Milk Samples Research Articles

The Effect of a Direct Fed Microbial on Liveweight and Milk Production in Dairy Cattle.

This longitudinal study aimed to quantify the effects of dietary supplementation of a direct-fed microbial (DFM) consisting of three lactobacilli isolates on milk yield, milk fat and protein yields, somatic cell count (SCC), and liveweight in a single dairy herd in Australia. A total of 150 dairy cows were randomly selected based on parity and days in milk and divided into two groups: control (n = 75) and DFM treatment (n = 75). Throughout the study, the two groups of cows were housed separately in a dry lot yard, and each group had their own feeding area. For the DFM treatment group, selected cows in mid-lactation were supplemented with 10 mL/cow/day of the DFM via top dressing of the feed for the remainder of the lactation and through the dry period, extending into subsequent lactation. The control group had no supplementation. The milk yield and liveweight were recorded daily. Milk samples were collected every two months for milk component analysis (fat, protein, and somatic cell count [SCC]). The DFM-treated cows gained more liveweight across the study (19.40 kg, 95% CI 0.44 kg; 38.30 kg, p = 0.05) compared to the control cows. In the second production year, the DFM-treated cows mobilized more liveweight (-6.06 kg, 95% CI -10.49 kg; -1.61 kg, p = 0.01) and produced more milk (0.39 L/d 95% CI 0.10; 0.89, p = 0.05). Over a full lactation, DFM cows yielded at least 258 L (95% CI 252 L; 265 L) more milk than controls. No significant differences were found in fat and protein yield or SCC. This study suggests that consistent and ongoing supplementation with a Lacticaseibacillus- and Lentilactobacillus-based DFM could have a positive effect on milk production, but further research is needed to understand the underlying mechanism.

Simultaneous detection of Cu2+ and amoxicillin based on the copper and nitrogen-regulated fluorescent probe

A novel fluorescence-based probe represented as Cu, N-CDs, was developed for the dual sensingdetection of Cu2+ and amoxicillin (AMO), which had a high quantum yield up to 60.08 %, along with excellent photoluminescent properties and remarkable stability. When detecting Cu2+, the fluorescence signal from Cu, N-CDs probe experienced quenching due to photoinduced electron transfer (PET). Subsequently, the introduction of AMO led to the recovery of the probe's fluorescence. This recovery was likely attributed to the superior affinity between Cu2+ and AMO, resulting in the displacement of Cu2+ from the Cu, N-CDs and blocked the PET process. Notably, the detection limits of Cu2+ and AMO were determined 18 nM and 9 nM, respectively. We further demonstrated the practical utility of the Cu, N-CDs probe through its successful application in the analysis of Cu2+ and AMO in Chinese herbal medicine and milk samples. Overall, our findings highlighted the potency of Cu, N-CDs as versatile and effective tools for the detection of Cu2+ and AMO, paving the way for their broad applications in the safety analysis of food and pharmaceutical.

Resistant S. aureus Isolates Capable of Producing Biofilm from the Milk of Dairy Cows with Subclinical Mastitis in Slovakia

Staphylococcus spp. is the most common cause of mastitis, with a significantly low cure rate. Bacterial characteristics like adhesion and biofilm formation, as well as extracellular factors, can affect the pathogenesis of staphylococcal mastitis. The study’s objectives were to confirm S. aureus, assess their antibiotic resistance, identify methicillin resistance genes, verify biofilm formation, and detect biofilm-associated genes from bovine mastitis samples using multiplex PCR (mPCR). From 215 milk samples, six were confirmed as S. aureus. Most isolates were sensitive to all measured antibiotics. One isolate was identified as an inducible form of MLSB resistance (macrolides, lincosamides, and streptogramin B resistance), while the other two isolates were resistant to penicillins and carboxypenicillins. In S. aureus cultures used for methicillin resistance genotypic analysis by PCR, the mecA and mecC genes were not found. Biofilm formation phenotypes were determined in four strains. An mPCR analysis revealed that all strains of S. aureus carried icaABCD, agrA, srtA, fnbA, clfA, and clfB genes. Only in one isolate was the fnbB gene detected; the bap gene was not detected in any of the isolates. This emphasizes the importance of using appropriate treatment and continuous monitoring of S. aureus to prevent the spread of antibiotic-resistant strains in dairy cow farms.

