Actinobacillosis outbreak in cattle with clinical manifestation of hippopotamus-like face / Actinobacilose em bovinos com manifestação clínica de cara de hipopótamo

Actinobacillosis outbreak with clinical manifestation of hippopotamus-like face observed in a property located in the municipality of Capão do Leão, Southern Brazil, in September 2016, is described. The cattle herd remained for most of the year in rice stubble. When these areas were occupied with new crops, they were transferred to areas where there were small native forests. Three cattle were affected. They presented a volume increase in the nasolabial and maxillary region, and there was also regional lymph node swelling. The evolution of the disease occurred in approximately six months. In tissue fragments collected for culture, Actinobacillus lignieresii was isolated. The diagnosis was based on clinical findings, histopathological evaluation characterized by the presence of piogranulomas with Splendore Hoepli reaction in its center, bacterial isolation, and identification of A. lignieresii by polymerase chain reaction (PRC) and genetic sequencing.(AU)

Descreve-se um surto de actinobacilose com manifestação clínica de cara de hipopótamo diagnosticado em uma propriedade localizada no município do Capão do Leão, Rio Grande do Sul em setembro de 2016. Os bovinos permaneciam durante a maior parte do ano em restevas de arroz e quando as áreas eram ocupadas com novas lavouras eram transferidos para áreas onde havia pequenas matas nativas. Foram afetados três bovinos adultos que apresentavam aumento de volume na região nasolabial e maxilar e havia, também, tumefação dos linfonodos regionais. A evolução da enfermidade era de aproximadamente seis meses. Nos fragmentos coletados para cultura houve isolamento de Actinobacillus lignieresii. O diagnóstico foi baseado nos achados clínicos, na avaliação histopatológica caracterizada pela presença de piogranulomas com reação de Splendori Hoepli no centro, no isolamento bacteriano, identificação de Actinobacillus lignieresii por reação em cadeia da polimerase (PRC) e sequenciamento genético.(AU)

Development of Actinobacillus pleuropneumoniae ApxI, ApxII, and ApxIII-specific ELISA methods for evaluation of vaccine efficiency

Among various vaccines against Actinobacillus pleuropneumoniae, subunit vaccines using recombinant proteins of ApxI, ApxII, and ApxIII as vaccine antigens have shown good efficacy in terms of safety and protection. Therefore, subunit vaccines are being applied worldwide and the development of new subunit vaccines is actively being conducted. To evaluate the efficacy of the subunit vaccines, it is important to measure immune responses to each Apx toxin separately. However, the cross-reactivity of antibodies makes it difficult to measure specific immune reactivity to each toxin. In the present study, specific antigen regions among the toxins were identified and cloned to solve this problem. The antigenicity of each recombinant protein was demonstrated by Western blot. Using the recombinant proteins, we developed enzyme-linked immunosorbent assay (ELISA) methods that can detect specific immune responses to each Apx toxin in laboratory guinea pigs. We suggest that the ELISA method developed in this study can be an important tool in the evaluation of vaccine efficiency and vaccine development.

In silico analysis of putative drug and vaccine targets of the metabolic pathways of Actinobacillus pleuropneumoniae using a subtractive/comparative genomics approach

Actinobacillus pleuropneumoniae is a Gram-negative bacterium that resides in the respiratory tract of pigs and causes porcine respiratory disease complex, which leads to significant losses in the pig industry worldwide. The incidence of drug resistance in this bacterium is increasing; thus, identifying new protein/gene targets for drug and vaccine development is critical. In this study, we used an in silico approach, utilizing several databases including the Kyoto Encyclopedia of Genes and Genomes (KEGG), the Database of Essential Genes (DEG), DrugBank, and Swiss-Prot to identify non-homologous essential genes and prioritize these proteins for their druggability. The results showed 20 metabolic pathways that were unique and contained 273 non-homologous proteins, of which 122 were essential. Of the 122 essential proteins, there were 95 cytoplasmic proteins and 11 transmembrane proteins, which are potentially suitable for drug and vaccine targets, respectively. Among these, 25 had at least one hit in DrugBank, and three had similarity to metabolic proteins from Mycoplasma hyopneumoniae, another pathogen causing porcine respiratory disease complex; thus, they could serve as common therapeutic targets. In conclusion, we identified glyoxylate and dicarboxylate pathways as potential targets for antimicrobial therapy and tetra-acyldisaccharide 4′-kinase and 3-deoxy-D-manno-octulosonic-acid transferase as vaccine candidates against A. pleuropneumoniae.

