Comparative assessment of emulsifiers for in vitro ruminal gas production and fermentation measurements: Tween 80 is a suitable emulsifier.

Emulsifiers are essential for achieving a homogenous distribution of lipophilic supplements in in vitro rumen fluid incubations. Since emulsifiers can alter rumen fermentation, it is crucial to select one that minimally impacts fermentation parameters to reduce potential biases. This study aimed to evaluate seven emulsifiers' impact on in vitro ruminal fermentation using the Hohenheim Gas Test in order to identify the most inert emulsifier. Rumen fluids were collected from three non-lactating Original Brown-Swiss cannulated cows before morning feeding and incubated for 24 h with a basal diet in triplicates. The emulsifiers tested were ethanol, ethyl acetate, propylene glycol, glycerol, ethylene glycol, soy lecithin, and Tween® 80, each in two dosages (0.5% or 1% v/v). The untreated basal diet served as control. Compared to control, in vitro organic matter digestibility was enhanced by ethyl acetate (by 36.9 and 48.2%), ethylene glycol (by 20.6 and 20.1%), glycerol (by 46.9 and 56.8%) and soy lecithin (by 19.7 and 26.8%) at 0.5 and 1% dosage, respectively. Additionally, the 24-h methane production increased for ethanol (by 41.9 and 46.2%), ethylene glycol (by 50.5 and 51.5%), and glycerol (by 63.1 and 65.4%) for the 0.5 and 1% dosage, respectively, and 0.5% dosage for ethyl acetate (by 31.6%). The acetate molar proportion was 17.2%pt higher for ethyl acetate, and 25.5%pt lower for glycerol at 1% dosage, compared to the control. The propionate concentration was 22.1%pt higher 1% glycerol, and 15.2%pt and 15.1%pt higher for 0.5 and 1% propylene glycol, respectively, compared to the control. In summary, Tween® 80 did not significantly affect in vitro rumen fermentation parameters, making it the most suitable choice for in vitro incubations involving lipophilic substances in rumen fluid. Ethanol may be considered as an alternative emulsifier if methane production is not the variable of interest.

Effect of diets enriched in n-6 or n-3 fatty acids on dry matter intake, energy balance, oxidative stress, and milk fat profile of transition cows.

The objective of this study was to determine the effect of dietary supplementation of n-3 polyunsaturated fatty acids (PUFA) and n-6 PUFA on dry matter intake (DMI), energy balance, oxidative stress, and performance of transition cows. Forty-five multiparous Holstein dairy cows with similar parity, body weight (BW), body condition score (BCS), and milk yield were used in a completely randomized design during a 56-d experimental period including 28 d prepartum and 28 d postpartum. At 240 d of pregnancy, cows were randomly assigned to one of the 3 isoenergetic and isoprotein dietary treatments, including a control ration containing 1% hydrogenated fatty acid (CON), a ration with 8% extruded soybean (HN6, high n-6 PUFA source), and a ration with 3.5% extruded flaxseed (HN3; high n-3 PUFA source). The HN6 and HN3 diets had an n-6/n-3 ratio of 3.05:1 and 0.64:1 in prepartum cows and 8.16:1 and 1.59:1 in postpartum cows, respectively. During the prepartum period (3, 2, and 1 wk before calving), DMI, DMI per unit of BW, total net energy intake, and net energy balance were higher in the HN3 than in the CON and NH6 groups. During the postpartum period (2, 3, and 4 wk after calving), cows fed HN3 and HN6 diets both showed increasing DMI, DMI as a percentage of BW, and total net energy intake compared with those fed the CON diet. The BW of calves in the HN3 group was 12.91% higher than those in the CON group. Yield and nutrient composition of colostrum (first milking after calving) were not affected by HN6 or HN3 but milk yield from 1 to 4 wk of milking was significantly improved compared with CON. During the transition period, BW, BCS, and BCS changes were not affected. Cows fed the HN6 diet had a higher plasma NEFA concentration compared with the CON cows during the prepartum period. Feeding HN3 reduced the proportion of de novo fatty acids and increased the proportion of preformed long-chain fatty acids in regular milk. In addition, the n-3 PUFA-enriched diet reduced the n-6/n-3 PUFA ratio in milk. In conclusion, increasing the n-3 fatty acids concentration in the diet increased both DMI during the transition period and milk production after calving, and supplementing n-3 fatty acids was more effective in mitigating the net energy balance after calving.

