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Postpartum reproductive infections in cows generate significant economic losses. The use of lactic acid bacteria in animal health is an alternative tool to avoid antibiotic therapy in the prevention/treatment of bovine reproductive infections. In previous studies, 6 lactic bacteria from bovine mammary glands and vagina with beneficial, safe and technological characteristics were selected, and included in probiotic/phytobiotic formulas (combined with Malva and Lapacho extracts). In this work, probiotic and phytobiotic formulations were designed and their long-term viability determined. They were administered intravaginally to 30 females pregnant bovine pre and postpartum. The modification of the native microbiota and permanence/colonization of cultivable bacteria was evaluated, and also the safety of the designed products through the application of nutritional, clinical, hematological and biochemical parameters. The microorganisms maintained their viability up to 9 months at refrigeration temperature. The number of cultivable bacteria showed different pattern: total aerobic mesophylls increased slightly in all experimental groups, while Enterobacteriaceae increased after delivery, except in beneficial acid lactic bacteria + vegetable extract cows. Control and vegetable extract females showed the highest numbers of Enterobacteriaceae at the end of the trial (30 days postpartum). The number of lactic acid bacteria increased significantly in all the groups between 15 days pre and postpartum. The different parameters evaluated demonstrate the safety and harmlessness of the designed formulas, without producing local and systemic adverse effects in the cows.
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As a first step for the use of probiotics in a formula for cattle, it is required to have available low-cost culture medium(s) and efficient production conditions for the growth of probiotic bacteria and high production of cell biomass. De Man-Rogosa-Sharpe medium, used frequently for lactic acid bacteria (LAB) contains adequate ingredients for their growth but is very expensive for industrial application. The nutrients required for LAB growth are strain-dependent. In this work, traditional culture media were evaluated omitting and/or modifying ingredients in their composition, as carbon or nitrogen source, on the basis of their low-cost industrial waste, to select those supporting the most efficient growth. The results showed that the formulation of culture media containing fructose (0.5%) and molasses (1.0%) was better for the growth and production of cell biomass for all the strains assayed, except Lactobacillus gasseri CRL1421 growing in 1.5% corn syrup. FM902 yeast extract at concentrations between 1.5% and 2.5% was the most adequate for most of the strains. The LAB grown in the designed media maintained the beneficial properties for which they selected. The use of the culture media designed to produce biomass decrease production costs, which is an important step for the feasible industrial production of probiotic pharmaceuticals.
Assuntos
Lactobacillales , Probióticos , Bovinos , Animais , Biomassa , Melaço , Meios de Cultura , FermentaçãoRESUMO
AIMS: The aim was to complement the evaluation of functional and safety properties of beneficial vaginal lactic acid bacteria to select the most adequate strains as potential probiotics for their inclusion in a vaginal probiotic formula. METHODS AND RESULTS: Twenty-four beneficial vaginal lactic acid bacteria strains previously isolated from healthy women were characterized by applying phenotypical and genetic techniques. The biofilm formation and exopolysaccharide production by the different strains showed a strong influence of the growth medium on the expression of these properties. The evaluated strains evidenced different antibiotic susceptibility patterns by phenotypic tests, while the resistance genes were not always correlated with the phenotypic profiles. None of the evaluated strains showed lecithinase, gelatinase, or ß-hemolytic activity. Based on the results obtained, Lactobacillus gasseri CRL 1320 and Lacticaseibacillus rhamnosus CRL 1332 were selected to advance in the design of the products. CONCLUSIONS: A wide variability in the properties expressed by beneficial vaginal lactic acid bacteria strains were shown, evidencing a strain-specific and independent behaviour of the species/metabolic group where classified. The selection criteria applied provided a valuable tool for the selection of the best vaginal lactobacilli strains for their inclusion in formulas for vaginal application. SIGNIFICANCE AND IMPACT OF THE STUDY: Since probiotics are an important tool for improving human health, the designed formula are an excellent alternative to reconstitute the vaginal microbiota and prevent/treat urogenital tract infections in women. The results obtained contribute to deepen the characterization of homologous beneficial vaginal lactic acid bacteria and advance in the design of vaginal probiotic products.
