A pico-injection-aided digital droplet detection platform is presented that integrates loop-mediated isothermal amplification (LAMP) with molecular beacons (MBs) for the ultrasensitive and quantitative identification of pathogens, leveraging the sequence-specific detection capabilities of MBs. The microfluidic device contained three distinct functional units including droplet generation, pico-injection, and droplet counting. Utilizing a pico-injector, MBs are introduced into each droplet to specifically identify LAMP amplification products, thereby overcoming issues related to temperature incompatibility. Our methodology has been validated through the quantitative detection of Escherichia coli, achieving a detection limit as low as 9 copies/μL in a model plasmid containing the malB gene and 3 CFU/μL in a spiked milk sample. The total analysis time was less than 1.5 h. The sensitivity and robustness of this platform further demonstrated the potential for rapid pathogen detection and diagnosis, particularly when integrated with cutting-edge microfluidic technologies.

Sheep\'s milk (SM) is known to differ from cow\'s milk (CM) in nutritional composition and physicochemical properties, which may lead to different digestion behaviours. This work aimed to investigate the impact of the species (cow vs sheep) and the structure (milk vs yogurt) on the digestion of dairy products. Using an in vitro static gastrointestinal digestion model, CM, SM, cow\'s milk yogurt (CY) and sheep\'s milk yogurt (SY) were compared on particle size evolution, microscopic observations, degree of lipolysis, degree of proteolysis, specific protein degradation and calcium bioaccessibility. Species and structure affected particle size evolution during the gastric phase resulting in smaller particles for yogurts compared to milks as well as for CM products compared to SM products. Species impacted lipid composition and lipolysis, with SM products presenting higher short/medium-chain fatty acids content and higher intestinal degree of lipolysis. Proteolysis was influenced by structure, with milks showing higher intestinal degree of proteolysis compared to yogurts. Caseins were digested faster in CM, ⍺-lactalbumin was digested faster in SM despite its higher concentration, and during gastric digestion β-lactoglobulin was more degraded in CM products compared to SM products and more in yogurts compared to milks. Lastly, SM products released more bioaccessible calcium than CM products. In conclusion, species (cow vs sheep) impacted more the digestion compared to the structure (milk vs yogurt). In fact, SM was different from CM mainly due to a denser protein network that might slow down the accessibility of the enzyme to its substrate which induce a delay of gastric disaggregation and thus lead to slower the digestion of the nutrients.

The Minas artisanal cheese from the Serra da Canastra (MAC-CM) microregion is a traditional product due to its production and ripening process. Artisanal chesses manufactured with raw cow\'s milk and endogenous dairy starters (\"also known as pingo\") have distinctive flavors and other sensory characteristics because of the unknown microbiota. The aim of this study was to evaluate the microbiota during 30 days of ripening, the physicochemical changes, and their relation in MACs produced in two different microregions located in the Serra da Canastra microregion through culture-dependent and culture-independent methods. The MACs were collected in the cities of Bambuí (MAC-CMB) and Tapiraí (MAC-CMT) in the Canastra microregion (n = 21). Cheeses uniqueness was demonstrated with the multivariate analysis that joined the microbiota and physicochemical characteristics, mainly to the proteolysis process, in which the MAC-CMT showed deeper proteolysis (DI -T0:14.18; T30: 13.95), while the MAC-CMB reached only a primary level (EI -T0:24.23; T30: 31.10). Abiotic factors were responsible for the differences in microbial diversity between the cheese farms. Different microbial groups: the prokaryotes, like Corynebacterium variabile, Lactococcus lactis, and Staphylococcus saprophyticus; and the eukaryotes, like Kluyveromyces lactis and Diutina catenulata dominated ripening over time. The microbial community and proteolysis were responsible for the predominance of volatile groups, with alcohols predominating in MAC-CMB and free fatty acids/acids and esters in MAC-CMT.

