Aptasensors application for cow's milk allergens detection and early warning: Progress, challenge, and perspective.

Cow's milk allergy (CMA) is considered one of the most prevalent food allergies and a public health concern. Modern medical research shows that the effective way to prevent allergic reactions is to prevent allergic patients from consuming allergenic substances. Therefore, the development of rapid and accurate detection technology for milk allergens detection and early warning is critical to safeguarding those with a cow milk allergy. As the oligonucleotide sequences with high specificity and selectivity, aptamers frequently assemble with transduction elements forming multifarious aptasensors for quantitative detection owing to their high-affinity binding to the target. Current aptasensors in the field of cow's milk allergen detection in recent years are explored in this review. This review takes a look back at a few common assays, including ELISA and PCR, before presenting a clear overview of the aptamer and threshold doses. It delves into a detailed discussion of the current aptamer-based detection techniques and related theories for milk allergen identification. Last but not least, we conclude with a discussion and outlook of the advancements made in allergen detection with aptamers. We sincerely hope that there will be more extensive applications for aptasensors in the future contributing to reducing the possibility of patients suffering from adverse reactions.

Changes in mammary infection status in dairy cows during the dry period using dry cow therapy approaches on three farms.

This case study evaluated the mammary infection status of dairy cows during the dry periods and explored the associated problems in their quarters with dry cow therapy (DCT). This study assessed intramammary infections, antibiotic efficacy, and antimicrobial resistance of pathogens in 464-quarter milk samples from 59 dairy cows during the dry periods after applying blanket DCT, non-DCT, and selective DCT approaches on three farms. The recovery rates of intramammary infections were 95% (19/20 quarters) with blanket DCT on farm A, 70% (14/20) with non-DCT on farm B, and 19% (4/21) with selective DCT on farm C. Analysis of mammary infections in cows with DCT revealed that mammary infections were controlled by blanket DCT, well controlled by non-DCT, and substantial problems remained in selective DCT. Lower intramammary infection prevalence in the quarters at postpartum appeared to be associated with higher recovery of mammary infections, fewer new infections, and lower uncured mammary infections within the herds. Antibacterial resistance in 14 coagulase-negative staphylococci isolated to six antimicrobial drugs was suggested to be linked to antibiotic use on the farm. Follow-up studies on the quarter-based infection status with DCT will assist in improving mastitis control in cows during the dry period.

Mechanism of collagen type IV regulation by focal adhesion kinase during retained fetal membranes in dairy cows.

Retained fetal membranes (RFM) is an important reproductive disease in dairy cows, caused by maternal and fetal placental tissue adhesion. The main collagen in maternal and fetal placenta tissues is collagen type IV (COL-IV) and its breakdown is the key to placental expulsion. Focal adhesion kinase (FAK) has been shown to regulate the hydrolysis of Col-IV by affecting the activity of MMP-2 and MMP-9 activity, but the regulation of the mechanisms involved in placenta expulsion in dairy cows after postpartum are still unclear. The aim of this study was to investigate the pathogenic mechanism of RFM by studying the relationship between the FAK signaling pathway and COL-IV regulation. Maternal placental tissues were collected from six healthy and six cows with RFM of similar age, parity, body condition and milk yield at 12 h postpartum. In vitro experiments were performed on bovine endometrial epithelial cells from three groups including a FAK inhibitor group, a FAK activator group and a control group without FAK inhibitor and activator. The abundance of molecules involved in the FAK signaling pathway and COL-IV was detected by immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. The immunohistochemical results showed that the key molecules of FAK signaling pathway FAK, Src, MMP-2 and MMP-9 and Col-IV were expressed in placental tissues. The expression level of FAK, Src, MMP-2, and MMP-9 were significantly down-regulated (P < 0.05) and the abundances of COL-IV were significantly up-regulated (P < 0.05) in maternal placental tissues of RFM cows compared with healthy cows. In the FAK inhibitor treatment group, the relative expression levels of FAK and other related proteins were significantly down-regulated (P < 0.05) and the relative expression levels of COL-IV were significantly up-regulated (P < 0.05) with the results of the FAK activation group the opposite. These results indicated that FAK in maternal endometrial epithelial cells could regulate the hydrolysis process of Col-IV through the expression of key factors of signaling pathways and promote collagen hydrolysis, which in turn facilitated the process of postpartum placenta expulsion in dairy cows.

Relationship between the effect of human chorionic gonadotropin treatment on the fifth day after artificial insemination and ovarian ultrasonographic findings and blood nutritional metabolic factors in dairy cows.

