Metagenomics approach to identify lignocellulose-degrading enzymes in the gut microbiota of the Chinese bamboo rat cecum

BACKGROUND: Lignocellulose is considered a renewable organic material, but the industrial production of biofuel from lignocellulose is challenging because of the lack of highly active hydrolytic enzymes. The guts of herbivores contain many symbiotic microorganisms that have evolved to hydrolyze plant lignocellulose. Chinese bamboo rats mainly consume high-fiber foods, indicating that some members of the intestinal tract microbiota digest lignocellulose, providing these rats with the energy required for growth. RESULTS: Here, we used metagenomics to analyze the diversity and functions of the gut microbiota in Chinese bamboo rats. We identified abundant populations of lignocellulose-degrading bacteria, whose main functions involved carbohydrate, amino acid, and nucleic acid metabolism. We also found 587 carbohydrate-active enzyme genes belonging to different families, including 7 carbohydrate esterase families and 21 glycoside hydrolase families. The glycoside hydrolase 3, glycoside hydrolase 1, glycoside hydrolase 43, carbohydrate esterase 4, carbohydrate esterase 1, and carbohydrate esterase 3 families demonstrated outstanding performance. CONCLUSIONS: The microbes and enzymes identified in our study expand the existing arsenal of proficient degraders and enzymes for lignocellulosic biofuel production. This study also describes a powerful approach for targeting gut microbes and enzymes in numerous industries.

Utilização da técnica in vitro semiautomática de produção de gases na avaliação da fermentação microbiana do ceco de suínos / Use of the semi-automatic in vitro gas production technique for the evaluation of swine cecal microbial fermentation

O presente estudo teve como objetivo avaliar o os padrões de fermentação microbiana do ceco de suínos, utilizando-se um modelo aplicado na técnica in vitro de produção de gases para ruminantes. Três tipos de inóculo foram utilizados: ceco de suínos criados ao ar livre (CSF), ceco de suínos criados confinados (CSC) e líquido ruminal de bovino (LRB). Os substratos utilizados derivaram de uma dieta de suínos à base de farelo de soja e de grãos de milho, atendendo as exigências nutricionais dos suínos. Para composição dos substratos, foram estabelecidos diferentes níveis de substituição da dieta basal pela torta da amêndoa da Acrocomia aculeata (AA), coproduto da produção do biodiesel, sendo: BAS - 100% de dieta basal; M10 - 90% de dieta basal e 10% AA; M20 - 80% de dieta basal e 20% AA e o TF - feno de Tifton 85 (Cynodon spp.). A produção de gases foi avaliada nos tempos zero, três, seis, nove, 12, 16, 24, 48, 72 e 96 horas após a incubação. Foram analisados o potencial máximo de produção de gás (A) e o tempo de colonização (L). O delineamento experimental utilizado foi inteiramente ao acaso, em arranjo fatorial 4 x 3. Os dados de A e L foram submetidos à análise de variância, e as médias comparadas pelo teste de Tukey a 5%. Comparando os inóculos para A (mL/gMS), LRB e CSF apresentaram valores similares, diferindo apenas para TF. O tempo de colonização (L) entre inóculos, o CSC e o CSF apresentaram os menores tempos de colonização quando a ração foi BAS. Com os substratos M10 e M20, o inóculo LRB apresentou o menor tempo, CSF o maior tempo e CSC não diferiu de ambos. Não houve diferença significativa entre os inóculos para o substrato TF. A técnica de produção de gases utilizada para o ceco de suínos apresentou resultados semelhantes aos encontrados para ruminantes. O modelo matemático usado foi adequado para descrever a curva de fermentação no ceco de suínos, mostrando semelhanças entre as microbiotas do ceco e do rúmen.(AU)

