The Efficacy and Safety of a Combination of Amino Acids, Vitamins and Probiotics in PROM and PPROM

The prevalence and severity of prelabor rupture of the membranes (PROM)/preterm PROM (PPROM) are a worldwide public health concern. PROM is the result of a cascade of events involving matrix metalloproteinase (MMP)-9, tissue inhibitor of metalloproteinases 1 (TIMP1), cytokines and proapoptotic genes, which is initiated by several factors such as infection, genotoxic agents or some unknown etiology. In PROM, there is an increased expression and activation of MMP-2, MMP-3 and MMP-9 and a reduction of TIMP1. p53 and tumor necrosis factor (TNF)-? mediate the major apoptotic pathway of PROM. p53 can transactivate some MMP genes, resulting in the overexpression of MMPs. This leads to apoptosis. MMP-2 and MMP-9 degrades type-IV collagen, which is the major structural component of chorioamnion. Understanding the fundamental pathology at the molecular level, it appears necessary to adjust the biologically protective mechanism to prevent spontaneous preterm labor. Our findings show that the novel combination of arginine, ascorbic acid, folic acid, glutamine, glutathione, thiamine, lactic acid bacillus spores, vitamin E acetate and pyridoxine is safe and effectively prevents PROM and PPROM (97% patients) and prolongs pregnancy term.

In Vitro Antimicrobial Activity and Aggregation Abilities of Probiotic Lactobacillus casei and Lactobacillus salivarius Against Oral Pathogens

ABSTRACT@#The antagonistic effect of probiotics against oral pathogens merits exploration because these bacteria are beneficial to the host’s health. The antimicrobial activity of two probiotic strains, Lactobacillus casei and Lactobacillus salivarius, as well as L. casei and L. salivarius combination (1:1), was investigated against Streptococcus mutans, Streptococcus sobrinus, Candida albicans, Candida glabrata and Candida tropicalis using agar-well diffusion, auto-aggregation and coaggregation assays. L. salivarius cell-free supernatant (CFS) alone exhibited greater inhibitory effect against Streptococci spp. compared to L. casei CFS alone and the combination. However, no inhibition was observed for Candida spp. L. salivarius alone exhibited significantly stronger auto-aggregation than L. casei alone (p ≤ 0.05) and L. casei and L. salivarius combination. L. salivarius exhibited strong coaggregation ability with Candida spp., followed by Streptococci spp. while L. casei exhibited coaggregation only with Streptococci spp. However, L. casei and L. salivarius combination did not display any coaggregation with all strains. L. salivarius alone exhibited a stronger antagonistic effect on the tested organisms than L. casei alone or in combination. Based on the results, both probiotic strains showed good antimicrobial activities against oral pathogens and should be further studied for their human health benefits.

Evaluation of the vaginal microflora of women of reproductive age / Шинэ санаа Шинэ нээлт

Background@#The knowledge about the normal and abnormal vaginal microbiome has changed over the last years. The normal and the abnormal vaginal microbiota are complex ecosystems of more than 200 bacterial species influenced by genes, ethnic background and environmental and behavioral factors. Major changes in the vaginal physiology and microbiota over a woman’s lifetime are largely shaped by transitional periods such as puberty, menopause and pregnancy. Many studies have shown that bacterial vaginitis (BV) has an increased risk of preterm birth, miscarriage, premature rupture of membran, and postpartum uterine inflammation and sepsis. To assess different bacterial and epidemiological factors associations with increased vaginal pH in the women of reproductive age.@*Methods@#In the 1st Health center of Bayanzurkh district hospital, 100 non-pregnant women were randomly selected between 18-45 years of age. Women were submitted to an interview, vaginal examination and vaginal specimen collection for pH measurement and microscopy. Descriptive statistics are reported for the vaginal pH according to a specially designed survey card, the survey respondents assessed the status of the mother’s health.@*Results@#The non pregnant group studied herein were mostly young adults with ages in the age range 25-29 (32.0; 32%, mean±SD: 29.1±5.7) years. The vaginal pH mean in non-pregnant women was 4.5±0.5. Elevated vaginal pH was signicantly associated with bacterial vaginosis (p < 0.001), and bacterial vaginosis flora (p < 0.001). 162 of 200 women (81%) had an increased vaginal pH. 65,8% of participants in research were normal nugent score.@*Conclusion@#In non-pregnant women, the vaginal pH mean was 4.5 ± 0.5. Changes in vaginal pH are statistically significant with age, and clinical manifestations. /р<0.05/. The human vaginal ecosystem is a dynamic environment in which microbes can affect host physiology but also where host physiology can affect the composition and function of the vaginal microbiota.

Correspondencia entre el índice de placa y el recuento de Streptococcus mutans y Lactobacillus sp. en una población infantil: Un estudio transversal / Correspondence between plaque index and Streptococcus mutans and Lactabacillus spp. counts in a child population: A cross-sectional study

Antecedentes: La identificación de microorganismos como Streptococcus mutans(SM) y Lactobacillus sp. (LB) es un predictor del desarrollo de caries, sin embargo los métodos de identificación son de acceso limitado por su alto costo. Objetivo:determinar la correlación entre la biopelícula dental medida por el índice de placa bacteriana de (IPB) Silness y Löe modificado y el recuento de SM y LB en niños colombianos en edad preescolar. Este es un estudio descriptivo de corte transversal y correlacional. Materiales y métodos: Se tomaron 202 muestras de niños entre los 3 y 4 años de edad de tres jardines Infantiles de Cali, Colombia, pertenecientes al estrato socioeconómico 1 y 2, evaluados en cuatro tiempos (0, 3, 6 y 9 meses). Resultados: Hubo una correlación entre el IPB más alto y el mayor recuento de unidades formadoras de colonia de SMsólo a los 3 meses (p = 0,003*). Los recuentos de LB fueron homogéneos en todos los niños y no presentaron correlación con el IPB. Conclusión: Se observaron resultados mixtos sobre la relación entre IPB y los recuentos de SM presentes en la saliva, por lo cual se requiere un análisis de múltiples factores.