Prediction of enteric methane production, yield, and intensity in dairy cattle using an intercontinental database.

Enteric methane (CH4 ) production from cattle contributes to global greenhouse gas emissions. Measurement of enteric CH4 is complex, expensive, and impractical at large scales; therefore, models are commonly used to predict CH4 production. However, building robust prediction models requires extensive data from animals under different management systems worldwide. The objectives of this study were to (1) collate a global database of enteric CH4 production from individual lactating dairy cattle; (2) determine the availability of key variables for predicting enteric CH4 production (g/day per cow), yield [g/kg dry matter intake (DMI)], and intensity (g/kg energy corrected milk) and their respective relationships; (3) develop intercontinental and regional models and cross-validate their performance; and (4) assess the trade-off between availability of on-farm inputs and CH4 prediction accuracy. The intercontinental database covered Europe (EU), the United States (US), and Australia (AU). A sequential approach was taken by incrementally adding key variables to develop models with increasing complexity. Methane emissions were predicted by fitting linear mixed models. Within model categories, an intercontinental model with the most available independent variables performed best with root mean square prediction error (RMSPE) as a percentage of mean observed value of 16.6%, 14.7%, and 19.8% for intercontinental, EU, and United States regions, respectively. Less complex models requiring only DMI had predictive ability comparable to complex models. Enteric CH4 production, yield, and intensity prediction models developed on an intercontinental basis had similar performance across regions, however, intercepts and slopes were different with implications for prediction. Revised CH4 emission conversion factors for specific regions are required to improve CH4 production estimates in national inventories. In conclusion, information on DMI is required for good prediction, and other factors such as dietary neutral detergent fiber (NDF) concentration, improve the prediction. For enteric CH4 yield and intensity prediction, information on milk yield and composition is required for better estimation.

Chiropractic effects on stride parameters and heart rate during exercise in sport horses.

BACKGROUND: Equine veterinarians performing chiropractic treatments are frequently asked to evaluate and treat sound horses to improve their performance and address pain associated with the axial skeleton. Studies describing the effects and mechanisms of chiropractic treatments in horses without overt lameness are scarce. OBJECTIVES: This study aimed to evaluate the effect of chiropractic treatments on stride rate, length, symmetry, heart rate and rider-perceived quality of the ridden work in sport horses. STUDY DESIGN: A blind randomised controlled trial with crossover design. METHODS: Thirty-eight horses ridden in the hunter-jumper discipline were enrolled. Exercise tests were recorded before and after chiropractic or sham treatment while horses were wearing a fitness tracker. Stride length, rate and symmetry, heart rate and the perceived quality of the ridden work were compared. RESULTS: There was a difference in the qualitative assessment of the ridden work by riders between treatments (odds ratio 33.8, 95% CI [4.68, 458.71], p < 0.01). Stride length, rate, symmetry and heart rate were not different between treatments. MAIN LIMITATIONS: The quantitative outcomes measured may not be sensitive enough to detect changes that improve the ridden work. Terrain, weather and rider were not standard across horses making small changes difficult to detect. CONCLUSIONS: Riders participating in a blind randomised controlled trial perceived a positive effect of chiropractic treatments on the quality of the ridden work. There were no differences in stride length, stride rate, stride symmetry or heart rate. The mechanisms, indications and potential benefits of chiropractic treatments in horses need further study.