Assuntos
Lactobacillales , Probióticos , Feminino , Humanos , Lactobacillales/genética , Vagina/microbiologia , Antibacterianos , Higiene , Fosfolipases , GelatinasesRESUMO
Modulation of animal gut microbiota is a prominent function of probiotics to improve the health and performance of livestock. In this study, a large-scale survey to evaluate the effect of lactic acid bacteria probiotics on shaping the fecal bacterial community structure of feedlot cattle during three experimental periods of the fattening cycle (163 days) was performed. A commercial feedlot located in northwestern Argentina was enrolled with cattle fed mixed rations (forage and increasing grain diet) and a convenience-experimental design was conducted. A pen (n = 21 animals) was assigned to each experimental group that received probiotics during three different periods. Groups of n = 7 animals were sampled at 40, 104 and 163 days and these samples were then pooled to one, thus giving a total of 34 samples that were subjected to high-throughput sequencing. The microbial diversity of fecal samples was significantly affected (p < 0.05) by the administration period compared with probiotic group supplementation. Even though, the three experimental periods of probiotic administration induced changes in the relative abundance of the most representative bacterial communities, the fecal microbiome of samples was dominated by the Firmicutes (72-98%) and Actinobacteria (0.8-27%) phyla, while a lower abundance of Bacteroidetes (0.08-4.2%) was present. Probiotics were able to modulate the fecal microbiota with a convergence of Clostridiaceae, Lachnospiraceae, Ruminococcaceae and Bifidobacteriaceae associated with health and growth benefits as core microbiome members. Metabolic functional prediction comparing three experimental administration periods (40, 104 and 163 days) showed an enrichment of metabolic pathways related to complex plant-derived polysaccharide digestion as well as amino acids and derivatives during the first 40 days of probiotic supplementation. Genomic-based knowledge on the benefits of autochthonous probiotics on cattle gastrointestinal tract (GIT) microbiota composition and functions will contribute to their selection as antibiotic alternatives for commercial feedlot.
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Lactobacillales , Microbiota , Probióticos , Animais , Bactérias/genética , Bovinos , Fezes/microbiologia , Probióticos/farmacologia , RNA Ribossômico 16S/análise , RNA Ribossômico 16S/genéticaRESUMO
Lactobacilli are the predominant microorganisms of the healthy human vagina. A novel alternative for the prevention and treatment of female urogenital tract infections (UGTI) is the inclusion of these microorganisms as active pharmaceutical ingredients in probiotic formulas, and more recently in female hygienic products. Probiotics are defined as "live microorganisms that, when administered in adequate amounts, confer a health benefit on the host." A list of requirements must be considered during the development of probiotic product/formula for the female urogenital tract (UGT). This review aims to resume the requirements, probiotic characteristics, and clinical trial applied to determine the effect of probiotic and potentially probiotic strains on different woman's physiological and pathological conditions, and in preterm birth prevention. A revision of female hygienic products available in the world market is included, together with novel studies applying nanotechnology for Lactobacillus incorporation in hygienic products. Further studies and well-designed clinical trials are urgently required to complement the current knowledge and applications of probiotics in the female UGT. The use of probiotic formulas and products will improve and restore the ecological equilibrium of the UGT microbiome to prevent and treat UGTI in women under different conditions.
