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Overexpression of cholinergic receptor nicotinic gamma subunit inhibits proliferation and differentiation of bovine preadipocytes

  • Jiawei, Du;Hui, Zhao;Guibing, Song;Yuan, Pang;Lei, Jiang;Linsen, Zan;Hongbao, Wang
    • Animal Bioscience
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    • v.36 no.2
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    • pp.200-208
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    • 2023
  • Objective: Muscle acetylcholine receptors have five alpha subunits (α, β, δ, ε, or γ), and cholinergic receptor nicotinic gamma subunit (CHRNG) is the γ subunit. It may also play an essential role in biological processes, including cell differentiation, growth, and survival, while the role of CHRNG has not been studied in the literature. Therefore, the purpose of this study is to clarify the effect of CHRNG on the proliferation and differentiation of bovine preadipocytes. Methods: We constructed a CHRNG overexpression adenovirus vector and successfully overexpressed it on bovine preadipocytes. The effects of CHRNG on bovine preadipocyte proliferation were detected by Edu assay, cell counting Kit-8 (CCK-8), real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), Western blot and other techniques. We also performed oil red O, RT-qPCR, Western blot to explore its effect on the differentiation of preadipocytes. Results: The results of Edu proliferation experiments showed that the number of EDU-positive cells in the overexpression group was significantly less. CCK-8 experiments found that the optical density values of the cells in the overexpression group were lower than those of the control group, the mRNA levels of proliferating cell nuclear antigen (PCNA), cyclin A2 (CCNA2), cyclin B1 (CCNB1), cyclin D2 (CCND2) decreased significantly after CHRNG gene overexpression, the mRNA levels of cyclin dependent kinase inhibitor 1A (CDKN1A) increased significantly, and the protein levels of PCNA, CCNB1, CCND2 decreased significantly. Overexpression of CHRNG inhibited the differentiation of bovine preadipocytes. The results of oil red O and triglyceride determination showed that the size and speed of lipid droplets accumulation in the overexpression group were significantly lower. The mRNA and protein levels of peroxisome proliferator activated receptor gamma (PPAR class="checkNonKBPoint">γ), CCAAT enhancer binding protein alpha (CEBPα), fatty acid binding protein 4 (FABP4), fatty acid synthase (FASN) decreased significantly. Conclusion: Overexpression of CHRNG in bovine preadipocytes inhibits the proliferation and differentiation of bovine preadipocytes.

Effects of β-glucan with vitamin E supplementation on the physiological response, litter performance, blood profiles, immune response, and milk composition of lactating sows

  • Tae Wook, Goh;Jinsu, Hong;Hong Jun, Kim;Sun Woo, Kang;Yoo Yong, Kim
    • Animal Bioscience
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    • v.36 no.2
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    • pp.264-274
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    • 2023
  • Objective: This study was conducted to evaluate the effects of β-glucan with vitamin E supplementation on the physiological response, litter performance, blood profiles, immune response, and milk composition of lactating sows. Methods: A total of 50 multiparous F1 sows (Yorkshire×Landrace) with an average body weight (BW) of 233.6±4.30 kg and an average parity of 4.00±0.307 and their litters were used in this experiment. All sows were allotted to one of five treatments, taking into consideration BW, backfat thickness, and parity in a completely randomized design with 10 replicates. The experimental diets included a corn-soybean meal-based basal diet with or without 0.1% or 0.2% β-glucan and 110 IU vitamin E/kg diet. Results: All treatments added with β-glucan or vitamin E were statistically higher in the average daily feed intake (ADFI) of lactating sows compared to those of the control (Diet, p<0.01). Additionally, the ADFI of lactating sows was significantly higher in the groups supplemented with 0.1% β-glucan compared to 0.2% β-glucan (BG, p<0.01). There was an increasing trend in piglet weight at weaning (BG, p = 0.07), litter weight at the 21st day of lactation (BG, p = 0.07) and litter weight gain (BG, p = 0.08) in groups supplemented with 0.1% β-glucan. The addition of 110 IU vitamin E/kg diet increased vitamin E concentration significantly in lactating sows (VE, p<0.01) and exhibited a trend for higher concentrations of vitamin E (VE, p = 0.09) in piglets. Adding 0.1% β-glucan compared to 0.2% β-glucan induced a decrease in the concentration of tumor necrosis factor-α in lactating sows (BG, p = 0.06) and in piglets (BG, p = 0.09) on the 21st day of lactation. There were no significant differences in the milk composition of sows. Conclusion: Adding 0.1% β-glucan and 110 IU vitamin E/kg to a lactating sow's diet was beneficial to the growth performance of piglets by leading to an increase in the feed intake of sows and efficiently supplying vitamin E to both the sows and piglets.

