Understanding how different livestock species and breeds respond to consumption of brackish water could improve usage of this resource. Therefore, Angora, Boer, and Spanish goat doelings and Dorper, Katahdin, and St. Croix ewe lambs (6 animals per animal type [AT]; initial age = 296 ± 2.1 days) consuming water with varying concentrations of minerals of a natural brackish water source (BR) and sodium chloride (NaCl; SL) were used to determine effects on water and feed intake, nutrient digestion, heat energy, methane emission, ruminal fluid conditions, and blood constituent concentrations. There were 6 simultaneous 6 (water treatments [WT]) × 6 (AT) Latin squares with 3-wk periods. The WT were fresh (FR), BR alone (100-BR), a similar total dissolved solids (TDS) concentration as 100-BR via NaCl addition to FR (100-SL), BR with concentrations of all minerals increased by approximately 50% (150-BR), a similar TDS level as 150-BR by NaCl addition to FR (150-SL), and a similar 150 TDS level achieved by addition of a 1:1 mixture of BR minerals and NaCl to 100-BR (150-BR/SL). Concentrations (mg/kg) in BR were 4928 TDS, 85.9 bicarbonate, 224.9 calcium, 1175 chloride, 60.5 magnesium, 4.59 potassium, 1387 sodium, 1962 sulfate, and 8.3 boron, and TDS in other WT were 209, 5684, 7508, 8309, and 7319 mg/kg for FR, 100-SL, 150-BR, 150-SL, and 150-BR/SL, respectively. There were very few significant effects of WT or AT × WT interactions, although AT had numerous effects. Water intake was affected by AT (P = 0.02) and WT (P = 0.04), with greater water intake for 150-SL than for FR, 100-BR, 100-SL, and 150-BR. Dry matter intake among AT was lowest (P < 0.05) for Angora. Digestion of organic matter and neutral detergent fiber and heat energy differed among AT (P < 0.05), but nitrogen digestion and ruminal methane emission were similar among AT. Blood aldosterone concentration was higher (P < 0.05) for FR than for other WT. In conclusion, all AT seemed resilient to these WT regardless of mineral source and concentrations, with TDS less than 8300 mg/kg, which did not influence nutrient utilization, ruminal fermentation, energy balance, or blood constituent levels.

Glucose plays a vital part in milk protein synthesis through the mTOR signaling pathway in bovine mammary epithelial cells (BMEC). The objectives of this study were to determine how glucose affects hexokinase (HK) activity in BMEC and investigate the regulatory effect of HK in kappa casein (CSN3) synthesis via the mechanistic target of rapamycin complex 1 (mTORC1) signaling pathway in BMEC. For this, HK1 and HK2 were knocked out in BMEC using the CRISPR/Cas9 system. The gene and protein expression, glucose uptake, and cell proliferation were measured. We found that glucose uptake, cell proliferation, CSN3 gene expression levels, and expression of HK1 and HK2 increased with increasing glucose concentrations. Notably, glucose uptake was significantly reduced in HK2 knockout (HK2KO) BMEC treated with 17.5 mM glucose. Moreover, under the same glucose treatment conditions, the proliferative ability and abundance of CSN3 were significantly diminished in both HK1 knockout (HK1KO) and HK2KO BMEC compared with that in wild-type BEMC. We further observed that the phosphorylation levels of ribosome protein subunit 6 kinase 1 (S6K1) were reduced in HK1KO and HK2KO BMEC following treatment with 17.5 mM glucose. As expected, the levels of glucose-6-phosphate and the mRNA expression levels of glycolysis-related genes were decreased in both HK1KO and HK2KO BMEC following glucose treatment. These results indicated that the knockout of HK1 and HK2 inhibited cell proliferation and CSN3 expression in BMEC under glucose treatment, which may be associated with the inactivation of the S6K1 and inhibition of glycolysis.

