Fresh Milk Research Articles

Milk Value Chain Analysis: The Case of Urban and Peri-Urban Areas of Ambo and Dendi Districts, West Shewa Zone, Oromia National Regional State, Ethiopia

This study was aimed at analyzing value chain of milk in Urban and Peri – Urban areas of Ambo and Dendi districts of West Shewa Zone. The study made use of both primary and secondary data collected from 146 smallholder farmers, 30 consumers, 10 Hotels and Cafeterias and 2 primary milk cooperatives through application of individual interview schedules, focus group discussions and key informant interview. The analysis was made using descriptive statistics and value chain analysis. The value chain analysis revealed that the major actors in the districts milk value chains are input suppliers, milk producers, collectors, processors and consumers. Accordingly, the value chain activities in the chain were input supply, production, processing, marketing and consumption. Four milk marketing channels were identified and relative to all other fresh milk marketing channels, producers get a higher proportion of total gross marketing margin when they sell milk to cooperatives. Profitability analysis result for dairy production indicated that, a dairy producer obtains average gross margin of 83.65per improved cow per day, in the study areas which shows that dairy production was a profitable venture for smallholder dairy producers. Milk value chain in the study areas was hindered by shortage of feeds, continuous increasing price of feeds, poor AI service provision, poor supply of genetically improved cattle breeds, limited availability of credit services, limited extension service provision, poor feeding and animal husbandry practices, low level of market linkage among the value chain actors, absence of milk processing plant and non-market oriented milk production. Therefore, policy aiming at increasing farmers’ access to modern inputs, feed conservation practices, adequate training provision to AI technicians, developing and improving infrastructure, cooperative development and improving extension services are recommended to accelerate the milk value chain’s development in the study areas.

Handheld NIR Spectroscopy Combined with a Hybrid LDA-SVM Model for Fast Classification of Retail Milk

The EU market offers different types of milk, distinguished by origin, production method, processing technology, fat content, and other characteristics, which are often detailed on product labels. In this context, ensuring the authenticity of milk is crucial for maintaining standards and preventing fraud. Various food authenticity techniques have been employed to achieve this. Among them, near-infrared (NIR) spectroscopy is valued for its non-destructive and rapid analysis capabilities. This study evaluates the effectiveness of a miniaturized NIR device combined with support vector machine (SVM) algorithms and LDA feature selection to discriminate between four commercial milk types: high-quality fresh milk, milk labeled as mountain product, extended shelf-life milk, and TSG hay milk. The results indicate that NIR spectroscopy can effectively classify milk based on the type of milk, relying on different production systems and heat treatments (pasteurization). This capability was greater in distinguishing high-quality mountain and hay milk from the other types, while resulting in less successful class assignment for extended shelf-life milk. This study demonstrated the potential of portable NIR spectroscopy for real-time and cost-effective milk authentication at the retail level.

Characteristics of Buffalo Milk Curd on Variations in Fermentation Time

Curd is a processed product from buffalo milk through a fermentation process. The clumping of fresh buffalo milk into curd occurs due to the presence of Lactic Acid Bacteria (LAB) in buffalo milk. Curd contains lactic acid bacteria (LAB), which are beneficial for health. These bacteria not only extend the shelf life of food by inhibiting the growth of spoilage and pathogenic bacteria but also produce bioactive compounds that are good for the body. LAB contained in curd even has probiotic properties. This research used a starter of 10% buffalo milk curd. In one production, 4500 ml of fresh buffalo milk and 500 ml of starter are used, yielding 3.8 kg of buffalo milk curd. The results of observations during 7 days of fermentation were water content (81.25-72.53%), ash content (0.93-0.89%), protein content (8.97-7.94%), fat content (7 .72-7.10%), carbohydrate content (1.13-11.54%), pH value (5.72-4.27), temperature (26.87-28.43oC), Total Lactic Acid Bacteria (1.6x109- 8.1x109 cfu/ml).

Characterization of psychrotrophic and thermoduric bacteria in raw milk using a multi-omics approach.