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Produtos de Higiene Feminina/microbiologia , Lactobacillus , Microbiota , Probióticos/uso terapêutico , Vagina/microbiologia , Candidíase Vulvovaginal/terapia , Portador Sadio/terapia , Cesárea , Parto Obstétrico , Feminino , Genitália Feminina/microbiologia , Humanos , Nanotecnologia , Nascimento Prematuro/microbiologia , Nascimento Prematuro/prevenção & controle , Infecções Estreptocócicas/terapia , Streptococcus agalactiae , Vaginite por Trichomonas/terapia , Sistema Urinário/microbiologia , Vaginose Bacteriana/terapiaRESUMO
In recent years, veterinary probiotic formulations constitute an interesting alternative to the use of antibiotics in animals for human consumption, but beneficial microorganisms must meet certain requirements to be included in these products. The objective of this work was to evaluate the safety and innocuity of beneficial autochthonous lactic bacteria (BALB) as well as to determine their beneficial, environmental, and technological characterization. Antibiotic resistance was assayed using phenotypic and genotypic methodology. A bovine vaginal fluid simulated medium (MSBVF) was designed where growth, pH changes, and expression of beneficial characteristics of lactic bacteria were evaluated; additionally, the optimal culture conditions in commercial media were determined in order to obtain the highest biomass production of the strains. Finally, the best strains were lyophilized and administered intravaginally to pregnant cows and their permanence in the vagina and adverse effects were evaluated. The results show that most of the strains were resistant to vancomycin, tetracycline, and streptomycin, with a high sensitivity to ampicillin, gentamicin, and clindamycin. The strains evaluated did not show gelatinase or hyaluronidase activity; however, 11 strains produced α-type hemolysis. The optimal growth of the microorganism was obtained in MRS broth, under slight agitation and without pH control. The strains grown in the MSBVF grew well and maintained the probiotic properties. Animals treated with probiotics bacteria did not show systemic or local inflammation. These strains can be included in a probiotic veterinary product to be applied to different bovine mucosa.
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Lactobacillales , Probióticos , Vagina/microbiologia , Administração Intravaginal , Animais , Antibacterianos/farmacologia , Bovinos , Meios de Cultura , Feminino , GravidezRESUMO
Introducción: Portulaca oleracea, conocida como verdolaga y considerada culturalmente una maleza, presenta múltiples propiedades benéficas que permiten emplearla como alimento debido a su fácil acceso y bajo costo. No se detectan muchos estudios sobre el tema. Objetivos: elaborar productos alimenticios empleando verdolaga como materia prima, con características organolépticas adecuadas; calcular su valor nutricional y determinar el porcentaje de ingesta diaria recomendada (IDR) que cubre; identificar el nivel de conocimiento sobre la verdolaga y sus propiedades nutricionales; determinar las características organolépticas de los alimentos elaborados, el grado de aceptabilidad y satisfacción en la población en estudio. Materiales y métodos: estudio de tipo descriptivo, no experimental y experimental (pre experimento), de corte transversal. Se elaboraron tres preparaciones con verdolaga: sopa (S), tortilla (T) y ensalada (E), y se sometieron a degustación. Resultados: el valor calórico de las preparaciones por porción fue: S: 24,2 kilocalorías (kcal)/200 g, T: 129,2 kcal/200 g, y E: 116,16 kcal/150 g. Los porcentajes de IDR que cubre una porción de cada alimento fueron: S: 1,21% kcal, 1,31% hidratos de carbono, 1,96% proteínas, 0,54% lípidos; T: 6,43% kcal, 3,48% hidratos de carbono, 9,54% proteínas, 11,81% lípidos, y E: 5,8% kcal, 5,06% hidratos de carbono, 4,28% proteínas y 8,6% lípidos. Estos valores se calcularon en base a los valores diarios de referencia de nutrientes (VDR) de declaración obligatoria que son: 2.000 kcal, 300 g de hidratos de carbono, 75 g de proteínas y 55 g de lípidos para un grupo de hombres y mujeres adultos de entre 18 y 60 años de edad. La muestra estuvo formada por 50 personas, de 42 años edad promedio, 58% femenino y 42% masculino, 40% con estudios secundarios, 34% universitarios (N=17), 18% terciarios (N=9) y 8% primarios. El 76% presentó un conocimiento bajo sobre la verdolaga y sus propiedades nutricionales. Las elaboraciones fueron satisfactorias para el 82% de los encuestados y aceptadas por el 90% Conclusiones: en base a las propiedades nutricionales que se sugieren y su alta versatilidad para incorporar en diversas recetas, la verdolaga se presenta como una excelente alternativa para la alimentación humana