The objective of the present study was to determine the ovarian ultrasonographic findings and metabolic factors that influence the effect of human chorionic gonadotropin (hCG) treatment on the fifth day after artificial insemination (AI) in dairy cows. Thirty-seven lactating Holstein cows were assigned to two groups: the hCG group (n = 25), which received 3000 IU of hCG intramuscularly on Day 5 after AI (day of AI = Day 0), and the control group (n = 12), which received no treatment. Ovarian ultrasonography measured luteal tissue area (LTA), luteal blood flow area (LBF), relative LBF (= LBF/LTA), and dominant follicle area on Day 5. Blood tests measured plasma insulin-like growth factor-I, insulin, and metabolite concentrations on Day 5 and plasma progesterone concentrations on Days 5 and 7. LBF was greater in pregnant cows than in non-pregnant cows, and plasma Glu concentration was lesser in pregnant cows than in non-pregnant cows, but in both cases there was no interaction between group and pregnancy outcome. For plasma insulin concentration, there was an interaction between group and pregnancy outcome, with pregnant cows in the hCG group having lesser concentrations than the other groups. Logistic regression analysis showed that group and the interaction between group and plasma insulin concentration were associated with pregnancy outcome. These results suggest that the effect of hCG treatment on Day 5 after AI is related to plasma insulin concentration and is more effective in cows with lesser plasma insulin concentrations.

Metagenomics reveals the characteristics and potential spread of microbiomes and virulence factor genes in the dairy cattle production system.

Virulence factor genes (VFGs) pose a potential threat to ecological security and animal health, and have attracted increasing attention in the livestock industry. As one of the primary livestock types, dairy cattle may be an important source of VFG transmission. However, the distribution, transmission, and evolution of VFGs in the gastrointestinal tract and surrounding environment of dairy cattle remain unclear. In the present study, a total of 263 samples were collected from cows, calves, colostrum, farm wastewater, and soil. Metagenomics was conducted to analyze changes in the microbiome and VFGs characteristics in these ecological niches. The VFGs of the cows showed distinct differences between the rumen and feces, and were influenced by the region. The dominant VFG hosts was regulated by their microbial structure. Colostrum administration of cows increased VFG abundance in their newborn calf feces sharply and Enterobacteriaceae became the primary host. While diet was the primary driving force for the temporal variation in calf VFGs. For samples of the surrounding environment, water and soil had higher VFG concentrations and were more structurally stable. Moreover, extensive interactions between the mobile genetic elements and VFGs and gene mobile analysis map based on metagenomic binning both displayed the potential horizontal transfer ability of VFGs in the cows and environment. Our study revealed the prevalence, diffusion, and regulatory factors of VFGs in dairy cattle production systems, providing novel insights into reducing livestock VFGs and limiting their spread.

Exploring the Gut Microbiota Landscape in Cow Milk Protein Allergy: Clinical Insights and Diagnostic Implications in Pediatric Patients.

Cow milk protein allergy (CMPA) is a significant health concern characterized by adverse immune reactions to cow milk proteins. Biomarkers for the accurate diagnosis and prognosis of CMPA are lacking. This study analyzed the clinical features of CMPA, and 16S RNA sequencing was used to investigate potential biomarkers through fecal microbiota profiling. Children with CMPA exhibit a range of clinical symptoms, including gastrointestinal (83% of patients), skin (53% of patients), and respiratory manifestations (26% of patients), highlighting the complexity of this condition. Laboratory analysis revealed significant differences in red cell distribution width (RDW) and inflammatory markers between the CMPA and control groups, suggesting immune activation and inflammatory responses in CMPA. Microbial diversity analysis revealed higher specific diversity indices in the CMPA group compared with those in control group, with significant differences at the genus and species levels. Bacteroides were more abundant in the CMPA group, whereas Bifidobacterium, Ruminococcus, Faecalibacterium, and Parabacteroides were less abundant. The control group exhibited a balanced microbial profile, with a predominant presence of Bifidobacterium bifidum and Akkermansia muciniphila. The significant abundance of Bifidobacterium in the control group (23.19% vs 9.89% in CMPA) was associated with improved growth metrics such as height and weight, suggesting its potential as a probiotic to prevent CMPA and enhance gut health. Correlation analysis linked specific microbial taxa such as Coprococcus and Bifidobacterium to clinical parameters such as family allergy history, weight and height, providing insights into CMPA pathogenesis. Significant differences in bacterial abundance suggested diagnostic potential, with a panel of 6 bacteria achieving high predictive accuracy (area under curve (AUC) = 0.8708). This study emphasizes the complex relationship between the gut microbiota and CMPA, offering valuable insights into disease mechanisms and diagnostic strategies.

Epidemiology of antimicrobial resistance in commensal E. coli from healthy dairy cattle on a Mediterranean pasture-based system of Australia: A cross-sectional study.

