Healthy Milk Research Articles

Impact of bounding attachments on breastfeeding success in primipara postpartum mothers

Breastfeeding is more than just generating healthy breast milk. It is a cooperative activity involving both parties. This study examined how ligature ties affect primiparous women's ability to nurse their babies after giving birth. This type of research is cross-sectional. The study sample consisted of 144 primiparous postpartum women whose children were aged between 0 and three months. The entire sample employed in this study consisted of 106 primiparous postpartum women, selected by a combination of simple random sampling and probability. Questionnaires were used to collect data on attachment factors and breastfeeding success. A correlation value of r=0.437 was obtained, indicating a very positive association between the two variables, and the results demonstrated a significance level of ≤0.05. This correlation implies that effective nursing is mostly dependent on the mother and infant developing strong emotional attachment. These results further emphasize the importance of providing primiparous postpartum mothers with sufficient assistance and direction to enhance their bonding and breastfeeding experiences. Teaching caregivers and medical professionals the value of emotional connection and how it affects nursing outcomes are also essential. Given the established link between effective nursing and emotional attachment, healthcare providers can provide customized interventions and support to improve mother-child bonds throughout the postpartum phase.

RELAÇÃO ENTRE BOAS PRÁTICAS DE ORDENHA, ÍNDICES DE QUALIDADE DE LEITE E BEM-ESTAR ANIMAL

Milk quality is directly related to milking management practices and herd health, with mastitis being one of the main diseases that compromise milk production. This study aimed to evaluate the impact of good milking practices on milk quality and animal welfare, with an emphasis on reducing mastitis and somatic cell count (SCC). The research was carried out at Fazenda Roxa, in Candeal-BA, with 70 cows, 40 of which were in lactation. Milk samples were collected before and after the implementation of good practices, which included reorganization of the milking line, disinfection of teats, cup testing for mastitis detection, and training of milkers. The results showed a significant reduction in SCC, from 1,100,000 cells/ml to 550,000 cells/ml, within the limits of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). Furthermore, there was a decrease in the incidence of clinical mastitis, indicating the effectiveness of good practices in improving milk quality. The study also showed that such practices contribute to animal welfare and the production of healthier milk, with economic benefits for producers. The implementation of these practices is essential to ensure the production of healthy milk, with positive impacts on the quality of food products and food safety.

Deciphering the conformational changes induced by high-risk nsSNPs in β-lactoglobulin

Whey protein from bovine milk is highly valued in the food and pharmaceutical industries because of its high protein content and abundance of essential amino acids. The relationship between whey protein and the β-lactoglobulin (BLG) gene has been extensively discussed because BLG is the most abundant whey protein, making up approximately 50 % of the total whey protein in bovine milk. In recent years, researchers have been interested in this gene because of its critical role in healthy milk production, and any genetic polymorphism in this gene may deteriorate the milk quality. In the current study, we identified several deleterious and damaging non-synonymous single nucleotide polymorphisms (nsSNPs) in BLG and analyzed their destabilizing effects using different computational algorithms. Cumulative results from all tools and evolutionary conservation profiles of BLG suggested that four nsSNPs, G17A, W19C, F136S, and C119R, were the most deleterious and could affect the structural integrity of the protein. Detailed molecular dynamics simulation analysis revealed that all variants induced major structural alterations, that affected the ability of the protein to interact with natural and synthetic ligands. Particularly, the G17A, F136S, and C119R variants induced large conformational changes in the EF loop and main α-helix of BLG, which may affect the access of natural and synthetic ligands to the central calyx of BLG. We hope that the suggested nsSNPs will guide future studies and assist researchers in improving the quality of bovine milk.