Introduction: portulaca oleracea, known as purslane (verdolaga), culturally considered an undergrowth, has multiple beneficial properties allowing to be used as a food, due to its easy access and low cost. There are not many studies on the subject. Objectives: the objective of present work was to prepare food products using purslane as raw material, with adequate organoleptic characteristics. To calculate their nutritional value and to determine the recommended dietary allowance (RDA) percentage they cover. Also, to identify the level of knowledge that a certain population has about purslane and its nutritional properties, and to define the organoleptic characteristics of the foods elaborated, evaluating the degree of acceptability and satisfaction in the population under study. Materials and methods: the study was descriptive, nonexperimental and experimental (pre-experiment), cross-sectional. Three preparations with purslane were elaborated and subjected to tasting: soup (S), omelette (O) and salad (E). Results: tthe caloric value per serving of the preparations was S: 24.2 kcal/200 g; O: 129.2 Kcal/200 g, and E: 116.16 kcal/150 g. The percentages of RDA covered by a serving of each food were: S: 1.21% kcal, 1.31% carbohydrates, 1.96% proteins, 0.54% fats; T: 6.43% kcal, 3.48% carbohydrates, 9.54% proteins, 1.81% fats, and E: 5.8% kcal, 5.06% carbohydrates, 4.28% proteins and 8.6% fats. The values were calculated on referred to the recommended daily intake (RDI), which are: 2,000 kcal, 300 g carbohydrates, 75 g proteins y 55 g fats, defined to a group of adult women and men between 18 and 60 years old. The sample included 50 people with 42 years average age, 58% female and 42%male, 40% with secondary studies, 34% university (N=17), 18% tertiary (N=9) and 8% primary. 76% of the participants presented a low knowledge about purslane and its nutritional properties. The preparations were satisfactory for 82% of the participants and accepted by 90%. Conclusions: due to the suggested nutritional properties and its versatility for its incorporation in various recipes, purslane can be indicated as an excellent alternative for human consumption
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Humanos , Adulto , Pessoa de Meia-Idade , Portulaca , Recomendações NutricionaisRESUMO
The pharmaceutical industry shows an emerging interest in formulas that contain live and beneficial microorganisms, also known as probiotics or pharmabiotics, which in many cases, are host-specific. The resistance to higher temperature is an essential feature of these microorganisms when working on the design of products for vaginal formula. In order to obtain a high number of viable cells and a prolonged shelf life in the designed product, it is required to apply technological procedures using high temperatures or abrupt changes of them, which result in conditions that are different from the optimal growth temperature and can affect the metabolic capabilities of the bacteria when administered to the host in order to reestablish the ecological mucosa. The aim of this work was to evaluate the behavior of 30 different species and strains of autochthonous beneficial vaginal lactobacilli (BVL) when exposed to high temperatures, determine their survival capabilities and analyze their pre-adaptation to those temperatures, in order that they still maintain their viability after technological processes and further conservation. BVL were exhibited to temperatures higher than optimal, with the purpose of evaluating their growth kinetics and parameters. Later, they were exposed to higher temperatures, and then, returned to their optimal, to determine if they were able to grow again. The strains that showed higher resistance were selected, and their viability and beneficial properties studied further. The growth kinetics of strains exposed to higher temperatures showed different patterns, which provided evidence that the thermal adaptation is strain-dependent and is not related to any particular species and/or metabolic group in which the strains were taxonomically classified. The pre-adaptive step allowed the growth of some of the strains, preserving their viability and probiotic properties after the high temperatures were applied. The results shows that BVL can be exposed to high temperatures used in different technological processes that are applied for pharmabiotic formulations, such as spray dried or vacuum rotary evaporation, and/or during the conservation period. The results obtained indicate that some specific BVL strains resist high temperatures and grow afterwards at optimal conditions.