This study aimed to determine the prevalence of antimicrobial resistance (AMR) in commensal E. coli from healthy lactating cows and calves in the Mediterranean pasture-based feeding dairy system of Western Australia (WA). Fecal samples were collected from healthy adult lactating cows and healthy calves from dairy farms in WA. Presumptive commensal E. coli was isolated from these samples and confirmed using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Broth microdilution was used to assess the prevalence and the phenotypic AMR profiles of the E. coli isolates to 8 antimicrobial agents of dairy industry and human importance. The minimum inhibitory concentration (MIC) for each isolate was interpreted using the Epidemiologic Cutoff (ECOFF) and Clinical and Laboratory Standards Institute (CLSI) breakpoints. Genomic characterization provided multi-locus sequence types and AMR genes for a selection of isolates categorised as non-wild type (NWT) by ECOFF values for the combination of ampicillin, trimethoprim-sulfamethoxazole, and tetracycline. From a total of 1,117 fecal samples (633 adult, 484 calf) collected across 26 randomly selected farms, 891 commensal E. coli isolates were recovered (541 adult, 350 calf). Commensal E. coli classified as NWT was highest for ampicillin for both adult (68.8%; 95% CI = 64.7 - 72.7) and calf feces (67.1%; 95% CI = 62.0 - 72.0). A large proportion of tetracycline NWT and trimethoprim-sulfamethoxazole NWT organisms were also identified from calf feces, being 44.0% (95% CI = 38.7 - 49.4) and 24.6% (95% CI = 20.2 - 29.4) respectively. Clinical resistance prevalence was low, being higher for calves than for adult feces (ampicillin (adult: 7.8% (95% CI = 5.7 - 10.3); calf: 30.0% (95% CI = 25.2 - 35.1), tetracycline (adult: 6.3% (95% CI = 4.4 - 8.7); calf: 40.3% (95% CI = 35.1 - 45.6), and trimethoprim-sulfamethoxazole (adult: 2.6% (95% CI = 1.4 - 4.3); calf: 22.0% (95% CI = 17.7 - 26.7)). Commensal E. coli originating from calf feces was significantly higher in NWT prevalence compared with adult feces for ciprofloxacin (P = 0.023), gentamicin (P = 0.02), tetracycline (P < 0.001), and trimethoprim-sulfamethoxazole (P < 0.001). The overall number of antimicrobials an isolate was classified as NWT toward varied among farms and was significantly higher for isolates originating from calf than adult feces (P < 0.001). The strain type and sampling source of the commensal E. coli investigated were both associated with the commonality of the resultant resistance genome. Clinical resistance and NWT classification were highest for ampicillin, tetracycline, and trimethoprim-sulfamethoxazole, all antimicrobials commonly used in the treatment of dairy cattle in Australia. Though highly variable across farms, commensal E. coli isolated from healthy dairy calf feces had significantly higher NWT and multidrug resistance (MDR) prevalence compared with feces from healthy adult lactating dairy cows. The resistant genome identified in MDR isolates, though not always consistent with the phenotype, included QnrS1 and genes encoding AmpC ß-lactamase and aminoglycoside phosphotransferase.

A systematic review and meta-analysis of cow-level factors affecting milk urea nitrogen and urinary nitrogen output under pasture-based diets.

With dairy cattle farming under pressure to lower its environmental footprint it is important to find effective on-farm proxies for evaluation and monitoring of management practices aimed at reducing the risk of nitrogen (N) losses and optimizing N use efficiency of dairy farm systems. Urinary N (UN) is regarded as the most potent source of N emissions. In contrast to confinement systems, there have been few studies from pasture-based systems associating on-farm animal and nutritional factors with UN output. Thus, the aims of this meta-analysis were to collate a database from pasture-based research to: (a) investigate the associations of management, dietary, and animal variables with MUN concentration, and daily UN output; (b) describe the MUN-UN association; and (c) assess whether animal, management, and dietary factors influence the relationship. We developed a data set consisting of 95 observations representing 919 lactating dairy cattle fed pasture-based diets, which was compiled from 32 unique research publications that reported both MUN and UN output. Multi-level, mixed meta-analysis regression techniques were used to analyze the data. Initially, all variables were assessed as the sole fixed effect in a 2-level random effects model, accounting for within publication heterogeneity. Meta-regression techniques were then used to assess the relationship of all variables with MUN and UN output, respectively, accounting for 3 sources of variability: the sampling error of the individual observation, within publication heterogeneity, and among publication heterogeneity. At the univariable level, despite more than 10 dietary, animal, or management variables being significantly associated with MUN, none explained a large amount of the MUN variation. The variables that explained the greatest amount of variation were dietary crude protein (CP) content and the nitrogen: metabolizable energy content ratio, which explained about 33% and 31% of the variation in MUN concentrations, respectively. Combining factors in multiple regressions improved the model fit, such that the variation within publications explained by dietary CP and N intake increased to 40.0% in the final multiple meta-regression model. For UN output, individual variables explained a greater proportion of variance reported among observations, compared with MUN, whereby diet CP content (pseudo R2 = 66.1%), N to metabolizable energy intake ratio (pseudo R2 = 64.0%), N intake (pseudo R2 = 58.3%), and MUN (pseudo R2 = 43.5%) explained the greatest amount of the total variation. Milk urea nitrogen, N intake and dry matter intake were associated with UN output in the final multiple meta-regression model. Substantial heterogeneity existed in both MUN and UN among publications, with among publication heterogeneity accounting for 73.4% of all the variation noted in MUN, and 88.6% of all the variation in UN output. As such, the meta-analyses could not predict MUN and UN to any great extent. It is recommended that a consistent approach to measuring and reporting MUN concentrations and UN output is carried out for all future research in pasture-based systems.