Bacterial pathogens destroy the structural integrity of functional organs in fish, leading to severe challenges in the aquaculture industry. Vitamin D₃ (VD₃) prevents bacterial infections and strengthens immune system function via vitamin D receptor (VDR). However, the correlation between VD₃/VDR and the structural integrity of functional organs remains unclarified. This study aimed to investigate the influence of VD₃ supplementation on histological characteristics, apoptosis, and tight junction characteristics in fish intestine during pathogen infection. A total of 540 healthy grass carp (257.24 ± 0.63 g) were fed different levels of VD₃ (15.2, 364.3, 782.5, 1,167.9, 1,573.8, and 1,980.1 IU/kg) for 70 d. Subsequently, fish were challenged with Aeromonas hydrophila, a pathogen that causes intestinal inflammation. Our present study demonstrated that optimal supplementation with VD₃ (1) alleviated intestinal structural damage, and inhibited oxidative damage by reducing levels of oxidative stress biomarkers; (2) attenuated excessive apoptosis-related death receptor and mitochondrial pathway processes in relation to p38 mitogen-activated protein kinase signaling (P < 0.05); (3) enhanced tight junction protein expression by inhibiting myosin light chain kinase signaling (P < 0.05); and (4) elevated VDR isoform expression in fish intestine (P < 0.05). Overall, the results demonstrated that VD₃ alleviates oxidative injury, apoptosis, and the destruction of tight junction protein under pathogenic infection, thereby strengthening pathogen defenses in the intestine. This finding supports the rationale for VD₃ intervention as an essential practice in sustainable aquaculture.

An experiment was conducted to determine the standardized ileal digestible (SID) calcium requirement of fast-growing broilers from hatch to d 14 post-hatch. Ross 308 (n = 360) male broilers were obtained on day of hatch and allocated to 1 of 5 treatments in battery cages. There were 6 birds per cage and 12 pens per treatment. Four treatments were formulated to contain 0.60%, 0.46%, 0.32% or 0.18% SID Ca. The final treatment was formulated using total Ca to meet or exceed all nutrient requirements, including 0.90% total Ca and 0.49% non-phytate P (nPP), using the same ingredients. This treatment was the reference diet for comparison and validation of the SID Ca diets. Birds and feed were weighed at placement and on d 14. Tibias and ileal contents were obtained on d 14 and excreta was collected per pen and pooled on d 14. Data were analyzed using JMP Pro and requirements were estimated using 3 different non-linear regression models. Increasing the SID Ca content in the diet from 0.18% to 0.60% improved (quadratic, P < 0.05) body weight gain and mortality corrected feed conversion ratio (mFCR). The estimated SID Ca requirement to optimize gain or mFCR was between 0.39% and 0.52%. Tibia ash percent and weight increased (quadratic, P < 0.05) as SID Ca content in the diet increased and the estimated SID Ca requirement was between 0.32% and 0.58%. Phosphorus utilization was improved in birds fed diets formulated using SID Ca compared with birds fed the reference diet. In conclusion, the SID Ca requirement of fast-growing broilers from hatch to d 14 was estimated between 0.534% and 0.398% when quadratic, straight-broken line, or quadratic-broken line regressions were used. These results agree with previously published data evaluating the SID Ca requirement of fast-growth broilers from hatch to d 10.