Atypical actinobacillosis in bulls in Argentina: granulomatous dermatitis and lymphadenitis / Actinobacilose atípica em touros na Argentina: dermatite granulomatosa e linfadenite

Actinobacillosis is a common cause of sporadic infection in cattle. It was mostly characterized as a pyogranulomatous inflammation of the tongue, but also soft tissues as lymph nodes, other digestive tract localization and skin. The aim of this study was to describe an episode of granulomatous dermatitis and lymphadenitis affecting a bull herd in Argentina during 2010. Actinobacillus lignieresii was isolated from samples collected from one of the affected bulls, and characteristic lesions were observed. Lesions other than 'wooden tongue' are usually uncommon; however, actinobacillosis should be included as a differential diagnosis for cutaneous diseases.

A actinobacilose é causa comum de infecções esporádicas em bovinos. Esta afeção tem sido caracterizada como uma infecção piogranulomatosa não somente da língua como também de tecidos moles tais como linfonodos, ou outras localizações no trato digestivo e na pele. O objetivo do presente trabalho é descrever um episódio de dermatite piogranulomatosa e linfadenite que afetou um rebanho de touros na Argentina em 2010. As amostras recolhidas de um dos animais afetados permitiram o isolamento de Actinobacillus lignieresii. Observaram-se as lesões características da doença. Habitualmente não são comuns outras lesões para além das descritas como "língua de pau", no entanto, a actinobacilose deve ser incluída como um possível diagnóstico diferencial de doenças cutâneas.

Progress in microbial production of succinic acid / 生物工程学报

Succinic acid is one of the key intermediates in the tricarboxylic acid cycle (TCA)and has huge potentials in biopolymer, food, medicine applications. This article reviews recent research progress in the production of succinic acid by microbial fermentation, including discovery and screening of the succinic-acid-producing microbes, the progress of genetic engineering strategy and metabolic engineering technology for construction of succinic acid-producing strains, and fermentation process control and optimization. Finally, we discussed the limitation of current progress and proposed the future research needs for microbial production of succinic acid.

Optimization of succinic acid fermentation with Actinobacillus succinogenes by response surface methodology / 生物工程学报

Succinic acid is an important C4 platform chemical in the synthesis of many commodity and special chemicals. In the present work, different compounds were evaluated for succinic acid production by Actinobacillus succinogenes GXAS 137. Important parameters were screened by the single factor experiment and Plackeet-Burman design. Subsequently, the highest production of succinic acid was approached by the path of steepest ascent. Then, the optimum values of the parameters were obtained by Box-Behnken design. The results show that the important parameters were glucose, yeast extract and MgCO3 concentrations. The optimum condition was as follows (g/L): glucose 70.00, yeast extract 9.20 and MgCO3 58.10. Succinic acid yield reached 47.64 g/L at the optimal condition. Succinic acid increased by 29.14% than that before the optimization (36.89 g/L). Response surface methodology was proven to be a powerful tool to optimize succinic acid production.