Ultra-sensitive fluorescent biosensor for multiple bacteria detection based on CDs/QDs@ZIF-8 and microfluidic fluidized bed.

An ultra-sensitive fluorescent biosensor based on CDs/QDs@ZIF-8 and microfluidic fluidized bed was developed for rapid and ultra-sensitive detection of multiple target bacteria. The zeolitic imidazolate frameworks (ZIF-8) act as the carrier to encapsulate three kinds of fluorescence signal molecules from the CDs/QDs@ZIF-8 signal amplification system. Besides, three kinds of target pathogenic bacteria were automatically, continuously, and circularly captured by the magnetic nanoparticles (MNPs) in the microfluidic fluidized bed. The neutral Na2EDTA solution was the first time reported to not only dissolve the ZIF-8 frameworks from the MNPs-bacteria-CDs/QDs@ZIF-8 sandwich complexes, but also release the CDs/QDs from sandwich complexes with no loss of fluorescence signal. Due to the advantages of signal amplification and automated sample pretreatment, the proposed fluorescent biosensor can simultaneously detect Escherichia coli O157:H7, Salmonella paratyphi A, and Salmonella paratyphi B as low as 101CFU/mL within 1.5h, respectively. The mean recovery in spiked milk samples can reach 99.18%, verifying the applicability of this biosensor in detecting multiple bacteria in real samples.

The content of aflatoxin M1 in the milk of cows from Turkish farms: the effect on milk quality and the effectiveness of mycotoxin binding by a claybased adsorbent

The aim of this study was to evaluate the concentration of aflatoxin M1 (AFM1) in milk from Holstein-Friesian cows on local farms located in two different cities in Turkey, to investigate the effect of this mycotoxin on milk quality and seasonal variation, and to investigate the efficacy of a commercial clay-based binder. As part of the study, milk samples from 144 dairy cows were analyzed for the presence of AFM1 prior to the use of a clay-based toxin binder. As a result of this analysis, 26.9% of milk samples from individual animals had AFM1 levels exceeding the U.S. permissible limit of 0.5 µg/kg. In addition, AFM1 was detected in 36.5% of winter and 18.6% of summer milk samples. The presence of AFM1 caused a decrease in milk fat, dry matter, and protein levels. The affected animals were further divided into two groups: control group (n = 14) and experimental group (n = 26). The animals of the experimental group were fed 25 g/animal of clay-based binder toxin daily. The treatment group showed a gradual decrease in AFM1 levels by 24.4% on day 4 and 48.9% on day 7. In addition, the fat, non-fat solids, and protein content of milk increased significantly. As a result, a high level of AFM1 contamination in milk was detected, which can be assumed to negatively affect the quality of milk production. The use of clay-based toxin binders may be an effective method to improve milk quality and protect animal and consumer health.

Colorimetry/fluorescence dual-mode detection of Salmonella typhimurium based on a “three-in-one” nanohybrid with high oxidase-like activity for AIEgen

A colorimetry/fluorescence dual-mode assay based on the aptamer-functionalized magnetic covalent organic framework-supported CuO and Au NPs (MCOF-CuO/Au@apt) was developed for Salmonella typhimurium (S. typhimurium) biosensing. The nanohybrid combined three functions in one: good magnetic separation characteristic, excellent oxidase-mimic activity for tetrap-aminophenylethylene (TPE-4A), and target recognition capability. The attachment of MCOF-CuO/Au@apt onto the surface of S. typhimurium resulted in a significant reduction in the oxidase-mimicking activity of the nanohybrid, which could generate dual-signal of colorimetry and fluorescence through the catalytic oxidation of TPE-4A. Based on this, S. typhimurium could be specifically detected in the linear ranges of 102– 106 CFU·mL−1 and 101– 106 CFU·mL−1, with LODs of 7.6 and 2.1 CFU·mL−1, respectively in colorimetry/fluorescence modes. Moreover, the smartphone and linear discrimination analysis-based system could be used for on-site and portable testing. In addition, this platform showed applicability in detecting S. typhimurium in milk, egg liquid and chicken samples.