Background:The identification of microorganisms such as Streptococcus mutans(SM) and Lactobacillusspp. (LB) is a predictor of the development of caries, however the identification methods are of limited access due to their high cost. Aim:To determine the correlation between the dental biofilm measured by the modified Silness and Löe dental plaque index (DPI)) and the SM and LB count, in preschool children from Colombia. Materials and methods: This is a descriptive cross-sectional and correlational study. We took202 samples from children between 3 and 4 years of age from three kindergartens in Cali, Colombia, belonging to socioeconomic stratum 1 and 2, evaluated at four times (0, 3, 6 and 9 months). Results: We found a correlation between the highest IPB and thehighest count of SM colony forming units only at 3 months (p = 0.003*). LB counts were homogeneous in all children and there was no correlation with IPB. Conclusion: We observed mixed results on the relationship between IPB and the counts of SM present in the saliva, therefore an analysis of multiple factors is required.KEY WORDSDental plaque;Streptococcus mutans;Lactobacillus spp;dental caries.1

Preliminary Studies on Isolation, Bile Tolerance and Antibiogram of Potential Probiotics (Probionts) from Locally Fermented Food Products at Beach Market, Nsukka Metropolis, Enugu State, Nigeria

Aims: The study aims to isolate, evaluate bile tolerance and antibiogram studies of potential probiotics (Lactobacillus spp) from locally fermented Food Products (Akamu, Aqua Rafa® Yoghurt, Ogiri, Okpeye) and Kunu at Beach Market, Nsukka. Study Design: A ten - fold serial dilution and spread plate method using De Man, Rogosa and Sharpe (MRS) medium was adopted for isolation of potential Probionts. Place and Duration of Study: Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences, University of Nigeria Nsukka, between March - September, 2018. Methodology: Only catalase negative and Gram positive isolates characteristic of lactobacilli were used. Bile tolerance test was performed by monitoring the bacterial growth at different Bile salt concentrations (0.2%, 0.3% and 2%). The antibiogram of the isolates was assessed using the Kirby-Bauer disc diffusion method against commercial antibiotic discs of ampicillin, vancomycin, gentamycin, ciprofloxacin, methicillin and erythromycin. Results: All the 18 screened isolates were tolerant to bile salt at 0.2 % and 0.3 % with inhibition of growth at 2 % bile concentration. All isolates were observed to be resistant to methicillin (100 %) but very sensitive to gentamycin (11%) and ciprofloxacin (22%) respectively. The isolates showed intermediate resistance to other antibiotics: vancomycin (33%), erythromycin (33%) and ampicillin (44%). The decreasing pattern of resistance was thus: methicillin > ampicillin > vancomycin and erythromycin > ciprofloxacin > gentamycin. Isolates from Yoghurt (66.67%) and Ogiri (53.33%) provided most of the resistant isolates. Methicillin would provide best antagonist potential as all the isolates exhibited very high level of resistance (100 %). Conclusion: These results suggest that all the eighteen potential Lactobacillus spp strain show potential for probiotic applications and the locally fermented food products are rich sources of probionts.

Effects of a food enriched with probiotics on Streptococcus mutans and Lactobacillus spp. salivary counts in preschool children: a cluster randomized trial

Abstract Probiotics have provided benefits to general health, but they are still insufficient to dental health. Objective: This study aimed to evaluate milk supplemented with probiotic bacteria and standard milk, measured by levels of Streptococcus mutans (S. mutans) and Lactobacillus spp., in 3-4-year-old children after 9 months of intervention. Material and Methods: The study was a triple-blind, placebo-controlled, randomized trial. The sample was composed of 363 preschoolers attending five child development centers in Cali, Colombia. They were randomized to two groups: children in the intervention group drank 200 mL of milk with Lactobacillus rhamnosus 5x106 and Bifidobacteruim longum 3x106, and children in the control group drank 200 mL of standard milk. Interventions occurred on weekdays and information was gathered through scheduled clinical examination. The primary result was the number of colony forming units (CFU) of S. mutans and Lactobacillus spp. in the saliva. Secondary results were dental caries, rated by the International Caries Detection and Assessment System (ICDAS), dental plaque, pH, and salivary buffer capacity. Results: The proportion of S. mutans was lower in the intervention group compared with the control group after 9 months; however, the differences did not reach statistical significance (p=0.173); on the other hand, statistically significant differences between groups were found in the CFU/mL of Lactobacillus spp. (p=0.002). There was not statistically significant difference in the prevalence of dental caries for both groups (p=0.767). Differences between groups were found in the salivary buffering capacity (p=0.000); neither salivary pH nor dental plaque were significantly different. Conclusions: Regular consumption of milk containing probiotics bacteria reduced CFU/mL of Lactobacillus spp. and increased salivary buffering capacity at 9 months of consumption.