This study aimed to evaluate the microbial fermentation patterns of the pig ceca using the technique of in vitro gas production for ruminants. Three types of inoculums were used: swine ceca raised in a free range system (CSF), swine ceca raised in a conventional confined system (CSC) and bovine rumen fluid (LRB). The substrates used were derived from pig's diet with soybean meal and corn grits, according to the nutritional requirements of swine. The substrates were composed by different set levels of substitution of basal diet by meal almond Acrocomia aculeata (AA), as: BAS - 100% of basal diet; M10 - 90% of basal diet and 10% AA (M10); M20 - 80% of basal diet and 20% AA (M20) and TF - Tifton 85 hay (Cynodon spp.).The gas production was evaluated at times 0, 3, 6, 9, 12, 16, 24, 48, 72 and 96 hours after incubation. The study analyzed the maximum potential of gas production (A) and lag time (L). The experimental design was completely randomized in a 4 x 3 factorial arrangement. The A and L data were submitted to analysis of variance and the averages compared by Tukey 5% test. Comparing the inoculum for A (ml / gDM), LRB, and CS, they showed similar values, differing only for TF. The lag time (L) between inoculum, the CSC and CSF had the lowest colonization times when the meal was BAS. With M10 and M20 substrates, the LRB inoculum showed the lowest time, CSF had the longest time and CSC did not differ from both. There was no significant difference between the inoculum for TF substrate. The gas production technique used for the ceca of pigs showed similar results to those for ruminants. The mathematical model used was adequate to describe the fermentation curve in the ceca of pigs, showing similarities between the microbiota of the ceca and the rumen.(AU)

The effect of amifostine on bacterial translocation after radiation induced acute enteritis

ABSTRACT PURPOSE: To investigate the effects of amifostine on bacterial translocation and overgrowth in colonic flora after acute radiation enteritis in a rat model. METHODS: Thirty-two female Wistar albino rats were divided into four groups: Group-1 (n=8): only normal saline was administered intraperitoneally. Group-2 (n=8): first serum saline was administered intraperitoneally and 30 minutes later 20 Gy radiation was applied to abdominopelvic region. Group-3 (n=8): only amifostine 200 ml/kg was administered intraperitoneally and radiation was not applied. Group-4 (n=8): first amifostine 200 ml/kg was administered intraperitoneally and 30 minutes later 20 Gy radiation was applied to abdominopelvic region. On the 5th day after radiation, samples of mesenteric lymph tissues and cecal contents were taken by laparotomy for microbiological culture. RESULTS: Intraperitoneal amifostine administration significantly decreased the bacterial overgrowth related to radiation in colon but did not significantly decrease the bacterial translocation. CONCLUSİON: Although not providing a full protection on the damaged mucosal barrier, amifostine significantly decreased the bacterial overgrowth in the cecal content after high dose radiation. There is a need to find out appropriate amifostine dose under different radiation applications avoiding bacterial translocation in gastrointestinal system.

Propionibacterium acnes-killed attenuates the inflammatory response and protects mice from sepsis by modulating inflammatory factors

BACKGROUND: Sepsis is a systemic inflammation associated with infection caused by pathogenic micro-organisms with high mortality rates. OBJECTIVE: In this study, we investigated the protective effect of Propionibacterium acnes-killed against polymicrobial sepsis induced by cecal ligation and puncture. METHODS: The mice were treated by intramuscular route in 1, 3, 5, and 7 days before the cecal ligation and puncture induction. The control group animals received vehicle (saline solution 0.9%) and the animals of the treated group received the P. acnes-killed (0.4 mg/animal). After anesthesia, midline laparotomy was performed with exposure of cecum followed by ligature and one transverse perforation of the same, with a 18 G needle, for induction of lethal sepsis. After surgery, the cecum of the animals was replaced into the peritoneal cavity, and it was closed with a 4.0 nylon suture. The survival of animals subjected to lethal sepsis was evaluated after cecal ligation and puncture induction. Six hours after the induction of sepsis, neutrophil migration, the number of bacteria, TNF-α, MCP-1, IL-6, and IL-10 were performed in the peritoneal lavage. RESULTS: Prophylactic treatment with P. acnes-killed increased the survival of the animals, followed by a significant decrease in the TNF-α, IL-10, and MCP-1 levels, 6 h after cecal ligation and puncture. Furthermore, P. acnes-killed administration reduced the number of bacteria in the peritoneal cavity with increased migration of leukocytes, especially neutrophils. CONCLUSION: P. acnes-killed promoted increased survival rate of animals with sepsis, in part attributed to its immunomodulatory properties against pathogenic microorganisms, as well as better control of infection by reducing bacterial counts.