Comprehensive proteomic analysis of buffalo milk extracellular vesicles

Extracellular vesicles are secretory vesicles involved in cell-to-cell communication via their encapsulated cargo of proteins, lipids, and nucleic acids. Bovine milk provides a rich source of extracellular vesicles (mEVs) that have been studied as therapeutics and drug delivery systems. Therefore, insight into the mEV cargo, such as its proteome, may help in understanding the molecular mechanism underlying the potential health benefits attributed to the mEVs. Hence, mEVs were isolated from healthy buffalo milk after screening the milk somatic cell count. The total proteins of mEVs were analyzed using LC-MS, and 331 proteins were found commonly present among three buffalo milk samples. These proteins were primarily derived from extracellular regions and lysosomes. The major biological roles associated with the proteins were immune response, metabolism, and cell cycle regulation. The molecular functions of the proteins were transporter activity, catalytic activity, and GTPase activity. Further, comparative analysis with the previously available bovine mEVs proteome data showed 114 proteins to be newly identified in the buffalo mEVs. The biological pathways associated with these proteins may play a major role in muscle development. These findings shed a light on the potential health benefits of buffalo mEVs as therapeutics as well as drug delivery vehicles.

Impact of a teat disinfectant based on Lactococcus cremoris on the cow milk proteome

BackgroundDairy cow milking practices require cleaning and disinfection of the teat skin before and after milking to ensure the safety and quality of milk and prevent intramammary infections. Antimicrobial proteins of natural origin can be valuable alternatives to traditional disinfectants. In a recent field trial, we demonstrated that a teat dip based on a nisin A-producing Lactococcus cremoris (L) had comparable efficacy to conventional iodophor dip (C) in preventing dairy cow mastitis. Here, we present the differential shotgun proteomics investigation of the milk collected during the trial.MethodsFour groups of quarter milk samples with low (LSCC) and high somatic cell count (HSCC) collected at the beginning (T0) and end (TF) of the trial were analyzed for a total of 28 LSCC (14 LSCC T0 and 14 LSCC TF) and 12 HSCC (6 HSCC T0 and 6 HSCC TF) samples. Milk proteins were digested into peptides, separated by nanoHPLC, and analyzed by tandem mass spectrometry (LC-MS/MS) on an Orbitrap Fusion Tribrid mass spectrometer. The proteins were identified with MaxQuant and interaction networks of the differential proteins were investigated with STRING. The proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD045030.ResultsIn healthy milk (LSCC), we detected 90 and 80 differential proteins at T0 and TF, respectively. At TF, the Lactococcus group showed higher levels of antimicrobial proteins. In mastitis milk (HSCC), we detected 88 and 106 differential proteins at T0 and TF, respectively. In the Lactococcus group, 14 proteins with antimicrobial and immune defense functions were enriched at TF vs. 4 proteins at T0. Cathelicidins were among the most relevant enriched proteins. Western immunoblotting validation confirmed the differential abundance.ConclusionsAt T0, the proteomic differences observed in healthy milk of the two groups were most likely dependent on physiological variation. On the other hand, antimicrobial and immune defense functions were higher in the milk of cows with mammary gland inflammation of the Lactococcus-treated group. Among other factors, the immunostimulatory action of nisin A might be considered as a contributor.

In vitro characterization of probiotic potential of Lactobacillus plantarum CM49 against selected cattle mastitogens

BackgroundBovine mastitis results in significant economic losses for the dairy industry globally due to milk production losses and decreased herd efficiency. This research aimed to isolate, select, and characterize indigenous lactobacilli with probiotic properties. A total of 40 lactobacilli were isolated from healthy milk samples of cattle and identified at the species level through 16S rDNA sequencing. All isolates were initially screened for antimicrobial activity, and selected isolates underwent in vitro assessment of probiotic properties.ResultsAmong the lactobacilli isolates, varying levels of activity (9 to 19 mm) against cattle mastitogens; Stapylococcus aureus (Staph. aureus), Escherichia coli (E. coli) and Streptococcus dysgalactiae (Strep. dysgalactiae) were observed in the well diffusion assay. These isolates demonstrated auto-aggregation (ranging from 14.29 ± 0.96% to 62.11 ± 1.09%) and co-aggregate (ranging from 9.21 ± 0.14% to 55.74 ± 0.74%) with mastitogens after 2 h. Lactobacillus (Lb.) plantarum CM49 showed sensitivity to most antibiotics tested and exhibited strong inhibitory effects, with mean log10 reductions of 3.46 for Staph. aureus, 2.82 for E. coli, and 1.45 for Strep. dysgalactiae in co-culture experiments. Furthermore, Lb. plantarum CM49 significantly decreased the adhesion rate of mastitogens on the bovine mammary cell line and mouse model, demonstrating its potential effectiveness in preventing mastitis.ConclusionIt is concluded that Lb. plantarum CM49 has remarkable probiotic potential with activity against cattle mastitogens in the laboratory and cell culture and competitively excludes mastitogens from bovine mammary cells and ameliorates Staph. aureus-induced mastitis in mice.

