Biosynthesis of 2,5-dimethylpyrazine from L-threonine by whole-cell biocatalyst of recombinant Escherichia coli / 生物工程学报

2,5-dimethylpyrazine (2,5-DMP) is of important economic value in food industry and pharmaceutical industry, and is now commonly produced by chemical synthesis. In this study, a recombinant Escherichia coli high-efficiently converting L-threonine to 2,5-DMP was constructed by combination of metabolic engineering and cofactor engineering. To do this, the effect of different threonine dehydrogenase (TDH) on 2,5-DMP production was investigated, and the results indicate that overexpression of EcTDH in E. coli BL21(DE3) was beneficial to construct a 2,5-DMP producer with highest 2,5-DMP production. The recombinant strain E. coli pRSFDuet-tdh(Ec) produced (438.3±23.7) mg/L of 2,5-DMP. Furthermore, the expression mode of NADH oxidase (NoxE) from Lactococcus cremoris was optimized, and fusion expression of EcTDH and LcNoxE led to balance the intracellular NADH/NAD⁺ level and to maintain the high survival rate of cells, thus further increasing 2,5-DMP production. Finally, the accumulation of by-products was significantly decreased because of disruption of shunt metabolic pathway, thereby increasing 2,5-DMP production and the conversion ratio of L-threonine. Combination of these genetic modifications resulted in an engineered E. coli Δkbl ΔtynA ΔtdcB ΔilvA pRSFDuet-tdhEcnoxELc-PsstT (EcΔkΔAΔBΔA/TDH(Ec)NoxE(Lc)-PSstT) capable of producing (1 095.7±81.3) mg/L 2,5-DMP with conversion ratio of L-threonine of 76% and a yield of 2,5-DMP of 28.8% in 50 mL transformation system with 5 g/L L-threonine at 37 °C and 200 r/min for 24 h. Therefore, this study provides a recombinant E. coli with high-efficiently catalyzing L-threonine to biosynthesize 2,5-DMP, which can be potentially used in biosynthesis of 2,5-DMP in industry.

Bacteriemia por Lactococcus garvieae: primer caso en Colombia / Lactococcus garvieae bloodstream infections: first case in Colombia

Lactococcus garvieae es una causa inusual de bacteriemia. No existen guías diagnósticas y terapéuticas actuales disponibles para tratar las infecciones causadas por estos organismos. Con base en un informe de caso, proporcionamos una revisión de la literatura sobre bacteriemia causada por L. garvieae y destacamos los desafíos de diagnóstico y tratamiento de estas infecciones y las implicaciones para el manejo. Comunicamos el primer caso de bacteriemia por Lactococcus garvieae en Colombia.

Lactococcus garvieae is an unusual cause of bloodstreams infections. Currently, there are no diagnostic and therapeutic guides available to treat infections caused by these organisms. Based on a case report, we provide a literature review on bloodstreams caused by L. garvieae and highlighted the challenges for diagnose and treatment and direct implications for its management. We report the first case of bloodstream infections due to Lactococcus garvieae in Colombia.

Gliotoxin is Antibacterial to Drug-resistant Piscine Pathogens

By activity-guided fractionation, gliotoxin was isolated as an antibacterial metabolite of the fungus Penicillium decumbens which was derived from the jellyfish Nemopilema nomurai. Gliotoxin was further evaluated for antibacterial activity against several piscine and human MDR (multidrug resistance) pathogens. Gliotoxin showed significant antibacterial activity against Gram-positive piscine pathogens such as Streptococcus iniae FP5228, Streptococcus iniae FP3187, Streptococcus parauberis FP3287, Streptococcus parauberis SPOF3K, S. parauberis KSP28, and Lactococcus garvieae FP5245. Gliotoxin showed strong activity especially against S. parauberis SPOF3K and S. iniae FP5228, which are resistant to oxytetracycline. It is noteworthy that gliotoxin effectively suppressed streptococci which are the major pathogens for piscine infection and mortality in aquaculture industry. Gliotoxin also showed strong antibacterial activity against multidrug-resistant human pathogens (MDR) including Enterococcus faecium 5270 and MRSA (methicillin-resistant Staphylococcus aureus) 3089.