HISTORIAL: A los veterinarios de equinos que realizan tratamientos quiroprácticos, se les pide frecuentemente evaluar y tratar caballos que están sanos para mejorar su desempeño y tratar el dolor asociado con el esqueleto axial. Estudios que describen los efectos y mecanismos de los tratamientos quiropráctico en caballos sin cojeras aparentes, son pocos. OBJETIVOS: Este estudio tiene por objetivo evaluar el efecto de los tratamientos quiroprácticos sobre frecuencia, largo y simetría de la zancada, la frecuencia cardiaca y la calidad del trabajo montado percibida por el jinete en caballos de deporte. DISEÑO DEL ESTUDIO: Prueba aleatoria cegada controlada con diseño cruzado. MÉTODOS: Se enrolaron 38 caballos montados en la disciplina de caza-salto. Pruebas de ejercicio fueron anotadas antes y después de tratamientos quiropráctico reales o simulados mientras los caballos llevaban un monitor físico. Se compararon el largo, frecuencia y simetría de la zancada, frecuencia cardiaca y calidad del trabajo montado percibida por el jinete. RESULTADOS: Se encontró una diferencia en la evaluación cualitativa del trabajo montado por los jinetes entre los tratamientos (odds ratio 33.8, 95% CI [4.68, 458.71], p < 0.01). Largo, frecuencia y simetría de zancada y frecuencia cardiacas no difirieron entre tratamientos. LIMITACIONES PRINCIPALES: Los resultados cuantitativos medidos, pueden no ser lo suficientemente sensibles para detectar cambios que mejoran el trabajo montado. El terreno, tiempo y jinete no fueron estandarizados a través de los caballos, lo que hizo que cambios pequeños fuesen difíciles de detectar. CONCLUSIONES: Los jinetes que participaron en una prueba aleatoria cegada controlada, percibieron un efecto positivo de los tratamientos quiroprácticos sobre la calidad del trabajo montado. No hubo diferencia en largo de zancada, frecuencia de zancada, simetría de zancada o frecuencia cardiaca. Los mecanismos, indicaciones y beneficios potenciales de los tratamientos quiroprácticos en caballos necesitan ser estudiados mas.

Physical pretreatment of three biowastes to improve black soldier fly larvae bioconversion efficiency.

Black soldier fly larvae (BSFL, Hermetia illucens (L.)) are recognized for efficient biowaste reduction while yielding valuable proteins and fats for animals. However, lignocellulosic fibers in biowastes are difficult to digest by biowaste and larval digestive tract microorganisms as well as the larvae themselves. This study investigated two biowaste physical pretreatments (thermal, mechanical) for improving BSFL processing of fibrous biowastes. Cow manure, spent grain, and grass clippings were thermally pretreated at 90 °C for three durations (0.5, 1 and 4 h). Contrary to expectations, thermal pretreatment resulted in either no improvement or decreased larval performance on all substrates, regardless of treatment duration. In contrast, mechanical pretreatment of spent grain and grass clippings, involving milling with three screen sizes (0.5, 1 and 2 mm) showed promising results. Specifically, bioconversion rates on 0.5 mm-milled spent grain and grass clippings increased by 0-53 % and 25-44 % dry mass, respectively compared to untreated. Additionally, larval protein conversion increased by 41 % and 23 % on spent grain and grass clippings, respectively. However, mechanical pretreatment did not affect fiber degradation by larval conversion, as hemicellulose decreased by 25 % and 75 % for spent grain and grass clippings, respectively, regardless of particle size. Particle size reduction influenced substrate microbial respiration (CO2 mg/min), with 0.5-mm milled grass clippings exhibiting higher respiration compared to untreated, although this effect was not observed for spent grain. This study highlights mechanical pretreatment's potential in enhancing BSFL bioconversion of fibrous biowastes and the importance of understanding substrate physical properties influencing substrate microorganisms and BSFL.

Preservation of agri-food byproducts by acidification and fermentation in black soldier fly larvae bioconversion.