Spore-forming bacteria are the most complex group of microbes to eliminate from the dairy production line due to their ability to withstand heat treatment usually used in dairy processing. These ubiquitous microorganisms have ample opportunity for multiple points of entry into the milk chain, creating issues for food quality and safety. Certain spore-formers, namely bacilli and clostridia, are more problematic to the dairy industry due to their possible pathogenicity, growth, and production of metabolites and spoilage enzymes. This research investigated the spore-forming population from raw milk reception at two Norwegian dairy plants through the cheesemaking stages until ripening. Samples were collected over two years and examined by amplicon sequencing in a culture independent manner and after an anaerobic spore-former enrichment step. In addition, a total of 608 isolates from the enriched samples were identified at the genus or species level using MALDI-TOF analysis. Most spore-forming isolates belong to the genera Bacillus or Clostridium, with the latter dominating the enriched MPN tubes of raw milk and bactofugate. Results showed a great variation among the clostridia and bacilli detected in the enriched MPN tubes. However, B. licheniformis and C. tyrobutyricum were identified in all sample types from both plants throughout the 2-year study. In conclusion, our results shed light on the fate of different spore-formers at different processing stages in the cheese production chain, which could facilitate targeted actions to reduce quality problems.

Omega-3 long-chain polyunsaturated fatty acids (LCPUFA) play critical roles in human development and health. Their intake is often effectively estimated solely based on seafood consumption, though the high intake of terrestrial animal-based foods with minor amounts of LCPUFA may be significant. Covalent adduct chemical ionization (CACI) tandem mass spectrometry is one approach for de novo structural and quantitative analysis of minor unsaturated fatty acids (FA), for which standards are unavailable. Here, CACI-MS and MS/MS are used to identify and quantify minor omega-3 LCPUFA of terrestrial animal foods based on the application of measured response factors (RFs) to various FA. American mean intakes of pork, beef, chicken, and eggs contribute 20, 27, 45, and 71 mg/day of docosahexaenoic acid (DHA), respectively. The estimated intake of omega-3 DHA, eicosapentaenoic acid, and docosapentaenoic acid from nonseafood sources is significant, at 164, 103, and 330 mg/day, greater than most existing estimates of omega-3 LCPUFA intake.

The study aimed to assess performance traits in Hardhenu cattle by analysing data from 445 animals born to 59 sires and 227 dams. The investigation focused on estimating (co)variance components and genetic parameters for reproduction and production traits in dairy cattle. Results from least-squares analysis indicated a significant effect (p < .01) of the period of calving (POC) on key production traits, including first lactation milk yield (FLMY), 300-day milk yield (FLMY300), first peak yield (FPY) and total lactation milk yield (TLMY) in studied population. The least squares means for these traits were reported as follows: FLMY (2665.68 ± 45.66 kg), FLMY300 (2425.52 ± 34.41 kg), FLL (312.95 ± 3.83 days), FPY (11.52 ± 0.15 kg) and TLMY (9282.44 ± 167.03 kg) in Hardhenu cattle. In the studied population, only additive genetic variability was found to be present and there was absence of any significant maternal effect with respect to targeted traits in the resource population. Direct heritability estimates (h2) for FLMY, FLMY300, FLL, FPY, TLMY and other traits ranged from 0.03 to 0.41 in Hardhenu cattle. These findings offer valuable insights into the genetic factors influencing performance traits, contributing to the enhancement of breeding and management practices in Hardhenu cattle.