Lactobacillus plantarum-derived Extracellular Vesicles Protect Atopic Dermatitis Induced by Staphylococcus aureus-derived Extracellular Vesicles

PURPOSE: The microbial environment is an important factor that contributes to the pathogenesis of atopic dermatitis (AD). Recently, it was revealed that not only bacteria itself but also extracellular vesicles (EVs) secreted from bacteria affect the allergic inflammation process. However, almost all research carried out so far was related to local microorganisms, not the systemic microbial distribution. We aimed to compare the bacterial EV composition between AD patients and healthy subjects and to experimentally find out the beneficial effect of some bacterial EV composition METHODS: Twenty-seven AD patients and 6 healthy control subjects were enrolled. After urine and serum were obtained, EVs were prepared from samples. Metagenomic analysis of 16s ribosomal DNA extracted from the EVs was performed, and bacteria showing the greatest difference between controls and patients were identified. In vitro and in vivo therapeutic effects of significant bacterial EV were evaluated with keratinocytes and with Staphylococcus aureus-induced mouse AD models, respectively. RESULTS: The proportions of Lactococcus, Leuconostoc and Lactobacillus EVs were significantly higher and those of Alicyclobacillus and Propionibacterium were lower in the control group than in the AD patient group. Therefore, lactic acid bacteria were considered to be important ones that contribute to the difference between the patient and control groups. In vitro, interleukin (IL)-6 from keratinocytes and macrophages decreased and cell viability was restored with Lactobacillus plantarum-derived EV treatment prior to S. aureus EV treatment. In S. aureus-induced mouse AD models, L. plantarum-derived EV administration reduced epidermal thickening and the IL-4 level. CONCLUSIONS: We suggested the protective role of lactic acid bacteria in AD based on metagenomic analysis. Experimental findings further suggest that L. plantarum-derived EV could help prevent skin inflammation.

Effect of vitamin C on azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced colitis-associated early colon cancer in mice

BACKGROUND/OBJECTIVES: The objective of this study was to investigate the effects of vitamin C on inflammation, tumor development, and dysbiosis of intestinal microbiota in an azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced inflammation-associated early colon cancer mouse model. MATERIALS/METHODS: Male BALB/c mice were injected intraperitoneally with AOM [10 mg/kg body weight (b.w)] and given two 7-d cycles of 2% DSS drinking water with a 14 d inter-cycle interval. Vitamin C (60 mg/kg b.w. and 120 mg/kg b.w.) was supplemented by gavage for 5 weeks starting 2 d after the AOM injection. RESULTS: The vitamin C treatment suppressed inflammatory morbidity, as reflected by disease activity index (DAI) in recovery phase and inhibited shortening of the colon, and reduced histological damage. In addition, vitamin C supplementation suppressed mRNA levels of pro-inflammatory mediators and cytokines, including cyclooxygenase-2, microsomal prostaglandin E synthase-2, tumor necrosis factor-α, Interleukin (IL)-1β, and IL-6, and reduced expression of the proliferation marker, proliferating cell nuclear antigen, compared to observations of AOM/DSS animals. Although the microbial composition did not differ significantly between the groups, administration of vitamin C improved the level of inflammation-related Lactococcus and JQ084893 to control levels. CONCLUSION: Vitamin C treatment provided moderate suppression of inflammation, proliferation, and certain inflammation-related dysbiosis in a murine model of colitis associated-early colon cancer. These findings support that vitamin C supplementation can benefit colonic health. Long-term clinical studies with various doses of vitamin C are warranted.