Specific Gut Microbiome Signatures in Children with Cow's Milk Allergy.

Although gut dysbiosis is associated with cow's milk allergy (CMA), causality remains uncertain. This study aimed to identify specific bacterial signatures that influence the development and outcome of the disease. We also investigated the effect of hypoallergenic formula (HF) consumption on the gut microbiome of milk-allergic children. 16S rRNA amplicon sequencing was applied to characterize the gut microbiome of 32 milk-allergic children aged 5-12 years and 36 age-matched healthy controls. We showed that the gut microbiome of children with CMA differed significantly from that of healthy children, regardless of whether they consumed cow's milk. Compared to that of healthy cow's milk consumers, it was depleted in Bifidobacterium, Coprococcus catus, Monoglobus, and Lachnospiraceae GCA-900066575, while being enriched in Oscillibacter valericigenes, Negativibacillus massiliensis, and three genera of the Ruminococcaceae family. Of these, only the Ruminococcaceae taxa were also enriched in healthy children not consuming cow's milk. Furthermore, the gut microbiome of children who developed tolerance and had received an HF was similar to that of healthy children, whereas that of children who had not received an HF was significantly different. Our results demonstrate that specific gut microbiome signatures are associated with CMA, which differ from those of dietary milk elimination. Moreover, HF consumption affects the gut microbiome of children who develop tolerance.

Seasonal assessment of mastitis in crossbred goats: A thermographic approach

Sub-clinical mastitis exhibits a higher prevalence in dairy goats than clinical mastitis, necessitating the adoption of non-invasive diagnostic techniques such as infrared thermography (IRT) to detect this economically significant production disease in the dairy sector. Accordingly, this study aims to employ IR imaging of the udder and teat quarters of lactating crossbred goats (Alpine × Beetal and Sanen × Beetal) across various seasons, utilising IRT, to discern cases of sub-clinical (SCM) and clinical mastitis (CM). Over a year, 100–110 lactating crossbred dairy goats underwent consistent IRT screenings, followed by a comprehensive evaluation of udder health status using the California mastitis test and somatic cell count (SCC). The receiver operating characteristic (ROC) analysis was conducted to establish the cut-off values for different thermographic parameters in this study. The results revealed that the SCC increased significantly (p < 0.01) in healthy, SCM, and CM milk samples across the seasons. The analysis of ROC revealed a comparatively higher sensitivity, specificity, and accuracy for udder thermograms during SCM than CM and vice versa for teat thermograms. IRT analysis reflected a difference (p < 0.01) in the udder and teat thermograms among quarters of healthy, SCM, and CM in summer, winter, autumn, and rainy seasons. A significant increase (p < 0.01) in udder thermograms was observed for quarters affected with SCM and CM relative to healthy, with an increase of 1.89 and 2.94 °C in winter, 0.85 and 1.63 °C in summer, 0.73 and 1.41 °C in rainy, and 1.33 and 2.38 °C in autumn, respectively. Similarly, for teat thermograms it was 1.79 and 2.81 °C in winter, 0.76 and 1.41 °C in summer, 0.70 and 1.37 °C in rainy, and 1.09 and 1.93 °C in autumn, respectively. Therefore, regardless of the seasons examined in this study, IRT proves to be an effective and supportive tool for early mastitis detection in lactating crossbred dairy goats.