Study of Bacteria Associated with Dental Caries Using a 3 Tone Disclosing Agent / 대한소아치과학회지

The aim of this study was to determine the efficacy of a 3 tone plaque disclosing gel in assessing the risk of caries related to the population of Streptococcus mutans, Streptococcus sobrinus, and Lactobacillus spp. quantified using a quantitative real-time polymerase chain reaction (qRT-PCR).15 healthy children of ages 9 – 12 years were randomly examined. The 3 tone plaque disclosing gel was applied on teeth surfaces, which changed the color to pink or red, blue or purple and light blue. Plaque was divided into 3 groups based on staining. Genomic DNA from each sample was subjected to a qRT-PCR assay for quantitative detection of target bacteria. The Kruskal-Wallis test was conducted for correlation between the color of plaque and the number of bacterial species.The levels of S. mutans, S. sobrinus, and Lactobacillus spp. were significantly different in the plaque samples of the 3 groups (p < 0.05). The proportion of S. sobrinus to S. mutans showed correlation to the color of plaque.The different color-dyed plaque was related to the number of acidogenic bacteria. The 3 tone plaque disclosing gel could be used as one of the indicators to assess the clinical risk of caries associated with the population of S. mutans, S. sobrinus, and Lactobacillus spp.

Efeitos imunomoduladores e atividade anti-Candida de lactobacilos com propriedades probióticas em macrófagos e em modelo invertebrado de Galleria mellonella / Immunomodulatory effects and anti-Candida activity of lactobacilli with probiotic properties in macrophages and invertebrate model Galleria mellonella

Muitos produtos probióticos são compostos pela associação de lactobacilos no intuito de potencializar seu efeito benéfico. Assim, o objetivo deste estudo foi avaliar a imunomodulação e atividade anti-Candida causada por diferentes combinações de suspensões probióticas em macrófagos e em Galleria mellonella. Lactobacillus rhamnosus (LR), L. acidophilus (LA), e L. paracasei (LP) foram suspendidos para obtenção das suspensões LR, LA, LP, LR+LA, LR+LP, LA+LP e LR+LA+LP. Foram utilizadas suspensões de Candida albicans (CA) ATCC 18804, C. krusei (CK) ATCC 6258 e C. glabrata (CG) ATCC 9030. Macrófagos de camundongo (RAW 264.7) foram ativados por cada suspensão de lactobacilos e desafiados por cada uma das cepas fúngicas. Foram investigadas taxa de fagocitose, produção de citocinas e óxido nítrico e a viabilidade celular. A melhor suspensão probiótica para cada cepa fúngica seria usada nos testes in vivo, com G. mellonella, no entanto, devido à baixa virulência das espécies não-albicans, os testes in vivo prosseguiram com CA e LR. Duas cepas clínicas de CA foram acrescentadas: 21 e 60. Suspensões de LR foram injetadas na hemolinfa das larvas 24 h antes do desafio com as cepas fúngicas. Foram determinadas curva de sobrevivência, contagem de UFC/mL das cepas de C. albicans e de hemócitos. Os resultados foram analisados estatisticamente de acordo com a distribuição normal (ANOVA e Tukey ou Kruskal-Wallis e Dunn) e a curva de sobrevivência pelo teste de Log-rank (Mantel-Cox), p<0,05. As melhores suspensões para CA, CK e CG foram LR (com melhores resultados no perfil de citocinas), LP e LA (com melhores resultados sobre a fagocitose), respectivamente. Em G. mellonella, a suspensão de LR proporcionou aumento da percentagem de sobrevivência e do número de hemócitos e diminuição da contagem de UFC/mL em todos os grupos com diferenças estatisticamente significantes. Sendo assim, pode-se concluir que as suspensões de lactobacilos possuem potencial imunomodulador cepa dependente mas sua mistura não favorece essa qualidade. Além disso, possuem atividade anti-Candida, demonstrada pela diminuição na contagem de Candida(AU)

Many probiotic products are composed by association of lactobacilli in order to potentiate their beneficial effect. Therefore, the objective of this study was evaluating the immunomodulation and anti-Candida activity caused by different combinations of probiotic suspensions in macrophages and in Galleria mellonella. Lactobacillus rhamnosus (LR), L. acidophilus (LA) and L. paracasei (LP) were suspended to obtain the suspensions LR, LA, LP, LR + LA, LR + LP, LA + LP and LR + LA + LP. Suspensions of Candida albicans (CA) ATCC 18804, C. krusei (CK) ATCC 6258 and C. glabrata (CG) ATCC 9030 were used. Mouse macrophages (RAW 264.7) were activated by each suspension of lactobacilli and then challenged by each fungal strain. Phagocytosis, cytokine and nitric oxide production and cell viability were investigated. The best lacobacilli suspension for each fungal strain would be used in the in vivo assays with G. mellonella, however, because of low virulence of the non-albicans species, in vivo tests were performed with CA and LR. Two clinical strains of CA were added: 21 and 60. LR suspensions were injected into the haemolymph of the larvae 24 h before challenging with fungal strains. Survival curve, CA strains CFU/mL counting and hemocytes were determined. The results were statistically analyzed according to their normal distribution (ANOVA and Tukey or Kruskal-Wallis and Dunn) and the survival curve by Log-rank test (MantelCox), p <0.05. The best suspensions for CA, CK and CG were LR (which obtained the best results in the cytokine profile), LP and LA (which obtained the best results on phagocytosis), respectively. In G. mellonella, LR suspension increased percentage survival and hemocyte counting and decreased CFU/mL counting in all groups, with statistically significant differences in the results. Thus, it can be concluded that the suspensions of lactobacilli have immunomodulatory potential strain-dependent but their association does not favor this quality. Besides, they showed anti-Candida activity, demonstrated by the decrease of Candida counting (AU)

Vaginosis citolítica en pacientes con diagnóstico clínico de vulvovaginitis candidiásica / Cytolytic vaginosis in patients with clinical diagnosis of vulvovaginal candidiasis