This study was conducted to explore the regulatory mechanism of leucine (Leu) on lipid metabolism of finishing pigs. Twenty-four Duroc × Landrace × Large cross pigs with an average body weight of 68.33 ± 0.97 kg were randomly allocated into 3 treatment groups with 8 replicates per group (1 pig per replicate). The dietary treatments were as follows: control group (CON), 0.25% Leu group and 0.50% Leu group. The experimental period was 42 d. The results showed as follows. (1) Compared with the CON, 0.25% and 0.50% Leu increased (P < 0.01) the average daily gain (ADG), while the average backfat thickness (ABT) and the ratio of feed intake to body weight gain (F:G ratio) were decreased (P < 0.05). (2) In the 0.25% Leu group, the relative mRNA expression levels of sterol regulatory element binding protein-1c (SREBP1c), recombinant fatty acid transport protein 1 (FATP1), chemerin and peroxisome proliferator-activated receptor γ (PPARγ) were decreased but the level of fatty acid binding protein 4 (FABP4) and fatty acid translocase (FAT/CD36) were increased in backfat tissue. In the 0.25% Leu group, the protein levels of p-Rictor, p-Raptor, p-eIF4E-binding protein 1 (p-4EBP1), p-silent mating type information regulator 2 homolog 1 (p-SIRT1) and acetylation ribosome s6 protein kinase 1 (Ac–S6K1) were increased (P < 0.05). (3) Compared to the CON, the diversity of gut microbiota in the 0.25% Leu group was increased. Principal component analysis showed that the relative abundance of Bacteroidetes, Lactobacillus and Desulfovibrio was higher in the 0.25% Leu group than the CON, but the relative abundance of Firmicutes, Treponema and Shigella was lower than in the CON (P < 0.05). (4) Four different metabolites were screened out from the serum of finishing pigs including allolithocholic acid (alloLCA), isolithocholic acid (isoLCA), ursodeoxycholic acid (UDCA) and hyodeoxycholic acid (HDCA), which correlate to various degrees with the above microorganisms. In conclusion, Leu could promote adipose tissue lipolysis of finishing pigs through the mTOR-SIRT1 signaling pathway, and S6K1 is acetylated at the same time, and the interaction between gut microbiota and bile acid metabolism is also involved.

Net energy (NE) enables the prediction of more accurate feed energy values by taking into account the heat increment which is approximately 25% of apparent metabolizable energy (AME) in poultry. Nevertheless, application of NE in poultry industry has not been practiced widely. To predict the NE values of broiler diets, 23 diets were prepared by using 13 major ingredients (wheat, corn, paddy rice, broken rice, cassava pellets, full-fat soybean, soybean meal, canola meal, animal protein, rice bran, wheat bran, palm kernel meal and palm kernel oil). The diets were formulated in order to meet the birds' requirements and get a wide range of chemical compositions (on DM basis; 33.6% to 55.3% for starch; 20.8% to 28.4% for CP, 2.7% to 10.6% for ether extract [EE] and 7.0% to 17.2% for NDF), with low correlations between these nutrients and low correlations between the inclusion levels of ingredients allowing for the calculation of robust prediction equations of energy values of diets or ingredients. These diets were fed to Ross 308 broilers raised in 12 open-circuit respiratory chambers from 18 to 23 d of age (4 birds per cage) and growth performance, diet AME content and heat production were measured, and dietary NE values were calculated. The trial was conducted on a weekly basis with 12 diets measured each week (1 per chamber), 1 of the 23 diets (reference diet) being measured each week. Each diet was tested at least 8 times. In total, 235 energy balance data values were available for the final calculations. Growth performance, AME (15.3 MJ/kg DM on average) and AME/GE (79.4% on average) values were as expected. The NE/AME value averaged 76.6% and was negatively influenced by CP and NDF and positively by EE in connection with efficiencies of AME provided by CP, EE and starch for NE of 73%, 87% and 81%, respectively. The best prediction equation was: NE = (0.815 × AME) — (0.026 × CP) + (0.020 × EE) — (0.024 × NDF) with NE and AME as MJ/kg DM, and CP, EE and NDF as % of DM. The NE prediction equations from this study agree with other recently reported equations in poultry and are suitable for both ingredients and complete feeds.

Low-fishmeal and protein-saving diets are two prominent nutritional strategies utilized to address challenges related to the scarcity and sustainability of protein sources in aquaculture. However, these diets have been associated with adverse effects on the growth performance, feed utilization, and disease resistance of aquatic animals. To mitigate these challenges, exogenous protease has been applied to enhance the quality of diets with lower protein contents or fishmeal alternatives, thereby improving the bioavailability of nutritional ingredients. Additionally, protease preparations were also used to enzymatically hydrolyze fishmeal alternatives, thus enhancing their nutritional utilization. The present review aims to consolidate recent research progress on the use of protease in aquaculture and conclude the benefits and limitations of its application, thereby providing a comprehensive understanding of the subject and identifying opportunities for future research.