Psychrotrophic and thermoduric bacteria are the main reasons for the spoilage of dairy products. This study aims to address the composition and function of psychrotrophic and thermoduric bacteria in eight groups of raw milk samples obtained from Heilongjiang Province and Inner Mongolia (China). Microbial enumeration showed an average total bacterial count of 4.63 log c.f.u. ml-1 and psychrotrophic bacterial counts of 4.82 log c.f.u. ml-1. The mean counts of mesophilic and thermophilic thermoduric bacteria were 3.68 log and 1.81 log c.f.u. ml-1, respectively. Isolated psychrotrophic bacteria (26 genera and 50 species) and mesophilic thermoduric bacteria (20 genera and 32 species) showed high microbial diversity. Through metagenomic and proteomic analyses, significant disparities in the concentration and community structure of psychrotrophic and thermoduric bacteria were observed among different locations. A large number of peptidases were annotated by metagenomics, which may result in milk spoilage. They mainly come from some typical psychrotrophic and thermoduric bacteria, such as Chryseobacterium, Epilithonimonas, Pseudomonas, Psychrobacter, Acinetobacter, Lactococcus, Escherichia and Bacillus. However, the main proteins detected in fresh raw milk were associated with bacterial growth, reproduction and adaptation to cold environments. This investigation provides valuable insights into the microbial communities and protein profiles of raw milk, shedding light on the microbial factors contributing to milk deterioration.

A Multiband Microwave Sensor Designed to Sense the Shelf Life of Raw Fresh Milk

A Multiband Microwave Sensor Designed to Sense the Shelf Life of Raw Fresh Milk

The making and unmaking of demand for dairy and chicken in Norway: a systems of provision perspective

This study examines the supply and demand for chicken meat and fresh cow’s milk in Norway and argues that understanding demand as being co-constituted and interrelated with supply is crucial for reducing consumption. This study takes a constructivist systems of provision perspective to examine the chain of activities that links production and consumption. Analyzing how the use of chicken meat and fresh cow’s milk has evolved through socially and historically contingent processes in Norway, the article describes how these products became commonplace through state involvement, nutritional advice, companies, and consumers. The demand for liquid milk decreased in the 1980s, providing a rare insight into a system of provision in decline, although both government and industry actors wanted the situation to be otherwise. This study highlights that the milk-provision system failed to address the factors that contributed to the decline in milk consumption, which worsened the situation for dairy farmers. This is contrasted with the case of chickens, which represents a story of expanded demand through industrial intensification and new food norms. Under the presumption that a theory that explains an increase in demand must also be able to explain a decrease in demand, the analysis highlights the factors that caused a reduction in fresh milk consumption and the factors that led to growth in the demand for chicken meat. The article concludes that a constructivist perspective allows for a more nuanced understanding of the making and unmaking of demand through processes of alignment/misalignment with established political, economic, and societal goals.

Measurement and Improvement of Fresh Milk Supply Chain Performance (Case Study of XYZ Farmers Group, Bogor Regency)

The challenges in the small-scale dairy industry often stem from outdated business management practices and a lack of comprehensive performance evaluation. This study adopts a structural approach by analyzing the performance of the fresh milk supply chain within Farmers Group XYZ, located inBogor Regency. We employed the SCOR and AHP methods for our performance analysis. Data were garnered through questionnaires, interviews, and observations. The findings reveal that the overall performance of the fresh milk supply chain was moderate, with performance scores of 84.25% forfeed suppliers, 77.47% for Farmers Group XYZ, and 82.05% for consumers. Notably, areas requiring enhancement include reliability, responsiveness, and cost-efficiency. Consequently, we have developed strategic recommendations aimed at augmenting supply chain effectiveness. These include enhancing the transportation system to ensure timely deliveries (Reliability), shortening order fulfillment times and improving the flexibility to adapt to demand fluctuations (Responsiveness), and optimizing logistics and storage to reduce costs (Cost). Implementing these strategies will bolster collaboration and fortify relationships among suppliers, producers, and consumers, contributing to a more robust dairy industry.