Comparison of miR-106b, miR-191, and miR-30d expression dynamics in milk with regard to its composition in Holstein and Ayrshire cows

  • Marina V. Pozovnikova;Viktoria B. Leibova;Olga V. Tulinova;Elena A. Romanova;Artem P. Dysin;Natalia V. Dementieva;Anastasiia I. Azovtseva;Sergey E. Sedykh
    • Animal Bioscience
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    • v.37 no.6
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    • pp.965-981
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    • 2024
  • Objective: Milk composition varies considerably and depends on paratypical, genetic, and epigenetic factors. MiRNAs belong to the class of small non-coding RNAs; they are one of the key tools of epigenetic control because of their ability to regulate gene expression at the post-transcriptional level. We compared the relative expression levels of miR-106b, miR-191, and miR-30d in milk to demonstrate the relationship between the content of these miRNAs with protein and fat components of milk in Holstein and Ayrshire cattle. Methods: Milk fat, protein, and casein contents were determined in the obtained samples, as well as the content of the main fatty acids (g/100 g milk), including: saturated acids, such as myristic (C14:0), palmitic (C16:0), and stearic (C18:0) acids; monounsaturated acids, including oleic (C18:1) acid; as well as long-, medium- and short-chain, polyunsaturated, and trans fatty acids. Real-time stem-loop one-tube reverse transcription polymerase chain reaction with TaqMan probes was used to measure the miRNA expression levels. Results: The miRNA expression levels in milk samples were found to be decreased in the first two months in Holstein breed, and in the first four months in Ayrshire breed. Correlation analysis did not reveal any dependence between changes in the expression level of miRNA and milk fat content, but showed a multidirectional relationship with individual milk fatty acids. Positive associations between the expression levels of miR-106b and miR-30d and protein and casein content were found in the Ayrshire breed. Receiver operating characteristic curve analysis showed that miR-106b and miR-30d expression levels can cause changes in fatty acid and protein composition of milk in Ayrshire cows, whereas miR-106b expression level determines the fatty acid composition in Holsteins. Conclusion: The data obtained in this study showed that miR-106b, miR-191, and miR-30d expression levels in milk samples have peculiarities associated with breed affiliation and the lactation period.

Effects of Monensin and Fish Oil on Conjugated Linoleic Acid Production by Rumen Microbes in Holstein Cows Fed Diets Supplemented with Soybean Oil and Sodium Bicarbonate

  • Jin, G.L.;Choi, S.H.;Lee, H.G.;Kim, Y.J.;Song, Man K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1728-1735
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    • 2008
  • The present study was conducted with four ruminally canulated Holstein cows to observe the effects of monensin or fish oil on diet fermentation and production of conjugated linoleic acids (CLAs) in the rumen when fed diets supplemented with soybean oil and sodium bicarbonate. Cows of the control treatment were fed a basal diet (CON) consisting of 60% commercial concentrate and 40% chopped rye grass hay. Cows of other treatments were fed the same diet as CON, but the concentrate was supplemented with 7% of soybean oil and 0.5% of sodium bicarbonate (SO-B), SO-B supplemented with monensin (30 ppm, SO-BM) or concentrate supplemented with 6.3% of soybean oil, 0.5% of sodium-bicarbonate, 30 ppm of monensin and 0.7% of fish oil (SO-BMF). Dry matter (DM) intake of the cows was significantly (p<0.011) reduced by feeding the SO-BMF diet compared to the other diets which did not differ in DM intake. Whole tract digestibility of major dietary components was significantly (p<0.004-0.027) higher for SO-BMF than the other supplement-containing diets. Dietary supplements did not clearly affect rumen pH and ammonia concentrations compared to the CON diet. Significantly reduced (p<0.05) total VFA concentration was obtained by the addition of fish oil to the diet (SO-BMF) compared to other diets. No differences, however, were obtained in major VFA proportions as well as in total VFA between the supplemented diets. The SO-BM diet increased (p<0.01-0.05) the concentrations of trans-11 $C_{18:1}$ and linoleic acid in rumen fluid. Total CLA concentration was also increased by the feeding of SO-B and SO-BM diets during early fermentation times (up to 3 h) post-feeding. Cis-9, trans-11 CLA concentration in rumen fluid was highest (p<0.05) for SO-B up to 1 h while the highest (p<0.01) value for SO-BM occurred at 3 h post-feeding. An increased trans-10, cis-12 CLA concentration was obtained from the SO-B and SO-BM diets at 1 and 3 h post feeding compared to the other diets. Supplementation of oils with monensin and sodium bicarbonate increased (p<0.05) the proportions of $C_{18:1}$ and CLA in the plasma of cows, but the effect of monensin and/or fish oil was limited to trans-10, cis-12 CLA.