La microbiota vaginal en la edad fértil está dominada por Lactobacillus spp. Su sobrecrecimiento provoca el aumento en la producción de ácido láctico, disminución del pH vaginal y lisis de células del epitelio vaginal, causando vaginosis citolítica, la cual se confunde con vulvovaginitis candidiásica (VVC) por la aparición de signos y síntomas similares. Se determinó la prevalencia de vaginosis citolítica, así como sus características clínicas y epidemiológicas en pacientes que consultaron con clínica sugestiva de VVC durante el período agosto-septiembre del año 2015. Se encontraron 12 pacientes con clínica compatible de VVC. En 11 (91,7%) se realizó el diagnóstico de vaginosis citolítica y en 1 (8,3%) de vulvovaginitis candidiásica. El 63,7% de las pacientes (7) tenían diagnóstico previo de VVC recurrente. Los síntomas más frecuentes fueron prurito vaginal o vulvar y secreción vaginal blanquecina de aspecto grumoso como leche cortada. La clínica iniciaba entre los 4-6 días posteriores a la menstruación, y desaparecía días antes o justo con el inicio de una nueva fase menstrual. En conclusión, se encontró que en las pacientes examinadas la sintomatología de ambas patologías resultó indistinguible, por tanto el diagnóstico clínico no es suficiente y debe recurrirse al diagnóstico de laboratorio para distinguir entre vaginosis citolítica y vulvovaginitis candidiásica.

The vaginal microbiota in fertile age is dominated by Lactobacillus spp. Its overgrowth causes increased production of lactic acid, decreased vaginal pH and lysis of cells of the vaginal epithelium, causing cytolytic vaginosis, which can be confused with vulvovaginal candidiasis (VVC) due to the similar signs and symptoms. The prevalence of cytolytic vaginosis, as well as its clinical and epidemiological characteristics was determined in patients who consulted with suggestive signs and symptoms of VVC during the period August to September of the year 2015. Twelve patients with signs and symptoms compatible with VVC were considered. Of those, 11 (91.7%) were diagnosed as cytolytic vaginosis and 1 (8.3%) corresponded to vulvovaginal candidiasis. From the 12 patients included, 7 (63.7%) had previous diagnosis of recurrent VVC. The most frequent symptoms recorded were vaginal or vulvar pruritus and whitish, cheesy or curd-like vaginal discharge. Signs and symptoms started between 4 to 6 days after menstruation, and disappeared days before or just with the onset of a new menstrual cycle. In conclusion, it was found that in the patients examined, signs and symptoms of both pathologies were indistinguishable. Therefore, the clinical diagnosis is not enough and laboratory studies should be used to distinguish between cytolytic vaginosis and vulvovaginal candidiasis.

Vaginosis citolítica: presentación de un caso clínico / Cytolytic vaginosis: presentation of a clinical case

La vaginosis citolítica, descrita hace 30 años como citólisis de Döderlein, es frecuente en mujeres en la edad reproductiva y, por las características de flujo vaginal blanquecino y síntomas clínicos, es indistinguible de la vulvovaginitis micótica. Se presenta el caso de una paciente, con diagnóstico clínico presuntivo de vulvovaginitis por Candida spp. a repetición, tratada empíricamente con antifúngicos por año y medio sin ninguna mejoría. Luego de estudios microbiológicos, la coloración de Gram, demostró la presencia de 50 bacilos grampositivos por campo y abundantes núcleos celulares desnudos. El cultivo resultó puro para Lactobacillus spp., lo que permitió confirmar el diagnóstico de vaginosis citolítica. La paciente fue tratada con ampicilina-sulbactam y no ha vuelto a presentar recidivas. En conclusión, es fundamental determinar el pH vaginal de las pacientes en la consulta, así como practicar una coloración de Gram de la secreción vaginal para poner en evidencia los cambios celulares por el exceso de ácido en la vagina y así evitar tratamientos antifúngicos innecesarios que acrecentarán los trastornos de la microbiota vaginal.

Cytolytic vaginosis, described 30 years ago as Döderlein cytolysis, is common in women of reproductive age and, due to the characteristics of whitish vaginal discharge and clinical symptoms, is indistinguishable from mycotic vulvovaginitis. We describe the case of a patient with presumptive clinical diagnosis of recurrent vulvovaginitis by Candida spp. treated empirically with antifungal agents for one and a half years without improvement. After microbiological studies, Gram staining demonstrated the presence of 50 Gram-positive bacilli per field and abundant nude cell nuclei. The culture recovered pure Lactobacillus spp. which permitted the diagnosis of cytolytic vaginosis. The patient was treated with ampicillin-sulbactam and since, has not had recurrences. In conclusion, it is essential to examine the pH of the patient vaginal discharge, as well as to practice a Gram staining of the vaginal secretion to demonstrate the cellular changes produced by the excess of acid in the vagina and therefore avoid unnecessary antifungal treatments that will produce undue changes of the vaginal microbiota.

Prevalence of lactic acid bacteria in sliced cooked ham as an indicator of its shelf life / Prevalencia de bacterias de ácido láctico en jamón cocido friccionado como indicador de su vida útil