Maintaining a consistent supply of feedstock for efficient bioconversion of black soldier fly larvae (BSFL) presents challenges due to the fluctuating availability of biowastes and agri-food products. To address the challenge of consistent feedstock supply for BSFL, this study investigated the influence of three preservation methods: wild fermentation, inoculated fermentation, and acidification on agri-food by-products applied over three storage durations (1, 7, and 14 days), evaluating their impact on BSFL bioconversion, and feedstock nutrient and microbiota composition. The preserved feedstocks were characterized for gross nutrient, sugar, fermentation metabolite, and bacterial community analyses. All feedstock preservation methods and storage durations had a high bioconversion rate (21-25 % dry mass) and wet larval mass (170-196 mg). Notably, 7-and-14-day acidified feedstock had a significantly higher bioconversion rate compared to fermented feedstock. Acidification preserved feedstock nutrients best with only a 10 % difference compared to initial nutrient values. Fermentation produced typical lactic acid fermentation metabolites with reducing sugar contents; however, adding a lactic acid bacterial inoculum (7 log10 CFU kg feedstock-1) had no benefit, presumably due to the high nutrient content and existing richness in lactic acid bacteria. Preservations had little influence on Enterobacteriaceae (6.2-7.5 log10 CFU g-1) in freshly harvested larvae. Future research should assess the acidification and fermentation of different BSFL feedstocks and investigate the roles of feedstock pH, organic acids, and fermentation metabolites in more detail. Therefore, this study advances toward reliable and efficient insect-based nutrient recovery from agri-food by-products within the food system.

LncRNA MIR181A2HG negatively regulates human keratinocytes proliferation by binding SRSF1.

Psoriasis is a common chronic inflammatory skin disease. Abnormal proliferation of keratinocytes plays an important role in the pathogenesis of psoriasis. Long non-coding RNAs (lncRNAs) are involved in the regulation of a variety of cell biological processes. The purpose of this study was to investigate the potential role of lncRNA MIR181A2HG in the proliferation of human keratinocytes. qRT-PCR and Western blotting were performed to measure the expression levels of MIR181A2HG, SRSF1, KRT6, and KRT16 in tissue specimens and HaCaT keratinocytes. The effects of MIR181A2HG on HaCaT keratinocytes proliferation were evaluated using Cell Counting Kit-8 (CCK-8) assays, 5-Ethynyl-2'-deoxyuridine (EdU) incorporation, and cell-cycle assays. RNA pulldown-mass spectrometry (MS) was applied to identify the proteins interacting with MIR181A2HG. RNA pull-down-Western blotting and RNA immunoprecipitation coupled with real-time quantitative reverse transcription-PCR (RIP-qRT-PCR) assays were used to determine the interactions between MIR181A2HG and its RNA-binding proteins (RBPs). MIR181A2HG was down-regulated in psoriasis tissues. MIR181A2HG overexpression induced G0/G1 and G2/M phase cell cycle arrest and decreased the protein levels of KRT6, KRT16, Cyclin D1, CDK4, and Cyclin A2 in HaCaT keratinocytes. MIR181A2HG knockdown showed the opposite effect. By using RNA pulldown-MS, 356 proteins were identified to interact with MIR181A2HG potentially. Bioinformatics analysis showed that NOP56 and SRSF1 may be RNA binding proteins (RBPs) that may be interact with MIR181A2HG. Furthermore, by using RNA pull-down-Western blotting and RIP-qRT-PCR, SRSF1 was determined to interact with MIR181A2HG. Moreover, silencing of SRSF1 inhibited keratinocytes proliferation, which could be reversed with the knockdown of MIR181A2HG. Our findings indicated that MIR181A2HG can negatively regulate HaCaT keratinocytes proliferation by binding SRSF1, suggesting that MIR181A2HG and SRSF1 may serve as potential targets for the treatment of psoriasis. Supplementary Information: The online version contains supplementary material available at 10.1007/s10616-024-00621-6.

LncRNA MIR181A2HG inhibits keratinocytes proliferation through miR-223-3p/SOX6 axis.

