Absceso en dedo por Escherichia vulneris tras traumatismo con una rama

Introducción: Es importante considerar la posible implicación de microorganismos poco frecuentes en infecciones de piel y partes blandas si la muestra obtenida para cultivo es de buena calidad, y además se aísla en cultivo puro, como ocurre con Escherichia vulneris. Caso Clínico: Presentamos en caso de una mujer de 34 años, sin antecedentes mórbidos, quien desarrolló un absceso en el 4° dedo de la mano tras un traumatismo con una rama y que requirió drenaje quirúrgico y tratamiento antibiótico para su resolución. En el contenido del absceso, se aisló E. vulneris en cultivo puro, con un perfil antibiótico multisensible. Discusión y Conclusión: E. vulneris es una enterobacteria cuya patogenicidad ha estado clásicamente discutida, pero que se ha visto que puede tener participación en infección de heridas, especialmente aquellas relacionadas con material vegetal. Este microorganismo, muy relacionado con E. harmannii, presenta buena sensibilidad a los aminoglucósidos, con excepción a la penicilina y al cotrimoxazol. En las infecciones de piel y tejidos blandos causadas por E. vulneris y que cursen como un absceso, es importante realizar desbridamiento quirúrgico, si es necesario para la resolución completa del cuadro, además del tratamiento con amoxicilina/ácido clavulánico que parece adecuado.

Introduction: It is important to assess the possible involvement of rare microorganisms in skin and soft tissue infections if the sample obtained for culture is of good quality, and is isolated in pure culture, as occurs with Escherichia vulneris. Case Report: We present the case of a 34-year-old woman, with no history of morbidity, who developed an abscess in the 4th finger of the hand after trauma with a branch and which required surgical drainage and antibiotic treatment for its resolution. In the content of the abscess, E. vulneris was isolated in pure culture, with a multisensitive antibiotic profile. Discusion: E. vulneris is an Enterobacteriaceae whose pathogenicity has been classically discussed, but it has been seen that it may have participated in the infection of wounds, especially those related to plant material. This organism, closely related to E. harmannii, shows good sensitivity to aminoglycosides, with the exception of penicillin, and cotrimoxazole. In skin and soft tissue infections caused by E. vulneris and that present as an abscess, it is important to perform surgical debridement if necessary for complete resolution of the condition, in addition to treatment with amoxicillin/clavulanic acid, which seems appropriate.

Caracterización de Escherichia coli D7111 productora de ß-LACTAMASA TEM-176 / Characterization of Escherichia coli D7111 producing the ß-LACTAMASA TEM-176

RESUMEN El presente reporte es la descripción original de bla TEM-176. Se caracterizaron los mecanismos de resistencia a antimicrobianos de un aislamiento de Escherichia coli enterotoxigénica, determinándose la resistencia a 22 antimicrobianos categorizados en 15 grupos diferentes mediante difusión en agar, estableciéndose grupo filogenético, mecanismos de resistencia y presencia de integrones de Clase 1 y 2 mediante PCR. Integrones y genes de resistencia a β-lactámicos fueron secuenciados. El aislamiento del grupo filogenético A, mostró resistencia o sensibilidad disminuida a ampicilina, amoxicilina más ácido clavulánico, ácido nalidíxico, ciprofloxacino, estreptomicina, kanamicina, tetraciclina, trimetoprim, sulfisoxazol, cotrimoxazol, azitromicina y nitrofurantoina, detectándose la presencia de bla TEM, aadA1/2, aphA1, sul3, tet(A) y un integron de Clase 2 conteniendo un gen dfrA1. La resistencia a quinolonas se relacionó con la substitución Ser83Ala. La secuencia de TEM mostró la substitución Ala222Val, la cual a la fecha no había sido descrita, reportándose como una nueva β-lactamasa, con el nombre de bla TEM-176.

ABSTRACT The present report is the original description of bla TEM-176. The mechanisms of resistance to antimicrobial agents were determined in an enterotoxigenic Escherichia coli, determining the susceptibility to 22 antimicrobials classified in 15 different groups by agar diffusion and establishing the phylogenetic group, mechanisms of resistance and presence of Class 1 and 2 integrons. Integrons and β-lactam resistance genes were sequenced. The isolate, belonging to phylogenetic group A, showed the presence of resistance or diminished susceptibility to a ampicillin, amoxicillin plus clavulanic acid, nalidíxic acid, ciprofloxacin, streptomycin, kanamycin, tetracycline, trimethoprim, sulfisoxazole, cotrimoxazole, azithromycin and nitrofurantoin, carrying bla TEM, aadA1/2, aphA1, sul3, tet(A) and a Class 2 integron containing a dfrA1 gene. Quinolone resistance was related to the substitution Ser83Ala. The TEM sequencing showed the presence of the new substitution Ala222Val, which led to the description of the new β-lactamase bla TEM-176.