Background: Ready to eat sliced-cooked-meat-products (RTE) are popular convenience foods. Slicing of such products at retail point is a common practice in supermarkets. Due to handling, as well as the supermarket environment, it has been suggested that the counts and presence of specific lactic acid bacteria (LAB) in these products may be associated with their rapid deterioration and short shelf life. Objectives: The aim was to quantify and identify LAB in commercial sliced cooked ham. Methods: Thirty samples of sliced cooked ham were collected from five supermarkets. Each sample was analyzed in terms of: Lactobacillus on De Man Rogosa, Sharpe agar (30°C), mesophilic (30°C) and thermophilic (37ºC) Streptococcus on M17 agar containing 1% lactose, Lactococcus on LM17 agar containing 100 µg per liter of cycloheximide (25°C) and Enterococci on Kanamicin Aesculin Azide agar containing kanamycin (37°C), all under anaerobic conditions (Gas-Pack System®, BBL) for 48-72h. Twenty-one colonies were randomly picked and physiologically and biochemically characterized. Results: No difference was observed in genuses between supermarkets; however, significant differences were observed between microbial genuses. Enterococci showed the lowest count (2.34 ± 0.05 log10 CFU/g) and Lactobacilli had the highest counts (5.98 ± 0.04 log10 CFU/g). Of the strains isolated 23.8% were identified as thermophillic Lactobacillus, 23.8% mesophilic Lactobacillus, 28.6% Enterococcus, 14.3% Lactococcus and 9.5% Streptococcus. From these, only six could be characterized to species level; one was L.lactis subsp. lactis and five were L. amylolyticus. Conclusions: High LAB counts are a common cause of spoilage in RTE meat products, since these are commonly found in meat environments. Therefore a study on the processing, distribution and in-site handling of RTE meat products at supermarkets may be necessary in order to increase its shelf life.

Antecedentes: Los productos cárnicos rebanados cocidos listos para el consumo (RTE) son alimentos populares de conveniencia. El rebanado de este tipo de productos es una práctica común en los supermercados. Debido a la manipulación, así como al entorno del supermercado, se ha sugerido que los recuentos y la presencia de bacterias ácido lácticas (LAB) en estos productos pueden estar asociados con su rápido deterioro y corta vida en anaquel. Objetivo: El objetivo de este trabajo fue cuantificar e identificar LAB en jamón cocido rebanado en el supermercado. Métodos: se recolectaron 30 muestras de jamón cocido rebanado de cinco supermercados. Cada muestra se analizó en términos de: Lactobacillus en agar De Man Rogosa, Sharpe (30°C), Streptococcus mesofílicos (30°C) y termofílicos (37°C) en agar M17 conteniendo 1% de lactosa, Lactococcus en agar LM17 contiendo 100 µg por litro de ciclohexamida (25°C), y Enterococci en agar Kanamicin Aesculin Azide conteniendo kanamicina (37°C), todos bajo condiciones anaeróbicas (Gas-Pack System®, BBL) por 48-72h. Veintiún colonias seleccionadas al azar fueron fisiológica y bioquímicamente caracterizadas. Resultados: No se observaron diferencias en el género entre los supermercados; sin embargo, se observaron diferencias significativas entre género microbiano. Los enterococos mostraron los recuentos más bajos (2,34 ± 0,05 log10 CFU/g) y los lactobacilos los más altos (5,98 ± 0,04 log10 CFU/g). De las cepas aisladas 23,8% fueron identificados como Lactobacillus termófilos, 23,8% Lactobacillus mesófilos, 28,6% Enterococcus, 14,3% Lactococcus y 9,5% Streptococcus. De éstos, sólo seis pudieron caracterizarse a nivel de especie; uno fue L.lactis subsp lactis y cinco fueron L. amylolyticus. Conclusiones: Conteos altos de LAB son una causa común del deterioro de los productos cárnicos listos para el consumo, ya que estos se encuentran comúnmente en los entornos donde se maneja carne. Por lo tanto, es necesario llevar a cabo un estudio sobre el procesamiento, distribución y manejo de productos cárnicos listos para el consumo en los supermercados con el fin de aumentar su vida en anaquel.

Avaliação dos efeitos de 5-hidroxitriptofano em-hidroxibenzilhidrazine associados a Lactobacillus spp. na morfometria intestinal e imunomarcação de serotonina em frangos de corte desafiados com Salmonella Enteridis / Assessment of the effects of 5-hydroxytryptophan and m-hydroxybenzyl hydrazine associated with Lactobacillus spp. in intestinal morphology and immunostaining of serotonin in broilers challenged with Salmonella Enteritidis.

As células enterocromafins são um dos componentes da mucosa intestinal que liberam serotonina para o lúmen, promovendo atividades secretórias e crescimento celular de vários tecidos, incluindo vilosidades intestinais. O presente estudo avaliou as influências do 5-hidroxitriptofano (5HTP) e do m-hidroxibenzilhidrazine (NSD1015), associados a Lactobacillus spp., sobre o peso corporal e o desenvolvimento das vilosidades intestinais na porção proximal do duodeno de frangos de corte desafiados com Salmonella Enteritidis. Verificou-se também se a presença de Lactobacillus spp. e Salmonella Enteritidis influenciaram a imunomarcação de serotonina no duodeno e, para isso, o estudo foi dividido em dois experimentos, com e sem desafio por S. Enteritidis. No Experimento 1, em aves sem desafio, os pesos corporais não diferiram significantemente (p>0,05) e, no Experimento 2, aves com desafio, os tratamentos com o precursor isolado e associado a Lactobacillus spp. determinaram maior peso corporal das aves. Nos dois experimentos, as aves tratadas com 5HTP apresentaram aumento na densidade e altura das vilosidades no duodeno, sugerindo a atuação de 5HTP como um agente trófico. A administração de Lactobacillus spp. também determinou altura maior de vilosidades duodenais. Quanto a imunomarcação de serotonina, as aves tratadas com Lactobacillus spp. no Experimento 1 e as aves tratadas com Lactobacillus spp. e desafiadas com S. Enteritidis no Experimento 2, apresentaram valores superiores aos demais tratamentos, sugerindo que a presença destas bactérias promove maior liberação de serotonina para o duodeno, porém o mecanismo exato de como este processo ocorre necessita ser mais elucidado...