BACKGROUND: Psoriasis is a complex and recurrent chronic inflammatory skin disease, and the abnormal proliferation of keratinocytes plays a crucial role in the pathogenesis of psoriasis. Long non-coding RNAs (lncRNAs) play an indispensable role in regulating cellular functions. This research aims to explore the potential impact of lncRNA MIR181A2HG on the regulation of keratinocyte proliferation. METHODS: The expression level of MIR181A2HG and the mRNA level of KRT6, KRT16, and SOX6 were assessed using qRT-PCR. The viability and proliferation of keratinocytes were evaluated using CCK-8 and EdU assays. Cell cycle analysis was performed using flow cytometry. Dual-luciferase reporter assays were applied to test the interaction among MIR181A2HG/miR-223-3p/SOX6. Protein level was detected by Western blotting analysis. RESULTS: The findings indicated that psoriasis lesions tissue exhibited lower levels of MIR181A2HG expression compared to normal tissue. The overexpression of MIR181A2HG resulted in the inhibition of HaCaT keratinocytes proliferation. The knockdown of MIR181A2HG promoted cell proliferation. The dual-luciferase reporter assay and rescue experiments provided evidence of the interaction among MIR181A2HG, SOX6, and miR-223-3p. CONCLUSIONS: The lncRNA MIR181A2HG functions as a miR-223-3p sponge targeting SOX6 to regulate the proliferation of keratinocytes, which suggested that MIR181A2HG/miR-223-3p/SOX6 might be potential diagnostic and therapeutic targets for psoriasis.

miR-181a/b-5p negatively regulates keratinocytes proliferation by targeting MELK.

Accumulating evidence indicates that microRNAs (miRNAs) have a vital effect on the pathogenesis of psoriasis. This study is conducted to investigate the potential involvement of miR-181a-5p and miR-181b-5p in the proliferation of HaCaT keratinocytes. Cell viability and proliferation were evaluated respectively in this study using the CCK-8 and the 5-ethynyl-2'-deoxyuridine (EdU) assays. The expression of Maternal Embryonic Leucine Zipper Kinase (MELK) and Keratin 16 (KRT16) mRNA and protein in tissues and cells was assessed using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The Luciferase reporter system analyzes the connection between miR-181a-5p/miR-181b-5p and MELK. The results showed that miR-181a/b-5p expression was downregulated in the psoriasis lesions and negatively regulated the proliferation of keratinocytes. MELK was directly targeted by miR-181a-5p/miR-181b-5p. In addition, HaCaT keratinocytes proliferation was inhibited by knockdown of MELK while promoted dramatically by MELK overexpression. Notably, miR-181a/b-5p mimics could attenuate the effects of MELK in keratinocytes. In conclusion, our research findings suggested miR-181a-5p and miR-181b-5p negatively regulate keratinocyte proliferation by targeting MELK, providing potential diagnostic biomarkers and therapeutic targets for psoriasis.

Evaluation of ammonia pretreatment of four fibrous biowastes and its effect on black soldier fly larvae rearing performance.

Biowaste treatment with black soldier fly larvae (BSFL, Hermetia illucens L.) can promote a more sustainable food system by reusing nutrients that would otherwise be wasted. However, many agri-food wastes and byproducts are typically high in lignocellulosic fibers (i.e., cellulose, hemicellulose, and lignin), making it resistant to efficient larval and/or microbial degradation. Ammonia pretreatment could be used to partially degrade lignocellulose, making the biowaste more easily degradable by the larvae and/or microorganisms. This study evaluated ammonia pretreatment for lignocellulose degradation and its effect on BSFL performance on four fibrous biowastes: brewers spent grain, cow manure, oat pulp, and grass clippings. First, the optimal ammonia dose (1 % or 5 % dry mass) and pretreatment time (three or seven days) were assessed by measuring fibers after treatment and further examined using Fourier transform infrared spectroscopy (FTIR) spectra and scanning electron microscopy (SEM) images. Second, BSFL rearing performance on ammonia-pretreated substrates was assessed with a 9-day feeding experiment. Three-day pretreatment with 5 % ammonia was chosen as it decreased the total fiber content by 8-23 % for all substrates except cow manure. Contrary to expectations, ammonia pretreatment with all substates decreased BSFL rearing performance metrics by more than half compared to the untreated control. Follow-up experiments suggested that ammonia pretreatment had a dose-dependent toxicity to BSFL. Interestingly, three-day fermentation of cow manure and oat pulp increased bioconversion rate by 25-31 %. This study shows that ammonia pretreatment is not suitable before BSFL rearing. Ammonia toxicity to BSFL and other pretreatments, such as fermentation, should be further studied.