Caracterización de Escherichia coli D7111 productora de ß-LACTAMASA TEM-176 / Characterization of Escherichia coli D7111 producing the ß-LACTAMASE TEM-176

RESUMEN El presente reporte es la descripción original de bla TEM-176. Se caracterizaron los mecanismos de resistencia a antimicrobianos de un aislamiento de Escherichia coli enterotoxigénica, determinándose la resistencia a 22 antimicrobianos categorizados en 15 grupos diferentes mediante difusión en agar, estableciéndose grupo filogenético, mecanismos de resistencia y presencia de integrones de Clase 1 y 2 mediante PCR. Integrones y genes de resistencia a β-lactámicos fueron secuenciados. El aislamiento del grupo filogenético A, mostró resistencia o sensibilidad disminuida a ampicilina, amoxicilina más ácido clavulánico, ácido nalidíxico, ciprofloxacino, estreptomicina, kanamicina, tetraciclina, trimetoprim, sulfisoxazol, cotrimoxazol, azitromicina y nitrofurantoina, detectándose la presencia de bla TEM, aadA1/2, aphA1, sul3, tet(A) y un integron de Clase 2 conteniendo un gen dfrA1. La resistencia a quinolonas se relacionó con la substitución Ser83Ala. La secuencia de TEM mostró la substitución Ala222Val, la cual a la fecha no había sido descrita, reportándose como una nueva β-lactamasa, con el nombre de bla TEM-176.

ABSTRACT The present report is the original description of bla TEM-176. The mechanisms of resistance to antimicrobial agents were determined in an enterotoxigenic Escherichia coli, determining the susceptibility to 22 antimicrobials classified in 15 different groups by agar diffusion and establishing the phylogenetic group, mechanisms of resistance and presence of Class 1 and 2 integrons. Integrons and β-lactam resistance genes were sequenced. The isolate, belonging to phylogenetic group A, showed the presence of resistance or diminished susceptibility to a ampicillin, amoxicillin plus clavulanic acid, nalidíxic acid, ciprofloxacin, streptomycin, kanamycin, tetracycline, trimethoprim, sulfisoxazole, cotrimoxazole, azithromycin and nitrofurantoin, carrying bla TEM, aadA1/2, aphA1, sul3, tet(A) and a Class 2 integron containing a dfrA1 gene. Quinolone resistance was related to the substitution Ser83Ala. The TEM sequencing showed the presence of the new substitution Ala222Val, which led to the description of the new β-lactamase bla TEM-176.

Simultaneous determination of amoxicillin and clavulanic acid by LC-MS/MS in human plasma and its application to a bioequivalence study / 中国药科大学学报

@#In this paper, we developed an accurate and sensitive LC-MS/MS method for the determination of amoxicillin and clavulanic acid in human plasma. A 50% aqueous acetic acid solution was used as a stabilizer, and the plasma samples were evaporated to dryness and resolved after protein precipitation on ice bathing and then were placed in an autosampler for injection. The gradient was eluted by Hedera ODS-2 column(2. 1 mm×150 mm). The aqueous phase was an aqueous solution containing 0. 2% acetic acid. The organic phase was methanol. The amoxicillin and clavulanic acid were detected under negative ion detection with electrospray ionization(ESI)in multiple reaction monitoring(MRM)mode of m/z 364. 1→223. 1 and 198. 1→135. 9 in the triple quagdrupole tandem mass spectrometer(Triple Quad TM 6500+). The concentration ranges of plasma from 20. 0 ng/mL to 5 000 ng/mL for amoxicillin and 10. 0 ng/mL to 2 500 ng/mL for clavulanic acid were good linear relationship. The accuracy deviation were ±15. 0% and precision were less than 15. 0% for the intra-assay and inter-assay. The matrix effect and recovery meeted the acceptance criteria, amoxicillin and clavulanic acid were stable under storage and processing conditions. Healthy subjects were given a test preparation of amoxicillin and clavulanate potassium granules 1 bag(125 mg/31. 25 mg/bag)and the reference preparation amoxicillin clavulanate potassium dry mix Suspension “Augmentin® ” 5 mL(125 mg/31. 25 mg/5 mL)was used to determine the plasma concentration of amoxicillin and clavulanic acid. The Phoenix WinNonlin 6. 4 software was used to estimate the pharmacokinetic parameters of non-compartmental models. The pharmacokinetic parameters of amoxicillin and clavulanic acid were statistically calculated and evaluated the bioequivalence. what′s more, we evaluated the diet on the pharmacokinetics of amoxicillin and clavulanic acid. The analytical method was rapid and sensitive, which was successfully employed in the bioequivalence study of amoxicillin(125 mg/bag)and clavulanate potassium granules(31. 25 mg/bag)for determining the concentration of amoxicillin and clavulanic acid.