Fresh Milk Supply Chain Analysis in Kelompok Ternak Mandiri Sejahtera using Food Supply Chain Network Approach

Supply chain management plays a role in ensuring the availability of raw materials from suppliers to the final product reaching consumers. Kelompok Ternak Mandiri Sejahtera (KTMS) is a cooperative that acts as a link between farmers and dairy processing consumers. However, research on theapplication of the supply chain concept at KTMS has not yet been explored. One commonly used approach to analyze supply chain conditions is the Food Supply Chain Network (FSCN). Therefore, the aim of this study is to analyze the fresh milk supply chain conditions at KTMS using the FSCNapproach to comprehensively understand the supply chain conditions. The research was conducted at the Kelompok Ternak Mandiri Sejahtera (KTMS) from June to July 2024. The method used was qualitative descriptive with observations and interviews using questionnaires relevant to the FoodSupply Chain Network (FSCN) framework with 17 respondents. The results of the study indicate that the fresh milk supply chain conditions at Kelompok Ternak Mandiri Sejahtera (KTMS), based on the Food Supply Chain Network framework approach, have been running smoothly, although transportation resources and technology resources remain weaknesses for both the farmers and KTMS.

Sosialisasi Manajemen Kesehatan Kambing Perah serta Pengaruhnya terhadap Kualitas Susu pada Peternak Kambing Perah di Kecamatan Ampelgading dan Tirtoyudo Kabupaten Malang

Goat's milk is a food of animal origin that is easily damaged and requires handling after the milking process. Ampelgading and Tirtoyudo sub-districts are areas in Malang Regency where some of the population are dairy goat breeders. This community service aims to provide socialization and education on good animal health management so that it can improve the quality of goat milk. The activity began with collecting farmer data and analyzing the situation of dairy goat farming. The socialization method was carried out by conducting focus group discussions about knowledge of animal health management and checking the quality of goat milk. Milk composition testing uses Lactoscan, while organoleptic testing is based on Indonesian National Standard 01-2346-2006 concerning organoleptic and sensory testing instructions. The results of community service show that the quality of dairy goat milk in Ampelgading and Tirtoyudo Districts is premium based on the Thailand Agriculture Standard 2008. The results of the organoleptic test of goat milk have remained the same based on SNI 3141-1-2011 regarding the quality of fresh milk. The results of this goat milk quality test can be used as a basis for farmers to improve livestock management and milking. Through this activity, farmers can implement good livestock health management so that the quality of goat milk and the selling price increase. This activity will encourage young farmers to be more interested in becoming dairy goat breeders who have high economic prospects and contribute to increasing goat milk productivity. Apart from that, this activity can improve the welfare of farmers by improving the quality of goat milk with higher selling power.

Determining the influence of raw milk β-casein polymorphism on the efficiency of making soft cheese

One of the ways to increase the profitability of the cheese industry is the genetic selection of dairy cows to obtain milk with excellent cheese-capacity characteristics. The object of this study is the technology of fresh soft cheeses made by the acid-rennet and thermo-acid method from the milk of cows with different β-casein genotypes (A1A1, A1A2, A2A2). The subjects of research are the physicochemical indicators of raw milk from cows with different genotypes for β-casein (A1A1, A1A2, A2A2); as well as the yield of soft cheeses. The study has established that the physicochemical parameters of milk from cows with different β-casein genotypes are typical for fresh cow's milk. The study showed that with the acid-rennet method, the composition of cheeses from A1A1 milk was 51.60 %, 21.63 %, and 23.62 % of moisture, protein, and fat, respectively. A1A2 milk cheeses contained 50.70 % moisture, 20.96 % protein, and 25.12 % fat. A2A2 milk cheeses consisted of 52.50 % moisture, 20.70 % protein, and 23.71 % fat. With the thermo-acid method, cheeses from A1A1 milk were characterized by the moisture content of 55.13 %, proteins – 23.31 %, and fat – 20.21 %. A1A2 milk cheeses contained 58.13 %, 22.62 %, and 17.98 % of moisture, protein, and fat, respectively. A2A2 milk cheeses consisted of 54.03 % moisture, 22.33 % protein, and 22.25 % fat. The calculation of the production efficiency of soft cheeses from the milk of cows with different genotypes according to β-casein with the acid-rennet method of production is 119.3 % on average, which is more compared to milk A1A2 (by 4 %) and A2A2 (by 7 %). With the thermo-acidic method, the efficiency of cheese production from A2A2 milk is 107.5 %, which is higher compared to A1A2 milk (by 9 %) and A1A1 (by 5 %). The conclusions show that changes in the β-casein genotype in raw milk can affect the yield and quality of cheese, and therefore, the profitability of production