Effect of Lactobacillus acidophilus & epidermal growth factor on experimentally induced Clostridium difficile infection.

Background & objectives: Clostridium difficile-associated disease (CDAD) remains an important nosocomial ailment. Antimicrobial therapy used for CDAD gives inconsistent results. This experimental study was planned to investigate the beneficial effects of Lactobacillus acidophilus and epidermal growth factor (EGF) for CDAD management. Methods: Among 10 groups of BALB/c mice (6 in each), group 1 served as controls receiving no inoculum. Animals in groups 2-10 received C. difficile, those in groups 3, 6 and 9 received L. acidophilus and those in groups 4, 7 and 10 received EGF after C. difficile inoculation. Animals in groups 5-7 were pre-treated with ampicillin and those in groups 8-10 with lansoprazole prior to C. difficile. The animals were killed and investigated for colonisation by C. difficile and toxin production, myeloperoxidase (MPO) activity and histopathology. Results: Colonisation by C. difficile was found to be significantly different (P<0.001) in the various groups. C. difficile toxin titres and MPO activity were significantly lower in animals given L. acidophilus and EGF after ampicillin (groups 6 and 7) and lansoprazole (groups 9 and 10). The severity of acute inflammation was also significantly less (P<0.05) in caecal and colonic segments of animals in groups 6 and 7 compared to those in group 5. Although the severity of acute inflammation was less in the caecal and colonic segment of animals in groups 9 and 10, the reduction was not significant compared to group 8. Interpretation & conclusions: Our findings showed that the administration of L. acidophilus and EGF reduced the severity of C. difficile infection in the experimental animals.

Effect of treatment with neem [azadirachta indica] compared with baycox drug on the caecum of chicken experimentally infected with eimeria tenella

The study evaluated the therapeutic efficacy of Neem herb in chicken experimentally infected with E. tenella compared to Baycox as a reference anticoccidial drug. 120 broiler chicks were enrolled, randomly divided to 4 groups, [A, B, C and D] non-infected non-treated [naive control], [B] infected with 10[4] E. tenella oocysts [infected control], [C] infected and treated with Baycox [7 mg/kg b.w. for 2 days] and [D] infected and treated with Neem leaves water extract [100 mg/kg b.w. for 9 days]. Evaluation was by clinical signs, performance data [gain weigh, food consumption oocyst shed/gram feaces [OPG]], in addition to histopathological changes in all chickens. The results revealed that chicks of GA had the best performance data compared to GB, GC and GD. In GC and GD there were a remarkable improvement in the data performance, clinical signs, gross and microscopically cecal lesions compared to GB. The efficacy of Baycox [GC] was shown to be superior to that of Neem [GD] compared to GB but an additive histopathological toxic effect besides those produced by E. tenella infection could be recorded. In contrast, Neem appeared to have a remarkable improvement on cecal integrity

Serovariedades de Salmonella enterica subespecie enterica en porcinos de faena y su resistencia a los antimicrobianos / Serovars of Salmonella enterica subspecies enterica and its antimicrobial resistance in slaughterhouse pigs