Screening of medium constituents for clavulanic acid production by Streptomyces clavuligerus

ABSTRACT Clavulanic acid is a β-lactam compound with potent inhibitory activity against β-lactamases. Studies have shown that certain amino acids play essential roles in CA biosynthesis. However, quantitative evaluations of the effects of these amino acids are still needed in order to improve CA production. Here, we report a study of the nutritional requirements of Streptomyces clavuligerus for CA production. Firstly, the influence of the primary nitrogen source and the salts composition was investigated. Subsequently, soybean protein isolate was supplemented with arginine (0.0-3.20 g L-1), threonine (0.0-1.44 g L-1), ornithine (0.0-4.08 g L-1), and glutamate (0.0-8.16 g L-1), according to a two-level central composite rotatable design. A medium containing ferrous sulfate yielded CA production of 437 mg L-1, while a formulation without this salt produced only 41 mg L-1 of CA. This substantial difference suggested that Fe2+ is important for CA biosynthesis. The experimental design showed that glutamate and ornithine negatively influenced CA production while arginine and threonine had no influence. The soybean protein isolate provided sufficient C5 precursor for CA biosynthesis, so that supplementation was unnecessary. Screening of medium components, together with experimental design tools, could be a valuable way of enhancing CA titers and reducing the process costs.

Improvement of clavulanic acid production in Streptomyces clavuligerus F613-1 by using a claR-neo reporter strategy

Background: Streptomyces clavuligerus was the producer of clavulanic acid, claR, a pathway-specific transcriptional regulator in S. clavuligerus, positively regulates clavulanic acid biosynthesis. In this study, the promoter-less kanamycin resistance gene neo was fused with claR to obtain strain NEO from S. clavuligerus F613-1. The claR-neo fusion strain NEO was mutated using physical and chemical mutagens and then screened under high concentrations of kanamycin for high-yield producers of clavulanic acid. Results: The reporter gene neo was fused downstream of claR and used as an indicator for expression levels of claR in strain NEO. After three rounds of continuous treatment and screening, the high-yield clavulanic acid-producing strain M3-19 was obtained. In the shaking flask model, the clavulanic acid titer of M3-19 reached 4.33 g/L, which is an increase of 33% over the titer of 3.26 g/L for the starting strains S. clavuligerus F613-1 and NEO. Conclusions: Our results indicate that neo can be effectively used as a reporter for the expression of late-stage biosynthetic genes when screening for high-yield strains and that this approach has strong potential for improving Streptomyces strains of industrial value.

Amoxicillin–clavulanic acid induced sperm abnormalities and histopathological changes in mice

Objective To explore the genotoxic potential and histopathological changes induced in liver, kidney, testis, brain and heart after using the antibiotic drug amoxicillin/clavulanic acid (4:1). Methods The study included chromosomal aberration analysis in bone-marrow and mouse spermatocytes, induction of sperm morphological abnormalities and histopathological changes in different body organs. The drug was administrated orally at a dose of 81 mg/kg body weight twice daily (Total = 162 mg/kg/day) for various periods of time equivalent to 625 mg/men (twice daily). Results The results revealed non-significant chromosomal aberrations induced after treatment with amoxicillin/clavulanic acid (AC) in both bone marrow and mouse spermatocytes after 7 and 10 days treatment. On the other hand, statistically significant percentages of sperm morphological abnormalities were recorded. Such percentage reached 8.10 ± 0.55, 9.86 ± 0.63 and 12.12 ± 0.58 at the three time intervals tested (7, 14 and 35 days after the 1st treatment respectively) (treatment performed for 5 successive days) compared with 2.78 ± 0.48 for the control. The results also revealed histopathological changes in different body organs after AC treatment which increased with the prolongation of the period of therapy. Congestion of central vain, liver hemorrhage and hydropic changes in hepatocytes were noticed in the liver. Degenerative changes were found in kidney glomerulus and tubules while testis showed atrophy of seminiferous tubules, and reduction of spermatogenesis. AC also induced neurotoxicity and altered brain neurotransmitter levels. Hemorrhage in the myocardium, disruption of cardiac muscle fibers and pyknotic nuclei in cardiomyocytes were recorded as side effects of AC in heart tissue. Conclusions The results concluded that AC treatment induced sperm morphological abnormalities and histopathological changes in different body organs. Clinicians must be aware of such results while describing the drug.