Effects of supplementing rumen-protected arginine on performance of transition cows.

An experiment was conducted to determine the effects of supplementing rumen-protected arginine (RPA) on productive performance in dairy cows. One-hundred and 2 cows were blocked by parity and then by energy-corrected milk (ECM) yield. Within block, cows were randomly assigned to control (CON) that received 200 g/d of a mixture of hydrogenated soybean oil and heat-treated soybean meal to supply 30 g of metabolizable protein (MP), or 200 g/d of a product containing 30 g of metabolizable arginine (RPA), which increased the dietary arginine from 5.7 to 7.5% of the MP from 250 d of gestation to 21 d postpartum. After 21 d postpartum, cows were fed the same diet and data were collected until 84 d postpartum. Cows fed RPA produced an additional 2.5 kg of colostrum (5.3 vs. 7.8 ± 1.0 kg) and 220 g more immunoglobulin G (526 vs. 746 ± 93 g) than CON cows. Supplementing RPA increased the yields of milk (32.8 vs. 34.9 ± 1.0 kg/d), ECM (37.8 vs. 40.9 ± 1.2 kg/d), and milk total solids (4.48 vs. 4.86 ± 0.14 kg/d) in the first 21 DIM. The benefits of RPA extended beyond the period of supplementation, with a 6.4% increase in yield of ECM per kg of dry matter consumed in all cows (1.88 vs. 2.00 ± 0.05 kg/kg) and an increase in ECM yield, but only in parous cows (44.2 vs. 48.5 ± 1.5 kg/d). Feeding RPA increased the concentrations of urea N in plasma pre- (12.5 vs. 13.9 ± 0.4 mg/dL) and postpartum (11.6 vs. 13.2 ± 0.4 mg/dL), and in milk during the first 21 d postpartum (11.0 vs. 12.0 ± 0.3 mg/dL). Treatment did not affect the concentrations of AA in plasma prepartum, but feeding RPA tended to increase citrulline (72.5 vs. 77.5 ± 2.7 µM), whereas RPA either tended to decrease isoleucine (129.5 vs. 120.9 ± 5.7 µM) or decreased the concentrations of leucine (181.3 vs. 170.2 ± 6.4 µM) and valine (293.2 vs. 276.7 ± 10.4 µM) postpartum. Feeding RPA increased the relative expression of transcripts involved in AA transport (SLC38A4), urea cycle (ARG1), and gluconeogenesis (PC, PEPCK, and G6PC) in hepatic tissue. Feeding diets to supply additional arginine as RPA during the transition period benefited productive performance in dairy cows that extended beyond the period of supplementation despite minor changes in plasma AA concentrations.

Molecular characteristics and antimicrobial susceptibility profiles of bovine mastitis agents in western Türkiye.

IMPORTANCE: Identifying bovine mastitis agents using molecular methods to reveal their phylogenetic relationships and antimicrobial resistance profiles is essential for developing up-to-date databases in mastitis cases that cause severe economic losses. OBJECTIVE: This study examined bacterial mastitis agents in cows with clinical and subclinical mastitis observed in various dairy cattle farms to reveal their phylogenetic relationships and antibiotic resistance properties. METHODS: Sixty-two clinical and subclinical bovine mastitis milk samples were collected from 15 dairy farms. The polymerase chain reaction (PCR) was used to amplify the 16S rRNA gene regions of the bacteria. The 16S rRNA gene sequences obtained from sequencing include the V4-V6 regions. The strains were compared using a similarity analysis method that produced phylogenetic trees using the Molecular Evolutionary Genetics Analysis 11 program. Antibiotic susceptibilities were determined using the Kirby-Bauer disk diffusion method. RESULTS: Sixty-three bacteria were isolated and identified in this study. The most isolated bacteria from all mastitis cases were Staphylococcus spp. (30.2%), Escherichia coli (25.4%), Streptococcus spp. (14.3%), and Aerococcus spp. (7.9%), respectively. The phylogenetic trees were drawn from the 16S rRNA sequences. Some of these bacteria showed resistance to different types of antibiotics at varying rates. CONCLUSIONS AND RELEVANCE: The bacteria isolated in this study originated from environmental sources. Regular cleaning of barns and proper hygiene practices are essential. Regular screenings for mastitis should be conducted in herds instead of the random or empirical use of antibiotics.