Enterochromaffin cells are components of the intestinal mucosa to release serotonin lumen, promoting cell growth and secretory activity of various tissues, including intestinal villi. This study evaluated the influence of 5-hydroxytryptophan (5HTP) and m-hidroxibenzilhidrazine (NSD1015) associated with Lactobacillus spp. on body weight and development of intestinal villi in the proximal duodenum of broilers challenged with Salmonella Enteritidis. It was found that the presence of Lactobacillus spp. and Salmonella Enteritidis influenced immunostaining of serotonin in the duodenum. The study was divided into two experiments with and without challenge by S. Enteritidis. In Experiment 1, birds without challenge, body weights did not differ significantly (p>0.05), and in Experiment 2, the treatments with precursor and precursor associated with Lactobacillus spp. determined higher body weight of the birds. In both experiments the birds treated with 5HTP showed increased density and villus height in the duodenum, suggesting the presence of 5HTP as a trophic agent. The use of Lactobacillus spp. also determined greater duodenal villus height. The immunostaining of serotonin, birds treated with Lactobacillus spp. in Experiment 1, and the birds treated with Lactobacillus spp. and challenged with S. Enteritidis in Eperiment 2 showed higher values, suggesting that the presence of these bacteria promotes greater release of serotonin into the duodenum. The exact mechanism of how this process occurs needs to be further elucidated...

Quality of fermented milks produced with Lactobacillus rhamnosus and Lactobacillus fermentum isolated from artisanal cheeses / Qualidade de leites fermentados produzidos com Lactobacillus rhamnosus e Lactobacillus fermentum isolados de queijos artesanais

Leites fermentados por Lactobacillus rhamnosus e Lactobacillus fermentum, isolados de queijos artesanais, foram produzidos e avaliados quanto às características físico-químicas e microbiológicas durante 45 dias de estocagem a 8-10ºC. Análises sensoriais foram realizadas aos 15 e 60 dias. Ambos os leites fermentados apresentaram contagens adequadas das bactérias láticas, superiores a 108 UFC/g, durante toda a estocagem. As médias das análises físico-químicas e microbiológicas dos produtos durante a estocagem foram iguais e todos atenderam às especificações da legislação brasileira. Melhores resultados de avaliações sensoriais (P<0.05) foram aos 15 dias de estocagem. Leites fermentados por L. fermentum obtiveram melhor aceitação sensorial aos 60 dias de estocagem, quando apresentavam acidez titulável inferior à encontrada no leite fermentado por L. rhamnosus. Portanto, a utilização dessas culturas pode ser viável para a elaboração de novos leites fermentados, que apresentariam prazo de validade de 45 dias de estocagem sob refrigeração.(AU)

Probióticos na avicultura / Probiotics on poultry production

Com a proibição do uso de antibióticos como melhoradores de desempenho, aumentaram as pesquisas para desenvolver produtos alternativos que fossem eficientes para melhorar o desempenho zootécnico dos animais de produção e manter a saúde intestinal deles, por meio da redução de patógenos e da imuno modulação do trato gastrointestinal. O uso de probióticos já é conhecido há muito tempo, entretanto os mecanismos de ação destes produtos e a interação deles com o organismo animal hospedeiro estão sendo mais profundamente pesquisados agora. Sabe-se que os probióticos auxiliam no equilíbrio da microbiota intestinal, podendo, por meio deste efeito, melhorar o desempenho dos animais. Esta atividade se dá pela produção de diferentes metabólitos com ação antimicrobiana e também pela competição por sítios de ligação com bactérias patogênicas na mucosa do trato gastrointestinal ou através de imunomodulação no organismo animal. Devido a todas as ações dos probióticos, eles são utilizados como melhoradores de desempenho animal e também podem ser uma ferramenta para controle de enterobactérias de importância em saúde pública.

As antibiotics growth promoters have been banned from animal feed, probiotics have received special attention as alternative products to enhance performance from livestock and keep their gastrointestinal health. Probiotics benefits are known since long time ago, however, the mechanisms of action of these products are being better researched now. Is known that probiotics could balance intestinal microbiot and due to this, they can improve animal performance. This activity is due to the production of different metabolites with antimicrobial activity and competition by bind sites on gastrointestinal mucosal. Due to all this probiotic actions, they are recommended as animal growth promoters and also may be an important tool to control enterobacterias considered risky to public health.

In vitro probiotic potential of Lactobacillus spp. isolated from fermented milks / Potencial probiótico in vitro de Lactobacillus spp. isolados de leites fermentados

The potential of in vitro probiotic Lactobacillus spp. was evaluated in fermented milks marketed in Belo Horizonte, MG, Brazil. Of the samples analyzed, 86.7% had at least 10(6) CFU/mL of Lactobacillus spp., complying with the Brazilian quality standards for fermented milks. Furthermore, 56.7% had minimum count ranging from 10(8) to 10(9) CFU/mL, which is in accordance with legal parameters. The remaining 43.3% would not be able to satisfactorily guarantee benefits to consumers. The amount of Lactobacillus spp. varied between batches of products, which may indicate failures in monitoring during manufacture, transport or storage. All strains of Lactobacillus spp. showed some inhibitory activity against the indicator microorganisms, being more pronounced against pathogenic microorganisms than against non-pathogenic (P<0.05). Samples of Lactobacillus spp. showed different profiles of antimicrobial susceptibility, with an occurrence of cases of multidrug resistance. All strains tested showed sensitivity to bile salts (0.3%) and resistance to gastric pH (2.0). Lactobacillus spp. of commercial fermented milks should be present in higher amounts in some brands, be resistant to bile salts and have no multiple resistance to antimicrobials.