An Uncommon Side Effect of a Commonly Used Antibiotic: Amoxicillin-Clavulanic Acid Induced Hepatitis / 가정의학회지

Amoxicillin-Clavulanic acid continues to be one of the most commonly used antibiotic combinations. Hepatic injury due to this antibiotic is rare. We report a case of amoxicillin-clavulanic acid induced hepatitis causing painless jaundice to bring to attention this rare side effect of this commonly used antibiotic. This is a case of a 62-year-old Caucasian female, who presented with acute onset severe painless jaundice, nausea, vomiting, and pruritus of less than 1-week duration. She had completed a course of amoxicillin-clavulanic acid 3 weeks prior to presentation. A careful history pointed to this simple diagnosis. It may be easily missed without an in-depth history and the patient may be subjected to unnecessary expensive tests. This case is reported to highlight cost conscious care by keeping in mind a rare side effect of the commonly used antibiotic.

Amoxicillin-clavulanic acid induced sperm abnormalities and histopathological changes in mice

Objective:To explore the genotoxic potential and histopathological changes induced in liver,kidney,testis,brain and heart after using the antibiotic drug amoxicillin/clavulanic acid (4∶1).Methods:The study included chromosomal aberration analysis in bone-marrow and mouse spermatocytes,induction of sperm morphological abnormalities and histopathological changes in different body organs.The drug was administrated orally at a dose of 81 mg/kg body weight twice daily (Total =162 mg/kg/day) for various periods of time equivalent to 625 mg/men (twice daily).Results:The results revealed non-significant chromosomal aberrations induced after treatment with amoxicillin/clavulanic acid (AC) in both bone marrow and mouse spermatocytes after 7 and 10 days treatment.On the other hand,statistically significant percentages of sperm morphological abnormalities were recorded.Such percentage reached 8.10 ± 0.55,9.86 ± 0.63 and 12.12 ± 0.58 at the three time intervals tested (7,14and 35 days after the 1st treatment respectively) (treatment performed for 5 successive days) compared with 2.78 ± 0.48 for the control.The results also revealed histopathological changes in different body organs after AC treatment which increased with the prolongation of the period of therapy.Congestion of central vain,liver hemorrhage and hydropic changes in hepatocytes were noticed in the liver.Degenerative changes were found in kidney glomerulus and tubules while testis showed atrophy of seminiferous tubules,and reduction of spermatogenesis.AC also induced neurotoxicity and altered brain neurotransmitter levels.Hemorrhage in the myocardium,disruption of cardiac muscle fibers and pyknotic nuclei in cardiomyocytes were recorded as side effects of AC in heart tissue.Contusions:The results concluded that AC treatment induced sperm morphological abnormalities and histopathological changes in different body organs.Clinicians must be aware of such results while describing the drug.

Clinical efficacy and safety of azithromycin versus amoxicillin-clavulanic acid in the treatment of some acute respiratory infections in children:systematic evaluation / 国际药学研究杂志

Objective To systematically evaluate the clinical efficacy and safety of azithromycin(Az)versus amoxicillin-cla?vulanic acid(A-Cva)in the treatment of some acute respiratory infections in children. Methods Pubmed,EMBase,Medline,Co?chrane Library and CJFD were retrieved to collect the randomized controlled trial(RCT)of their clinical efficacy and safety in the treat?ment of acute respiratory infections in children. The methodological quality of included studies was evaluated.The RevMan 5.2 software was chosen for data analysis. Results Twenty RCTs involving 4980 pediatric patients were included for assessment of the clinical effi?cacy. Meta-analysis showed that Az had more significant effect on the treatment of some bacterial repiratory infections in children〔OR=0.78,95%CI(0.65,0.93),P=0.007〕than A-Cva. In the treatment of upper respiratory infections,acute otitis media and so on,Az had more significant effect〔OR=0.75,95%CI(0.62,0.91),P=0.003〕;in the treatment of lower respiratory infections,such as community acquired pneumonia and so on,Az and A-Cva acid had the similar effect〔OR=1.20,95%CI(0.62,2.33),P=0.58〕. Thirteen RCT in?volving 3474 pediatric patients were included for assessment of the clinical safety. Meta-analysis shows that the difference between Az and A-Cva is statistic significant in the treatment of some bacterial repiratory infections in children〔OR=0.49,95%CI(0.40,0.60),P<0.000 01〕. Conclusion Overall,Meta-analysis shows that Az is more effective and safer in the treatment of some bacterial repiratory infections in children than A-Cva.