Reduction of the environmental impact of the determination of l-carnitine and acetyl-l-carnitine in milk products by microchip electrophoresis

A novel miniaturized analytical method for the simultaneous determination of l-carnitine and its ester, acetyl-l-carnitine in fresh milk and milk products by microchip electrophoresis (MCE) with conductivity detection was developed. Under optimized separation conditions, separation voltage of 4 kV and 50 mmol L−1 acetic acid and 0.1 % methylhydroxyethylcellulose at pH 3.0 as background electrolyte, carnitines were sufficiently separated in less than 10 min. An elimination of adsorption of the cationically migrated analytes on a poly(methyl methacrylate) microchip was achieved by addition of 1 mmol L−1 triethylenetetramine to all analyzed samples. The detector response was linear in the range 0.2–1.5 µg mL−1 for l-carnitine and 0.4–2.0 µg mL−1 for acetyl-l-carnitine. The LOQ values were 0.1 µg mL−1 and 0.4 µg mL−1 for l-carnitine and acetyl-l-carnitine, respectively. Recoveries of the analytes in the analyzed samples were in the range 87.5–97.8 %. The developed MCE method has been successfully applied to the analysis of carnitines in eight milk products after a sample pretreatment which included precipitation, centrifugation, and filtration. The developed MCE method has a low hazardous impact on human health and the environment.

Exposure to aflatoxins and ochratoxin A from the consumption of selected staples and fresh cow milk in the wet and dry seasons in Ghana

Across sub-Saharan Africa, the heavy reliance on mycotoxin-susceptible staple foods means that populations in the region are particularly vulnerable to chronic mycotoxin exposure. This study assessed the exposure risk to ochratoxin A (OTA) and aflatoxins (AFs) from 18 samples of selected staple foods (maize, millet, groundnut) and 56 fresh cow milk samples collected from across Ghana. The foods were sampled simultaneously to maximise comparability, and at two timepoints in March/April (during the dry season) and July/August (during the rainy season) to assess the effects of duration of storage and seasonal conditions on the mycotoxin levels as measured by high-performance liquid chromatography. The margin of exposure (MOE) approach was used to assess the exposure risk from consumption of the sampled foods. Each of the sampled staples was contaminated with OTA (0.19–3.11 ng/g) and at least one AF (0.75–13.05 ng/g B1, ND-12.12 ng/g B2, 0.1–9.95 ng/g G1, ND-16.78 ng/g G2). Up to 67% had contamination above European Food Safety Authority (EFSA) maximum limits, and 50% were above Ghana Standards Authority (GSA) limits. The fresh cow milk samples were contaminated with AFM1 in the range of 0.05–1.49 ng/g, with 95% above EFSA limits and 36% above GSA limits. Aflatoxin contamination in the staples was high, particularly in July/August when the wet conditions may have adversely impacted the handling and storage of farm produce. Variation in AFM1 between the two sampling periods mirrored total aflatoxin in the staples, suggesting that even if dairy cattle were grazing in open pasture and not being rationed on stored feed, then there was a high environmental presence of aflatoxigenic fungi. The MOE estimates were ≤533.09, far below the safe cut-off of 10,000 for suspected carcinogenic compounds. The high mycotoxin levels indicate a priority risk to child nutrition which relies heavily on cereal mixes based on one or all the three sampled staples. The data from this study underscore the urgent need for interventions to better appreciate and address mycotoxin exposure for enhanced food security and public health in Ghana and across sub-Saharan Africa.

Dielectric spectroscopy technology combined with machine learning methods for nondestructive detection of protein content in fresh milk.

To quickly achieve nondestructive detection of protein content in fresh milk, this study utilized a network analyzer and an open coaxial probe to analyze the dielectric spectra of milk samples at 100 frequency points within the 2-20GHz range, focusing on the dielectric constant ε' and the dielectric loss factor ε''. Feature variables were extracted from the full dielectric spectra using the successive projections algorithm (SPA), uninformative variables elimination (UVE), and the combined UVE-SPA method. These variables were then used to develop partial least squares regression (PLSR), support vector machine (SVM), decision tree (DT), random forest (RF), and least squares boosting (LSBOOST) models for predicting protein content. The results showed that ε' decreased monotonically with increasing frequency, while ε'' increased monotonically. The UVE-SPA method for feature extraction demonstrated superior performance, with the UVE-SPA-PLSR model being the best for predicting milk protein content, achieving the highest RC 2=0.998 and RP 2=0.989 and the lowest RMSEC=0.019% and RMSEP=0.032%. This study provides a theoretical reference for evaluating milk quality and developing intelligent detection equipment for natural milk.

