Aeromonas is a Gram-negative bacterium that lives in aquatic habitats. It can be infective in humans. One of its remarkable attributes is the ability of biofilm formation. Many factors are involved in the construction...Aeromonas is a Gram-negative bacterium that lives in aquatic habitats. It can be infective in humans. One of its remarkable attributes is the ability of biofilm formation. Many factors are involved in the construction of biofilms as has been described for Pseudomonas, Klebsiella, and Vibrio, among others. The aim of this work was to study the bacterial morphology during the establishment of biofilm through scanning electron microscopy (SEM) and transmission electron microscopy (TEM) with a modified microtiter plate assay and to determine the best conditions for the establishment of Aeromonas caviae Sch3 biolfilm in vitro. We observed several phenotypic changes, including surface appearance, size, presence of extracellular vesicles from 100 to 250 nm in diameter, and flagella. The best conditions for biofilm formation were to grow cultures at 28℃ at pH 6, as determined by the crystal violet assay. This is, to the best of our knowledge, the first study that describes the cell’s biological events involved in the establishment of biofilm formation of Aeromonas caviae Sch3 in vitro.展开更多
Recently, the rarely reported tet(31) tetracycline resistance determinant was commonly found in Aeromonas salmonicida, Gallibacterium anatis, and Oblitimonas alkaliphila isolated from farming animals and related envir...Recently, the rarely reported tet(31) tetracycline resistance determinant was commonly found in Aeromonas salmonicida, Gallibacterium anatis, and Oblitimonas alkaliphila isolated from farming animals and related environment. However, its distribution in other bacteria and potential molecular dissemination mechanism in environment are still unknown. The purpose of this study was to investigate the potential mechanism underlying dissemination of tet(31) by analysing the tet(31)-carrying fragments in A. caviae strains isolated from an aerobic biofilm reactor treating oxytetracycline bearing wastewater. Twenty-three A. caviae strains were screened for the tet(31) gene by polymerase chain reaction(PCR). Three strains(two harbouring tet(31), one not) were subjected to whole genome sequencing using the PacBio RSII platform. Seventeen A. caviae strains carried the tet(31) gene and exhibited high resistance levels to oxytetracycline with minimum inhibitory concentrations(MICs)ranging from 256 to 512 mg/L. tet(31) was comprised of the transposon Tn6432 on the chromosome of A. caviae, and Tn6432 was also found in 15 additional tet(31)-positive A. caviae isolates by PCR. More important, Tn6432 was located on an integrative conjugative element(ICE)-like element, which could mediate the dissemination of the tet(31)-carrying transposon Tn6432 between bacteria. Comparative analysis demonstrated that Tn6432 homologs with the structure ISCR2-ΔphzF-tetR(31)-tet(31)-ΔglmM-sul2 were also carried by A. salmonicida, G. anatis, and O. alkaliphila, suggesting that this transposon can be transferred between species and even genera. This work provides the first report on the identification of the tet(31) gene in A. caviae, and will be helpful in exploring the dissemination mechanisms of tet(31) in water environment.展开更多
This study investigates the gastroprokinetic effects of motilin and erythromycin A （EM-A） and its potential mechanism in guinea pigs Carla porcellus in vitro. Guinea pig stomach strips were mounted under organ baths...This study investigates the gastroprokinetic effects of motilin and erythromycin A （EM-A） and its potential mechanism in guinea pigs Carla porcellus in vitro. Guinea pig stomach strips were mounted under organ baths containing Krebs solution. Motilin, EM-A, Nω-Nitro-L-arginine （L-NNA）, L-arginine （L-AA） were added to the bathing solution in a non-cumulative way. Then the effects of motilin and EM-A was studied during electrical field stimulation （EFS） in the absence and presence of L-NNA and L-AA in the gastric anturm and fundus of guinea pigs. In addition, we observed the co-expression of motilin receptors and neuronal nitric oxide synthase （nNOS） in the gastric myenteric plexus of guinea pigs by fluo-immunohistochemistry. The results showed that the circular muscle tissues of the gastric fundus generated on-relaxations and off-contractions with the frequency of 1 - 16 Hz. The on-responses induced a relaxation, partially mediated by the release of nitric oxide （NO） because addition of L-NNA turned the relaxations into cholinergically mediated contractions. The off-contractions were also cholinergically mediated as they disappeared under non-adrenergic, non-cholinergic （NANC） conditions using atropine and guanethidine. In fundic strips, motilin and EM-A induced on-relaxation and off-contraction and both motilin （1 μmol/L） and EM-A （100 μmol/L） may significantly increased on-response and reduced