Clinical efficacy and safety of azithromycin versus amoxicillin-clavulanic acid in the treatment of some acute respiratory infections in children: Systematic evaluation / 国际药学研究杂志

Objective To systematically evaluate the clinical efficacy and safety of azithromycin (Az) versus amoxicillin-clavulanic acid (A-Cva) in the treatment of some acute respiratory infections in children. Methods Pubmed, EMBase, Medline, Cochrane Library and CJFD were retrieved to collect the randomized controlled trial (RCT) of their clinical efficacy and safety in the treatment of acute respiratory infections in children. The methodological quality of included studies was evaluated. The RevMan5.2 software was chosen for data analysis. Results Twenty RCTs involving 4980 pediatric patients were included for assessment of the clinical efficacy. Meta-analysis showed that Az had more significant effect on the treatment of some bacterial repiratory infections in children (OR=0.78, 95%CI (0.65,0.93), P=0.007) than A-Cva. In the treatment of upper respiratory infections, acute otitis media and so on, Az had more significant effect (OR=0.75, 95%CI (0.62,0.91), P=0.003); in the treatment of lower respiratory infections, such as community acquired pneumonia and so on, Az and A-Cva acid had the similar effect (OR=1.20, 95%CI(0.62, 2.33), P=0.58). Thirteen RCT involving 3474 pediatric patients were included for assessment of the clinical safety. Meta-analysis shows that the difference between Az and A-Cva is statistic significant in the treatment of some bacterial repiratory infections in children (OR=0.49, 95%CI (0.40, 0.60), P<0.000 01). Conclusion Overall, Meta-analysis shows that Az is more effective and safer in the treatment of some bacterial repiratory infections in children than A-Cva.

Simultaneous determination of amoxicillin and clavulanic acid in pharmaceutical preparations by capillary zone electrophoresis

ABSTRACT Clavulanic acid enhances the antibacterial spectrum of amoxicillin by rendering most β-lactamase producing isolates susceptible to the drug. A fast, simple and efficient capillary electrophoresis method was developed for the simultaneous determination of amoxicillin and clavulanic acid from complex mixtures. Using a 25 mM sodium tetraborate as background electrolyte at a pH of 9.30, + 25 kV applied voltage, 25 °C system temperature, UV determination at 230 nm; we succeeded in simultaneous separation of amoxicillin and clavulanic acid in approximately 2 minutes. The analytical performance of the method was evaluated in terms of reproducibility, precision, accuracy, and linearity. The optimized analytical method was applied for the determination of the two analytes from combined commercial pharmaceutical preparations. This CE method is fast, inexpensive, efficient, and environmentally friendly when compared with the more frequently used high performance liquid chromatography methods described in the literature.

RESUMO O ácido clavulânico acentua o espectro antibacteriano de amoxicilina, tornando a maioria dos isolados produtores de β-lactamase sensíveis ao fármaco. Desenvolveu-se um método rápido, simples e eficiente de electroforese capilar (EC) para a determinação simultânea de amoxicilina e de ácido clavulânico a partir de misturas complexas. Usando tetraborato de sódio 25 mM como electrólito em pH de 9,30, voltagem aplicada de + 25 kV, em sistema a 25 ° C e determinação por UV a 230 nm, a foi bem-sucedida a separação simultânea de amoxicilina e ácido clavulânico em, aproximadamente, 2 minutos. O desempenho analítico do método foi avaliado em termos de reprodutibilidade, precisão, exatidão e linearidade. O método analítico otimizado foi aplicado para a determinação dos dois analitos em associação, a partir de preparações farmacêuticas comerciais. Este método de EC é rápido, barato, eficiente e ecologicamente correto, quando comparado aos métodos de cromatografia líquida de alta eficiência mais frequentemente descritos na literatura.