Preliminary study of molecular identification of Mycobacterium bovis from cow's milk in Lorestan (Iran).

Bovine tuberculosis is one of the most important common infectious diseases between humans and livestock. Cow's milk can be investigated as one of the transmission reservoirs of the disease. Our study was conducted to investigate the presence of Mycobacterium bovis (M. bovis) DNA in cow's milk from different regions of Lorestan province using Touch-down PCR (TD-PCR) method. So, 100 milk samples from industrial and traditional cattle farms were collected and evaluated according to the animal breed, the average age of the animal and the region where the animal was kept. Seven (26.9%) out of the 26 cow's milk samples contaminated with Mycobacterium spp., were diagnosed as M. bovis positive. The cattle with an average age of more than 5 years were most infected with Mycobacterium. Also, the crossbred cattle with an average age of 3 to 5 years, which were kept and raised in tropical areas, showed the highest rate of M. bovis infection. Based on our knowledge, this is the first study regarding the presence of Mycobacterium in cow's milk in Iran.

Investigation of the regulatory mechanisms of Guiqi Yimu Powder on dairy cow fatty liver cells using a multi-omics approach.

Introduction: Fatty liver disease in dairy cows is a metabolic disorder that significantly affects their health and productivity, imposing a notable economic burden on the global dairy industry. Traditional Chinese medicine (TCM), characterized by its multi-component and multi-target features, has shown unique advantages in the prevention and treatment of various diseases. Guiqi Yimu Powder, a traditional TCM formula, enhances growth, boosts production efficiency, and strengthens immune function in livestock by regulating antioxidant along with anti-inflammatory pathways. However, its specific regulatory mechanisms on fatty liver in dairy cows remain unclear. This study aims to investigate the molecular-level effects and potential regulatory mechanisms of Guiqi Yimu Powder in a Trimethylamine N-oxide (TMAO) induced fatty liver cell model of dairy cows. Methods: We employed a comprehensive analysis integrating transcriptomics, proteomics, metabolomics, and network pharmacology. An in vitro dairy cow fatty liver cell model was established using TMAO to induce lipid accumulation. Cells were treated with the optimal TMAO concentration identified through preliminary experiments, and further divided into a lipid accumulation group and Guiqi Yimu Powder treatment groups. The treatment groups received varying concentrations of Guiqi Yimu Powder (10, 20, 30, 40, or 50 g/L). High-throughput omics sequencing technologies were utilized to perform a comprehensive analysis of the treated cells. Bioinformatics methods were applied to explore the regulatory effects, aiming to elucidate the specific impacts of Guiqi Yimu Powder on lipid metabolism, liver function, and related signaling pathways, thereby providing scientific evidence for its potential application in the prevention and treatment of fatty liver in dairy cows. Results: Guiqi Yimu Powder treatment significantly affected 1,536 genes, 152 proteins, and 259 metabolites. KEGG enrichment analysis revealed that the significantly altered molecules are involved in multiple pathways related to the pathology of fatty liver, including metabolic pathways, glutathione metabolism, hepatitis B, and AMPK signaling pathway (p < 0.05). Notably, joint analysis highlighted the regulatory mechanisms of Guiqi Yimu Powder on glutathione cycling, with L-5-Oxoproline identified as an important metabolic compound. These findings indicate its impact on oxidative stress, energy metabolism, and liver function, suggesting potential therapeutic applications for fatty liver in dairy cows. Discussion: This study elucidated the regulatory mechanisms of Guiqi Yimu Powder on fatty liver cells in dairy cows, providing new scientific evidence for its potential application in the prevention and treatment of fatty liver disease.

Mechanical positional information guides the self-organized development of a polygonal network of creases in the skin of mammalian noses.