An investigative study on Yersinia enterocolitica in animals, humans and dried milk in New Valley Governorate, Egypt

BackgroundYersiniosis is one of the most significant intestinal disorders caused by Yersinia enterocolitica and affects both humans and animals. This study aimed to investigate the prevalence of Y. enterocolitica in New Valley Governorate, Egypt in animals, humans, fresh milk and dried milk. Additionally, this study analyzed the presence of virulence genes, including ail and Yst in tested isolates and conducted a phylogenetic analysis to determine the genetic similarity between human, and animal Y. enterocolitica isolates. Finally, the antimicrobial resistance patterns of the isolates were examined.ResultsAmong the 982 samples examined, the prevalence of Y. enterocolitica based on ISO10273-2017 was 11.7% in animal samples including 12.8% of animal faeces, and 10.4% in milk samples. Moreover, the prevalence of Y. enterocolitica was 13.2% in human stool, and 9.5% in dried milk samples. The molecular characterization of the six randomly selected isolates showed that the 16S rRNA, ail and Yst genes were found in 50, 33.3 and 100% of the examined Y. enterocolitica isolates, respectively. Phylogenetic analysis of animal and human isolates based on the 16S rRNA gene revealed a high degree of similarity between the isolates. All the tested animal and human Y. enterocolitica isolates (100%) were resistant to ampicillin and cefotaxime, but highly sensitive to norfloxacin.ConclusionsThe high prevalence of Y. enterocolitica in animal and human samples with high degrees of genetic similarity poses a threat to public and animal health. Animal faeces, milk and milk powder represent the main sources of Y. enterocolitica infection in humans. Additionally, high levels of antibiotic resistance of Y. enterocolitica can cause public health hazards by leading to the failure of disease prevention and treatment programs in humans and animals.

Outcomes after intranasal human milk therapy in preterm infants with intraventricular hemorrhage.

Intraventricular hemorrhage (IVH) is a common cause of brain injury in preterm infants. Fresh human milk (HM) contains stem cells (SCs) that could potentially be delivered via intranasal HM (IHM). In this IHM pilot study, we describe outcomes. Infants <33 weeks gestation with IVH were given IHM until maximum 28 days of age. Short-term neurologic outcomes and follow-up testing were compared to historic HM-fed infants. Longitudinal outcomes were plotted using linear mixed models. Weighted G-computation quantified treatment effects. Propensity score models calculated inverse probability weights for IVH grade, gestational age, and sex. 37 infants (35.1% grade 3-4 IVH) were compared to 191 historic controls (17.8% grade 3-4 IVH). Post-hemorrhagic ventricular dilatation was common (25.7% IHM patients). Most weighted outcomes, although not significant, favored IHM at 4-12 and 18 months corrected age. This phase 1 study suggests powered trials of IHM for brain injury are needed. CLINICAL TRIAL REGISTRY NAME: clinicaltrials.gov identifier NCT04225286.

Perspectives of healthcare workers on the acceptability of donor human milk banking in Southwest Nigeria