Overproduction of clavulanic acid by extractive fermentation

Background Clavulanic acid is an important beta-lactamase inhibitor produced as a secondary metabolite by the actinomycete Streptomyces clavuligerus. Clavulanic acid is chemically unstable; therefore, it is degraded during bacterial cultivation. In this work, the adsorbents clinoptilolite, activated carbon, calcined hydrotalcite, and Amberlite IRA 400 anionic exchange resin were studied in terms of their ability to adsorb clavulanic acid during extractive fermentation, in order to prevent product degradation and avoid product concentrations reaching inhibitory levels. Adsorption assays were used to investigate the effect of pH, and the decrease in the clavulanic acid concentration in the culture broth was measured for each adsorbent. Results IRA 400 was found to be most effective, with 78% adsorption of clavulanic acid. The maximum production of clavulanic acid in Erlenmeyer flask cultures increased 86% in terms of mass of CA, and 248% in cumulative CA concentration, with the use of Amberlite IRA 400 as adsorbent in extractive fermentation, compared to control fermentation performed without product removal. Conclusions The results indicated that extractive fermentation using a solid phase could be an important way of enhancing clavulanic acid titers. It was also possible to show that clavulanic acid acts as an inhibitor of its own synthesis.

Studies on the formation and synthetic mechanism of related substance G in potassium clavulanate production

The objective of this study was to investigate the formation and synthetic mechanism of related substance G in potassium clavulanate production. The impurity in the potassium clavulanate final product, with a retention time of 13.5 min, was confirmed as related substance G by high performance liquid chromatography-mass spectrometry/mass spectrometry. Related substance G analysis during the production of clavulanic acid showed that this impurity could be synthesized during fermentation, and the amount increased with the fermentation time. Studies on its synthetic mechanism showed that L-tyrosine and succinic acid were the precursors for biosynthesis of related substance G in vivo. The reaction was deduced to be catalyzed by an enzyme. The enzyme was a type of extracellular enzyme present in the fermentation supernatant.

O objetivo deste estudo foi investigar a formação e o mecanismo sintético da substância G relacionada na produção de clavulanato de potássio. A impureza do produto final clavulanato de potássio, com tempo de retenção de 13,5 min, foi confirmada como substância G relacionada por cromatografia líquida de alta eficiência-espectrometria de massas/espectrometria de massas. A análise da substância G relacionada durante a produção do ácido clavulânico mostrou que essa impureza poderia ser sintetizada durante a fermentação e que a quantidade aumenta com o tempo de fermentação. Estudos do seu mecanismo sintético mostraram que a L-tirosina e o ácido succínico foram os precursores in vivo para a biossíntese da substância G relacionada. Deduziu-se que a reação foi catalisada por uma enzima. A enzima foi do tipo extracelular, presente no sobrenadante da fermentação.

Screening of wild type Streptomyces isolates able to overproduce clavulanic acid

The selection of new microorganisms able to produce antimicrobial compounds is hoped for to reduce their production costs and the side effects caused by synthetic drugs. Clavulanic acid is a β-lactam antibiotic produced by submerged culture, which is widely used in medicine as a powerful inhibitor of β-lactamases, enzymes produced by bacteria resistant to antibiotics such penicillin and cephalosporin. The purpose of this work was to select the best clavulanic acid producer among strains of Streptomyces belonging to the Microorganism Collection of the Department of Antibiotics of the Federal University of Pernambuco (DAUFPE). Initially, the strains were studied for their capacity to inhibit the action of β-lactamases produced by Klebsiella aerogenes ATCC 15380. From these results, five strains were selected to investigate the batch kinetics of growth and clavulanic acid production in submerged culture carried out in flasks. The results were compared with the ones obtained by Streptomyces clavuligerus ATCC 27064 selected as a control strain. The best clavulanic acid producer was Streptomyces DAUFPE 3060, molecularly identified as Streptomyces variabilis, which increased the clavulanic acid production by 28% compared to the control strain. This work contributes to the enlargement of knowledge on new Streptomyces wild strains able to produce clavulanic acid by submerged culture.

Amoxycillin Clavulanic Acid Induced Hepatotoxicity.

Drug-related hepatotoxicity is a serious health problem, with broad implications for patients, healthcare providers, the pharmaceutical industry and governmental regulatory agencies. Herein we report a rare case of amoxycillinclavulanic acid combination induced liver injury of cholestatic pattern in 40 years old, well educated male patient. Patient gave history that though other drugs were given to him by his physician for fever with chills & rigors, malaise, bodyache, except amoxycillin-clavulanic acid combination all other drugs were well tolerated previously by the patient, without appearance of jaundice. So jaundice in this patient was most probably due to amoxycillinclavulanic acid combination. Though severe liver injury is rare, proper history should be taken while prescribing amoxycillin-clavulanic acid combination. Attention must be paid to potential side-effects of the drugs and close follow-up with patients is a medical necessity to evaluate adverse reactions, especially in case of amoxycillinclavulanic acid combination.