Curry-Odorants and Their Metabolites Transfer into Human Milk and Urine.

The excretion of dietary odorants into urine and milk is evaluated and the impact of possible influencing factors determined. Furthermore, the metabolic relevance of conjugates for the excretion into milk is investigated. Lactating mothers (n=20) are given a standardized curry dish and donated one milk and urine sample each before and 1, 2, 3, 4.5, 6, and 8h after the intervention. The concentrations of nine target odorants in these samples are determined. A significant transition is observed for linalool into milk, as well as for linalool, cuminaldehyde, cinnamaldehyde, and eugenol into urine. Maximum concentrations are reached within 1h after the intervention in the case of milk and within 2-3h in the case of urine. In addition, the impact of glucuronidase treatment on odorant concentrations is evaluated in a sample subset of twelve mothers. Linalool, eugenol, and vanillin concentrations increased 3-77-fold in milk samples after treatment with β-glucuronidase. The transfer profiles of odorants into milk and urine differ qualitatively, quantitatively, and in temporal aspects. More substances are transferred into urine and the transfer needs a longer period compared with milk. Phase II metabolites are transferred into urine and milk.

Assessing the Microbial Contamination Levels in Milk Samples from Rural and Urban Areas: A Focus on Raisen and Bhopal Districts

AbstractThis study aimed to assess the level of microbial contamination in raw cow milk samples collected from farms in rural and urban regions of Raisen and Bhopal districts, Madhya Pradesh, India. Recent data are gathered to investigate the microbial quality of the milk samples directly, rather than relying on reported findings from other sources. The results reveal that both rural and urban milk samples exhibit significantly higher bacterial colony counts compared to acceptable limits for total viable count. The average total viable count for urban samples is 8.699 log CFU mL−1, with a standard deviation of 0.821 log CFU mL−1, using a sample size of 1 mL. Moreover, urban samples show a higher initial incubation count (8.728 ± 0.431 log CFU mL‐1), indicating potential unsanitary milk handling practices in urban areas. In contrast, rural milk samples demonstrate significantly lower bacterial counts (8.0778 ± 0.079 log CFU mL−1) and exhibit superior laboratory sanitization measures. Enteric bacteria such as Escherichia coli, Enterobacter species, Klebsiella species, and Shigella are found in both urban and rural milk samples after isolation. However, strains of bacterial culture specific to Pseudomonas and Salmonella species are exclusively detected in urban samples, suggesting a higher contamination level in urban areas compared to rural regions. These findings highlight the urgent need for improved milk production and handling practices in both rural and urban areas, particularly in urban settings where the contamination levels are found to be significantly higher. The results of this study contribute to the understanding of microbial quality in raw cow milk and emphasize the importance of implementing appropriate interventions to ensure the safety and quality of milk consumed by the population.

Estimation of udder emptying based on milk constituents of strip samples after milking