Serotype and antimicrobial susceptibility of Actinobacillus pleuropneumoniae isolates from pigs in Korea / 대한수의학회지

Actinobacillus (A.) pleuropneumoniae is the causative agent of pleuropneumonia which is one of the most important respiratory diseases in pigs worldwide. A total of 32 A. pleuropneumoniae isolates from diseased pigs during 2008 to 2010 were serotyped by polymerase chain reaction method. The susceptibility of the isolates to 13 antimicrobial agents were determined by disk diffusion test. In all the 32 isolates examined in this study, serotype 5 (16 isolates: 50%), 1 (7 isolates: 21.9%), 2 (5 isolates: 15.6%) and 12 (1 isolate: 3.1%) were found. Of all tested antimicrobial agents, resistance to oxytetracycline was found in 96.9% of isolates, followed by resistance to amikacin (81.2%), neomycin (68.7%), kanamycin (53.1%), penicillin (50.0%), gentamicin (43.7%), florfenicol (25.0%), ampicillin (18.7%), colistin (9.4%), trimethoprim/sulfamethoxazole, ceftiofur (8.3%), amoxicillin/clavulanic acid (3.1%) and enrofloxacin (0%). Oxytetracycline or florfenicol-resistant isolates were examined for the presence of resistance gene. Among the 31 oxytetracycline-resistant isolates, tetB, tetH and tetO genes were detected in 22 (71%), 8 (26%) and 1 (3%) isolates, respectively. The floR genes were detected in 8 (100%) of the 8 florfenicol-resistant A. pleuropneumoniae isolates.

Effect of actinobacillus actinomycetem on the secretion of interleukin-6 and apoptosis rate of polymorphonuclear leukocyte in type 2 diabetes patients / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effect of Actinobacillus actinomycetem (A. actinomycetem) on the secretion and apoptosis rate of polymorphonuclear leukocytes (PMN) in type 2 diabetes patients.</p><p><b>METHODS</b>Peripheral PMN from healths and type 2 diabetes patients were isolated by Percoll gradient centrifugation. The PMN were stimulated with filtrate of ultrasonic pulverization from A. actinomycetem as the experiment group. As the control group, PMN suspension was incubated with PBS. The release of interleukin-6 (IL-6) was measured at 20, 40, 60 min by enzymelinked immune sorbent assay (ELISA) technique. The apoptosis rate of PMN was tested at 6 and 12 hours by flow cytometry.</p><p><b>RESULTS</b>Incubated with filtrate of ultrasonic pulverization from A. actinomycetem, the PMN of type 2 diabetes patients released significantly higher levels of IL-6 compared with the healthy subjects (P < 0.001). The apoptosis rate of PMN from the healthy subjects was higher than that from type 2 diabetes patients (P < 0.001). Regardless of body condition, interaction with filtrate of ultrasonic pulverization from A. actinomycetem could induce the seretion of IL-6 and reduce the apoptosis rate.</p><p><b>CONCLUSION</b>The PMN of type 2 diabetes patients may possess hyper-reactive inflammatory response trait.</p>

Breeding of ammonium-tolerant mutants of Actinobacillus succinogenes for succinic acid production and effect of ammonium / 生物工程学报

An ammonium-tolerant mutant of Actinobacillus succinogenes, YZ25, was obtained in the medium containing 61-242 mmol/L NH4+ after DES mutagenesis. Succinic acid produced by the mutant YZ25 reached 32.68 g/L when the medium contains 50 g/L glucose and 121 mmol/L ammonium, which was increased by 180.5% compared with that of the parent strain. The effects of different ammonium salts on the growth of the mutant and its metabolic response to high ammonium concentrations were investigated. The results showed that low ammonium concentration could improve the specific growth rates of the mutants, while high ammonium concentration inhibited cell growth. The ammonia-nitrogen half-inhibition constants (Ki) for different ammonium salts were as follows: 215 mmol/L for (NH4)2SO4, 265 mmol/L for NH4HCO3, 235 mmol/L for NH4Cl, and 210 mmol/L for NH4NO3. The process of ammonium inhibition on the mutant YZ25 was investigated in 3.0 L stirred fermenter. When NH4OH was used to buffer the pH, cell growth was not inhibited. However, production of succinic acid and consumption of glucose gradually decreased when cells entered the stationary phase, and the glucose could not be utilized completely at the end of fermentation. The possible ammonium inhibition mechanism was discussed based on the metabolic pathway of A. succinogenes.