Endocarditis por Lactococcus garvieae en un paciente sometido a hemodialisis crónica. Primer caso reportado en Chile y revisión de la literatura / Lactococcus garvieae endocarditis in a patient undergoing chronic hemodialysis. First case report in Chile and review of the literature

Resumen Los casos reportados de infección por Lactococcus garvieae son escasos y sólo uno asociado a hemodiálisis. Comunicamos el caso de endocarditis infecciosa de curso fatal por L. garvieae en un paciente con una enfermedad renal crónica sometido a hemodiálisis y portador de diverticulosis colónica no complicada. Se realiza una revisión de los casos publicados y se discuten los actuales desafíos diagnósticos y terapéuticos de este patógeno, capaz de producir infecciones graves y potencialmente fatales en pacientes susceptibles. Este sería el segundo caso de infección asociada a hemodiálisis y el primero reportado en Chile.

Reports of Lactococcus garvieae infections in humans are scarce, and only one of them in a patient under-going hemodialysis. We report the first case of Lactococcus garvieae infection in Chile, presenting as an infective endocarditis, ultimately fatal, in a patient with uncomplicated colonic diverticulosis and end stage renal failure undergoing chronic hemodialysis. We review the published cases and discuss the diagnostic and therapeutic challenges associated with this new, increasingly diagnosed pathogen, capable of producing serious infections in susceptible patients.

A Metagenomic Analysis Provides a Culture-Independent Pathogen Detection for Atopic Dermatitis

PURPOSE: Atopic dermatitis (AD) is an inflammatory skin disease, significantly affecting the quality of life. Using AD as a model system, we tested a successive identification of AD-associated microbes, followed by a culture-independent serum detection of the identified microbe. METHODS: A total of 43 genomic DNA preparations from washing fluid of the cubital fossa of 6 healthy controls, skin lesions of 27 AD patients, 10 of which later received treatment (post-treatment), were subjected to high-throughput pyrosequencing on a Roche 454 GS-FLX platform. RESULTS: Microbial diversity was decreased in AD, and was restored following treatment. AD was characterized by the domination of Staphylococcus, Pseudomonas, and Streptococcus, whereas Alcaligenaceae (f), Sediminibacterium, and Lactococcus were characteristic of healthy skin. An enzyme-linked immunosorbent assay (ELISA) showed that serum could be used as a source for the detection of Staphylococcus aureus extracellular vesicles (EVs). S. aureus EV-specific immunoglobulin G (IgG) and immunoglobulin E (IgE) were quantified in the serum. CONCLUSIONS: A metagenomic analysis together with a serum detection of pathogen-specific EVs provides a model for successive identification and diagnosis of pathogens of AD.

A Case of Septic Shock Following Catheter-related Infection Caused by Lactococcus lactis subsp. lactis in an Adult

Lactococcus lactis is a gram-positive cocci used extensively in the dairy industry, but considered an unusual pathogen in humans. Among its five subspecies, L. lactis subsp. lactis in particular has rarely been reported as a pathogen. We report a case of septic shock caused by L. lactis subsp. lactis in an adult patient. A 64-yr-old male patient was admitted to outpatient clinics, with chief complaints of fever and chills for one week after convalescent hospital admission. He had severe ileus requiring surgery. He had a peripherally inserted central catheter from convalescent hospital, which was immediately removed. From two sets of blood and catheter tip cultures, we identified L. lactis subsp. lactis using the Vitek 2 system (bioMerieux Inc., USA), and confirmed this result by 16S rRNA sequencing. The patient was empirically treated with ciprofloxacin, and he recovered and was discharged.

Antimicrobial activity of essential oil of Eucalyptus globulus against fish pathogenic bacteria / 한국실험동물학회지

The antibacterial activities of the essential oil of Eucalyptus globulus (EOEG) was determined against 7 fish pathogenic bacteria (Edwardsiella tarda, Streptococcus iniae, S. parauberis, Lactococcus garviae, Vibrio harveyi, V. ichthyoenteri and Photobacterium damselae) obtained from farmed olive flounder. The inhibitory activity was evaluated by three methods: Disc diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). According to the disc diffusion test, as the concentration of EOEG (5-40 µg) rises, the inhibitory zone increases in size. Compared with amoxicillin, tetracycline and chloramphenicol, EOEG showed similar antibacterial activity. The MIC of EOEG ranged from 7.8 to 125 mg/mL and MBC values ranged from 62 to 250 mg/mL. These results show that EOEG has antimicrobial activity against all seven bacteria, but there was no marked difference between each genus. From these results, it is suggested that EOEG can be used as an antimicrobial agent against fish bacterial diseases in the fish industry.