BackgroundThe World Health Organization prioritizes Mother’s Own Milk (MOM) or donor human milk (DHM) when MOM is unavailable or insufficient. It is also important for healthcare workers (HCWs) to provide adequate support, information, and education to mothers to help improve their milk production and breastfeeding experience. DHM is scarce in developing countries, prompting a need for understanding health workers’ perspectives.MethodsThis cross-sectional study, conducted in 2021 in Ekiti State, Nigeria, examined the knowledge and attitudes of HCWs regarding human milk banking. A sample of 321 participants from government-owned hospitals completed a self-administered questionnaire.ResultsOf the 321 participants (84.7% response rate), the majority were females (69.2%), aged 30–39 (32.1%), Christian (91.9%), and employed in tertiary hospitals (91.9%). About 65% of the HCWs believe that HMB is a safe practice and 42% believe that the DHM has the same quantity of immunological factors as fresh human milk. While 80.4% displayed good DHM knowledge, attitudes varied. The HCWs profession influenced their attitudes and a higher proportion of all female HCWs (71%), except for health assistants (65.4%), expressed willingness to donate their breast milk if needed. Among males HCWs, the majority of those who were doctors (82.8%) and pharmacists (62.5%) expressed willingness to support their spouses to donate breast milk, and they also had higher acceptance of DHM for their infants. In all, more than 80% of the HCWs will encourage mothers to donate their milk and feed babies under their care with DHM, but only 47% would accept DHM to feed their own children. Health assistants had less favourable views, and negative perceptions were linked to the internet and media sources.ConclusionsThe study highlights health workers’ awareness of DHM but indicates a reluctance to fully embrace it, especially among health assistants. The need for targeted education programs, to address knowledge gaps and negative perceptions, is crucial for the successful implementation of human milk banks in Nigeria. Overcoming challenges, such as safety concerns and sociocultural influences, requires focused efforts from policymakers and healthcare institutions.

Non-destructive detection of water adulteration level in fresh milk based on combination of dielectric spectrum technology and machine learning method

To rapidly realize the identification of fresh milk for water adulteration and to predict the amount of water adulteration, this study adopts a coaxial probe and a vector network analyzer to analyze the variation laws of dielectric constant ε', dielectric loss factor ε'' and dielectric loss angle tangent tanδ in the range of 2–20 GHz under 100 frequency points. Soft independent modeling of class analogy (SIMCA), Naive Bayes (NB), K-nearest neighbors (KNN), and support vector machine (SVM) models were built based on full dielectric spectra for qualitative testing of adulteration levels in milk. Feature variables of FDS were extracted by using the successive projections algorithm (SPA) and uninformative variables elimination (UVE). Partial least squares regression (PLSR), support vector regression (SVR), and particle swarm optimization least square support vector regression (PSO-LSSVR) models were built for quantitative testing of adulteration levels in milk. The results demonstrate an increasing trend of ε'' and tanδ with increasing frequency and a decreasing trend of ε'. The ε'-SIMCA model achieves the best effect in distinguishing water adulteration in milk, showing accuracy (ACC) of 1, Sensitivity (SNS) of 1, Specificity (SPC) precision (PRE) of 1, and an F1-score (F1) of 1. The tanδ-SPA-PSO-LSSVR model optimally predicts the optimal prediction effect of moisture content of milk, showing an RP2 of 0.994 and an RMSEP of 0.016 %. This study is conducive to building a more comprehensive knowledge system of water adulteration in fresh milk. Its results can provide a theoretical reference in the development of non-destructive detection instruments for natural milk.

Exonic mutations of POU class 1 homeobox 1 are associated with milk pH in high-producing Holstein Friesian cows

Background and Aim: Milk physicochemical properties play essential role in the milk processing industry, which are moderately to highly affected by genetic factors. This study aimed to evaluate the association between single nucleotide polymorphisms in POU class 1 homeobox 1 (POU1F1) and the physicochemical properties of milk in high-producing Holstein Friesian (HF) cows. Materials and Methods: A total of 149 high-producing dairy cows from PT Ultra Peternakan Bandung Selatan was included in this study. The physicochemical properties of milk, including density, freezing point, pH, lactose, solid non-fat, protein, and ash content, were determined. Moreover, three polymorphisms within the exon regions of POU1F1 (c.195G&gt;A, c.300G&gt;T, and c.828G&gt;A) were analyzed using polymerase chain reaction-restriction fragment length polymorphism. The association between these polymorphisms and the physicochemical properties of milk was determined using a mixed-effects model analysis, in which the lactation period was used as a covariate. Results: This study found that two polymorphisms, c.195G&gt;A and c.828G&gt;A, significantly affected the pH of fresh milk. Cows with both the GG genotypes c.195G&gt;A and c.828G&gt;A had lower milk pH values than those with the other genotypes. In addition, a non-significant effect of POU1F1 was observed on the other physicochemical properties of milk. Conclusion: Two POU1F1 polymorphisms determined the pH of fresh milk in the Indonesian HF population. These are potential marker candidates for milk pH that directly affect the development of dairy products in the milk processing industry. Keywords: exonic mutation, Holstein Friesian, milk pH, physicochemical properties, POU1F1.