off-response （P 〈 0.05）. And the effects of motilin on strip responses were significantly enhanced compared with EM-A. The on-responses could be reversed into a cholinergically mediated contraction by addition of NOS inhibitors L-NNA. In contrast, administration of substrate of NOS, L-AA, significantly increased on-relaxations and reduced cholinergic motor responses which were induced by motilin or EM-A. However D-arginine （D-AA） did not change the above response induced by motilin or EM-A. In the antral strips, motilin and EM-A only increased off-contractions. The response to motilin and EM-A in the presence of L-NNA did differ from that obtained with L-NNA alone. It showed that both motilin and EM- A could enhance off-contractions induced by L-NNA （P 〈 0.05）. Immunohistochemistry study showed that motilin receptor immunoreactive positive neurons were co-localized with nNOS positive neurons in the gastric myenteric plexus of the guinea pigs. These results suggested that motilin or EM-A modulates gastrointestinal motility which was mediated by activating gastric nervous and NO pathways in guinea pigs gastro-intestinal tract.展开更多
A subacute toxicity study of a potent tetanus toxoid (250 Lf) was carried out in guinea pigs.The toxoid was injected subcutaneously at the nape of the neck at dose levels of 1.0, 1.5.and 2.0 ml in Groups Ⅱ. Ⅲ. and ...A subacute toxicity study of a potent tetanus toxoid (250 Lf) was carried out in guinea pigs.The toxoid was injected subcutaneously at the nape of the neck at dose levels of 1.0, 1.5.and 2.0 ml in Groups Ⅱ. Ⅲ. and Ⅳ, respectively.In the controls (Group Ⅰ) 2.0 ml of aluminum phosphate suspension was given in each injection.Periodic evaluations of body weight, food/water intake, general observable behavior, hematology.and blood chemistry in toxoid-injected guinea pigs were similar to those in control guinea pigs.Thus, the toxoid did not cause any side effects up to four times the dose proposed for humans.1990 Academic Press, Inc.展开更多
文摘Aeromonas is a Gram-negative bacterium that lives in aquatic habitats. It can be infective in humans. One of its remarkable attributes is the ability of biofilm formation. Many factors are involved in the construction of biofilms as has been described for Pseudomonas, Klebsiella, and Vibrio, among others. The aim of this work was to study the bacterial morphology during the establishment of biofilm through scanning electron microscopy (SEM) and transmission electron microscopy (TEM) with a modified microtiter plate assay and to determine the best conditions for the establishment of Aeromonas caviae Sch3 biolfilm in vitro. We observed several phenotypic changes, including surface appearance, size, presence of extracellular vesicles from 100 to 250 nm in diameter, and flagella. The best conditions for biofilm formation were to grow cultures at 28℃ at pH 6, as determined by the crystal violet assay. This is, to the best of our knowledge, the first study that describes the cell’s biological events involved in the establishment of biofilm formation of Aeromonas caviae Sch3 in vitro.
基金supported by the National Natural Scientific Foundation of China (No.21437005)
文摘Recently, the rarely reported tet(31) tetracycline resistance determinant was commonly found in Aeromonas salmonicida, Gallibacterium anatis, and Oblitimonas alkaliphila isolated from farming animals and related environment. However, its distribution in other bacteria and potential molecular dissemination mechanism in environment are still unknown. The purpose of this study was to investigate the potential mechanism underlying dissemination of tet(31) by analysing the tet(31)-carrying fragments in A. caviae strains isolated from an aerobic biofilm reactor treating oxytetracycline bearing wastewater. Twenty-three A. caviae strains were screened for the tet(31) gene by polymerase chain reaction(PCR). Three strains(two harbouring tet(31), one not) were subjected to whole genome sequencing using the PacBio RSII platform. Seventeen A. caviae strains carried the tet(31) gene and exhibited high resistance levels to oxytetracycline with minimum inhibitory concentrations(MICs)ranging from 256 to 512 mg/L. tet(31) was comprised of the transposon Tn6432 on the chromosome of A. caviae, and Tn6432 was also found in 15 additional tet(31)-positive A. caviae isolates by PCR. More important, Tn6432 was located on an integrative conjugative element(ICE)-like element, which could mediate the dissemination of the tet(31)-carrying transposon Tn6432 between bacteria. Comparative analysis demonstrated that Tn6432 homologs with the structure ISCR2-ΔphzF-tetR(31)-tet(31)-ΔglmM-sul2 were also carried by A. salmonicida, G. anatis, and O. alkaliphila, suggesting that this transposon can be transferred between species and even genera. This work provides the first report on the identification of the tet(31) gene in A. caviae, and will be helpful in exploring the dissemination mechanisms of tet(31) in water environment.