Survey of canine tick-borne diseases in Lábrea, Brazilian Amazon: ‘accidental’ findings of Dirofilaria immitis infection / Pesquisa de agentes transmitidos por carrapatos em cães de Lábrea, Amazonas: achados “acidentais” de Dirofilaria immitis

Blood samples were collected from 99 domestic dogs from the urban and rural areas of the Lábrea municipality, state of Amazonas, Brazil. Canine serum samples were tested by immunofluorescence assay against Rickettsia spp., which revealed that only 3.0% (1/33) and 7.6% (5/66) of the dogs from urban and rural areas, respectively, reacted positively to at least one Rickettsia species. DNA was extracted from canine blood and tested by a battery of PCR assays targeting protozoa of the genera Babesia and Hepatozoon, and bacteria of the genera Rickettsia and Ehrlichia and family Anaplasmataceae. All samples were negative in the PCR assays targeting the genera Babesia, Hepatozoon, Ehrlichia and Rickettsia. For Anaplasmataceae, 3% (1/33) and 39.4% (26/66) of the urban and rural dogs, respectively, yielded amplicons that generated DNA sequences 100% identical to the corresponding sequence of Wolbachia endosymbiont of Dirofilaria immitis. Because of these results, all canine DNA samples were further tested in a PCR assay targeting filarial nematodes, which was positive for 18.2% (6/33) and 57.6% (38/66) urban and rural dogs, respectively. Filarial-PCR products generated DNA sequences 100% identical to D. immitis. While tick-borne infections were rare in Lábrea, D. immitis infection rates were among the highest reported in South America.

Amostras de sangue foram coletadas de 99 cães domésticos de áreas urbana e rural do município de Lábrea, estado do Amazonas. Soros caninos foram testados pela técnica de imunofluorescência indireta contra Rickettsia spp., resultando em apenas 3,0% (1/33) e 7,6% (5/66) de cães soropositivos nas áreas urbana e rural, respectivamente. DNA foi extraído do sangue canino e testado por diferentes protocolos da PCR para detecção de protozoários dos gêneros Babesia e Hepatozoon, e bactérias dos gêneros Rickettsia e Ehrlichia e da família Anaplasmataceae. Todas as amostras foram negativas nos protocolos de PCR para os gêneros Babesia, Hepatozoon, Ehrlichia e Rickettsia. Para Anaplasmataceae, 3% (1/33) e 39,4% (26/66) dos cães de áreas urbana e rural, respectivamente, geraram sequências de DNA 100% idênticas ao endosimbionte Wolbachia de Dirofilaria immitis. Posteriormente, as amostras foram testadas pela PCR para nematódeos filarídeos, resultando em 18,2% (6/33) e 57,6% (38/66) de amostras positivas nas áreas urbana e rural, respectivamente. Os produtos geraram sequências de DNA 100% idênticas a D. immitis. Em contraste com várias outras regiões do Brasil, infecções transmitidas por carrapatos foram raras em Lábrea. Por outro lado, as frequências de infecção por D. immitis estiveram entre as mais altas relatadas na América do Sul.