The glabrous skin of the rhinarium (naked nose) of many mammalian species exhibits a polygonal pattern of grooves that retain physiological fluid, thereby keeping their nose wet and, among other effects, facilitating the collection of chemosensory molecules. Here, we perform volumetric imaging of whole-mount rhinaria from sequences of embryonic and juvenile cows, dogs, and ferrets. We demonstrate that rhinarial polygonal domains are not placode-derived skin appendages but arise through a self-organized mechanical process consisting of the constrained growth and buckling of the epidermal basal layer, followed by the formation of sharp epidermal creases exactly facing an underlying network of stiff blood vessels. Our numerical simulations show that the mechanical stress generated by excessive epidermal growth concentrates at the positions of vessels that form rigid base points, causing the epidermal layers to move outward and shape domes-akin to arches rising against stiff pillars. Remarkably, this gives rise to a larger length scale (the distance between the vessels) in the surface folding pattern than would otherwise occur in the absence of vessels. These results hint at a concept of "mechanical positional information" by which material properties of anatomical elements can impose local constraints on an otherwise globally self-organized mechanical pattern. In addition, our analyses of the rhinarial patterns in cow clones highlight a substantial level of stochasticity in the pre-pattern of vessels, while our numerical simulations also recapitulate the disruption of the folding pattern in cows affected by a hereditary disorder that causes hyperextensibility of the skin.

Photothermal-Induced Multimodal Antibacterial Dressing Comprising N-Halamine Hydrogel Loaded with Cow Dung Biochar for Infected Wound Healing.

Disposal of the cow dung pollutants arising from cattle farming threatens the environment and public safety in diverse ways. To date, researchers have worked on developing new pathways to control and manage cattle farming wastes, but most do not involve the reuse of these wastes. Herein, a cow dung biochar-modulated photothermal N-halamine hydrogel (i.e., PAN/CA/CoBC/pMAG-Cl) was designed for converting cow dung into biochar (CoBC), which was then coupled with a 3D interpenetrating hydrogel network to treat infected wounds. The PAN/CA/CoBC/pMAG-Cl hydrogel exhibited excellent synergistic antibacterial performance against 106 CFU·mL-1 Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) within 60 min. The bactericidal effect was multimodal, involving CoBC-based photothermal killing (i.e., temperature as high as 80.5 °C) after 808 nm near-infrared light irradiation for 10 min, contact killing through the strong oxidative characteristic of N-halamine (pMAG-Cl), and release killing via active halogens (i.e., Cl+) reinforced by the photothermal action of CoBC. The S. aureus-infected wound model in vivo demonstrated that the PAN/CA/CoBC/pMAG-Cl hydrogel worked as an ideal wound dressing, capable of resisting bacterial infection, accelerating the healing process, and promoting epithelial regeneration. This proposed strategy could indicate a new way for the disposal of cow dung pollutant and its reuse in antibacterial-associated applications.

Spatial distribution and risk assessment of perchlorate in raw cow milk from China.

Perchlorate is a ubiquitous environmental contaminant worldwide, recognized as an emerging thyroid toxicant. This study focused on the pollution status, spatial distribution, possible sources of perchlorate in raw cow milk collected from 155 dairy farms in China, as well as the health risk of exposure to perchlorate through dairy products. The results showed that the detection rate of perchlorate in raw milk was 100% with the mean of 15.9 µg/kg, indicating the ubiquitous contamination of perchlorate in raw milk from China. The simulation of spatial distributions indicated that the levels of perchlorate in raw milk were spatially correlated, and relatively high levels of perchlorate exist in certain parts of Beijing-Tianjin-Hebei, Shanxi, Henan, and Zhejiang, suggesting potential environmental perchlorate contamination in these regions. A positive correlation was found between the perchlorate level in milk and the perchlorate level in feed, indicating the transfer process of perchlorate from feed to milk. The hazard quotient (HQ) values of exposure to perchlorate by dairy products ranged between 0 and 2.14, with the mean of 0.0188 and P95 of 0.101, indicating relatively low health risk to perchlorate through dairy products. To our knowledge, this is the first nationwide study on the spatial distribution and risk assessment of perchlorate in raw cow milk from China.

Progesterone increases metabolism via the pentose phosphate pathway in bovine uterine epithelial cells.

BACKGROUND: During early pregnancy, glucose is essential for the uterine epithelium and the developing embryo. In cows, progesterone increases the secretion of glucose into the uterine lumen. The uterine epithelium can convert glucose to fructose, but other fates of glucose in the uterine epithelium have been sparsely investigated. Therefore, our objective was to investigate how progesterone influences glucose metabolism in immortalized bovine uterine epithelial (BUTE) cells. METHODS: BUTE cells were grown to 80% confluence and treated with vehicle (DMSO) or 10 µM progesterone for 24 h. Cells were collected and analyzed. Immunohistochemistry was performed on endometrial samples collected from the bovine endometrium on days 1 and 11 of the reproductive cycle. RESULTS: Progesterone treatment increased glucose consumption of BUTE cells. RNAseq identified 3,072 genes regulated by progesterone. KEGG analysis indicated that progesterone altered genes associated with metabolic pathways and glutathione metabolism. Manually examining genes unique to specific glucose metabolic pathways identified an increase in the rate-limiting enzyme in the pentose phosphate pathway-glucose-6-phosphate dehydrogenase. Functionally, a major product of the pentose phosphate pathway is NADPH, and progesterone treatment increased NADPH levels in BUTE cells. In cows, immunohistochemistry confirmed that glucose-6-phosphate dehydrogenase levels were higher in the uterine epithelium in the luteal phase when progesterone concentrations are high. CONCLUSIONS: Progesterone increased glucose-6-phosphate dehydrogenase expression and metabolism via the pentose phosphate pathway in the bovine uterine epithelium. This metabolism could provide substrates for cell proliferation, molecules to be secreted into the uterine lumen, or maintain reduction/oxidation balance in the uterine epithelium.