Milk ejection disorders were induced by oxytocin receptor blockade. We tested the hypothesis that the degree of udder emptying at incomplete milk ejection can be estimated based on the concentration of various milk constituents in different milk fraction samples. To induce different levels of spontaneous udder emptying (SUE) 10 Holstein dairy cows were milked either with or without i.v. injection of the oxytocin receptor blocking agent atosiban (ATO). In ATOearly, 12 µg/kg BW ATO was injected immediately before and in ATOlate directly after a 1 min manual udder preparation. The normal milking routine served as Control treatment. In all 3 treatments the udder was completely emptied by the i.v. injection of 10 IU oxytocin (OT) at the end of spontaneous milk flow. During all experimental milkings 4 milk samples were taken in all treatments: at the start of udder preparation (foremilk; FM), immediately after cessation of spontaneous milk flow and cluster detachment by hand stripping (strip milk; SM), from spontaneous removed milk in the bucket 1 (milk before OT; MBOT) and from the milk obtained after OT injection in the bucket 2 (milk after OT; MAOT). Fat, protein, lactose and electrolytes (Na, Cl and K) were measured in each milk sample. In addition, electrical conductivity (EC) was determined in parallel to continuous milk flow recording. The treatments induced individual degrees of SUE; therefore, the final evaluations of data were based on SUE classes instead of treatments. The most pronounced differences of milk constituents at different degrees of SUE were found for the milk fat content. The fat content of SM and MBOT remained almost unchanged up to 60% SUE, but was considerably higher if >80% of the milk was spontaneously removed. The concentrations of Na and Cl were highest and of K lowest if less than 20% on milk was received in the different samples. The EC was higher in SM and MBOT if <20% of milk was received. In conclusion, the blockade of the OT effect influences primarily the fat content, which confirmed an OT-induced fat secretion during milking. Similar effects are likely found in situations of disturbed milk ejections, caused by a lacking or reduced release of OT in response to different degrees of tactile udder stimulation. Our results show that the measurement of fat content and the EC in strip milk samples collected after cluster detachment can be used to estimate the completeness of udder emptying.

Plasma and milk metabolomics profiles in dairy cows with subclinical and clinical ketosis

Ketosis, a commonly observed energy metabolism disorder in dairy cows during the peripartal period, is distinguished by increased concentrations of β-hydroxybutyrate (BHB) in blood. This condition has a negative impact on milk production and quality, causing financial losses. An untargeted metabolomics approach was performed on plasma samples from cows between 5 and 7 DIM diagnosed as controls (CON, BHB <1.2 mM, n = 30), subclinically ketotic (SCK, 1.2 < BHB <3.0 mM, n = 30), or clinically ketotic (CK, BHB >3.0 mM, n = 30). Cows were selected from a commercial farm of 214 Holstein cows (average 305-d yield in the previous lactation of 35.42 ± 7.23 kg/d; parity, 2.41 ± 1.12; body condition score, 3.1 ± 0.45). All plasma and milk samples (n = 90) were subjected to Liquid Chromatography-Mass Spectrometry (LC-MS)-based metabolomic analysis. Statistical analyses was performed using the Graph Pad Prism 8.0, MetaboAnalyst 4.0 and R packages (version 4.1.3). Compared with the CON group, both SCK and CK groups had greater milk fat, freezing point, and fat-to-protein ratio and lower milk protein, lactose, solids-nonfat, and milk density. Within 21 d after calving, compared with CON, the SCK group experienced a reduction of 2.65 kg/d in milk yield, while the CK group experienced a decrease of 7.7 kg/d. Untargeted metabolomics analysis facilitated the annotation of a total of 5,259 and 8,423 metabolites in plasma and milk. Differentially affected metabolites were screened in CON vs. SCK, CON vs. CK, and SCK vs. CK (unpaired t-test, False discovery rate <0.05; and absolute value of log(2)-fold change >1.5). A total of 1,544 and 1,888 differentially affected metabolites were detected in plasma and milk. In plasma, glycerophospholipid metabolism, pyrimidine metabolism, tryptophan metabolism, sphingolipid metabolism, amino sugar and nucleotide sugar metabolism, phenylalanine metabolism, steroid hormone biosynthesis were identified as significant pathways. Weighted gene co-expression network analysis (WGCNA) indicated that tryptophan metabolism is a key pathway associated with the occurrence and development of ketosis. Increases in 5-Hydroxytryptophan and decreases in kynurenine and 3-indoleacetic acid in SCK and CK were suggestive of an impact at the gut level. The decrease of most glycerophospholipids indicated that ketosis is associated with disordered lipid metabolism. For milk, pyrimidine metabolism, purine metabolism, pantothenate and CoA biosynthesis, amino sugar and nucleotide sugar metabolism, nicotinate and nicotinamide metabolism, sphingolipid metabolism, fatty acid degradation were identified as significant pathways. The WGCNA indicated that purine and pyrimidine metabolism in plasma was highly correlated with milk yield during the peripartal period. Alterations in purine and pyrimidine metabolism characterized ketosis, with lower levels of these metabolites in both milk and blood underscoring reduced efficiency in nitrogen metabolism. Our results may help to establish a foundation for future research investigating mechanisms responsible for the occurrence and development of ketosis in peripartal cows.