Quantification of methane emitted by ruminants: a review of methods.

The contribution of greenhouse gas (GHG) emissions from ruminant production systems varies between countries and between regions within individual countries. The appropriate quantification of GHG emissions, specifically methane (CH4), has raised questions about the correct reporting of GHG inventories and, perhaps more importantly, how best to mitigate CH4 emissions. This review documents existing methods and methodologies to measure and estimate CH4 emissions from ruminant animals and the manure produced therein over various scales and conditions. Measurements of CH4 have frequently been conducted in research settings using classical methodologies developed for bioenergetic purposes, such as gas exchange techniques (respiration chambers, headboxes). While very precise, these techniques are limited to research settings as they are expensive, labor-intensive, and applicable only to a few animals. Head-stalls, such as the GreenFeed system, have been used to measure expired CH4 for individual animals housed alone or in groups in confinement or grazing. This technique requires frequent animal visitation over the diurnal measurement period and an adequate number of collection days. The tracer gas technique can be used to measure CH4 from individual animals housed outdoors, as there is a need to ensure low background concentrations. Micrometeorological techniques (e.g., open-path lasers) can measure CH4 emissions over larger areas and many animals, but limitations exist, including the need to measure over more extended periods. Measurement of CH4 emissions from manure depends on the type of storage, animal housing, CH4 concentration inside and outside the boundaries of the area of interest, and ventilation rate, which is likely the variable that contributes the greatest to measurement uncertainty. For large-scale areas, aircraft, drones, and satellites have been used in association with the tracer flux method, inverse modeling, imagery, and LiDAR (Light Detection and Ranging), but research is lagging in validating these methods. Bottom-up approaches to estimating CH4 emissions rely on empirical or mechanistic modeling to quantify the contribution of individual sources (enteric and manure). In contrast, top-down approaches estimate the amount of CH4 in the atmosphere using spatial and temporal models to account for transportation from an emitter to an observation point. While these two estimation approaches rarely agree, they help identify knowledge gaps and research requirements in practice.

There is a need to accurately and precisely quantify greenhouse gas (GHG) emissions, specifically methane (CH4), to ensure correct reporting of GHG inventories and, perhaps more importantly, determine how to best mitigate CH4 emissions. The objective of this study was to review existing methods and methodologies to quantify and estimate CH4 emissions from ruminants. Historically, most techniques were developed for specific purposes that may limit their widespread use on commercial farms and for inventory purposes and typically required frequent calibration and equipment maintenance. Whole animal and head respiration chambers, spot sampling techniques, and tracer gas methods can be used to measure enteric CH4 from individual animals, but each technique has its own inherent limitations. The measurement of CH4 emissions from manure depends on the type of storage, animal housing, CH4 concentration inside and outside the boundaries of the area of interest, and ventilation rate, which is likely the most complex variable creating many uncertainties. For large-scale areas, aircraft, drones, and satellites have been used in association with the tracer flux method, inverse modeling, imagery, and LiDAR (Light Detection and Ranging), but research is lagging in validating these methods. Bottom-up approaches to estimating CH4 emissions rely on empirical or mechanistic modeling to quantify the contribution of individual sources. Top-down approaches estimate the amount of CH4 in the atmosphere using spatial and temporal models to account for transportation from an emitter to an observation point.