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Técnicas Bacteriológicas , Temperatura Alta , Lactobacillus/crescimento & desenvolvimento , Vagina/microbiologia , Feminino , Humanos , Cinética , Lactobacillus/isolamento & purificação , Probióticos , TemperaturaRESUMO
BACKGROUND: Lactobacillus spp. dominating the vaginal microbiota of healthy women contribute to the prevention of urogenital and sexually transmitted infections. Their protective role in the vagina can be mediated by Lactobacillus cells themselves, metabolites or bacterial components, able to interfere with pathogen adhesion and infectivity. Vulvovaginal candidiasis (VVC) is a common genital infection, caused by the overgrowth of opportunistic Candida spp. including C. albicans, C. glabrata, C. krusei and C. tropicalis. Azole antifungal drugs are not always efficient in resolving VVC and preventing recurrent infections, thus alternative anti-Candida agents based on vaginal probiotics have gained more importance. The present work aims to chemically characterize the biosurfactant (BS) isolated from a vaginal Lactobacillus crispatus strain, L. crispatus BC1, and to investigate its safety and antiadhesive/antimicrobial activity against Candida spp., employing in vitro and in vivo assays. RESULTS: BS isolated from vaginal L. crispatus BC1 was characterised as non-homogeneous lipopeptide molecules with a critical micellar concentration value of 2 mg/mL, and good emulsification and mucoadhesive properties. At 1.25 mg/mL, the BS was not cytotoxic and reduced Candida strains' ability to adhere to human cervical epithelial cells, mainly by exclusion mechanism. Moreover, intravaginal (i.va.) inoculation of BS in a murine experimental model was safe and did not perturb vaginal cytology, histology and cultivable vaginal microbiota. In the case of i.va. challenge of mice with C. albicans, BS was able to reduce leukocyte influx. CONCLUSIONS: These results indicate that BS from vaginal L. crispatus BC1 is able to interfere with Candida adhesion in vitro and in vivo, and suggest its potential as a preventive agent to reduce mucosal damage occasioned by Candida during VVC.
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Antifúngicos/farmacologia , Candida albicans , Candidíase Vulvovaginal , Lactobacillus crispatus/química , Tensoativos/farmacologia , Vagina/microbiologia , Animais , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Candidíase Vulvovaginal/microbiologia , Candidíase Vulvovaginal/prevenção & controle , Feminino , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos BALB C , MicrobiotaRESUMO
Vulvovaginal candidiasis (VVC) is one of the most frequent infections affecting women worldwide. Healthy vaginal microbiota is dominated by lactobacilli, which form a strong defense line against pathogens. In this work, in vitro antimicrobial properties of thirty vaginal Lactobacillus strains were evaluated against eleven Candida vaginal clinical isolates, employing three different methods. Also, the effect of intravaginal (i.va.) administrations (preventive, therapeutic and preventive-therapeutic) of L. reuteri CRL1324 or L. rhamnosus CRL1332 strains against the i.va. challenge with Candida albicans C2 (C.a.) was evaluated in a murine experimental model. From the results of agar overlay and liquid medium assays the selected lactobacilli strains have shown to inhibit the growth of at least one Candida strain. The inhibition was mainly due to the effect of organic acids. Anti-Candida activity was not evidenced in the agar plate diffusion method. In the experimental murine model, only preventive-therapeutic administration of both lactobacilli was able to significantly reduce viable C.a. numbers recovered in vaginal washes and the leukocyte influx induced by the fungi. In conclusion, lactobacilli exhibited in vitro and in vivo antimicrobial effects on Candida, suggesting that they could be promising candidates for protection against VVC.