Feeding rumen-protected methionine during the peripartum period improved milk fat content and reduced the culling rate of Holstein cows in a commercial herd

Researchers have reported the benefits of feeding rumen-protected methionine (RPM) during the peripartum on the health parameters of dairy cows. Rumen-protected methionine has reportedly improved milk yield, milk components and liver health, but the literature is scarce on its effects in commercial herds. Therefore, we aimed to determine the effects of feeding RPMet (Smartamine M®, Adisseo Inc., Antony, France) prepartum (8 g per cow per day) and postpartum (15 g per cow per day) on performance, metabolic profile, and culling rate of Holstein cows in a commercial herd. One-hundred and 66 (n = 166) Holstein cows, 58 nulliparous and 108 parous, were randomly assigned to 1 of 2 dietary treatments, consisting of TMR top-dressed with RPMet (2.35 and 2.24% Met of MP for close-up and fresh cows, respectively) or without (control, CON, (2.03 and 1.89% Met of MP for close-up and fresh cows, respectively), fed from 21 ± 6 d prepartum until 16 ± 5 d postpartum. From 17 d in milk (DIM) until dry-off, all cows received RPMet. Daily milk yield was recorded, and milk samples were collected in the first and second weeks after calving to determine their composition. Blood samples were collected before the morning feeding on -14, -7, +1, +7, and +14 d relative to calving. Mortality and morbidity were recorded during the first 60 DIM. Cows supplemented with RPMet had greater milk yield during the first 16 DIM (31.76 vs. 30.37 kg/d; SEM = 1.04, respectively), and had greater milk fat content (4.45 vs. 4.10%; SEM = 0.11, respectively), but not milk total protein (3.47 vs. 3.39%; SEM = 0.04, respectively) and casein contents (2.74 vs. 2.66%; SEM = 0.04, respectively) than CON cows. Cows in RPMet had increased plasma Met concentrations than cows in CON (24.9 vs. 21.0 µmol/L; SEM = 1.2, respectively). Although morbidity was similar between treatments, the culling rate from calving until 60 DIM was lower for RPMet cows than for CON cows (2.4 vs. 12.1%; SEM = 0.02). In conclusion, cows receiving RPMet have greater milk yield, improved milk fat content, and a lower culling rate at 60 DIM than CON cows.

A Smartphone-Enabled Colorimetric Platform Based on Enzyme Cascade Amplification Strategy for Detection of Staphylococcus aureus in Milk

Staphylococcus aureus (S. aureus) is a pathogenic bacterium-contaminating milk and dairy foods causing food poisoning and foodborne pathogens. In this work, a smartphone-enabled enzyme cascade-triggered colorimetric platform was constructed using cascade bio-nanozyme formed by immobilized glucose oxidase (GOx) on the Fe3O4@Ag for rapid detection of S. aureus. Benefiting from reasonable experimental design, a bio-nanozyme cascade-triggered reaction was achieved through H2O2 produced by GOx oxidation of glucose, followed by in situ catalysis of 3,3',5,5'-tetramethylbenzidine (TMB) by the inherent peroxidase-like activity of Fe3O4@Ag to produce color signals. S. aureus detection could be performed through naked-eye observation and smartphone measurement, the developed assay can achieve quantitative and qualitative detection of S. aureus. The on-site nanoplatform had satisfactory specificity and sensitivity with a low detection limit of 6.9 cfu·mL-1 in 50 min. Moreover, the nanoplatform has good practicality in the detection of S. aureus in milk samples. Therefore, the assay has potential application prospects in food safety inspection.