Enhancing Health Outcomes for Breastfeeding Mothers and Children Through Optimal Nutrition – A Narrative Review

Breastfeeding is one of the best ways to provide nutrition to babies. Apart from that, breastfeeding also provides many benefits for health, such as reducing the risk of breast and ovarian cancer. However, not all mothers can breastfeed optimally, this can be caused by various factors. This review aims to discuss the role of nutrition in improving the health of breastfeeding mothers and children. Research results show that ensuring breastfeeding mothers receive sufficient nutrition to support healthy breast milk production can improve health outcomes for both mother and child. There are various factors that affect the nutritional intake of breastfeeding mothers, including knowledge about breastfeeding nutrition, dietary habits, the mother's busy activity, and social support. Adequate nutrition during the breastfeeding period can significantly reduce the likelihood of mothers suffering from fatigue, anemia, and depression. Similarly, infants who receive sufficient breast milk are less likely to experience stunting, infections, and other chronic illnesses.

Development of novel lactose‐free Tigernut milk pudding with no added sugar and determination of physicochemical and organoleptic attributes

SummaryDeveloping health‐oriented novel products (e.g. no sugar added for diabetes and lactose‐free for lactose intolerance) supports diverse consumer needs, promotes healthier eating habits and fosters inclusivity in the food sector. This study aimed to develop a Tigernut milk pudding (TNP), an alternative to dairy‐based pudding and assess its physicochemical and sensory attributes. TNP was formulated by incorporating hydrocolloids κ‐carrageenan (KC), konjac powder (KP) and corn starch (CS), then compared with a commercial milk pudding (CMP) and traditionally prepared raw cow milk pudding (RMP). The crude protein, fat, ash and moisture levels ranged from 5.62% to 8.04%, 5.25% to 7.03%, 2.81% to 3.98% and 68.71% to 74.76% respectively. Lightness (L*) ranged from 69.87 to 77.89, while a* ranged from 0.36 to 0.86 and b* ranged from 8.13 to 9.97. Syneresis ranged from 4.98% to 8.00%, slightly higher than RMP (4.98%) and CMP (4.87%), while pH levels ranged from 6.31 to 6.89, nearly acidic to neutral pH range. Total soluble solids ranged from 2.90 °Brix to 3.67 °Brix. The bacterial count was in the marginal range below 105 CFU g−1, with no presence of Salmonella spp. or E. coli. Total antioxidant properties showed total flavonoid and phenolics, ranging from 306.24 mg QE/100 g to 358.58 mg QE/100 g and 39.06 mg GAE/100 g to 54.81 mg GAE/100 g respectively. An overall organoleptic rate of 3.50 was attained with 0.25% KC, 0.50% KP and 2.50% CS, achieving a desired texture comparable to CMP. The novel study on using Tigernut milk in pudding explores its plant‐based, vegan‐friendly, lactose‐free attributes as a healthy dairy milk alternative. With desirable physicochemical properties, promising improved nutrition, potential health benefits and possible contribution to food security and enhanced resource efficiency, TNP could be considered for commercialisation.

The effects of creamy and skimmed dairy products (milk, yoghurt and cheese) on lipid profile of healthy rats

Milk and dairy products contain saturated fatty acids that have different effects on human health status and are risk factors for many diseases including cardiovascular disease and diabetes mellitus. The present study aimed to evaluate the impact of creamy and skimmed milk administration and certain dairy products (yoghurt and cheese) on healthy rats. A total of forty-nine healthy male Sprague Dawley rats were divided to seven groups. The treatment diet includes creamy and skimmed milk, creamy and skimmed yoghurt, creamy and skimmed cheese. After 60 days of consumption, body weight (BW) and lipids profile were measured. The final BW was higher in rats fed with creamy milk (256.40±23.81), and rats fed with creamy yoghurt (262.71±25.41) and creamy cheese (266.57±22.71). A significant elevation in the liver (aspartate aminotransferase and serum alanine) and kidney functions (urea, creatinine, and uric acid) have been observed in rats fed with fresh creamy whole milk and skimmed milk groups. The measured lipid profile has increased, especially rats fed with creamy milk, creamy, and skimmed cheese. In which, the total cholesterol has reached 159.70, 170.11 and 160.11. Meanwhile, the triglycerides increased to 125.51, 117.35 and 110.41. The low-density lipoprotein and the very low-density lipoprotein have also elevated with their atherogenic indexes at 5.2±0.55, 5.74±0.68 and 5.31±0.44 respectively. On the other hand, notable decreases were seen in the high-density lipoprotein cholesterol and liver glycogen compared to rats fed with control diet. In conclusion, both skimmed and creamy yoghurt indicated healthy nutritional properties, especially with lipid and atherogenic indexes by means of healthy liver and renal functions. Thus, yoghurt (creamy or skimmed) improved the healthy rats' lipid profile while creamy milk additions increased the BW. However, cheese (creamy or skimmed) is suggested to be avoided due to hyperlipidemic conditions in rats.