Effect of Gestational Direct-Fed Microbials Supplementation on the Metabolic Profile in Periparturient Dairy Cows.

This study was conducted at a commercial dairy farm in Queensland, Australia to evaluate the effects of feeding a lactic acid bacteria-based direct-fed microbial (DFM) during gestation on the metabolic profile of periparturient dairy cows and its effects on milk production and body weight. A total of 150 multiparous Holstein cows were randomly selected based on parity (2.3) and days in milk (130 DIM) and divided into two groups of 75 cows each (control and DFM). The control cows were assigned to a basal diet consisting of a silage-based partial mixed ration (PMR), concentrate fed in the dairy twice a day, and ad libitum pasture. The DFM group received the same basal diet supplemented with three strains of Lactobacillus top-dressed in the feed. The DFM supplementation continued during both the dry period and the subsequent lactation. A subset of 82 cows (40 control and 42 DFM) were monitored during the calving season (March to July 2022) to assess the metabolic profile and postpartum performance. Blood samples were collected during the periparturient period (-4 to -2 w prepartum, around calving, and at weeks 1, 3, and 6 postpartum) to measure the levels of metabolites, enzymes, and minerals. Overall, the serum glucose, NEFA, and chloride levels were higher, while protein and urea were lower in cows supplemented with the DFM (p < 0.005). The pre-calving levels of glucose were higher and the total bilirubin, urea, and BHB were lower in cows supplemented with DFM than in the control (p < 0.05). The post-calving levels of glucose and Mg were also higher in the DFM cows than in the control cows (p < 0.05). Average milk production at 110 DIM was significantly higher in the DFM cows compared to control cows (p = 0.03). Although the total milk production over 305 days was numerically greater in the DFM cows, the difference was not statistically significant (p = 0.3), whereas the milk protein percentage was higher in the control cows (p = 0.03). The body weight of the DFM cows was greater during the periparturient period (p = 0.001) than that of the control cows. In the DFM cows, glucose levels had a positive correlation (r = 0.16) with milk yield, at 110 DIM, while serum total protein had a positive correlation with body weight (r = 0.32) (p < 0.05). In conclusion, feeding Lactobacillus-based DFM during gestation can positively influence the metabolic profile of periparturient cows, which, in turn, may affect the milk production and body weight of postpartum dairy cows.

Impact of Feed Bunk Space on the Prevalence of Torsion and Foreleg Distal Asymmetry in Holstein Cows.

Leg torsion and distal asymmetry (LTDA) among cows reared on intensive farms in the Comarca Lagunera region of northern Mexico may be indicative of underlying health concerns. To ascertain whether the incidence of LTDA is associated with trough measurements and with productive, reproductive, and disease variables, the prevalence of LTDA was determined in lactating dairy cows. The data were derived from two intensive dairy farms in northern Mexico (G60: 2043 cows in 13 pens with 142.0 m of linear feed bunk space and 65.0 m of pen depth; G100: 2227 cows in 12 pens with 215.0 m of linear feed bunk space and 49.0 m of pen depth). The cows were observed over the course of a week to identify any macroscopic lesions indicative of LTDA. Cows exhibiting lesions were assigned a value of "1", while those without lesions were assigned a value of "0". Furthermore, data regarding other limb diseases (LDs) were collected and subjected to analysis. A comparison was conducted to ascertain the impact of reproductive, productive, and feed bunk size variables on the prevalence of LTDA and other LDs. To ascertain the prevalence of LTDA and LDs, a chi-squared test was employed. The prevalence of LTDA was found to be 27.6% (G60: 31.4%, G100: 22.4%). No association was identified between LTDA and the variables included in the study. However, a correlation was identified between LDs and the following variables: total width, distance from the edge, width from the feed bunk to the tramp, and the presence of a manure crust on the walls. Principal component analysis (PCA) was conducted to examine the correlation between LDs and various factors at the farm and pen levels. The findings indicated that the distance from the feed bunk to the trough, the presence of manure crusts on the walls, and the breeding time were associated with LDs in G100, as well as in three of the twelve pens (302, 306, and 308) within G100. The findings suggest that the prevalence of LDs is associated with an increase in the width of the feed bunk and the width of the manure crust on the walls, particularly in pregnant cows. The results permit the conclusion that LTDA and LDs are associated with the measurements of the feed bunks, the absence of manure cleaning of the feed bunks, and gestation. This association gives rise to significant health issues for Holstein cows on these farms, with more than one in four cows affected. To reduce the prevalence of LTDA in lactating dairy cows, it is recommended that the standard measurements for feed bunk design be adhered to. This will reduce the prevalence of LTDA and LDs, promote the cleaning protocols to avoid the accumulation of manure crusts, and facilitate close monitoring of pregnant cows, thereby alleviating the impacts of these foot pathologies on animal welfare.