O potencial probiótico in vitro de Lactobacillus spp. foi avaliado em leites fermentados comercializados em Belo Horizonte, MG, Brasil. Das amostras analisadas, 86,7% apresentaram quantidade mínima de 106UFC/mL de Lactobacillus spp., enquadrando-se no padrão brasileiro de qualidade de leites fermentados. Além disso, 56,7% apresentaram quantidade mínima de 108a 109UFC/mL, estando em conformidade com os parâmetros legais vigentes. As demais 43,3% não estariam aptas a garantir satisfatoriamente efeitos benéficos aos consumidores. Observou-se variação na quantidade de Lactobacillus spp. entre lotes dos produtos, o que pode indicar falhas no monitoramento durante a fabricação, transporte ou estocagem destes. Todas as amostras de Lactobacillus spp. demonstraram alguma atividade inibitória frente aos microrganismos indicadores, sendo mais intensa contra microrganismos patogênicos que não patogênicos (P<0,05). As amostras de Lactobacillus spp. apresentaram diferentes perfis de susceptibilidade aos antimicrobianos, ocorrendo casos de multirresistência. Todas as amostras testadas apresentaram sensibilidade in vitro a sais biliares (0,3%) e resistência in vitro ao pH gástrico (2,0). Lactobacillus spp. dos leites fermentados testados deveriam estar presentes em quantidades mais elevadas em algumas marcas comerciais, possuir resistência aos sais biliares e não apresentar resistência múltipla aos antimicrobianos.

Anaerobically fermented colostrum: an alternative for feeding calves / Colostro fermentado anaerobicamente: uma alternativa para alimentação de bezerras

Milk or commercial milk replacer blends are the most expensive components in final costs of calves breeding. Colostrum is available and it is the appropriate sources for calves' nutrition, being an excellent option as milk substitute. Besides having both nutritional and immunological characteristics that are superior to milk, colostrum represents no costs to the producer. However, difficulties in preservation of colostrum generate controversy results. The aim of this study was to evaluate the anaerobically fermented colostrum (colostrum silage) as liquid diet for dairy calves. We evaluated the microbiological and physicochemical properties of silage, and performance of 31 animals up to 60 days age. From 21 days until 360 days of fermentation we isolated only bacteria of the genus Lactobacillus spp. The physicochemical evaluation of colostrum silage revealed a tendency to maintain the protein, dry matter and fat values during the evaluation period. The average weight gain of calves fed with milk was 0.6kg day-1 for female and 0.6g day-1 for males while those fed with colostrum silage was significantly higher (P<0.05) with a mean of 0.7kg day-1 for female and 0.7kg day-1 for males. The results of this study demonstrated that colostrum silage keeps the properties necessary for the development of the calves and is a suitable replacement for calves' milk.

O leite ou substitutos comerciais do leite são os componentes mais caros do custo final da criação de bezerras. O colostro é uma fonte adequadada de nutrientes, sendo uma excelente opção como substituto do leite. Além de ter características nutricionais e imunológicas superiores ao leite, o colostro não representa custos para o produtor. No entanto, as dificuldades de preservação do colostro têm gerado resultados controversos. O objetivo deste estudo foi avaliar o colostro fermentado anaerobicamente (silagem de colostro) como dieta líquida para bezerras leiteiras. Foram avaliadas as propriedades microbiológicas e físico-químicas da silagem de colostro e o desempenho de 31 animais até 60 dias de idade. A partir de 21 dias até 360 dias de fermentação, foram isoladas apenas bactérias do género Lactobacillus spp. A avaliação físico-química da silagem de colostro revelou uma tendência para manter os valores de proteína, gordura e matéria seca durante o período de avaliação. O ganho médio de peso dos bezerros alimentados com leite foi de 0,6kg dia-1 para as fêmeas e de 0.6kg dia-1 para os machos, enquanto que o ganho médio de peso para os animais alimentados com silagem colostro foi significativamente maior (P<0,05), com média de 0,7kg dia-1 para as fêmeas e 0,7kg dia-1 para os machos. Os resultados deste estudo mostram que a silagem de colostro mantém as propriedades necessárias para o desenvolvimento dos animais, sendo um substituto adequado para o leite.

Cuantificación de bacterias relacionadas con la caries dental en saliva de adultos y adultos mayores / Quantification of caries-associated bacteria from saliva of adults and older adults

Introducción: Se considera que Streptococcus mutans (S. mutans) y Lactobacillus spp. se asocian con la caries. Otras especies del biofilm oral, como Streptococcus sanguinis (S. sanguinis) han sido sindicadas como protectoras, pero principalmente en niños. Existe escasa evidencia sobre el nivel de estas bacterias en adultos mayores. Objetivo: Determinar si existen diferencias en los recuentos microbianos de tres especies relacionadas con la caries en pacientes adultos y adultos mayores. Materiales y Métodos: Una muestra de pacientes por conveniencia compuesta de 63 pacientes (18 a 79 años) proporcionó saliva estimulada con la que se sembraron placas de agar MSB, MM10 SB y Agar Rogosa para el cultivo de S. mutans, S. sanguinis y Lactobacillus spp., respectivamente. Los recuentos bacterianos fueron expresados como UFC/mL. Resultados: Los recuentos de S. mutans y Lactobacillus spp. no mostraron variaciones relacionadas con la edad (p>0.05). Los adultos mostraron mayores recuentos de S. sanguinis que los adultos mayores, 3.7 x 105 +/- 3.8 x 105 UFC/mL y 5.9 x 104 +/- 9.4 x 104 UFC/mL, respectivamente (p<0.05). Conclusiones: La edad no parece afectar los niveles de especies tradicionalmente consideradas como cariogénicas. Estos resultados sugieren que la edad puede relacionarse con los patrones de colonización de S. sanguinis en el biofilm oral.