Studies on the formation and forming mechanism of the related substance E in potassium clavulanate production by HPLC-MS/MS

The objective of this study was to investigate the formation and forming mechanism of the related substance E in potassium clavulanate production. The impurity with retention time of 11.1 min in potassium clavulanate final product was confirmed as the related substance E by high performance liquid chromatography with tandem mass spectrometric detection (HPLC-MS/MS).The related substance E analysis during the production of clavulanic acid showed that this impurity could be formed during both the fermentation and purification processes, especially in the later fermentation stage, filtration concentration and back-extraction procedure. Clavulanic acid was the precursor of the related substance E. Studies on its forming mechanism showed that the related substance E was formed by the combination of the imino group of one molecule of clavulanic acid with the carboxyl group of another molecule of clavulanic acid with the opening of β-lactam ring. Results of a multi-factor orthogonal test confirmed that the concentration of clavulanic acid was the dominant factor to accelerate the reaction, while the temperature was another contributing factor. The pH 5.0-6.5 had little impact on the generation of the related substance E.

O objetivo deste estudo foi investigar a formação da substância E e o respectivo mecanismo na produção de clavulanato de potássio. Confirmou-se a impureza com tempo de retenção de 11,1 min no produto final, clavulanato de potássio, como substância E, por meio de cromatografia líquida de alta eficiência, em conjunto com detecção por espectrometria de massas (CLAE-MS-MS). A análise da substância relacionada E durante a produção do ácido clavulânico mostrou que essa impureza pode ser formada tanto durante a fermentação quanto durante os processos de purificação, especialmente no estágio final de fermentação, filtração, concentração e procedimento de extração. O ácido clavulãnico foi o precursor da substância E. Estudos no mecanismo de sua formação mostraram que a substância E formou-se pela combinação do grupo imina da molécula do ácido clavulânico com o grupo carboxílico de outra molécula de ácido clavulânico, com a abertura do anel β-lactâmico. Resultados do teste ortogonal multifatorial confirmaram que a concentração do ácido clavulânico foi o fator dominante para acelerar a reação, enquanto a temperatura foi outro fator que contribuiu. O pH de 5,0 a 6,5 teve pouco impacto na geração da substância E.

Clavulanic acid production by the MMS 150 mutant obtained from wild type Streptomyces clavuligerus ATCC 27064

Clavulanic acid (CA) is a powerful inhibitor of the beta-lactamases, enzymes produced by bacteria resistants to penicillin and cefalosporin. This molecule is produced industrially by strains of Streptomyces clavuligerus in complex media which carbon and nitrogen resources are supplied by inexpensive compounds still providing high productivity. The genetic production improvement using physical and chemical mutagenic agents is an important strategy in programs of industrial production development of bioactive metabolites. However, parental strains are susceptible to loss of their original productivity due genetic instability phenomenona. In this work, some S. clavuligerus mutant strains obtained by treatment with UV light and with MMS are compared with the wild type (Streptomyces clavuligerus ATCC 27064). The results indicated that the random mutations originated some strains with different phenotypes, most divergent demonstrated by the mutants strains named AC116, MMS 150 and MMS 54, that exhibited lack of pigmentation in their mature spores. Also, the strain MMS 150 presented a larger production of CA when cultivated in semi-synthetics media. Using other media, the wild type strain obtained a larger CA production. Besides, using the modifed complex media the MMS 150 strain showed changes in its lipolitic activity and a larger production of CA. The studies also allowed finding the best conditions for a lipase activity exhibited by wild type S. clavuligerus and the MMS150 mutant.