A Novel Bifidobacterium longum Subsp. longum T1 Strain from Cow's Milk: Homeostatic and Antibacterial Activity against ESBL-Producing Escherichia coli.

Background/Objectives: The global emergence of antibiotic-resistant zooanthroponotic Escherichia coli strains, producing extended-spectrum beta-lactamases (ESBL-E) and persisting in the intestines of farm animals, has now led to the development of a pandemic of extra-intestinal infectious diseases in humans. The search for innovative probiotic microorganisms that eliminate ESBL-E from the intestines of humans and animals is relevant. Previously, we received three isolates of bifidobacteria: from milk of a calved cow (BLLT1), feces of a newborn calf (BLLT2) and feces of a three-year-old child who received fresh milk from this calved cow (BLLT3). Our goal was to evaluate the genetic identity of BLLT1, BLLT2, BLLT3 isolates using genomic DNA fingerprinting (GDF), to study the tolerance, adhesion, homeostatic and antibacterial activity of BLLT1 against ESBL-E. Methods: We used a complex of microbiological, molecular biological, and immunological methods, including next generation sequencing (NGS). Results: GDF showed that DNA fragments of BLLT2 and BLLT3 isolates were identical in number and size to DNA fragments of BLLT1. These data show for the first time the possibility of natural horizontal transmission of BLLT1 through with the milk of a calved cow into the intestines of a calf and the intestines of a child. BLLT1 was resistant to gastric and intestinal stresses and exhibited high adhesive activity to calf, pig, chicken, and human enterocytes. This indicates the unique ability of BLLT1 to inhabit the intestines of animals and humans. We are the first to show that BLLT1 has antibacterial activity against ESBL-E strains that persist in humans and animals. BLLT1 produced 145 ± 8 mM of acetic acid, which reduced the pH of the nutrient medium from 6.8 to 5.2. This had an antibacterial effect on ESBL-E. The genome of BLLT1 contains ABC-type carbohydrate transporter gene clusters responsible for the synthesis of acetic acid with its antibacterial activity against ESBL-E. BLLT1 inhibited TLR4 mRNA expression induced by ESBL-E in HT-29 enterocytes, and protected the enterocyte monolayers used in this study as a bio-model of the intestinal barrier. BLLT1 increased intestinal alkaline phosphatase (IAP) as one of the main molecular factors providing intestinal homeostasis. Conclusions: BLLT1 shows promise for the creation of innovative functional nutritional products for humans and feed additives for farm animals that will reduce the spread of ESBL-E strains in the food chain.

Study the effect of lactoperoxidase activation in combination with heat treatment or nisin on the microbiological quality of raw milk

The aim of the current study was to improve the microbiological quality of raw cow milk by activating the lactoperoxidase system using either nisin or heat treatment. A central composite experimental design with three factors was used to evaluate the effect of thiocyanate (SCN), hydrogen peroxide (H202) and nisin on the microbiological quality of raw milk. The experimental response was measured in terms of the logarithmic reduction in total microbial, total coliforms count, yeasts and molds. Our results indicate that the combination of both treatments was highly effective in destroying various microorganisms. In fact, the optimal values of each parameter (SCN, H202 and nisin) for a 6log, 5log and 4log reductions of the initial number of total aerobic mesophilic bacteria, coliforms, yeasts and molds were around 7 mg/L of SCN, 15 mg/L of H202 and 50 IU/mL of nisin, obtained after 72 h of incubation at 25 °C. Concerning the LPS activation at moderate temperature, our data suggest that this combination was very effective in destroying different microorganisms. Certainly, the total destruction of coliforms, yeasts and molds was noted after 96 h incubation at 25 °C after LPS activation for 4 h and 6 h followed by heat treatment (63 °C for 30 min). These results highlight a synergistic effect between the LPS activation and nisin or heat treatment in preserving microbiological quality of raw milk for 72 h or 96 h at 25 °C, respectively. Such treatments could be implemented by the dairy industry to enhance the quality of fresh milk and consequently of dairy products.