BACKGROUND: Poor infant and child feeding practices, in combination with increased rates of infectious diseases, are the main immediate causes of malnutrition during the first two years of life. Non-breastfed children require milk and other dairy products, as they are rich sources of calcium and other nutrients. As far as our search is concerned, there is no evidence on the pooled magnitude and determinants of minimum milk feeding frequency among non-breastfed children in sub-Saharan Africa conducted using the most recent indicators for assessing infant and young child feeding practices published in 2021. Therefore, this study is intended to determine the magnitude and associated factors of minimum milk feeding frequency among non-breastfed children aged 6-23 months in sub-Saharan Africa using the most recent guideline and demographic and health survey dataset.
METHODS: Data from the most recent health and demographic surveys, which were carried out between 2015 and 2022 in 20 sub-Saharan African countries, were used. The study comprised a weighted sample consisting of 13,315 non-breastfed children between the ages of 6 and 23 months. STATA/SE version 14.0 statistical software was used to clean, recode, and analyze data that had been taken from DHS data sets. Utilizing multilevel mixed-effects logistic regression, the factors associated with the outcome variable were identified. Model comparison and fitness were assessed using deviance (-2LLR), likelihood ratio test, median odds ratio, and intra-class correlation coefficient. Finally, variables with a p-value < 0.05 and an adjusted odds ratio with a 95% confidence interval were declared statistically significant.
RESULTS: The pooled magnitude of minimum milk feeding frequency among non-breastfed children aged 6-23 months in sub-Saharan African countries was 12.39% (95% CI: 11.85%, 12.97%). Factors like maternal educational level [AOR = 1.61; 95% CI (1.35, 1.91)], marital status of the mother [AOR = 0.77; 95% CI (0.67, 0.89)], maternal working status [AOR = 0.80; 95% CI (0.71, 0.91)], media exposure [AOR = 1.50; 95% CI (1.27, 1.77)], wealth index [AOR = 1.21; 95% CI (1.03, 1.42)], place of delivery [AOR = 1.45; 95% CI (1.22, 1.72)], ANC visit attended during pregnancy [AOR = 0.49; 95% CI (0.39, 0.62)], PNC checkup [AOR = 1.57; 95% CI (1.40, 1.76)], child\'s age [AOR = 0.70; 95% CI (0.53, 0.93)], and residence [AOR = 2.15; 95% CI (1.87, 2.46)] were significantly associated with minimum milk feeding frequency.
CONCLUSIONS: In sub-Saharan Africa, the proportion of minimum milk feeding frequency among non-breastfed children aged between 6 and 23 months was low. The likelihood of minimum milk feeding frequency increases with high levels of education, unemployment, media exposure, rich wealth status, being unmarried, having a child born in a health facility, getting PNC checks, being between 6 and 8 months old, and living in an urban area. Hence, promoting women\'s education, increasing the economic status of the household, disseminating nutrition information through media, strengthening maternal health service utilization like health facility delivery and PNC services, and giving prior attention to mothers with older children and from rural areas are strongly recommended.

The objective was to evaluate the effects of separate offering of feed ingredients (SF) and frequency of concentrate feeding versus offering a TMR, on lactational performance, ruminal fermentation, enteric CH4 emissions, nutrient digestibility, N use efficiency, milk fatty acid profile, and blood variables in mid-lactation dairy cows. Twenty-four Holstein cows (12 primi- and 12 multiparous) averaging (±SD) 141 ± 35 DIM and 43 ± 6 kg/d of milk yield (MY) at the beginning of the study were used in a replicated 3 × 3 Latin square design experiment with 3 periods of 28 d each, composed of 7 d for adaptation to the diets, 11 d for estimation of net energy and metabolizable protein requirements, and 10 d for data and samples collection. Cows were grouped based on parity, DIM, and MY into 4 Latin squares. Treatment allocation was balanced for carryover effects, and cows within square were assigned to (1) basal diet fed ad libitum as TMR; (2) basal diet fed as SF with forages fed ad libitum and concentrates fed 3×/d (SF×3); or (3) basal diet fed as SF with forages fed ad libitum and concentrates fed 6×/d (SF×6). Compared with TMR, SF decreased total DMI by 1.2 kg/d. Treatments did not affect MY, milk components, or ECM yield, except for a decrease in milk fat concentration and an increase in milk urea N by SF×3, compared with TMR. Feed efficiency (kg of MY/kg of DMI) was increased by 7% in SF, compared with TMR. Ruminal molar proportion of acetate and acetate-to-propionate ratio were decreased, whereas molar proportion of propionate was increased by SF×3, compared with TMR and SF×6. There was a 9% decrease in daily CH4 production by SF, compared with TMR. Enteric CH4 yield (per kg of DMI) was not affected by treatments in the current study. Methane intensity per kilogram of MY tended to be decreased by 10% in SF, compared with TMR. The sums of odd- and branched-chain, odd-chain, and anteiso milk fatty acids tended to be or were increased by SF, compared with TMR. Intake of nutrients tended to be or were decreased by SF, compared with TMR. The digestibility of amylase-treated NDF tended to be decreased and ADF digestibility was decreased by 3% in SF, compared with TMR. Urinary and fecal N excretions were not affected by treatments. As a percentage of total N intake, separate offering of feed ingredients increased milk N secretion, indicating an increased N use efficiency by SF, compared with TMR. Blood total fatty acid concentration was decreased by SF relative to TMR. Compared with both TMR and SF×6, SF×3 increased blood urea N concentration. Overall, feed and N use efficiencies were increased by separate offering of feed ingredients, and increasing the frequency of concentrate feeding promoted ruminal fermentation effects similar to those obtained by feeding a TMR.