Effect of subclinical mastitis on milk taurine concentration in dairy cows

The objective of this study was to evaluate whether taurine has any role in bovine subclinical mastitis, and the relationship between milk taurine concentration and somatic cell count. Forty milk samples from dairy cows aged 3 to 6 during lactation were used for this research. The California Mastitis Test (CMT) and somatic cell counts (SCC) were determined in the diagnosis of subclinical mastitis. The subclinical mastitis group consisted of 20 milk specimens positive for CMT and SCC&gt;200.000/ml of milk. The control group consisted of 20 milk specimens negative for CMT and SCC&lt;200.000/ml of milk. A bovine-specific taurine enzyme-linked immunosorbent assay kit was used to determine the taurine concentration in the milk. The milk serum taurine concentrations were determined as 232.1±89.8 pg/ml in the control group and 158.0±55.6 pg/ml in the subclinical mastitis group. These results determined that the taurine concentration in milk with subclinical mastitis decreased approximately 1.5 times compared to healthy milk (P&lt;0.05). A significant negative correlation was determined between the taurine concentration in cow’s milk with subclinical mastitis and the number of somatic cells (r= -0.933; P&lt;0.001). From the findings of this study, it was concluded that both somatic cell counts and the measurement of taurine in cow’s milk could be used to monitor mammary health.

A study on the development and the application strategy of FT-MIRS-based models for the diagnosis of subclinical mastitis and milk quality classification in buffaloes

Buffalo mastitis detection and buffalo milk quality are of great importance to the world dairy industry. Somatic cell count (SCC) can be employed to assess mammary gland health and milk quality in dairy cows, but SCC detection is costly, and the detection instrument is expensive. The purpose of this study was to develop high and low SCC identification models using Fourier-transform mid-infrared spectrum (FT-MIRS), which could be used to diagnose subclinical mastitis (SCM) in buffalo and to determine the milk quality grade at a low cost. The dataset contained 899 buffalo milk samples collected from two regions in China. Firstly, the samples were divided into positive group above the threshold (SCM or unqualified milk) and negative group below the threshold (healthy or qualified milk) with SCC = 200 × 103 cells/mL (SCM in buffalo), 400 × 103 cells/mL (EU standard for raw cow milk, ES), 500 × 103 cells/mL (Indian standard for raw buffalo milk, IS), and 750 × 103 cells/mL (US standard for raw cow milk, US) as thresholds, respectively. Then, with FT-MIRS as predictive variables, predictive models were developed using Partial Least Squares Discriminant Analysis (PLSDA), Random Forest (RF), and Gradient Boosting Machine (GBM). The main results were as follows: the AUCval of the diagnostic model of SCM in buffalo using SCM criteria as a threshold was 0.84 (PLSDA). The AUCval values of the three buffalo milk quality classification models with ES, IS, and US as thresholds were 0.76 (PLSDA), 0.78 (GBM), and 0.84 (PLSDA), respectively. The predictive models established in this paper had a weak predictive ability for positive samples, and the “stepwise discriminant analysis” was recommended to improve the model application effect: The models were applied to classify samples as positive and negative, and then the samples with higher predictive probability were selected. Finally, the remaining samples were further differentiated using the reference methods of SCC detection. To conclude, FT-MIRS has the potential to predict buffalo mammary gland health status and buffalo milk quality grade, reduce the cost of SCC detection, and improve work efficiency, which will lay the foundation for rapid buffalo milk quality classification by raw milk regulatory authorities and rapid diagnosis of SCM in buffalo by farms.