Molecular detection and antibiogram of Shiga toxin-producing Escherichia coli (STEC) from raw milk in and around Bahir Dar town dairy farms, Ethiopia

Illnesses associated with consuming infected milk and milk products are a widespread problem in low and middle-income countries. Shiga toxin-producing Escherichia coli (STEC) is a bacterium commonly found in raw milk and causes foodborne diseases ranging from mild diarrhea to severe hemorrhagic colitis and hemolytic uremic syndrome. This study aimed to investigate the virulence gene and antimicrobial resistance profiles of Shiga toxin-producing E. coli strains isolated from raw milk in dairy farms in and around Bahir Dar town. Raw milk samples (n = 128) collected from December 2021 to July 2022 were cultured, and E. coli strains were isolated using standard methods. Shiga toxin-producing E. coli strains were identified genotypically by the presence of the virulence markers using a single-plex polymerase chain reaction. The antibiotic susceptibility testing of Shiga toxin-producing E. coli isolates was done by the agar disk diffusion method. In total, 32 E. coli isolates were recovered from milk samples from lactating animals. PCR screening of these isolates resulted in 19 (59.3%) positives for Shiga toxin-producing E. coli. The stx2 gene was detected in 53% of cases, followed by stx1 (31%) and eae (16%. The STEC isolates were highly sensitive to ciprofloxacin (94.7%) and kanamycin (89.5%), while exhibiting significant resistance to amoxicillin (89.5%) and streptomycin (73.7%). The present study points out the occurrence of virulent and antibiotic-resistant Shiga toxin-producing E. coli strains in raw milk that could pose a potential risk to public health. Further analysis by whole genome sequencing is necessary for an in-depth assessment and understanding of their virulence and resistance factors. Moreover, large-scale studies are needed to identify the prevalence and potential risk factors and to prevent the spread of antibiotic-resistant STEC strains in the milk production chain.

Total bacterial count and somatic cell count in bulk and individual goat milk around kidding: two longitudinal observational studies.

Total bacterial count (TBC) and somatic cell count (SCC) are important quality parameters in goat milk. Exceeding the bulk milk TBC (BMTBC) thresholds leads to price penalties for Dutch dairy goat farmers. Controlling these milk quality parameters can be challenging, especially around kidding. First, we describe the variation and the peaks around kidding of TBC and SCC in census data on Dutch bulk milk over the last 22 years. Second, to explore causes of these elevations, we studied the variation of TBC and SCC in individual goat milk from 3 weeks before to 5 weeks after kidding and their association with systemic response markers interferon-γ (IFN-γ), calprotectin, β-hydroxybutyrate (BHB), body condition score (BCS) and fecal consistency. We visited 4 Dutch dairy goat farms weekly for 10 to 16 weeks around kidding. Some of the goats had been dried off, other goats were milked continuously throughout pregnancy. A total of 1,886 milk samples from 141 goats were collected for automated flowcytometric quantification of TBC and SCC measurement. IFN-γ, calprotectin and BHB were determined twice in blood of the same goats, most samples were collected after kidding. The BCS and fecal consistency were scored visually before and after kidding. We found a strong correlation between TBC and SCC (Spearman's rho = 0.87) around kidding. Furthermore, in the third week before kidding, the average TBC (5.67 log10 cfu/mL) and SCC (6.70 log10 cells/mL) were significantly higher compared with the fifth week after kidding, where the average TBC decreased to 4.20 log10 cfu/mL and the average SCC decreased to 5.92 log10 cells/mL. In multivariable linear regression models, farm and stage of lactation were significantly associated with TBC and SCC, but none of the systemic response markers correlated with TBC or SCC. In conclusion, TBC and SCC in dairy goats were high in late lactation and decreased shortly after parturition. For SCC, the dilution effect might have caused the decrease, but this was not plausible for TBC. Moreover, the excretion of bacteria and cells in goat milk was not associated with the selected systemic response markers that were chosen as a read out for general immunity status, intestinal health and metabolic diseases. Therefore, we assume that the TBC increase before kidding and the decrease after parturition is caused by other systemic, possibly hormonal, processes. To reduce BMTBC and BMSCC, it would be advisable to keep milk of goats with highest numbers of bacteria and cells in their milk out of the bulk milk during end lactation. Further studies are needed to investigate the effects of withholding this end lactation milk from the bulk tank.