Influence of polymer-coated slow-release urea on total tract apparent digestibility, ruminal fermentation and performance of Nellore steers

  • Gardinal, R.;Calomeni, G.D.;Consolo, N.R.B.;Takiya, C.S.;Freitas, J.E. Jr;Gandra, J.R.;Vendramini, T.H.A.;Souza, H.N.;Renno, F.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.1
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    • pp.34-41
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    • 2017
  • Objective: Two experiments were performed to evaluate the effects of coated slow-release urea on nutrient digestion, ruminal fermentation, nitrogen utilization, blood glucose and urea concentration (Exp 1), and average daily gain (ADG; Exp 2) of steers. Methods: Exp 1: Eight ruminally fistulated steers [$503{\pm}28.5kg$ body weight (BW)] were distributed into a d $4{\times}4$ Latin square design and assigned to treatments: control (CON), feed grade urea (U2), polymer-coated slow-release urea A (SRA2), and polymer-coated slow-release urea B (SRB2). Dietary urea sources were set at 20 g/kg DM. Exp 2: 84 steers ($350.5{\pm}26.5kg$ initial BW) were distributed to treatments: CON, FGU at 10 or 20 g/kg diet DM (U1 and U2, respectively), coated SRA2 at 10 or 20 g/kg diet DM (SRA1 and SRA2, respectively), and coated SRB at 10 or 20 g/kg diet DM (SRB1 and SRB2, respectively). Results: Exp 1: Urea treatments (U2+SRA2+SRB2) decreased (7.4%, p = 0.03) the DM intake and increased (11.4%, p<0.01) crude protein digestibility. Coated slow-release urea (SRA2+-SRB2) showed similar nutrient digestibility compwared to feed grade urea (FGU). However, steers fed SRB2 had higher (p = 0.02) DM digestibility compared to those fed SRA2. Urea sources did not affect ruminal fermentation when compared to CON. Although, coated slow-release urea showed lower (p = 0.01) concentration of $NH_3-N$ (-10.4%) and acetate to propionate ratio than U2. Coated slow-release urea showed lower (p = 0.02) urinary N and blood urea concentration compared to FGU. Exp 2: Urea sources decreased (p = 0.01) the ADG in relation to CON. Animals fed urea sources at 10 g/kg DM showed higher (12.33%, p = 0.01) ADG compared to those fed urea at 20 g/kg DM. Conclusion: Feeding urea decreased the nutrient intake without largely affected the nutrient digestibility. In addition, polymer-coated slow-release urea sources decreased ruminal ammonia concentration and increased ruminal propionate production. Urea at 20 g/kg DM, regardless of source, decreased ADG compared both to CON and diets with urea at 10 g/kg DM.

Supplementation of Holstein Cows with Low Doses of Bovine Somatotropin (bST) Prepartum and Postpartum Affects Physiological Adaptations and Milk Production

  • Liboni, M.;Gulay, M.S.;Hayen, M.J.;Belloso, T.I.;Head, H.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.404-413
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    • 2008
  • Major objectives were to evaluate effects of three schemes of bST-supplementation of Holstein cows (142.8 mg/14 d, POSILAC) during the prepartum and/or postpartum periods through 63 d (${\pm}3d$) of lactation. Measures evaluated the potential of treatments to improve body weight (BW) and body condition score (BCS), provoke changes in plasma concentrations of somatotropin (ST) and IGF-I, and improve milk yield, milk composition (percentages of protein and fat, and somatic cell counts), and several calving variables. Multiparous Holstein cows were randomly assigned to a $2{\times}2$ factorial arrangement of treatments (TRT) to give four groups (I = no bST, n = 26; II = bST postpartum, n = 25; III = bST prepartum, n = 27; IV = bST prepartum and postpartum, n = 25). During the prepartum period, cows in groups I and II were not supplemented but those in groups III and IV were supplemented every 2-wk beginning 21 d before expected calving date through calving. During the first 63 DIM only cows in groups II and IV were supplemented with bST. From 64 DIM through the end of lactation cows in all groups were supplemented with the full lactation dose of bST (500 mg/14 d). The BW and BCS were recorded weekly throughout the prepartum and postpartum periods and every 2-wk beyond 70 DIM. Blood samples were collected 3-times a week for analyses of ST and IGF-I. Milk yields were recorded daily though 150 DIM. Prepartum supplementation of bST did not affect BW or BCS, but mean concentrations of ST were increased 12.2% and were 15.5% greater at calving. Overall, mean concentration of IGF-I was not affected by treatment but concentrations were greater at 1 and 2 wk before calving in bST-supplemented cows. During the first 63 DIM the BW and BCS were not affected by treatment. Significant effects of bST-supplementation were detected on concentrations of ST, IGF-I and on milk yield compared to non-supplemented cows in group I. Postpartum concentrations of ST were greater in bST-supplemented cows (TRT II and IV; +41.9 and 54.6%). However, concentrations of IGF-I were greater only in cows in group IV (+25.9%) during the postpartum period. Overall, the three bST-supplemented groups had greater actual milk yield than the control group (I) during the first 63 and 150 DIM. The actual milk yields during 63 and 150 DIM were 6.5 and 4.6 kg/d greater for cows in group IV than cows in group I and the 305-d ME milk yield also was 15.6% greater. No adverse effects of TRT were observed on calf birth weight, colostrum immunoglobulins, ease of calving or other measures evaluated.