Breast milk microbiota of healthy women living in the Russian Federation

Introduction. Mother’s breast milk is considered to be the best source of infant nutrition and, paradoxical as it may sound, the only product of the infant diet, which composition is still poorly known to us. This article presents the findings of a study on the breast milk microbiota conducted in the Russian Federation for the first time.Aim. To study the composition of the breast milk microbiota of healthy women living in the Russian Federation and identify the impact of various factors on its diversity.Materials and methods. A molecular genetic study on 56 colostrum samples and 12 mature breast milk samples from 56 healthy donors using a method based on sequencing of bacterial 16S rRNA gene was conducted.Results. In a batch of 56 colostrum samples, 22 species and 242 genera of bacteria were identified. Firmicutes, Proteobacteria, Actinobacteriota were the most predominant species of bacteria in healthy women with a normal body mass index, from a physiological pregnancy, who did not take antibiotics, after natural delivery at term; Streptococcus, Staphylococcus and Gemella were the most predominant at the level of delivery. Statistical analysis of the impact of nationality, age, number of children in the family and duration of hospitalization (PERMANOVA) showed no significant differences in the proportions of bacteria in the colostrum of women (p &gt; 0.05). The gender of the child was the only factor that turned out to be significant. The biodiversity of colostrum of girl-delivered mothers were higher (Shannon index) than that of boy-delivered mothers. In addition, the mature breast milk microbiota (1 month after delivery) has lower alpha diversity as compared to colostrum.Discussion. The analysis of our results and reports from foreign colleagues showed significant similarities and differences, which, certainly, were explained by a number of reasons related to methodological and geographical differences, the method, timing and even the time of day when breast milk samples were collected.Conclusion. In general, the colostrum of healthy women had a fairly stable bacterial composition, and its rich biodiversity fully demonstrated high quality of the first microbial inoculum for the newborn intestine. Our results provide valuable insights into the healthy breast milk microbiota in women living in the Russian Federation and can be used as reference ranges, as well as for comparison with similar indicators in women from other countries.

Sisvan food intake markers from six to 23 months: critical analysis and validation.

To explore the internal structure and analyze evidence of measurement invariance of the Sisvan form of food intake markers of the Food and Nutrition Surveillance System (Sisvan) for children aged six to 23 months. Sisvan microdata from 2015 to 2019 were used. After analyzing sample adequacy, exploratory factor analysis was carried out considering factor loadings (≥ 0.30 and < 0.85), communality (> 0.20), model fit indices - Tucker-Lewis index (TLI) > 0.90, and root mean square error of approximation (RMSEA) < 0.08. A critical analysis of the form items was performed to adjust the parameters. Measurement stability was investigated between age groups, macro-regions and longitudinally by multi-group confirmatory factor analysis in configural, metric, and scalar invariance models. Configural invariance was accepted if RMSEA < 0.08 and TLI and comparative fit index (CFI) > 0.90. Metric and scalar invariances were accepted if ΔRMSEA < 0.015 and ΔCFI < 0.01, compared to the previous model. After critical analysis, items were grouped (vegetables and leafy greens; meat or eggs and liver) and excluded (salty food; porridge with milk; rice, potatoes, yams, cassava, flour, or pasta). The exploratory model comprised three factors: healthy complementary feeding (fruit; vegetables; orange or dark green leafy vegetables or fruits; meat, offal, or egg; beans), ultra-processed (yogurt; hamburger and/or sausages; sweetened drinks; instant noodles, packet snacks, or salty cookies; sandwich cookies, sweets, or treats), and milk feeding (breast milk; other milk), with satisfactory factor loadings, communalities and fit (TLI: 0.918, RMSEA: 0.071). The reformulated instrument was stable in the invariance models tested. With the incorporation of the modifications to the Sisvan form, the food intake markers subsidize a more adequate interpretation of the feeding situation of children aged six to 23 months.

Antibiogram profiling and detection of icaA and blaZ genes from Staphylococcus aureus and coagulase-negative Staphylococcus spp. of healthy bovine raw milk sample origin.