Effects of feeding whole-cracked rapeseeds, nitrate, and 3-nitrooxypropanol on protein composition, minerals, and vitamin B in milk from Danish Holstein cows

The present study was conducted to assess the individual or combined effects of feeding dietary fat (whole-cracked rapeseed), nitrate, and 3-nitrooxypropanol (3-NOP) on protein profile, mineral composition, B vitamins, and nitrate residues in milk from dairy cows. Forty-eight Danish Holstein cows used in an 8 × 8 incomplete Latin square design were fed 8 factorially arranged diets ((30 or 63 g crude fat/kg DM) × (0 or 10 g nitrate/kg DM) × (0 or 80 mg 3-NOP/kg DM)) over 6 periods of 21 d each. In each period, milk samples were collected from individual cows during the third week by pooling milk obtained from 4 consecutive milkings, and analyzed for protein profile including protein modifications, mineral composition, riboflavin, cobalamin, and presence of nitrate residues. Fat supplementation led to an increase in the phosphorylation degree of αS1-CN by 8.5% due to a decreased relative proportion of αS1-CN 8P and an increased relative proportion of αS1-CN 9P and further to a decrease in the relative proportion of αS2-CN by 2.4%. Additionally, fat supplementation decreased the relative proportions of glycosylated and unglycosylated forms of κ-CN, consequently leading to a 3.6% decrease in total κ-CN. In skim milk, K, Ca, P, and Mg concentrations were altered by individual use of fat, nitrate, and 3-NOP. Feeding nitrate resulted in a 5.4% increase in riboflavin concentration in milk while supplementing 3-NOP increased cobalamin concentration in milk by 21.1%. The nitrate concentration in milk was increased upon feeding nitrate however, this increased concentration was well below the maximum permissible limit of nitrate in milk (<50 mg/L). In conclusion, no major changes were observed in milk protein, and mineral compositions by feeding fat, nitrate, and 3-NOP to dairy cows while the increased riboflavin and cobalamin by nitrate and 3-NOP, respectively, could be of beneficial nutritional value for milk consumers.

Facile construction of sandwich ELISA based on double-nanobody for specific detection of α-hemolysin in food samples

α-hemolysin (Hla), a toxin secreted by Staphylococcus aureus (S. aureus), has been proved to be involved in the occurrence and aggravation of food poisoning. Hence, it is quite essential to establish its rapid detection methods to guarantee food safety. Sandwich ELISA based on nanobody is well known to be viable for toxins, but there is absence of nanobody against Hla, let alone a pair for it. Therefore, in this paper, we screened specific nanobodies by bio-panning and obtained the optimal nanobody pair for sandwich ELISA firstly. Then, RANbody, a novel nanobody owning both recognition and catalytic capability, is generated in a single step and at low cost through molecular recombination technology. Subsequently, sandwich ELISA was developed to detect Hla based on the nanobody and RANbody, that not only eliminated the use of secondary antibodies and animal-derived antibody, but also reduced detection time and cost, compared with traditional sandwich ELISA. Lastly, the performance has been evaluated, especially for specificity which showed no response to other hemolysins and a low limit of detection of 10 ng/mL. Besides, the proposed sandwich ELISA exhibits favorable feasibility and was successfully employed for the detection of Hla in milk and pork samples.

Effects of high moisture ear corn on production performance, milk fatty acid composition, serum antioxidant status, and immunity in primiparous dairy cows.

