The first objective of this study was to investigate the effects of subacute ruminal acidosis (SARA) on fermentation, ruminal free lipopolysaccharides (LPS), and expression of the cluster of differentiation 14 (CD14), toll-like receptor 4 (TLR4), and myeloid differentiation protein 2 (MD2) complex in white blood cells involved in the systemic immune response in dairy cows. The second objective was a study of whether increased expression of the LPS receptor complex led to increases in the concentrations of plasma high-density lipoprotein (HDL) and serum Ca. Three hundred five dairy cows located in 13 Polish high-yielding dairy commercial farms were selected according to their days in milk (40-150 d; average = 75), 305-d milk yield (10,070-12,041 kg; average = 10,940), and number of lactations (primiparous, n = 139 and multiparous, n = 166). Next, the herds were segregated into 3 groups based on the percentages of cows with an assigned value of ruminal fluid pH: SARA-positive, SARA-risk, and SARA-negative herds. Moreover, 305 selected dairy cows were divided according to the classification based on ruminal fluid pH into 3 groups as healthy (pH >5.81), risk (pH 5.8-5.6) and acidotic cows (pH <5.6). Rumen fluid samples were collected via rumenocentesis. In the AC group, we recorded higher concentrations of ruminal free LPS [4.57 Log endotoxin units (EU)/mL; 42,206 EU/mL] compared with the healthy group (4.48 Log EU/mL; 34,179 EU/mL). Similarly, the concentration of ruminal free LPS was higher in SARA-positive herds (4.60 Log EU/mL; 43,000 EU/mL) compared with SARA-negative herds (4.47 Log EU/mL; 32,225 EU/mL). The relative mRNA abundance of genes associated with the function of LPS receptors, such as CD14, TLR4, and MD2, in white blood cells differed between all experimental groups on both cow and herd levels. In the acidotic group, we recorded higher concentrations of HDL (78.16 vs. 68.32 mg/dL) and serum amyloid A (10.80 vs. 9.16 µg/mL) and lower concentrations of Ca (8.26 vs. 10.16 mg/dL) and haptoglobin (470.19 vs. 516.85 ng/mL) compared with the healthy group. Similar results were obtained in the SARA herd status analysis, but the concentration of lipopolysaccharide-binding protein differed statistically. Moreover, the pH of ruminal fluid was negatively correlated with relative mRNA abundance of genes such as CD14, TLR4, MD2, and concentrations of serum HDL and serum amyloid A, although positively correlated with serum Ca. The results indicated that decreases in ruminal fluid pH increased the release of free LPS into the rumen and stimulated the expression of the LPS receptor complex and immune response. Moreover, an increase in the expression of the LPS receptor led to higher concentrations of plasma HDL and lower serum Ca, which may be a protective mechanism against endotoxemia. However, the biological significance of these results needs to be investigated further in larger field trials.

The aim of this study was to estimate the prevalence of subacute ruminal acidosis (SARA) in Polish high-yielding dairy herds. Also, the relationships between the chemical composition of the diet and the feed particle size, ruminal pH and the occurrence of this metabolic disease and the fermentation profile were determined. Rumen fluid samples were obtained from a total of 213 cows from nine dairy herds (≥20 cows per herd) via rumenocentesis. Almost 14% (30/213) of cows as acidotic (pH<5.6) were found, which is indicative of SARA. Moreover, 44% of the herds were classified as SARA-positive and 56% as SARA-negative. Results of the current study suggested that the physically effective NDF (NDF -to-starch ratio could be better indices than peNDF to preventing the occurrence of SARA, and their level should not be lower than 1.00. Also, linear negative relationships between rumen fluid pH and concentration of propionate, valerate and total VFA were determined. According to the herd's SARA status and rumen fluid biochemical indices, there were significant differences between the pH of rumen fluid (p ≤ 0.01), concentrations of acetate (p ≤ 0.05), propionate (p ≤ 0.05), n-butyrate (p ≤ 0.01), n-valerate (p ≤ 0.01), the sum of VFA (p ≤ 0.01) and ammonia (p ≤ 0.05) in SARA-positive herds compared to SARA-negative herds. The better understanding the strategy of ruminant nutrition to coordinate energy conversion and the role of the ruminal pH in regulating N-NH production, absorption through rumen mucosa, urea secretion, the more successfully we can utilize these processes with due recognition of animal needs and welfare, as well as prevention of SARA occurrence.

