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Cows with ketosis display severe metabolic stress and immune dysfunction which renders them more susceptible to infections. Monocytes, one of the major subtypes of white blood cells, play an important role in innate immune defense against infections. Thus, the aim of this study was to investigate alterations in immune function, reactive oxygen species (ROS) production and activity of the NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway in monocytes (CD14+) of cows with clinical ketosis (CK). Twelve healthy multiparous Holstein cows [blood β-hydroxybutyrate (BHB) concentration < 1.2 mM] and 12 cows with CK (BHB > 3.0 mM) at 3 to 14 days in milk were used for blood sample collection. To determine effects of BHB on phagocytosis, ROS and protein abundance of the NLRP3 inflammasome pathway in vitro, monocytes isolated from healthy cows were treated with 3.0 mM BHB for 0, 6, 12 or 24 h. Dry matter intake (22.7 vs. 19.0 kg) was lower in cows with CK. Serum concentrations of fatty acids (0.30 vs. 0.88 mM) and BHB (0.52 vs. 3.78 mM) were greater in cows with CK, whereas concentration of glucose was lower (4.09 vs. 2.23 mM). The adhesion, migration and phagocytosis of monocytes were lower in cows with CK, but apoptosis and ROS content were greater. Protein abundance of NLRP3, cysteinyl aspartate specific proteinase 1 (caspase 1) and interleukin-1B p17 (IL1B p17) were greater in monocytes of cows with CK, while abundance of NADPH oxidase isoform 2 (NOX2) was lower. Compared with 0 h BHB, ROS content and apoptosis were greater in the monocytes challenged for 6, 12 or 24 h BHB. Compared with 0 h BHB, protein abundance of NLRP3, caspase 1, IL1B p17 and concentration of IL1B in medium were greater in the monocytes challenged for 6, 12 or 24 h BHB. However, compared with 0 h BHB, protein abundance of NOX2 and phagocytosis of monocytes were lower in the monocytes challenged for 6, 12 or 24 h BHB. Overall, the data suggested that exogenous BHB activated the ROS-NLRP3 pathway, which might be partly responsible for immune dysfunction of dairy cows with CK.

Most arthropod guts harbor diverse microbiota for symbiotic digestion. The European corn borer (ECB), Ostrinia nubilalis (Hübner), is a devastating pest that feeds the lignocellulose-rich tissues of maize plants. However, the potential role of ECB gut microbes in degrading maize cellulose remains largely unexplored. Here, we investigated the gut microbiota of ECB fed with different diets and their potential function in maize lignocellulose degradation. The diversity and composition of gut bacterial communities varied dramatically between the ECB larva fed with artificial diets (ECB-D) and maize plants (ECB-M). Draft genomes of the microbial consortia from ECB-D and ECB-M showed that the principal degraders of cellulose mainly belonged to Firmicutes or Proteobacteria and they were primarily found in the midgut. The cellulolytic microbial consortia contained genes encoding various carbohydrate-active enzymes (CAZyme). Furthermore, scanning electron microscopy revealed significant breakdown of lignocellulose in maize treated by the two microbial consortia for 9 days in vitro. Metabolomic analyses show that maize particles treated by two microbial consortia generate distinctive metabolomic profiles, with enrichment for different monosaccharides (i.e., Glucose, Rhamnofuranose, Isomaltose, and Cellobiose) and amino acids (i.e., Threonine, Histidine, and Lysine). The results indicated that the diet of the host impacted the composition and function of its gut microbiota and ECB exploited specific gut microbes to digest maize lignocellulose with distinctive products. Our study provides valuable microbiota resources for lignocellulose bioconversion.

This study aimed to investigate the effect of epiphytic microbiota from alfalfa and red clover on the fermentative products, bacterial community compositions, and their predicted functional characteristics in Italian ryegrass silage. By microbiota transplantation and γ-ray irradiation sterilization, the irradiated Italian ryegrass was treated as follows: (1) sterile distilled water (STIR); (2) epiphytic microbiota on Italian ryegrass (IRIR); (3) epiphytic microbiota on alfalfa (IRAL); and (4) epiphytic microbiota on red clover (IRRC). The irradiated Italian ryegrass was ensiled for 1, 3, 7, 15, 30, and 60 days. STIR had similar chemical components with fresh Italian ryegrass. IRAL had higher lactic acid concentrations [64.0 g/kg dry matter (DM)] than IRIR (22.3 g/kg DM) and IRRC (49.4 g/kg DM) on day 3. IRRC had the lowest lactic acid concentrations (59.7 g/kg DM) and the highest pH (4.64), acetic acid (60.4 g/kg DM), ethanol (20.4 g/kg DM), and ammonia nitrogen (82.6 g/kg DM) concentrations and Enterobacteriaceae [9.51 log10 cfu/g fresh weight (FW)] populations among treatments on day 60. On days 3 and 60, Lactobacillus was dominant in both IRIR (42.2%; 72.7%) and IRAL (29.7%; 91.6%), while Hafnia-Obesumbacterium was predominant in IRRC (85.2%; 48.9%). IRIR and IRAL had lower abundances of “Membrane transport” than IRRC on day 3. IRIR and IRAL had lower abundances of phosphotransacetylase and putative ATP-binding cassette transporter and higher abundances of arginine deiminase on day 3. IRAL had the highest abundance of fructokinase on day 3. Overall, inoculating epiphytic microbiota from different legume forages changed the fermentative products, bacterial community compositions, and their predicted functional characteristics in Italian ryegrass silage. The microbial factors that result in the differences in fermentative profiles between legume forage and grass were revealed. Knowledge regarding the effect of epiphytic microbiota could provide more insights into the improvement of silage quality.