This study evaluated the effects of high moisture ear corn (HMEC) on production performance, milk fatty acid composition, serum antioxidant status, and immunity in primiparous dairy cows. A total of 45 healthy primiparous Holstein cows (36.50±4.30 kg of milk/day, 201±9.00 lactating days in milk) were sorted into 3 groups: control group (CG, n = 15), 50% HMEC (replacing 50% steam-flaked corn with HMEC, n = 15), and 100% HMEC (replacing steam-flaked corn with HMEC, n = 15) on an equal dry matter (DM) basis. The study consisted of adaptation period of 14 days, followed by a formal period of 60 days. Feed intake and milk yield were recorded daily. Milk and blood samples were collected on 1, 30, and 60 d of the experimental period. The 50% HMEC group and 100% HMEC group significantly increased (p<0.05) milk yield and dry matter intake (DMI) in dairy cows compared to the control group (CG). The 100% HMEC group showed an increase (p<0.05) in 4% fat-corrected milk (4% FCM). Both the 50% HMEC group and 100% HMEC group exhibited significant decreases (p<0.05) in the content of C10:0, C12:0, and C14:0 fatty acids, along with a significant increase (p<0.05) in cis-9C18:1 content. The saturated fatty acid (SFA) content was significantly lower (p<0.05) in the 50% HMEC and 100% HMEC groups than that of CG. Conversely, the monounsaturated fatty acid (MUFA) content was higher (p<0.05) in the 50% HMEC and 100% HMEC groups than that in CG. Notably, the 100% HMEC group significantly increased (p<0.05) the serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) content, while also decreasing the serum malondialdehyde (MDA) content (p<0.05). Moreover, the 100% HMEC group significantly increased (p<0.05) the content of immunoglobulin G (IgG) and immunoglobulin M (IgM). High moisture ear corn could improve production performance and milk fatty acid levels and enhance immunity and antioxidant capacity in dairy cows. These results lay the foundation for the wider application of HMEC in ruminant animal diets.

Study protocol to assess aflatoxin M1 health risks versus benefits of dairy consumption in Ethiopian children: an epidemiological trial and risk-benefit analysis

IntroductionIn Sidama, Ethiopia, animal-source foods can be difficult to access. Milk has important nutrients for child growth, but carries the risk of aflatoxin M1 (AFM1) contamination. AFM1 is a metabolite...

Establishment of a real‐time fluorescence and visual colorimetric detection method for Staphylococcus aureus based on loop‐mediated isothermal amplification

AbstractA loop‐mediated isothermal amplification (LAMP) method combined with calcein was established for the detection of Staphylococcus aureus. LAMP primers were designed targeting the ASL18_04625 species‐specific gene screened through bioinformatics and PCR analysis. The target gene and detection method demonstrated 100% specificity for S. aureus and no cross‐reactions with other pathogens. For LAMP reactions, the results could be directly observed with the naked eye or evaluated by S‐shaped amplification curves of fluorescence signals. The assay's detection limit was 3.395 × 10−4 ng/μL genomic DNA or 3.21 CFU/mL pure bacterial culture. Furthermore, the target bacteria of 8.5 × 105–8.5 × 100 CFU/mL could be accurately detected in spiked milk samples, and the overall detection process could be completed within 40 min. This method improved the accuracy and convenience for S. aureus detection and provied a dependable tool for the rapid screening of S. aureus.

Melamine‐imprinted quartz crystal microbalance sensor for its detection in milk

AbstractA unique and selective molecularly imprinted polymer (MIP) was fabricated to detect melamine using a quartz crystal microbalance (QCM) surface. The MIP was synthesized by sol–gel technique utilizing 3‐(trimethoxysilyl) propyl methacrylate (TMSPMA) as the functional monomer bearing the methacrylate head group. To identify the melamine, we applied a coating of the as‐synthesized MIP over the silicon crystal of the quartz crystal microbalance (QCM) detector. High‐affinity adsorption of template molecules from the aqueous solution is the basis of the detection. Adsorption experiments performed in real‐time on MIP surfaces indicate that melamine forms strong bonds with its template molecule. We have evaluated the MIP surface using contact angle, BET, RAMAN and AFM investigations regarding wettability, reactivity, surface area, and smoothness. The real‐time adsorption experiments of template molecules on the MIP substrate reveal enhanced attachments with MIP, whereas the NIP demonstrates no interactions with the template molecules. Melamine detection in tainted milk samples further validated the MIP QCM sensor effectiveness. Melamine at a concentration ranging from 1–20 mg/L is tested. Findings conclude that the polymerization method and the QCM nanosensor for selective melamine detection were adequate substitutes for the technologies currently used for detecting melamine.