Association Between Dairy Production System and Milk Functionality Based on Analysis of miRNAs in Exosomes from Milk.

Dairy farming practices significantly affect the nutritional and functional properties of milk. This study compares miRNAs in milk exosomes from extensive and intensive dairy systems and explores their potential implications for human consumers. Extensive systems are believed to produce milk of higher quality with better animal welfare compared to intensive systems. Milk samples from eight extensive and nine intensive dairy farms were analysed. Milk-derived extracellular vesicles were isolated through sequential ultracentrifugation and characterised through Dynamic Light Scattering and Nanosight to determine the size and the concentration of the extracellular vesicles, in addition to immunoblotting to ensure the presence of exosome-specific proteins in their membrane. miRNA levels were quantified using RT-qPCR, and metabolic pathways associated with miRNAs showing significant differences between farm groups were analysed. EVs from extensive farms had higher concentrations. Notably, bta-miR-451 levels were significantly higher in milk from extensive farms (p = 0.021). Like human miRNA hsa-miR-451, it is linked to pathways related to Parkinson's disease and cancer. Our research suggests that milk production in extensive systems not only provides socioeconomic and environmental benefits but may also have positive effects on human health. Further research is warranted to explore the bioactive potential of these miRNAs and their implications for human health.

CAMLLA-YOLOv8n: Cow Behavior Recognition Based on Improved YOLOv8n.

Cow behavior carries important health information. The timely and accurate detection of standing, grazing, lying, estrus, licking, fighting, and other behaviors is crucial for individual cow monitoring and understanding of their health status. In this study, a model called CAMLLA-YOLOv8n is proposed for Holstein cow behavior recognition. We use a hybrid data augmentation method to provide the model with rich Holstein cow behavior features and improve the YOLOV8n model to optimize the Holstein cow behavior detection results under challenging conditions. Specifically, we integrate the Coordinate Attention mechanism into the C2f module to form the C2f-CA module, which strengthens the expression of inter-channel feature information, enabling the model to more accurately identify and understand the spatial relationship between different Holstein cows' positions, thereby improving the sensitivity to key areas and the ability to filter background interference. Secondly, the MLLAttention mechanism is introduced in the P3, P4, and P5 layers of the Neck part of the model to better cope with the challenges of Holstein cow behavior recognition caused by large-scale changes. In addition, we also innovatively improve the SPPF module to form the SPPF-GPE module, which optimizes small target recognition by combining global average pooling and global maximum pooling processing and enhances the model's ability to capture the key parts of Holstein cow behavior in the environment. Given the limitations of traditional IoU loss in cow behavior detection, we replace CIoU loss with Shape-IoU loss, focusing on the shape and scale features of the Bounding Box, thereby improving the matching degree between the Prediction Box and the Ground Truth Box. In order to verify the effectiveness of the proposed CAMLLA-YOLOv8n algorithm, we conducted experiments on a self-constructed dataset containing 23,073 Holstein cow behavior instances. The experimental results show that, compared with models such as YOLOv3-tiny, YOLOv5n, YOLOv5s, YOLOv7-tiny, YOLOv8n, and YOLOv8s, the improved CAMLLA-YOLOv8n model achieved increases in Precision of 8.79%, 7.16%, 6.06%, 2.86%, 2.18%, and 2.69%, respectively, when detecting the states of Holstein cows grazing, standing, lying, licking, estrus, fighting, and empty bedding. Finally, although the Params and FLOPs of the CAMLLA-YOLOv8n model increased slightly compared with the YOLOv8n model, it achieved significant improvements of 2.18%, 1.62%, 1.84%, and 1.77% in the four key performance indicators of Precision, Recall, mAP@0.5, and mAP@0.5:0.95, respectively. This model, named CAMLLA-YOLOv8n, effectively meets the need for the accurate and rapid identification of Holstein cow behavior in actual agricultural environments. This research is significant for improving the economic benefits of farms and promoting the transformation of animal husbandry towards digitalization and intelligence.