Effects of maternal toxic substance consumption during breastfeeding on lactic acid bacteria abundance and nutritional content.

Breast milk is widely recognized as the primary source of nourishment for newborns, making it an unparalleled and indispensable provider of essential nutrients, microbiological components, immunological factors, and energy content. To investigate this further, a cohort comprising 254 breastfeeding women participated in interviews, and milk samples were aseptically collected for subsequent analysis involving bromatological, microbiological, and clinical analysis. The investigation focused on the identification of specific microorganisms in breast milk and their susceptibility to the exposure of toxic substances and controlled medications. Notably, this study places particular emphasis on the significant decline in lactic acid bacteria observed in breast milk when influenced by substances such as cocaine, cannabis, crystal, and morphine. These detrimental agents have been found to adversely affect the growth of microorganisms within breast milk. On the contrary, the outcomes of this study indicate that the utilization of toxic substances does not exert a noteworthy impact on the nutritional quality of breast milk.

Human lactoferrin from breast milk: characterization by HPLC and its in vitro antibiofilm performance.

Preterm infants are at high risk of infection due to opportunistic bacteria as Pseudomonas aeruginosa, causing infections among infants in neonatal intensive care units. Human lactoferrin (hLf) is a multifunctional protein and one of the most abundant in breast milk, and plays an important role in prevention of different infections in neonates. This work offers a strategy to obtain a lyophilisate of purified lactoferrin from breast milk. In addition, a reliable HPLC method for quantification of lactoferrin with a linear quantification range of 0.040-0.140 mg/mL with selectivity, accuracy and repeatability, is described. Lyophilized hLf was obtained by purification through a heparin affinity column followed by ultrafiltration with a 30 kDa membrane. The final solution was lyophilized and the product was analyzed using HPLC method, recovering about 70% of initial lactoferrin in the sample. This molecule was elucidated through FTIR spectroscopy and SDS-PAGE electrophoresis. In addition, the capacity against biofilm formation of P. aeruginosa was demonstrated with 75% of inhibition at 6 mg/mL. These results suggest that lyophilized hLf can be obtained by purification of breast milk and that it can provide antibiofilm activity against P. aeruginosa.

Improvement and Validation of a Genomic DNA Extraction Method for Human Breastmilk.

The human milk microbiota (HMM) of healthy women can vary substantially, as demonstrated by recent advances in DNA sequencing technology. However, the method used to extract genomic DNA (gDNA) from these samples may impact the observed variations and potentially bias the microbiological reconstruction. Therefore, it is important to use a DNA extraction method that is able to effectively isolate gDNA from a diverse range of microorganisms. In this study, we improved and compared a DNA extraction method for gDNA isolation from human milk (HM) samples to commercial and standard protocols. We evaluated the extracted gDNA using spectrophotometric measurements, gel electrophoresis, and PCR amplifications to assess its quantity, quality, and amplifiability. Additionally, we tested the improved method's ability to isolate amplifiable gDNA from fungi, Gram-positive and Gram-negative bacteria to validate its potential for reconstructing microbiological profiles. The improved DNA extraction method resulted in a higher quality and quantity of the extracted gDNA compared to the commercial and standard protocols and allowed for polymerase chain reaction (PCR) amplification of the V3-V4 regions of the 16S ribosomal gene in all the samples and the ITS-1 region of the fungal 18S ribosomal gene in 95% of the samples. These results suggest that the improved DNA extraction method demonstrates better performance for gDNA extraction from complex samples such as HM.

Cold Plasma Treatment as an Alternative for Ochratoxin a Detoxification and Inhibition of Mycotoxigenic Fungi in Roasted Coffee.

Ochratoxin A (OTA) produced by mycotoxigenic fungi (Aspergillus and Penicillium spp.) is an extremely toxic and carcinogenic metabolite. The use of cold plasma to inhibit toxin-producing microorganisms in coffee could be an important alternative to avoid proliferation of mycotoxigenic fungi. Roasted coffee samples were artificially inoculated with A. westerdijikiae, A. steynii, A. versicolor, and A. niger, and incubated at 27 °C over 21 days for OTA production. Samples were cold plasma treated at 30 W input power and 850 V output voltage with helium at 1.5 L/min flow. OTA production in coffee was analyzed by high performance liquid chromatography coupled to a mass spectrometer (HPLC-MS). After 6 min of treatment with cold plasma, fungi were completely inhibited (4 log reduction). Cold plasma reduces 50% of OTA content after 30 min of treatment. Toxicity was estimated for extracts of artificially contaminated roasted coffee samples using the brine shrimp (Artemia salina) lethality assay. Toxicity for untreated roasted coffee was shown to be "toxic", while toxicity for cold plasma treated coffee was reduced to "slightly toxic". These results suggested that cold plasma may be considered as an alternative method for the degradation and reduction of toxin production by mycotoxigenic fungi in the processing of foods and feedstuffs.