El efecto antihipertensivo de las leches fermentadas / The antihypertensive effect of fermented milks

Existe una gran variedad de leches fermentadas con bacterias lácticas, con propiedades que promueven la salud. Recientemente se ha comunicado que las proteínas de los alimentos pueden, además, ejercer otras funciones in vivo, por medio de sus péptidos con actividad biológica. Estos péptidos se encuentran encriptados dentro de la estructura primaria de las proteínas y pueden ser liberados por fermentación de la leche, hidrólisis enzimática, o bien durante el tránsito gastrointestinal. Las funciones que presentan son diversas, ya que pueden actuar en diferentes sistemas del cuerpo humano: el cardiovascular, el digestivo, el endocrino, el inmune y el nervioso. Los péptidos bioactivos que presentan un efecto en el sistema cardiovascular (antihipertensivo, antitrombótico, antioxidante o hipocolesterolémico) pueden reducir los factores de riesgo para la manifestación de enfermedades crónicas y ayudar a mejorar la salud humana. Los péptidos bioactivos más estudiados son aquellos que ejercen un efecto antihipertensivo a través de la inhibición de la enzima convertidora de angiotensina (ACE). Este documento se enfoca en la producción de péptidos antihipertensivos inhibidores de la ACE en leches fermentadas, en su historia, y en las pruebas in vivo realizadas en ratas y en humanos, donde se ha demostrado su efecto hipotensor.

There is a great variety of fermented milks containing lactic acid bacteria that present health-promoting properties. Milk proteins are hydrolyzed by the proteolytic system of these microorganisms producing peptides which may also perform other functions in vivo. These peptides are encrypted within the primary structure of proteins and can be released through food processing, either by milk fermentation or enzymatic hydrolysis during gastrointestinal transit. They perform different activities, since they act in the cardiovascular, digestive, endocrine, immune and nervous systems. Bioactive peptides that have an antihypertensive, antithrombotic, antioxidant and hypocholesterolemic effect on the cardiovascular system can reduce the risk factors for chronic disease manifestation and help improve human health. Most studied bioactive peptides are those which exert an antihypertensive effect by inhibiting the angiotensin-converting enzyme (ACE). Recently, the study of these peptides has focused on the implementation of tests to prove that they have an effect on health. This paper focuses on the production of ACEinhibitory antihypertensive peptides from fermented milks, its history, production and in vivo tests on rats and humans, on which its hypotensive effect has been shown.

Polymicrobial Peritonitis with Lactococcus lactis in a Peritoneal Dialysis Patient / 전남의대학술지

Lactococcus lactis (L. lactis) is an important gram-positive bacterium in dairy products. It is a rare cause of opportunistic infections with only four cases of Lactococcus peritoneal dialysis (PD) peritonitis reported in the literature. In Korea, L. lactis infection was first reported in a liver abscess patient in 2010; however, PD peritonitis with Lactococcus has not been reported in Korea. Recently, we experienced a case of Lactococcus-associated polymicrobial PD peritonitis. The patient was initially managed with broad-coverage antibiotics; however, owing to a poor response, the PD catheter was removed and the patient was switched to hemodialysis. We discuss this case and review the literature.

Polymicrobial Peritonitis with Lactococcus lactis in a Peritoneal Dialysis Patient / 전남의대학술지

Lactococcus lactis (L. lactis) is an important gram-positive bacterium in dairy products. It is a rare cause of opportunistic infections with only four cases of Lactococcus peritoneal dialysis (PD) peritonitis reported in the literature. In Korea, L. lactis infection was first reported in a liver abscess patient in 2010; however, PD peritonitis with Lactococcus has not been reported in Korea. Recently, we experienced a case of Lactococcus-associated polymicrobial PD peritonitis. The patient was initially managed with broad-coverage antibiotics; however, owing to a poor response, the PD catheter was removed and the patient was switched to hemodialysis. We discuss this case and review the literature.