Effect of adding intermediate metabolites on succinate production by Actinobacillus succinogenes / 生物工程学报

We investigated the effect of adding intermediate metabolites on cell growth and succinate production. The yield of succinic acid achieved to the highest when 0.5 g/L phosphoenolpyruvic acid (PEP) was added. According to the metabolic network of Actinobaccilus succinogenes NJ113, the metabolic flux was calculated by metabolic flux analysis. The ratio of hexose monophosphate pathway to glycolytic pathway increased from 39.4:60.3 to 76.8:22.6 after adding 0.5 g/L PEP, thus the reducing power was better balanced. The flux of PEP to oxaloacetate was 23.8% higher, which made the succinic acid flux improve from 99.8 mmol/(g DCW x h) to 124.4 mmol/(g DCW x h) and the flux of acetic acid and formic acid decreased by 22.9% and 15.4%, respectively. The key enzyme activity analysis showed that the specific activity of PEP carboxykinase reached to 1910 U/mg with 0.5 g/L PEP addition, which was 74.7% higher than the control; and the specific activity of pyruvate kinase decreased by 67.5%. Finally, the concentration of succinic acid was 29.1 g/L with the yield of 76.2%.

Recycle of spent cells from anaerobic succinate fermentation / 生物工程学报

Spent cells recovered from anaerobic fermentation by Actinobacillus succinogenes were used as nitrogen source for succinic acid production. Three methods were investigated for cell wall-breaking. The results showed that enzymatic hydrolysis was more effective for higher succinic acid yield. When the enzymatic hydrolysate of spent cells was added to reach a total nitrogen concentration 1.11 g/L (equivalent to 10 g/L yeast extract), the succinic acid concentration was 42.0 g/L, but it increased slightly when enhancing the level of enzymatic hydrolysate. However, when 5 g/L yeast extract was supplemented with the enzymatic hydrolysate of spent cells, the succinic acid concentration reached 75.5 g/L after 36 hours and, the succinic acid productivity was 2.10 g/(L x h), which increased by 66.7% compared with the fermentation using 10 g/L yeast extract. Therefore, enzymatic hydrolysate of spent cells could replace 50% yeast extract in the original medium for succinic acid production.

Effects of different neutralizing agents on succinate production by Actinobacillus succinogenes NJ113 / 生物工程学报

Different neutralizing agents were used as pH controller to investigate their effects on the growth and succinic acid production of Actinobacillus succinogenes NJ113. The fermentation results showed that Ca(OH)2, CaCO3 and NH4OH were not suitable for succinic acid production by A. succinogenes NJ113 because of their negative effects on cell growth. When Na-base was used, cells would flocculate and lump, and due to the sodium ion concentration reaching to a high level, OD660 dropped sharply after 12 h of fermentation. Mg-base was better because there was no significant inhibition by magnesium ion. Two combined neutralizing agents were used to maintain pH level, one with NaOH and Mg(OH)2 while the other with Na2CO3 and Mg(OH)2. The optimum ratios of the combined neutralizing agents were both 1:1 (g:g) when using 100 g/L glucose. When NaOH and Mg(OH)2 were chosen with the ratio of 1:1(g:g), 69.8 g/L of the succinic acid and 74.5% of the yield was obtained.