Nine Holstein dairy cows were fed diets with increasing proportions of rapidly fermentable carbohydrates (RFCH) to investigate the effect on reticular pH, milk fat content (MFC), 18-carbon fatty acid proportions in blood plasma and milk, and bacterial community in buccal swab samples. Inter-animal variation was expected in terms of reticular pH response upon higher RFCH proportions, which would be reflected in the occurrence or not of milk fat depression (MFD). Moreover, this variation in occurrence of MFD was hypothesized to be related to differences in blood and milk fatty acid proportions and in the bacterial community in buccal samples. Cows were fed a total mixed ration throughout the experiment, which consisted of 4 periods: adaptation (d 0-4) and low (d 5-18), increasing (d 19-24), and high . During the increasing RFCH period, the standard concentrate (211 g of starch/kg of dry matter) was gradually and partly replaced by a concentrate high in RFCH (486 g of starch/kg of dry matter). The reticular pH was measured using a bolus and the time below pH 6.00 was calculated on a daily basis. On d 13, 14, 25, 27, and 28, plasma and milk samples were collected and analyzed for 18-carbon fatty acid proportions, and buccal swabs were collected for bacterial community analysis based on 16S rRNA gene amplicon sequencing. Inter-animal variation was observed in terms of reticular pH, which allowed us to divide the cows into 2 groups: tolerant (time below pH 6.00 ≤ 0.1 h/d) and susceptible cows (time below pH 6.00 ≥ 1.26 h/d). The lower reticular pH of susceptible cows was accompanied by lower MFC. Both groups already differed in reticular pH and MFC during the low-RFCH period. Furthermore, higher RFCH amounts did not decrease the reticular pH in either of the 2 groups. Nevertheless, MFD was observed in both groups during the high-RFCH period compared with the low-RFCH period. Lower MFC in animals with lower reticular pH or during the high-RFCH period was associated with a shift in 18-carbon fatty acids toward trans-10 at the expense of trans-11 intermediates, which was observed in plasma as well as in milk samples. Moreover, lower MFC was accompanied by shifts in the relative abundance of specific bacteria in buccal samples. Genera Dialister, Sharpea, Carnobacterium, Acidaminococcus, and uncultured genera belonging to the Betaproteobacteria were more abundant in situations with greater trans-10 proportions.

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The aim of the study was to investigate the presence of opioid receptor types in human breast adenocarcinoma MCF-7 cells and to characterize the changes in MOR expression induced by opioid agonist and antagonist treatment. We have shown that all three types of opioid receptors, but predominantly MOR, are expressed in MCF-7 cells. Selective MOR agonists, morphine, endomorphin-1, and endomorphin-2 downregulated MOR mRNA levels in a concentration- and time-dependent manner, but the effect produced by endomorphins was much stronger. Downregulation was blocked by the opioid antagonist naloxone. Naloxone alone produced a slight increase in MOR gene expression. Immunoblotting with antiserum against MOR-1 confirmed these results at the protein level. The results of our study indicate that, in MCF-7 cells, MOR gene expression is downregulated by opioid agonists and upregulated by opioid antagonists. We propose that the opioid-induced regulation of MOR mRNA expression is mediated by reduced binding of the transcription factors NFkappaB and AP-1 to the promoter region on the MOR gene.

The comparison of the effects of all forage offering methods would be particularly useful information in modeling growth performance and rumen fermentation of dairy calves. Therefore, this study attempted to evaluate the effects of methods of oat hay provision on growth performance, rumen fermentation and biochemical blood indices of dairy calves during preweaning and postweaning periods. At birth, 40 female Polish Holstein-Friesian calves (3 days of age; 39.6 ± 0.39 kg BW) were randomly assigned to four treatment groups differing in the access to chopped oat hay: CON (control, starter without oat hay), OH (starter feed containing 10% DM basis oat hay), OH-FC (starter feed containing 10% DM basis oat hay and oat hay fed as free-choice provision in different buckets) and FC (starter feed and oat hay fed as free-choice provision in different buckets). The calves were weaned on day 56, and then the study continued until day 84. Intakes of starter feed and oat hay were recorded daily, whereas BW and hip height (HH) on day 3 and then every 14 days. Samples of blood were collected on the initiation of experiment and then every 14 days, and rumen contents on day 28, 56 and 84. No treatment effects were found for starter, starch, CP, total DM intake, average daily gain, feeding efficiency, change in HH, ruminal fluid pH, concentrations of ruminal propionate and NH3-N, concentrations of urea nitrogen and non-esterified fatty acids in the blood. There were differences between treatments in terms of ruminal total volatile fatty acids and molar concentrations of acetate, butyrate and acetate to propionate ratio; highest in OH and OH-FC groups, especially during the postweaning period. On the other hand, lower concentrations of iso-valerate were found in OH and OH-FC groups on day 56 and 84. The concentrations of IGF-I throughout the experiment and β-hydroxybutyrate during the postweaning period in the blood were influenced by treatment, with the greatest values observed in OH and OH-FC calves. Results of this study indicate that starter feed containing chopped oat hay improves rumen fermentation parameters, which might allow successful transition from preruminant to mature ruminant state. Also, providing chopped oat hay with pelleted starter feed seems to be a better method than free-choice supplementation.

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