Chlorine (Cl) is indispensable for the growth of plants. While rarely systematic reports are available for the effect of Cl-containing fertilizers on citrus production. This study aimed to investigate the impacts of various Cl-containing fertilizers on the nutrients in the leaves, the yield and quality of sweet orange, and the Cl migration in the plant–soil–leaching system. A 5-year field experiment (2016–2020) with five Cl treatments (0, 75, 150, 450, and 900 kg ha−1), and soil core lysimeter test with five Cl levels (0, 150, 225, 300, and 450 kg ha−1) were carried out. The results showed that 77.0% of Cl leached into above 60 cm deeper soil layer, with calcium as the main accompanying ions, resulting in less Cl being absorbed by the citrus plants. The content of Cl in the leaves and soil was enhanced by the increasing input of Cl-containing fertilizer, without yearly increased characteristics, under a mean annual rainfall of 1,474 mm. Chlorine significantly increased the yield (13.24–37.8 9%), fruit weight, and vitamin C (Vc), in addition to enhancing the flavor and the juice yield of sweet orange via improving the absorption of N and K. Moreover, the long-term application of potassium sulfate has elevated the accumulation of sulfur in the soil and in leaves; it is becoming a potential risk factor for citrus production. Taken together, the application of Cl-containing fertilizer in sweet orange is feasible, and trace absorbance of Cl could improve the yield and fruit quality of sweet orange.

Fatty liver disease is one of the most common disorders afflicting dairy cows during the postpartum period, and is associated with increased blood non-esterified fatty acid (NEFA) uptake by the liver. Major long-chain fatty acids (LCFA) in NEFA are palmitic (PA), palmitoleic (POA), stearic (SA), oleic (OA), and linoleic (LA) acid. In order to investigate the characteristics of lipid accumulation and injury caused by these NEFA, primary calf hepatocytes were isolated and challenged for 12 h with 1.2 mmol/L PA, POA, SA, OA, LA, or a mixture of these LCFA (NEFA). Compared with POA, OA, and LA, culture with PA and SA led to greater abundance of CCAAT-enhancer binding protein, glucose-regulated protein 78 mRNA, and stearoyl-CoA desaturase 1 mRNA along with greater concentrations of H2O2, malondialdehyde and reactive oxygen species (ROS). Although culture with POA, OA, and LA led to lower very low density lipoprotein (VLDL) concentration in cell culture medium, POA and OA led to greater concentrations of triacylglycerol, protein abundance of sterol regulatory element-binding protein 1c, fatty acid synthase, acetyl coenzyme A carboxylase 1, ApoB100, and sortilin 1 (SORT1). Compared with individual fatty acids, culture with NEFA led to an intermediate degree of lipid accumulation and hepatocytes damage. Overall, the data suggest that saturated fatty acids cause more severe oxidative and ER stress. However, unsaturated fatty acids cause serious lipid accumulation. Furthermore, a fatty acid balanced nutrient regulation was suggested useful improve liver health of transition period dairy cows.

Freezing damages in forages represents a major economic loss to agriculture. This study was conducted to investigate the effects of freeze–thaw (FT) event on microbial community dynamics of red clover silage. Results showed that the FT-treated material displayed higher proportions of Weissella and aerobic bacteria, while lower Pantoea and Enterobacter compared with the control material. The FT event promoted the development of Lactobacillus in silage microflora, inducing more intense lactic fermentation after an initial short lag. The aerobic bacteria were suppressed immediately after the onset of ensiling. Microbiomes of the two silages tended to be almost similar after 2 days of ensiling. However, a small number of aerobic bacteria tended to revitalize in the FT silage with prolonged ensiling time, indicated by apparent abundances of Acinetobacter and Pseudomonas at the end of ensiling. The results obtained here suggest that the FT event could promote the development of Lactobacillus during ensiling and the control of aerobe revitalization need to be concerned with silages made from the freeze-damaged forages.