The First Case of Psoas Muscle Abscess and Sepsis Caused by Actinobacillus ureae in a Chronic Hepatitis B Patient in Korea

Actinobacillus ureae, formerly known as Pasteurella ureae, is a rare human pathogen. Twenty-eight cases of A. ureae infections in humans have been reported since its first description in 1960. Various predisposing conditions such as skull fracture, alcohol abuse, neurosurgery, schizophrenia, odontal infection, diabetes, HIV infection/AIDS, Waldenstrom macroglobulinemia, COPD, malnutrition, rheumatoid arthritis, HCV hepatitis, etanercept, or methotrexate have been associated with infections caused by A. ureae. We report the first case, in the medline-based literature, of A. ureae psoas muscle abscess and sepsis in a HBV carrier patient.

The First Case of Psoas Muscle Abscess and Sepsis Caused by Actinobacillus ureae in a Chronic Hepatitis B Patient in Korea

Actinobacillus ureae, formerly known as Pasteurella ureae, is a rare human pathogen. Twenty-eight cases of A. ureae infections in humans have been reported since its first description in 1960. Various predisposing conditions such as skull fracture, alcohol abuse, neurosurgery, schizophrenia, odontal infection, diabetes, HIV infection/AIDS, Waldenstrom macroglobulinemia, COPD, malnutrition, rheumatoid arthritis, HCV hepatitis, etanercept, or methotrexate have been associated with infections caused by A. ureae. We report the first case, in the medline-based literature, of A. ureae psoas muscle abscess and sepsis in a HBV carrier patient.

An atypical case of respiratory actinobacillosis in a cow

A not pregnant 4-year-old Jersey cow was presented with the sudden appearance of respiratory noise, nasal discharge and moderate respiratory difficulty. Upon physical examination a snoring-like noise, extended head and neck position, exaggerated abdominal effort, bilateral nasal discharge and left prescapular lymph node enlargement were noted. Sub-occlusion of the initial portion of the respiratory tract was suspected. Radiographic and endoscopic examinations revealed a pedunculate mass on the dorsal aspect of the rhinopharynx, which was removed with endoscopically assisted electrosurgery. Histologic examination revealed a chronic pyogranulomatous inflammation with eosinophilic club-like bodies surrounding small colonies of rod-shaped bacteria. Results of histochemical staining were consistent with Actinobacillus-like bacteria and a diagnosis of respiratory actinobacillosis was reached. Surgery and antibiotic therapy were resolutive, as demonstated by an endoscopic check at the second month after surgery, even without the association of the traditional iodine cure, which is regarded as the treatment of choice for actinobacillosis.

Environmental factors affecting the succinic acid production by Actinobacillus succinogenes CGMCC 1593 / 生物工程学报

Actinobacillus succinogenes is a promising candidate for the production of bio-based succinic acid. Previously, we isolated a succinic acid-producing strain Actinobacillus succinogenes CGMCC 1593 from bovine rumen. In this paper, the influence of the environmental factors such as gas phase, pH, ORP, on succinic acid production by A. succinogenes CGMCC 1593 was studied. The results showed that CO2 was the optimum gas phase for anaerobic fermentation ofA. succinogenes CGMCC 1593 as well as one of the substrate for the succinic acid synthesis. Using MgCO3 as a pH regulator, the pH was maintained within 7.1-6.2 during the anaerobic fermentation for the cell growth and acid production of A. succinogenes CGMCC 1593. Our results showed that low initial ORP was disadvantageous for the growth of A. succinogenes CGMCC 1593 and an ORP of -270 mV was demonstrated to be beneficial to the succinic acid production. By adding Na2S.9H2O to decrease ORP to -270 mV at the end of exponential growth phase in batch culture of A. succinogenes CGMCC 1593, the succinic acid concentration reached 37 g/L and the yield of succinic acid was 129% at 48 h. This work might provide valuable information for further optimization of succinic acid fermentation by A. succinogenes CGMCC 1593.

