self-limiting peritonitis, while those of groups 1 and 3 demonstrated clinical signs of higher intensity and duration... (complete abstract, access undermentioned eletronic address)