Se realizó un estudio para determinar la prevalencia de Salmonella y sus serovariedades en cerdos de faena, para evaluar sus perfiles de resistencia a los antimicrobianos y para conocer la presencia de integrones de clase 1 como posibles reservorios de resistencia. A partir de un total de 386 muestras de porcinos provenientes de cuatro frigoríficos de las provincias de Buenos Aires y de Santa Fe (Argentina), se identificaron 93 (24,1%) cepas de Salmonella enterica subespecie enterica, 52 (55,9%) de contenido cecal y 41 (44,1%) de nódulo linfático ileocecal. Se hallaron 13 serovariedades de S. enterica, las más prevalentes fueron S. Schwarzengrund, S. Heidelberg, S. subespecie I 6,8:e,h:-, S. Derby y S. Bredeney. Se probaron 15 antimicrobianos por el método de dilución en agar: amikacina, gentamicina, ciprofloxacina, cefalotina, cefotaxima, enrofloxacina, fosfomicina, polimixina-B, tetraciclina, cloranfenicol, estreptomicina, trimetoprima-sulfametoxazol, ampicilina, nitrofurantoína y ácido nalidíxico. Según se estableció mediante la determinación de la CIM, el 73% de las cepas de S. enterica subespecie enterica fueron sensibles a todos los antimicrobianos probados. Se observó resistencia a tetraciclina en 24 (25,8%) de las 93 cepas, a cloranfenicol en 22 (23,7%), a estreptomicina en 22 (23,7%) a trimetoprima-sulfametoxazol en 20 (21,5%), a ampicilina en 18 (19,4%), a nitrofurantoína en 3 (3,2%) y a ácido nalidíxico en 3 (3,2%). Algunos aislamientos de S. Typhimurium, S. Heildelberg, S. Derby y S. Orion presentaron multirresistencia y portaban el gen de la integrasa clase 1. Los mayores porcentajes de resistencia correspondieron a los antimicrobianos habitualmente utilizados en veterinaria y en las explotaciones porcinas.

A study was carried out in order to determine the prevalence of Salmonella and its serovars among porcine slaughterhouses, to evaluate the antimicrobial resistance profiles and to know the presence of class 1 integrons as possible reservoir of resistance. From a total of 386 samples from four porcine slaughterhouses of Buenos Aires and Santa Fe Provinces (Argentina), 93 (24,1%) Salmonella enterica subspecies enterica strains were identified, 52 (55,9%) from cecal contents and 41 (44,1%) from ileocecal lymph nodes. Thirteen serovars of S. enterica were found, the most prevalent were: S. Schwarzengrund, S. Heidelberg, S. subspecie I 6,8:e,h:-, S. Derby and S. Bredeney. Fifteen antimicrobials by the agar dilution method were tested: amikacin, gentamicin, ciprofloxacin, cephalotin, cefotaxime, enrofloxacin, fosfomycin, polimixin-B, tetracycline, chloramphenicol, streptomycin, trimethoprim-sulfamethoxazole, ampicillin, nitrofurantoin, and nalidixic acid. According to the CIM determination, 73% Salmonella enterica subspecies enterica strains were sensible to all the antimicrobials tested. Antimicrobial resistance was observed to tetracycline in 24 (25,8%) of 93 strains, to chloramphenicol in 22 (23,7%), to streptomycin in 22 (23,7%), to trimethoprim-sulfamethoxazole in 20 (21,5%), to ampicillin in 18 (19,4%), to nitrofurantoin in 3 (3,2%) and to nalidixic acid in 3 (3,2%). Some isolates of S. Typhimurium, S. Heidelberg, S. Derby, S. Orion showed multidrug resistance and carried the class 1 integrase gene. The highest percentage of resistance corresponded to the antimicrobials currently used in veterinary and porcine farms.

Uso de microbiota cecal congelada com crioprotetores em pintos infectados experimentalmente com Salmonella enterica sorovar Enteritidis / Use of frozen cecal microbiota with cryoprotectors in chicks experimentally infected with Salmonella enterica serotype Enteritidis