Utilização de sistemas poliméricos de duas fases aquosas (SPDFA) compostos por polietileno glicol/ácido poliacrílico (PEG/APA) para extração de ácido clavulânico / Utilization of aqueous two phase systems (ATPS) composed of polyethylene glycol/polyacrylic acid (PEG/APA) in the extraction of clavulanic acid

A viabilidade da produção em escala industrial de produtos biotecnológicos de interesse comercial e terapêutico, como os fármacos, depende significativamente das técnicas de separação e purificação utilizadas. A aplicação do sistema de duas fases aquosas (SDFA) é proposta como alternativa para a purificação, pois permite a separação e análise de biomoléculas, de modo que estas não percam sua atividade ou propriedades desejadas. Esta técnica é interessante para a purificação em larga escala, pois permite partição seletiva, com potencial de obtenção de altos rendimentos, além de apresentar boa relação custo-benefício. O presente trabalho estudou a purificação por extração líquido-líquido do ácido clavulânico em SDFA utilizando um novo sistema polimérico aquoso, formado pelos polímeros polietileno glicol (PEG) e ácido poliacrílico (APA). Foram estudadas diferentes composições do sistema polimérico aquoso PEG/APA, empregando diferentes massas molares e concentrações para o PEG e utilizando a massa molar 8000g/mol para o APA. Com base nas informações obtidas o melhor ponto de extração para o ácido clavulânico na presença de Na2SO4 foi definido como MPEG=400 g/mol, CPEG=17,5% (m/m) e CNaPA=22,5% (m/m) com K= 19,14, ηT=91,21%, BM=101,69 e R=0,45. Enquanto que na presença de NaCl, o melhor ponto encontrado foi: MPEG=400 g/mol, CPEG=35% (m/m) e CNaPA=10% (m/m) com K=11,96 ηT=80,04%, BM=90,18 e R=0,66. No trabalho será avaliada, também, a influência da temperatura, pH e força iônica nesse sistema. Estabeleceram-se os melhores parâmetros de separação do ácido clavulânico presente em meio fermentado produzido por Streptomyces clavuligerus utilizando a metodologia de fermentação extrativa com SDFA PEG/APA. O efeito do ácido clavulânico no diagrama de fases do sistema PEG-APA, bem como sua partição na forma pura e na presença de homogeneizado celular, foi estudado principalmente através da determinação do coeficiente de partição e recuperação do respectivo fármaco

The viability of industrial scale production of commercial and therapeutical biotechnological products, such as drugs, is significantly dependent on the separation and purification techniques applied. The use of two-aqueous phase systems (ATPS) is proposed as an alternative to purification because it allows the separation and analysis of biomolecules, so that they do not lose their activities or desired properties. This technique is interesting for large scale purification because it allows selective partition with high potential yield and good cost/benefit ratio. The present work studied the purification of clavulanic acid (CA) by liquid-liquid extraction in ATPS applying a new aqueous polymeric system composed of two polymers, namely polyethylene-glicol (PEG) and sodium polyacrylate (NaPA). Different compositions of the aqueous polymeric system (PEG/PAA) were utilized, employing different PEG molar masses (MPEG) and concentrations (CPEG) and a molar mass of PAA of 8000 g/mol. In the light of the results obtained, the best conditions for clavulanic acid extraction, in the presence of Na2SO4, were MPEG = 400 g/mol, CPEG = 17.5% (m/m) and CNaPA = 22.5% (m/m), which allowed obtaining a partition coefficient (K) of 19.14, a yield in the top phase (ηT) of 91.21%, a mass balance (MB) of 101.69 and a volume ratio (R) of 0.45. On the other hand, in the presence of NaCl, the best results (K = 11.96, ηT = 80.04%, MB = 90.18 and R = 0.66) were found at: MPEG = 400 g/mol, CPEG = 35% m/m and CNaPA = 10% m/m. The effect of clavulanic acid in the PEG-PAA system phase diagram and its partition either in its pure form or in the cell homogenate were studied mainly through both the determination of the partition coefficient and the recovery of the drug selected for this study

The comparison of the treatment effect of continuous intravenous infusion and sequential therapy in pneumonia in children / 中国基层医药

Objective To investigate the clinical effects and significance of continuous intravenous infusion and sequential therapy in pneumonia in children.Methods 90 patients with pneumonia were randomly divided into the observation group and the control group,with 45 patients in each group,observation group received continuous intravenous infusion,the control group received sequential therapy.The expectoration and rales disappear time,chest X-ray and body temperature return to normal time,clinical efficacy were observed.Results The expectoration and rales disappear time,chest X-ray and body temperature return to normal time were longer than control group,which were significantly different (x2 =6.21,P ＜ 0.05),the total effective rate of observation group was 77.8％,the total effective rate of control group was 97.8％,the differences were statistically significant (t =3.58,4.62,6.18,7.01,all P ＜ 0.05).Conclusion The sequential therapy treatment pneumonia in children has better clinical effect,shorter hospital stay and worthy of clinical application.