This study investigated the effects of feeding an amylase-enabled corn silage (ACS) on the performance and enteric gas emissions in lactating dairy cows. Following a 2-wk covariate period, 48 mid-lactation Holstein cows were assigned to 1 of 3 treatments in a 10-wk randomized complete block design experiment. Treatments were diets containing the same proportion of corn silage (40% of dietary DM) as follows: (1) a conventional hybrid corn silage control (CON), (2) ACS replacing the control silage (ADR), and (3) the ADR diet replacing soybean hulls with ground corn grain to achieve the same dietary starch concentration as CON (ASR). Control corn silage and ACS were harvested on the same day and contained 40.3% and 37.1% DM and (% of DM): 37.2% and 41.0% NDF and 37.1% and 30.0% starch, respectively. Enteric gas emissions were measured using the GreenFeed system. Two cows were culled due to health-related issues during the covariate period. Ruminal fluid was collected from 24 cows (8 per treatment) using the orogastric ruminal sampling technique. When compared with CON, cows fed ADR had increased DMI during experimental wk 3, 4, and 9, but treatment did not affect milk or ECM milk yields (39.0 kg/d on average; SEM = 0.89). Compared with CON, feed efficiency (per unit of milk, but not ECM) tended to be lower for ADR, whereas milk true protein concentration (a tendency) and yield were lower for ASR. Milk urea N was decreased by both ADR and ASR diets relative to CON. Compared with CON, daily CH4 emission and emission intensity were increased by ADR but not ASR. Total protozoal count tended to be increased by both diets formulated with ACS when compared with control corn silage. Total-tract digestibility of dietary NDF was greater for ASR, and that of ADF was greater for both ADR and ASR versus CON. The molar proportion of acetate (a tendency) and acetate-to-propionate ratio were increased by ADR, but not ASR, when compared with CON. Replacement of CON with ACS (having lower starch concentration) in the diet of dairy cows increased DMI during the initial weeks of the experiment, maintained ECM, tended to decrease feed efficiency, and increased enteric CH4 emissions, likely due to increased intake of digestible fiber, compared with CON.

Methane, both enteric and from manure management, is the most important greenhouse gas from ruminant livestock, and its mitigation can deliver substantial decreases in the carbon footprint of animal products and potentially contribute to climate change mitigation. Although choices may be limited, certain feeding-related practices can substantially decrease livestock enteric CH4 emission. These practices can be generally classified into 2 categories: diet manipulation and feed additives. Within the first category, selection of forages and increasing forage digestibility are likely to decrease enteric CH4 emission, but the size of the effect, relative to current forage practices in the United States dairy industry, is likely to be minimal to moderate. An opportunity also exists to decrease enteric CH4 emissions by increasing dietary starch concentration, but interventions have to be weighed against potential decreases in milk fat yield and farm profitability. A similar conclusion can be made about dietary lipids and oilseeds, which are proven to decrease CH4 emission but can also have a negative effect on rumen fermentation, feed intake, and milk production and composition. Sufficient and robust scientific evidence indicates that some feed additives, specifically the CH4 inhibitor 3-nitrooxypropanol, can substantially reduce CH4 emissions from dairy and beef cattle. However, the long-term effects and external factors affecting the efficacy of the inhibitor need to be further studied. The practicality of mass-application of other mitigation practices with proven short-term efficacy (i.e., macroalgae) is currently unknown. One area that needs more research is how nutritional mitigation practices (both diet manipulation and feed additives) interact with each other and whether there is synergism among feed additives with different mode of action. Further, effects of diet on manure composition and greenhouse gas emissions during storage (e.g., emission trade-offs) have not been adequately studied. Overall, if currently available mitigation practices prove to deliver consistent results and novel, potent, and safe strategies are discovered and are practical, nutrition alone can deliver up to 60% reduction in enteric CH4 emissions from dairy farms in the United States.