Introduction: Streptococcus mutans (S. mutans) and Lactobacillus spp. have been traditionally associated with caries, regardless of the subject’s age. Other oral biofilm species have been linked as caries protective, including Streptococcus sanguinis (S. sanguinis), but mainly in children. Scarce evidence exists on the levels of these bacteria in older adults. Aim: To determine whether there are differences in the microbial counts of three caries-associated bacterial species in adults and older adults. Methodology: A convenience sample of sixty three patients, aged 18 to 79 years, participated in the study. Stimulated saliva samples were obtained and in MSB, MM10 and Rogosa agar plates for the culture of S. mutans, S. sanguinis and Lactobacillus spp., respectively. Bacterial counts were obtained by microscopic observation (10x) of the colonies and expressed in CFU/mL. Results: Bacterial counts of S. mutans and Lactobacillus spp. did not reveal age-related differences (p>0.05). Adults showed higher S. sanguinis counts than older adults with 3.7 x 105 +/- 3.8 x 105 CFU/mL and 5.9 x 104 +/- 9.4 x 104 CFU/mL, respectively (p<0.05). Conclusions: Age does not seem to affect the levels of bacterial species traditionally associated with caries. The results suggest that age may be related to colonization patterns of S. sanguinis in the oral biofilm.

Molecular identification of Lactobacillus spp. associated with puba, a Brazilian fermented cassava food

Puba or carimã is a Brazilian staple food obtained by spontaneous submerged fermentation of cassava roots. A total of 116 lactobacilli and three cocci isolates from 20 commercial puba samples were recovered on de Man, Rogosa and Sharpe agar (MRS); they were characterized for their antagonistic activity against foodborne pathogens and identified taxonomically by classical and molecular methods. In all samples, lactic acid bacteria were recovered as the dominant microbiota (7.86 ± 0.41 log10 CFU/g). 16S-23S rRNA ARDRA pattern assigned 116 isolates to the Lactobacillus genus, represented by the species Lactobacillus fermentum (59 isolates), Lactobacillus delbrueckii (18 isolates), Lactobacillus casei (9 isolates), Lactobacillus reuteri (6 isolates), Lactobacillus brevis (3 isolates), Lactobacillus gasseri (2 isolates), Lactobacillus nagelii (1 isolate), and Lactobacillus plantarum group (18 isolates). recA gene-multiplex PCR analysis revealed that L. plantarum group isolates belonged to Lactobacillus plantarum (15 isolates) and Lactobacillus paraplantarum (3 isolates). Genomic diversity was investigated by molecular typing with rep (repetitive sequence)-based PCR using the primer ERIC2 (enterobacterial repetitive intergenic consensus). The Lactobacillus isolates exhibited genetic heterogeneity and species-specific fingerprint patterns. All the isolates showed antagonistic activity against the foodborne pathogenic bacteria tested. This antibacterial effect was attributed to acid production, except in the cases of three isolates that apparently produced bacteriocin-like inhibitory substances. This study provides the first insight into the genetic diversity of Lactobacillus spp. of puba.

Resistência bacteriana e ação das bacteriocinas de Lactobacillus spp em Staphylococcus aureus isolados de mastite bovina / Bacterial resistance and bacteriocins action in Staphylococcus aureus isolated from bovine mastitis

Staphylococcus aureus is the major pathogen causing intramammary infections in dairy cattle worldwide. Among the factors that contribute to its spread and infectious potential is the ability to overcome the mechanisms of antimicrobials activity. The present work investigated the antimicrobial resistance pattern and sensibility to bacteriocins produced by strains of Lactobacillus spp of 30 isolates of S. aureus from mastitis. From this, 29 are beta-lactamase producers. Eight isolates (26.6 percent) showed resistance to at least four antibiotics being considered multiresistent. All of them were mecA-positive. Otherwise, all isolates tested showed sensibility to at least one of the four bacteriocin producer strains. Due to the significant depletion of the efficacy of antimicrobials, pathogen growth inhibition by bacteriocins seems an alternative of biological control in infectious processes.

Vaginal lactobacilli as potential probiotics against Candida spp

Urogenital infections affect millions of people every year worldwide. The treatment of these diseases usually requires the use of antimicrobial agents, and more recently, the use of probiotic lactic acid bacteria (LAB) cultures for the management of vaginal infections has been extensively studied. In this work, 11 vaginal lactobacilli isolates, previously obtained from healthy patients, were studied to screen microorganisms with probiotic properties against Candida spp. The LAB were tested for their ability of auto-aggregation, coaggregation with C. albicans, C. glabrata, C. krusei, and C. tropicalis, adhesion to Caco-2 epithelial cells and production of lactic acid and hydrogen peroxide (H2O2). All lactobacilli isolates tested were able to auto-aggregate (ranging from 25.3 percent to 75.4 percent assessed at 4 hours of incubation) and to co-aggregate with the four Candida species into different degrees; among them L. crispatus showed the highest scores of coaggregation. The highest amount of lactic acid was produced by L. salivarius (13.9 g/l), followed by L. johnsonii (6.5 g/l), L. acidophilus (5.5 g/l), and L. jensenii (5.4 g/l). All isolates produced H2O2 , but the highest levels (3 -10 mg/l) were observed for L. acidophilus, L. crispatus, L. gasseri, L. johnsonii, and L. vaginalis. Only L. agilis, L. jensenii, L. johnsonii and L. ruminus were able to adhere to epithelial Caco-2 cells. Among the isolates evaluated, L agilis, L. jensenii, L. johnsonii, and L. ruminus exhibited simultaneously several desirable properties as potential probiotic strains justifying future studies to evaluate their technological properties in different pharmaceutical preparations for human use.