Caracterização de bactérias láticas em queijo de coalho e antagonismo a alguns patógenos / Characterization of lactic acid bacteria in coalho cheese andantagonism to some pathogenic food-related bacteria

O queijo de coalho é um produto tradicional do nordeste brasileiro e o conhecimento da sua microbiotaautóctone é de fundamental importância. Neste trabalho foi caracterizada a microbiota do queijo de coalhode três propriedades rurais (A, B e C), situadas respectivamente nas mesorregiões do Agreste, Borboremae Sertão paraibanos, com o intuito de avaliar a importância do perfil das bactérias láticas encontradasneste tipo de queijo e verificar a atividade antimicrobiana destas bactérias frente a agentes patogênicos. Ascontagens de bactérias láticas do queijo de coalho dos produtores A e B foram na ordem de 106 e as maiorescontagens (109 UFC/g) foram observadas no queijo do produtor C. Foram selecionadas 49 bactérias láticastípicas das três propriedades e os gêneros predominantes foram: Enterococcus, Lactococcus, Streptococcuse Leuconostoc. Dos 20 isolados, 19 apresentaram halos de inibição sobre as três bactérias patogênicas, comzonas de inibição de 2 a15 mm, e os maiores halos foram formados por Lactococcus lactis ssp lactis sobre S.enterica e S. aureus. A atividade antimicrobiana revelada por alguns isolados de bactérias láticas sugerema possibilidade de seu uso contra agentes patogênicos e podem atuar como barreira no desenvolvimentodestes micro-organismos e como conservantes biológicos.

Endocardite por lactococcus garvieae: primeiro relato de caso da América Latina / Lactococcus garvieae endocarditis: first case report in Latin America / Endocarditis por lactococcus garvieae: primer relato de caso de América Latina

Lactococcus garvieae, patógeno zoonótico emergente, é responsável por mastite em ruminantes e septicemia em peixes. Embora seja considerado oportunista e raramente causar infecções em humanos, sua incidência deve estar subestimada devido à dificuldade do diagnóstico. Há pouquíssimos relatos de osteomielite, abscesso hepático e peritonite, e apenas nove casos descritos na literatura mundial de endocardite. Relatamos o primeiro caso de endocardite por Lactococcus garvieae da América Latina em paciente portadora de prótese valvar metálica, com quadro de febre diária, calafrios, nodos de Osler e seis hemoculturas positivas para Lactococcus garvieae, que preenchiam os critérios de Duke para o diagnóstico de "endocardite infecciosa definitiva".

Lactococcus garvieae, an emerging zoonotic pathogen, is responsible for mastitis in rodents and sepsis in fish. Although deemed opportunistic and hardly ever causing infections in humans, its incidence is probably underestimated due to the difficulty in diagnosis. There are very few reports of osteomyelitis, liver abscess, and peritonitis, and only nine cases of endocarditis described in worldwide literature. We describe the first case of Lactococcus garvieae endocarditis in Latin America, in a female patient with metallic prosthetic heart valve who presented with daily fever, chills, Osler nodes and six positive blood cultures for Lactococcus garvieae, which met Duke's criteria for the diagnosis of "definitive infective endocarditis".

Lactococcus garvieae, patógeno zoonótico emergente, es responsable por mastitis en rumiantes y septicemia en peces. Aunque sea considerado oportunista y raramente cause infecciones en humanos, su incidencia debe estar subestimada debido a la dificultad del diagnóstico. Hay poquísimos relatos de osteomielitis, absceso hepático y peritonitis, y apenas nueve casos descriptos en la literatura mundial de endocarditis. Relatamos el primer caso de endocarditis por Lactococcus garvieae de América Latina en paciente portadora de prótesis valvar metálica, con cuadro de fiebre diaria, escalofríos, nódulos de Osler y seis hemocultivos positivos para Lactococcus garvieae, que llenaban los criterios de Duke para el diagnóstico de "endocarditis infecciosa definitiva".

Effect of Probiotics on Intestinal Infections

Probiotics are defined as live microorganisms that confer a health benefit on the host when consumed in adequate amounts, such as Lactobacillus, Streptococcus, Enterococcus, Lactococcus, Bifidobacterium, Bacillus, Clostridium, Saccharomyces, Aspergillus, and Escherichia coli species. Previous studies have reported beneficial effects of probiotics on intestinal infections, including Clostridium difficile antibiotic-associated diarrhea, rotavirus infection, traveler's diarrhea, and other bacterial enteric diseases. Although few studies have reported serious adverse effects from probiotics, more evidence-based research is required for an in-depth evaluation of probiotics in medical science.