This study focused on the antibiogram profiling of Staphylococcus aureus and coagulase-negative Staphylococcus spp. (CoNS) and the detection of icaA and blaZ genes from bovine raw milk samples. Bovine milk samples were collected from dairy farms, and Staphylococcus spp. were isolated and identified via conventional and molecular screening. Disk diffusion test (DDT) was implemented to determine the resistance pattern. Biofilm and β-lactamase-producing Staphylococcus spp. were identified via amplification of the icaA and blaZ genes. Methicillin-resistant Staphylococcus aureus and CoNS were identified by DDT and PCR of the mecA gene. From 63 samples, 35 were confirmed as Staphylococcus spp., of which 16 (25.39%) S. aureus isolates were coagulase-positive, while 19 (30.16%) were negative. PCR confirmed that 50% (8/16) of S. aureus and 36.84% (7/19) of CoNS possessed the icaA gene. All S. aureus isolates were found resistant to penicillin-G (P) both phenotypically and genotypically. The isolates were also resistant to erythromycin (ERY) and oxytetracycline (TET). While CoNS showed high to reduced resistance against P, TET, ERY, and azithromycin, no S. aureus isolates were resistant to sulfamethoxazole, while 10.53% of CoNS isolates were. All S. aureus and CoNS isolates were susceptible to vancomycin and gentamicin. MR was exhibited by 37.5% of S. aureus and 42.10% of CoNS isolates. Moreover, S. aureus and CoNS had 56.25% and 52.63% multidrug-resistant (MDR) isolates, respectively. The present study revealed the presence of a biofilm-producing, MDR staphylococcal strain in milk that might endanger consumers. Routine surveillance and monitoring, along with antimicrobial resistance learning, can reduce risks.

Safety Assessment and Probiotic Potential Comparison of Bifidobacterium longum subsp. infantis BLI-02, Lactobacillus plantarum LPL28, Lactobacillus acidophilus TYCA06, and Lactobacillus paracasei ET-66.

Bifidobacterium longum subsp. infantis BLI-02, Lactobacillus paracasei ET-66, Lactobacillus plantarum LPL28, and Lactobacillus acidophilus TYCA06, isolated from healthy breast milk, miso, and the healthy human gut, were assessed for safety in this study. BLI-02, LPL28, TYCA06, and ET-66 exhibited no antibiotic resistance and mutagenic activity in the Ames test at the highest dosage (5000 μg/plate). No genotoxicity was observed in micronucleus and chromosomal aberration assays in rodent spermatogonia at the maximum dosage of 10 g/kg body weight (BW). No acute and sub-chronic toxicity occurred in mice and rats at the maximum tested dosage of 10 g/kg BW and 1.5 g/kg BW, respectively. The lyophilized powder of these strains survived a low pH and high bile salt environment, adhering strongly to Caco-2 cells. Unique antimicrobial activities were noted in these strains, with BLI-02 demonstrating the best growth inhibition against Vibrio parahaemolyticus, LPL28 exhibiting the best growth inhibition against Helicobacter pylori, and ET-66 showing the best growth inhibition against Aggregatibacter actinomycetemcomitans. Based on the present study, the lyophilized powder of these four strains appears to be a safe probiotic supplement at tested dosages. It should be applicable for clinical or healthcare applications.

Screening, Identification and Production of L-asparagine from Lactobacillus reuteri of Healthy Mother Milk

Human breast milk (HBM) may benefit Lactobacillus reuteri, a probiotic strain isolated from a mother's breast milk. It has been shown to inhibit pathogen growth, modulate the immune system, and have strong anti-inflammatory effects. This study aimed to isolate and screen Lactobacillus reuteri for L-asparagine production. Lactobacillus reuteri were grown in submerged fermentation and liquid media and used for secondary screening by the Nesslerization method. The isolates were identified based on morphological, cultural, and biochemical tests and API 50 to identify Lactobacillus reuteri. According to the findings, L-asparaginase isolated from bacteria showed positive L-asparaginase activity on a modified medium using phenol red as an indicator dye. However, bacterial isolates showed the highest enzyme activity, exhibiting a pink zone of hydrolysis of around 7.0 cm after 48h. and 4.92 IU/mg of specific activity. These results illustrated the importance of these isolates to produce and control L-asparagine for biomedical production in the future. Keywords: Lactobacillus reuteri, L-asparagine, L-asparaginase