Pintos de corte com um dia de idade foram tratados com microbiota cecal cultivada em condição de aerobiose, nos tempos de congelamento de 90, 200, 290 e 360 dias, e associada aos crioprotetores sacarose, trealose, dimetilsulfóxido (DMSO) e glicerol. Posteriormente as aves foram desafiadas com Salmonella Enteritidis, visando determinar a eficácia dos tratamentos em relação à quantidade de bactérias viáveis da microbiota que foi maior aos 90 dias (10,58 Log10 UFC/ml), quando as aves foram tratadas com sacarose, e menor aos 290 dias, quando tratadas com glicerol (7,73 Log10 UFC/ml). No tempo zero, todas as aves apresentaram Salmonella (100 por cento) quando tratadas com DMSO e glicerol, com colonização cecal de 4,9 e 5,2 Log10 UFC/g do conteúdo cecal, respectivamente; aos 360 dias nenhuma ave foi infectada, independente do tratamento. A microbiota cecal, independente de tratamento, sempre determinou menor quantidade de S. Enteritidis em qualquer um dos parâmetros pesquisados, quando comparada com a das aves não tratadas. O congelamento em nitrogênio líquido foi eficaz na manutenção da viabilidade da microbiota cecal até 360 dias.

One-day-old broiler chicks were treated with cecal microbiota cultivated under aerobiose conditions, frozen during 90, 200, 290 and 360 days and associated with different cryoprotectors such as sucrose, trehalose, DMSO and glycerol. Subsequently, the birds were challenged with Salmonella Enteritidis in order to determine the efficacy of the different treatments in relation to the quantity of viable bacteria, which was higher at 90 days when treated with sucrose (10.58 log10 CFU/ml) and lower at 290 days when treated with glycerol (7.73 log10 CFU/ml). The quantity of infected birds was 100 percent in 0 time, when the cecal colonization by S. Enteritidis was 4.9 and 5.2 log10 CFU/g of cecal content, respectively treated with DMSO and glycerol. No bird was infected at 360 days, irrespectively of the treatment. In all treatments, the cecal microbiota always determined a lesser quantity of S. Enteritidis for all the studied parameters compared to non-treated birds. Frozen in liquid nitrogen was effective in maintaining the viability of cecal microbiota during the experimental period of 360 days.

Bio-metabolic disposition of metanil yellow, orange II and their blend by caecal microflora of rats.

Caecal microflora were employed to study the degradation pattern, with time course of Metanil yellow and Orange II-two extensively used non-permitted food colours. Metanil yellow and Orange II showed the respective Degradation Index 50 (DI 50) values of 369 and 288 min. However, the blend of Metanil yellow and Orange II (1:1) resulted in the D1 50.value of 288 min. Metanil yellow, Orange II and their blend were resolved into respective metabolites in different solvent systems.

Virulence of Entamoeba histolytica according to the Strains in Korea II. Studies on the Pathogenicity of Entamoeba histolytica Strains in Rats

The experimental infection of rats with seven strains of Entamoeba histolytica were carried out according to animal ages, number of inoculated amebae, rat strain differences and rat-culture passages. The rat cecal scoring technique of Neal (1951) was utilized to measure the invasiveness of the parisite. The results are summarized and concluded as follows; 1. In the infection of Sprague-Dawley strain rat with YS9-strain and NAMRU II-stran amebae, which wem confirmed highly invasive to the membrane of cecum of rabbits in the previous reports (Cho,1968; Cross,1968), remarkab1e invasiveness was observed in the 30-day-old rat groups with the average cecal score above 5.0. Although no statistical differences of virulence by the number of inoculations showed in rat groups, the cecal scores were markedly reduced in the 50,000 amebae inoculated rats. 2. The hybrid albino rats were considered unsuitable for virulence study of E. histolytica, since the invasiveness of the amabae was inconsistant. 3. The virulences of YS 14 and YS16-strains from cyst carrier showed no virulence, YS 15 from cyst carrier and YS 24 from liver abscess were moderately invasive, and only YS25 from liver abscess showed highly invasive as with YS 9 and NAMRU II-strain amebae. By rat-culture passage, YS14-strain and YS24-strain amebae showed marked increase of invasiveness. It was presumed that the rat-culture passage should be indispensably supplememed in the studies on the virulence of E. histolytica.