Influencia de la dieta de mujeres mexicanas sobre la calidad nutricional y la presencia de microorganismos benéficos en la leche humana / Influence of the diet of Mexican women on the nutritional quality and the presence of beneficial microorganisms in human milk

Introducción: la leche materna es el primer alimento fundamental de los neonatos y proporciona todas las fuentes de energía, nutrientes y protección inmunológica que necesitan durante sus primeros meses de vida. Lamentablemente, existen circunstancias específicas que implican que la madre no pueda alimentar correctamente a su bebé, ya que las necesidades nutricionales de la madre difieren en cierta medida durante los diversos periodos de la vida, especialmente en la lactancia, ya que se aumentan las necesidades nutricionales debido a la pérdida de nutrientes, primero por el calostro y luego a través de la leche materna. Objetivo: demostrar la influencia de la dieta de mujeres mexicanas sobre la calidad nutricional y la presencia de microorganismos benéficos en la leche humana. Métodos: se llevaron a cabo 70 encuestas descriptivas de nutrición y toma de muestras de leche a mujeres en estado lactante. Las leches fueron sometidas a diversos análisis bromatológicos y microbiológicos para evaluar su calidad nutricional y posible actividad probiótica. Resultados: se demostró que la ingesta de alimentos de la madre influye en la calidad nutricional de la leche. Asimismo, afecta el desarrollo y crecimiento de las bacterias lácticas. Se aislaron e identificaron diversas cepas en leche humana del género Lactobacillus, además de bacterias patógenas como el caso de Lodderomyces elongisporus, entre otros. Conclusiones: la alimentación de las madres se refleja directamente en la calidad nutricional de la leche. Se observó que la cantidad de nutrientes esenciales de la leche, como son los hidratos de carbono, lípidos y proteínas, varían conforme a la alimentación y el ritmo de vida de las madres, así como la disminución significativa de bacterias lácticas con potencial probiótico

Introduction: breast milk is the first fundamental food of newborns and it provides all the sources of energy, nutrients and the immunological protection they need during their first months of life. Unfortunately, there are specific circumstances that imply that the mother cannot feed her baby correctly, since the mother's nutritional needs differ to a certain extent during different periods of life. Especially in breastfeeding, since nutritional needs are increased, due to the loss of nutrients, first by colostrum and then by breast milk. Objective: to demonstrate the influence of the diet of Mexican women on the nutritional quality and the presence of beneficial microorganisms in human milk. Methods: seventy descriptive surveys of nutrition and sampling of milk to women in nursing state were carried out. The milks were subjected to various bromatological and microbiological analyzes to evaluate their nutritional quality and possible probiotic activity. Results: it was shown that the mother's food intake influences the nutritional quality of the milk. Likewise, it affects the development and growth of lactic acid bacteria. Several strains were isolated and identified in human milk of the genus Lactobacillus, as well as pathogenic bacteria such as Lodderomyces elongisporus among others. Conclusions: the mothers' nutrition is directly reflected in the nutritional quality of the milk. It was observed that the amount of essential nutrients of milk such as carbohydrates, lipids and proteins vary according to the diet and life rhythm of the mothers, as well as the significant decrease of lactic bacteria with probiotic potential

Impacto de la pasteurización/liofilización en el contenido disponible de inmunoglobulinas en leche humana madura. Estudio de aplicación en bancos de leche humana en hospitales / Impact of pasteurization/freeze-drying on available immunoglobulin content of the mature human milk. Use in human milk banking of hospitals

Introducción: este estudio analiza el efecto sobre el contenido de inmunoglobulinas y complemento C3 de la liofilización posterior a la pasteurización por tres métodos diferentes en leche humana madura (LHM). Objetivo: la liofilización es propuesta como método complementario para el mantenimiento de las propiedades terapéuticas de la LHM con mayor vigencia. Métodos: estudio descriptivo en el que se obtuvieron muestras de LHM. Alícuotas de las muestras obtenidas se pasteurizaron por tres métodos: 62,5 °C/30 minutos, 72 °C/15 minutos 85 °C/5 minutos, seguido de un enfriamiento rápido a 5 °C. Después, volúmenes de 30 ml de muestra pasteurizada fueron liofilizados durante un periodo de 36 horas. La determinación de proteínas totales fue realizada por el método Lowry. Las concentraciones de inmunoglobulinas A, G y M y el complemento C3 fueron determinadas por nefelometría convencional, siguiendo las instrucciones del fabricante. La significancia estadística se definió como p < 0,05. Resultados: el método de pasteurización de LHM con mayor retención de proteína e inmunoglobulinas fue a la temperatura de 62,5 °C, sin embargo, la pasteurización a 72 °C antes de la liofilización mostró mayor retención de inmunoglobulinas. Conclusiones: nuestros resultados sugieren que la liofilización de LHM pasteurizada es un método eficiente para la conservación en bancos de leche humana. Tanto la composición nutricional como la extensión de su vida útil y la aplicación de los dos procesos juntos proporcionan la ventaja de mantener las propiedades terapéuticas de la leche humana para mejorar la salud del recién nacido en estado vulnerable, desmedro o inmunosuprimido (AU)

Introduction: This study analyzes the effect on the content of immunoglobulins and C3 complement of freeze drying after pasteurization by three different methods in mature human milk (MHM). Objective: Freeze drying is proposed as a complementary method for the maintenance of MHM therapeutic properties with greater validity. Methods: This was a descriptive study in which MHM samples were obtained. Next, aliquots of the samples obtained were pasteurized by three methods: 62.5 °C/30 minutes, 72 °C/15 minutes, 85 °C/5 minutes, followed by a rapid cooling at 5 °C. Then, 30 ml volumes of pasteurized sample were freeze-dried over a period of 36 hours. Total protein determination was performed by the Lowry method. The concentrations of immunoglobulins A, G and M, and complement C3, were determined by conventional nephelometric technique following the manufacturer’s instructions. Statistical significance was defi ned as p < 0.05. Results: The method of pasteurization of MHM with increased protein and immunoglobulin retention was at 62.5 °C, however, pasteurization at 72 °C before freeze-drying showed better retention of immunoglobulins. Conclusions: Our results suggest that the freeze-drying of pasteurized MHM is a suitable method for the conservation in human milk banks. Both the nutritional composition and the extension of its validity and the application of the two processes together provide the advantage of maintaining the therapeutic properties of human milk to improve the health of the newborn in a vulnerable, impaired or immunosuppressed state (AU)