Isolation and characterization of lactococcus garvieae from diseased rainbow trout [Oncorhynchus mykiss, Walbaum] cultured in Iran

A total of 200 moribund rainbow trout with clinical signs of a hyperacute haemorrhagic septicemia were collected from rainbow trout farms in Fars, Kohkiloyeh-Boyer Ahmad and Charmohal-Bakhtiari provinces in the south and southwest of Iran during summer 2002 to winter 2008 for detection of Lactococcus garvieae, the causative agent of lactococcosis. Fish kidney samples were cultured aseptically onto brain heart infusion agar plates and incubated at 25°C for 48 h. Using conventional biochemical tests, L. garvieae was detected from 32 fish [16% of total fish samples]. Additionally, isolates were confirmed as L. garvieae using a specific PCR assay based on 16S rDNA gene by producing a single band of 1107 bp. Partial analysis of 16S rDNA revealed 100% sequence similarity for all Iranian isolates and there was a close genetic relationship among these isolates and those previously reported from mullet in Taiwan [AF352166] and yellowtail in Japan [AB267897] based on GenBank data. Results of antibiogram tests on L. garvieae isolates showed a high susceptibility to erythromycin, enrofloxacin, chloramphenicol and clarithromycin. In pathogenicity tests, immersion of fish in a bacterial suspension of 6x10[5] colony forming unit/ml of L. garvieae in challenge experiments showed 60% mortality during 14 days post-infection. Experimentally, infected fish showed typical blackening of skin haemorrhages, exophthalmia and wide haemorrhages on viscera. The present study provides useful molecular and biochemical information for L. garvieae isolates in Iran compared with those from different hosts and geographic locations

Meta analysis of lactic acid bacteria as probiotics for the primary prevention of infantile eczema / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To determine whether lactic acid bacteria as probiotics is efficacious in the primary prevention of infantile eczema or atopic eczema.</p><p><b>METHODS</b>For this meta analysis of randomized controlled trials (RCT) describing the efficacy of probiotics in infants with eczema or atopic eczema at ages of ≤2 years, a comprehensive search in the databases was performed up to January 2010. Three reviewers independently evaluated the studies for methodological qualities. RevMan 5.0.2 software was used for meta analysis.</p><p><b>RESULTS</b>Twelve RCTs on the preventive effects of lactic acid bacteria as probiotics on infantile eczema were included, and 7 of the 12 RCTs reported the preventive effect of lactic acid bacteria on atopic eczema. The meta analysis showed that there was an overall significant reduction in infantile eczema and atopic eczema favoring lactic acid bacteria compared with placebo. The relative risk (RR) ratios for eczema and atopic eczema were 0.80 (95%CI: 0.70-0.90; P<0.01) and 0.78 (95%CI: 0.64-0.97; P<0.01), respectively. Lactic acid bacteria combined with other probiotics decreased significantly the incidence of eczema, with a RR ratio of 0.79 (95%CI: 0.68-0.93; P<0.01). The use of lactic acid bacteria alone did not result in a reduction in the incidence of eczema, with a RR ratio of 0.85 (95%CI: 0.69-1.05; P>0.05).</p><p><b>CONCLUSIONS</b>The data from this meta analysis suggest that lactic acid probiotics combined with other probiotics play a role in the prevention of infantile eczema. There is insufficient evidence to recommend single use of lactic acid bacteria for prevention of eczema. Further studies are required to determine whether the findings are reproducible.</p>

Lactococcus garvieae: isolamento ecaracterização fenotípica / Lactococcus garvieae: isolation and phenotypic characterization