Buffalo yogurt fermented with commercial starter and Lactobacillus plantarum originating from breast milk lowered blood pressure in pregnant hypertensive rats

Nutritional therapy, which may have advantages over medication, is being investigated as a novel treatment for pregnancy-induced hypertension. Several studies have shown that probiotic yogurt supplementation during pregnancy has beneficial effects on maternal and fetal health. In this study, fermented buffalo milk was produced with yogurt culture and Lactobacillus plantarum B, a probiotic isolated from healthy breast milk with high angiotensin-converting enzyme inhibitory activity. The fermentation conditions under which the angiotensin-converting enzyme (ACE) inhibitory activity reached 84.51% were optimized by the response surface method as follows: 2 × 106 cfu/mL of L. plantarum B, yogurt culture 2.5 × 105 cfu/mL, and 8 h at 37°C. The distribution of ACE inhibitory peptides from fermented buffalo milk and fermented cow milk were further analyzed by liquid chromatography-mass spectrometry. By searching according to the structural features of ACE inhibitory peptides, 29 and 11 peptides containing ACE inhibitory peptide features were found in fermented buffalo milk and fermented cow milk, respectively. To investigate the in vivo antihypertensive activity of fermented buffalo milk, 18 pregnant rats were divided into 3 groups (n = 6 in each group) and administered 10 mL of normal saline, yogurt (20 mg/kg), or labetalol hydrochloride (4 mg/kg) daily from the beginning of pregnancy to parturition. To induce hypertension, methyl nitrosoarginine (125 mg/kg) was injected subcutaneously every day from d 15 of pregnancy to the day of delivery. Blood pressure was not significantly changed in the yogurt and labetalol groups after induction of hypertension and was lower compared with the normal saline group, but there was no difference between the yogurt and labetalol groups. This implied that the buffalo yogurt had a preventive and antihypertensive effect in the pregnancy-induced hypertensive rat model. Further studies to determine the mechanism of action, as well as a randomized control trial, are warranted.

Dilution Reduces Sample Matrix Effects for Rapid, Direct, and Miniaturised Phenotypic Antibiotic Susceptibility Tests for Bovine Mastitis.

The time-consuming nature of current methods for detecting antimicrobial resistance (AMR) to guide mastitis treatment and for surveillance, drives innovation towards faster, easier, and more portable technology. Rapid on-farm testing could guide antibiotic selection, reducing misuse that contributes to resistance. We identify challenges that arise when developing miniaturized antibiotic susceptibility tests (AST) for rapid on-farm use directly in milk. We experimentally studied three factors: sample matrix (specifically milk or spoiled milk); the commensal bacteria found in fresh bovine milk; and result time on the performance of miniaturised AST. Microfluidic "dip-and-test" devices made from microcapillary film (MCF) were able to monitor Gram-negative bacterial growth colourimetrically even in the presence of milk and yoghurt (used to simulate spoiled milk samples), as long as this sample matrix was diluted 1:5 or more in growth medium. Growth detection kinetics using resazurin was not changed by milk at final concentrations of 20% or lower, but a significant delay was seen with yoghurt above 10%. The minimum inhibitory concentration (MIC) for ciprofloxacin and gentamicin was increased in the presence of higher concentrations of milk and yoghurt. When diluted to 1% all observed MIC were within range, indicating dilution may be sufficient to avoid milk matrix interfering with microfluidic AST. We found a median commensal cell count of 6 × 105 CFU/mL across 40 healthy milk samples and tested if these bacteria could alter microfluidic AST. We found that false susceptibility may be observed at early endpoint times if testing some pathogen and commensal mixtures. However, such errors are only expected to occur when a susceptible commensal organism is present at higher cell density relative to the resistant pathogen, and this can be avoided by reading at later endpoints, leading to a trade-off between accuracy and time-to-result. We conclude that with further optimisation, and additional studies of Gram-positive organisms, it should be possible to obtain rapid results for microfluidic AST, but a trade-off is needed between time-to-result, sample dilution, and accuracy.