Breeding of monofluoroacetate-resistant strains of Actinobacillus succinogenes and the mechanism based on metabolic flux analysis / 生物工程学报

Succinic acid has received a great deal of attention as an important green chemical stock for the manufacture of synthetic resins, biodegradable polymers and chemical intermediates. In this paper, the breeding mechanism of Actinobacillus succinogenes based on metabolic flux analysis was demonstrated to improve the yield of succinic acid by fermentation. After the NTG treatment, mutants from A. succinogenes CGMCC 1593 which were able to grow in medium containing concentrations of about 50-100 mmol/L of sodium monofluoroacetate were obtained. Among them, a mutant SF-9 was selected for producing more succinic acid and less acetic acid. When fermentations were conducted in a 5 L bioreactors, the final succinic acid concentration of SF-9 (34.8 g/L) increased 23.4%, and the mass ratio of succinic acid/acetic acid increased from 3.3 to 9 compared with those of the parent strain. Based on the metabolic flux analysis of A. succinogenes, PEP was found to be a key node which has an important effect on the production of succinic acid, and the flux ratio of by-productions (acetic, formic, lactic acid) was influenced by PYR node. Compared with the parent strain, the flux to succinic acid of mutant (A. succinogenes SF-9) was significantly increased, while the flux to by-productions had an obvious decline. Therefore, PEP and PYR are not rigid nodes in the metabolic regulation of A. succinogenes.

Breeding of Actinobacillus succiniogenes mutants with improved succinate production based on metabolic flux analysis / 生物工程学报

It is very important to obtain high yield mutant strains on the base of metabolic flux analysis of Actinobacillus succinogenes S.JST for the industrial bioconversion of succinic acid. The metabolic pathway was analized at first and the flux of the metabolic networks was calculated by matrix. In order to decrease acetic acid flux, the strains mutated by soft X-ray of synchronous radiation were screened on the plates with high concentration of fluoroacetic acid. For decreasing the metabolic flux of ethanol the site-directed mutagenesis was carried out for the reduction of alcohol dehydrogenase(Adh) specific activity. Then the enzyme activity determination and the gene sequence analysis of the mutant strain was compared with those of the parent strain. Metabolic flux analysis of the parent strain indicated that the flux of succinic acid was 1.78(mmol/g/h) and that the flux of acetic acid and ethanol were 0.60 (mmol/g/h) and 1.04( mmol/g/h), respectively. Meanwhile the metabolic pathway analysis showed that the ethanol metabolism enhanced the lacking of H electron donor during the synthesis of succinic acid and that the succinic acid flux was weakened by the metabolism of byproducts ethanol and acetic acid. Compared with the parent strain, the acetic acid flux of anti-fluoroacetic mutant strain S.JST1 was 0.024 (mmol/g/h), decreasing by 96%. Then the enzyme determination showed that the specific activity unit of phosphotransacetylase(Pta) decreased from 602 to 74 and a mutated site was founded in the pta gene of the mutant strain S.JST1. Compared with that of the parent strain S.JST1 the ethanol flux of adh-site-directed mutant strain S.JST2 was 0.020 (mmol/g/h), decreasing by 98%. Then the enzyme determination showed that the specific activity unit of Adh decreased from 585 to 62 and the yield of end product succinic acid was 65.7 (g/L). The interdiction of Adh and Pta decreased the metabolism of byproducts and the H electron donor was well balanced, thus the succinic acid flux was strengthened by the redundant carbon flux from these byproducts. The mutant strain S.JST2 obtained in this paper deserves being extended to application of industrial fermentation.

The research progress of succinic acid fermentation strains / 生物工程学报

The potential of succinic acid as an important chemical intermediates had been realized and fermentation is one of the best ways to make it possible in economical aspect. Fermentation organism is the key part of the fermentation method. The updated research developments of fermentation organisms and the fermentation characteristics and problems of them were reviewed and analyzed in this paper. Finally,the development future of fermenation organism was forecasted.

Influence of the Sonic Power Toothbrush on Reduction of Gingival inflammation and on the Amount of Interleukin-6, Prevotella intermedia and Actinobacillus actinomycetemcomitans in Periodontal Pocket / 대한치주과학회지

No abstract available.