基金supported by the National Natural Science Foundation of China(No.30470642 and No.30670780)Shandong Province Tackle Key Problems in Science and Technology Program( No.2008GG10002006 )+1 种基金Shandong Province Health Department ( No.2007HZ026)Qingdao Municipal Science and Technology Commission (No.05-1-JC-93)
文摘This study investigates the gastroprokinetic effects of motilin and erythromycin A （EM-A） and its potential mechanism in guinea pigs Carla porcellus in vitro. Guinea pig stomach strips were mounted under organ baths containing Krebs solution. Motilin, EM-A, Nω-Nitro-L-arginine （L-NNA）, L-arginine （L-AA） were added to the bathing solution in a non-cumulative way. Then the effects of motilin and EM-A was studied during electrical field stimulation （EFS） in the absence and presence of L-NNA and L-AA in the gastric anturm and fundus of guinea pigs. In addition, we observed the co-expression of motilin receptors and neuronal nitric oxide synthase （nNOS） in the gastric myenteric plexus of guinea pigs by fluo-immunohistochemistry. The results showed that the circular muscle tissues of the gastric fundus generated on-relaxations and off-contractions with the frequency of 1 - 16 Hz. The on-responses induced a relaxation, partially mediated by the release of nitric oxide （NO） because addition of L-NNA turned the relaxations into cholinergically mediated contractions. The off-contractions were also cholinergically mediated as they disappeared under non-adrenergic, non-cholinergic （NANC） conditions using atropine and guanethidine. In fundic strips, motilin and EM-A induced on-relaxation and off-contraction and both motilin （1 μmol/L） and EM-A （100 μmol/L） may significantly increased on-response and reduced off-response （P 〈 0.05）. And the effects of motilin on strip responses were significantly enhanced compared with EM-A. The on-responses could be reversed into a cholinergically mediated contraction by addition of NOS inhibitors L-NNA. In contrast, administration of substrate of NOS, L-AA, significantly increased on-relaxations and reduced cholinergic motor responses which were induced by motilin or EM-A. However D-arginine （D-AA） did not change the above response induced by motilin or EM-A. In the antral strips, motilin and EM-A only increased off-contractions. The response to motilin and EM-A in the presence of L-NNA did differ from that obtained with L-NNA alone. It showed that both motilin and EM- A could enhance off-contractions induced by L-NNA （P 〈 0.05）. Immunohistochemistry study showed that motilin receptor immunoreactive positive neurons were co-localized with nNOS positive neurons in the gastric myenteric plexus of the guinea pigs. These results suggested that motilin or EM-A modulates gastrointestinal motility which was mediated by activating gastric nervous and NO pathways in guinea pigs gastro-intestinal tract.
文摘A subacute toxicity study of a potent tetanus toxoid (250 Lf) was carried out in guinea pigs.The toxoid was injected subcutaneously at the nape of the neck at dose levels of 1.0, 1.5.and 2.0 ml in Groups Ⅱ. Ⅲ. and Ⅳ, respectively.In the controls (Group Ⅰ) 2.0 ml of aluminum phosphate suspension was given in each injection.Periodic evaluations of body weight, food/water intake, general observable behavior, hematology.and blood chemistry in toxoid-injected guinea pigs were similar to those in control guinea pigs.Thus, the toxoid did not cause any side effects up to four times the dose proposed for humans.1990 Academic Press, Inc.