The global trend towards raising broiler chickens without the use of in-feed antibiotics (IFAs) means that there is an ongoing need to develop alternative treatments capable of delivering the benefits that IFAs previously provided. IFAs supported the productivity performance of chickens and played a key role in maintaining their health. Necrotic enteritis (NE) is an important disease of broilers that affects health, productivity, and welfare, and was previously well controlled by IFAs. However, with the reduction in IFA use, NE is resurgent in some countries. Vaccines and various feed additives, including pre-, pro-, and postbiotics, phytobiotics, fatty acids, and phage therapies have been introduced as alternative methods of NE control. While some of these feed additives have specific activity against the NE pathogen, Clostridium perfringens, most have the more general goal of reinforcing gut health. Extensive reviews of the effects of many of these feed additives on gut health have been published recently. Hence, rather than cover previously well reviewed areas of research this review focuses on the challenges and pitfalls in undertaking experimental assessment of alternative NE treatments and translating laboratory research to real world commercial production settings. The review is based on the author's particular experience, reading, thoughts, and analysis of the available information and inevitably presents a particular understanding that is likely to be at odds with others thinking on these issues. It is put forward to stimulate thinking and discussion on the issues covered.

Fat is one of the three macronutrients and a significant energy source for piglets. It plays a positive role in maintaining intestinal health and improving production performance. During the weaning period, physiological, stress and diet-related factors influence the absorption of fat in piglets, leading to damage to the intestinal barrier, diarrhea and even death. Signaling pathways, such as fatty acid translocase (CD36), pregnane X receptor (PXR), and AMP-dependent protein kinase (AMPK), are responsible for regulating intestinal fat uptake and maintaining intestinal barrier function. Therefore, this review mainly elaborates on the reasons for diarrhea induced by insufficient fat absorption and related signaling pathways in weaned-piglets, with an emphasis on the intestinal fat absorption disorder. Moreover, we focus on introducing nutritional strategies that can promote intestinal fat absorption in piglets with insufficient fat absorption-related diarrhea, such as lipase, amino acids, and probiotics.

Vitamin E (VE) is an essential lipid-soluble vitamin that improves the fish flesh quality. However, the underlying molecular mechanisms remain unclear. This study aimed to investigate the effects of VE on growth performance and flesh quality in sub-adult grass carp (Ctenopharyngodon idella). A total of 450 fish (713.53 ± 1.50 g) were randomly divided into six treatment groups (three replicates per treatment) and fed for nine weeks with different experimental diets (dietary lipid 47.8 g/kg) that contained different levels of VE (5.44, 52.07, 96.85, 141.71, 185.66, and 230.12 mg/kg diet, supplemented as dl-α-tocopherol acetate). Notably, the treatment groups that were fed with dietary VE ranging from 52.07 to 230.12 mg/kg diet showed improvement in the percent weight gain, special growth rate, and feed efficiency of grass carp. Moreover, the treatment groups supplemented with dietary VE level of 141.71, 185.66, and 230.12 mg/kg diet showed enhancement in crude protein, lipid, and α-tocopherol contents in the muscle, and the dietary levels of VE ranging from 52.07 to 141.71 mg/kg diet improved muscle pH_24h and shear force but reduced muscle cooking loss in grass carp. Furthermore, appropriate levels of VE (52.07 to 96.85 mg/kg diet) increased the muscle polyunsaturated fatty acid content in grass carp. Dietary VE also increased the mRNA levels of fatty acid synthesis-related genes, including fas, scd-1, fad, elovl, srebp1, pparγ, and lxrα, and up-regulated the expression of SREBP-1 protein. However, dietary VE decreased the expression of fatty acid decomposition-related genes, including hsl, cpt1, acox1, and pparα, and endoplasmic reticulum stress-related genes, including perk, ire1, atf6, eif2α, atf4, xbp1, chop, and grp78, and down-regulated the expression of p-PERK, p-IRE1, ATF6, and GRP78 proteins. In conclusion, dietary VE increased muscle fatty acid synthesis, which may be partly associated with the alleviation of endoplasmic reticulum stress, and ultimately improves fish flesh quality. Moreover, the VE requirements for sub-adult grass carp (713.53 to 1590.40 g) were estimated to be 124.9 and 122.73 mg/kg diet based on percentage weight gain and muscle shear force, respectively.