Os microrganismos do genero Lactococcus apresentam caracteristicas fenotipicas semelhantes as do genero Enterococcus podendo ser erroneamente identificados como tal no laboratorio clinico. Durante um estudo epidemiologico visando a deteccao de enterococos resistentes a vancomicina, foram avaliadas 155 amostras, inicialmente suspeitas de pertencerem ao genero Enterococcus, isoladas da microbiota intestinal e de diversos materiais clinicos oriundos de pacientes atendidos em Instituicoes de Saude do Municipio de Niteroi, no periodo entre janeiro de 2005 e janeiro de 2006. Duas amostras, que apresentaram caracteristicas fisiologicas naototalmente compativeis com o genero Enterococcus, foram identificadas como Lactococcus garvieae atraves da utilizacao de testes adicionaiscomo a analise do perfil proteico total atraves de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sodio. Esta metodologia pode ser uma alternativa para identificar corretamente L. garvieae visto que e uma tecnica relativamente simples e demenor custo, quando comparada aos testes baseados na analise do DNA.

The members of the genus Lactococcus have phenotypic characteristics that resemble those of the genus Enterococcus and therefore can be erroneously identified in the clinical laboratory. During an epidemiological study aiming the detection of vancomycin-resistant enterococci, were evaluated 155 isolates, initially suspected of belonging to the genus Enterococcus, recovery from the intestinal microflora and different clinical materials obtained from patients at Health Institutions of Niteroi city, in the period between January 2005 and January 2006. Two isolates, which showed physiological characteristics not fully compatible with the genusEnterococcus, 2006. Two isolates, which showed physiological characteristics not fully compatible with the genus Enterococcus, were identified as Lactococcus garvieae by using additional physiological tests and analysis of total protein profiles by polyacrylamide gel electrophoresis, containing sodium dodecyl sulfate. This is an alternative methodology to correctly identify L. garvieae provided it is relatively easy to perform and of lower cost when compared to DNA based tests.

PCR-based assembly of the DNA sequence coding for tetanus toxin C fragment / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a PCR-based method for gene assembly of tetanus toxin C fragment (TETC) DNA sequence from a large number of oligodeoxyribonucleotides (oligos).</p><p><b>METHODS</b>To allow for its cloning and expression in Lactococcus lactis, the TETC gene sequence was designed according to the known TETC gene sequence (GenBank accession number M12739, 367-1719) and the amino acid coding in Lactococcus lactis. The sequence contained 1383 nucleotides (nt) with Sal I site added to its 5' end and Xho I and Hind III sites to its 3' end. There were 209 synonymous codon substitutions in the designed gene sequence as compared with the sequence reported in GenBank for amino acid coding in Lactococcus lactis and elimination of the restriction site of EcoR I and Kpn I. The 1380 nt of the sequence was divided into 68 oligos designated as TETC 1 to TETC 68, each containing 40 nt. A 16 nt oligos designated as TETC 69 was designed as the downstream primer. The TETC 1-24 fragment was acquired using the oligos TETC 1 to TETC 24 by PCR-based gene assembly method, and the TETC 23-46 and TETC45-68 fragments were assembled similarly. The full-length TETC gene was assembled using TETC 1 and TETC 69 as the primers when the 3 fragments were mixed. The target gene was gel-purified and digested with Sal I and Hind III, followed by ligation to the pBluescript II SK(+) and digestion with the same enzymes. The positive clones were confirmed by restriction enzyme excision and sequencing.</p><p><b>RESULTS</b>Three 500-bp fragments were acquired by PCR-based gene assembly, and the full-length TETC gene was obtained from the 3 fragment mixed at a equal concentration by a second PCR-based gene assembly using TETC 1 and TETC 69 as the primers. The target gene was cloned to pBluescript II SK(+) vector, and sequence analysis of the positive clones indicated that the assembled sequence was identical to the designed coding sequence of TETC gene.</p><p><b>CONCLUSION</b>PCR-based assembly of the synthesized constitutive gene fragments into the complete sequence can be an effective strategy for synthesis of long DNA sequences in vitro.</p>