Effects of Saccharomyces cerevisiae Supplementation and Anhydrous Ammonia Treatment of Wheat Straw on In-situ Degradability and, Rumen Fermentation and Growth Performance of Yearling Lambs

  • Comert, Muazzez;Sayan, Yilmaz;Ozelcam, Hulya;Baykal, Gulsah Yegenoglu
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.5
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    • pp.639-646
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    • 2015
  • The effects of Saccharomyces cerevisiae supplementation ($6.6{\times}10^8cfu$) and anhydrous ammonia treatment (3%) of wheat straw (WS) were investigated on in-situ dry matter (DM) degradability, and on rumen fermentation and growth performance of lambs. Rumen-fistulated Menemen sheep fed a diet with and without live yeast were used to assess the DM degradability characteristics of WS and ammonia-treated wheat straw ($WS_{NH3}$). Twenty-six yearling Menemen male lambs were fed in four groups. Lambs of control group (WS) received untreated WS without supplemental yeast, whereas other three groups were fed WS treated with anhydrous ammonia ($WS_{NH3}$ group), untreated WS and yeast (WS+YEAST group) or WS treated with anhydrous ammonia and yeast ($WS_{NH3}$+YEAST group). Supplemented live yeast (4 g/d) was added in the diet. Lambs were offered untreated or ammonia treated WS ad-libitum and concentrate was fed at 1% of live body weight. The degradability of the water-insoluble (fraction B) was significantly increased by all of the treatment groups. Potential degradability (A+B), effective DM degradability's (pe2, pe5, and pe8) and average daily weight gain increased only in $WS_{NH3}$+YEAST group (p<0.05). Voluntary DM intake was not increased by the treatments (p>0.05), but voluntary metabolizable energy and crude protein intake were increased by $WS_{NH3}$ and by $WS_{NH3}$+YEAST (p<0.05). Average daily rumen pH was not affected by any of the treatments, but average daily $NH_3$-N was significantly higher in the $WS_{NH3}$ and $WS_{NH3}$+YEAST groups, and total volatile fatty acids were significantly higher in the WS+YEAST and $WS_{NH3}$+YEAST groups. In conclusion, the improvement of feed value of WS was better by the combination of ammonia-treatment and yeast supplementation compared to either treatment alone.

Chemical-nutritional characteristics and aromatic profile of milk and related dairy products obtained from goats fed with extruded linseed

  • Bennato, Francesca;Ianni, Andrea;Innosa, Denise;Grotta, Lisa;D'Onofrio, Andrea;Martino, Giuseppe
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.1
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    • pp.148-156
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    • 2020
  • Objective: This study aimed to evaluate the effect of dietary integration with extruded linseed (EL) on fatty acid (FA) and aromatic profile of goat cheese after 60 (T60) days of ripening. Methods: Thirty goats were divided in two groups. The control group (CG) was fed with conventional diet, whereas the experimental group (EL+) was fed with conventional diet supplemented with 10% of EL. Milk samples were collected on 30 and 60 days of trial to determinate chemical-nutritional composition and FA profile. At the end of experiment, six cheese-making sessions (3 for each group) were carried out using a pooled milk sample obtained from the 15 goats of each group. At 60 days of ripening, cheeses were analyzed for chemical-nutritional composition, FA and aromatic profile. Results: An increase in the milk production, protein, fat and lactose were evidenced in the EL+ goats. Conversely, a reduction of somatic cells was observed in the EL+ compared with the CG. However, no variation was observed for urea and casein levels content in milk samples, and no changes in protein and lipid content were found for cheeses in the two experimental groups. Dietary supplementation with EL modified the FA profile of milk. There was a decrease in saturated FAs and an increase in polyunsaturated FAs. Chemical composition of T60 cheese did not differ between the two groups but a different FA profile was observed. In T60 cheese obtained from EL+ milk, an increase in short-chain FA and a decrease in medium and long-chain FA were observed. The EL diet led to cheeses with butanoic acid 2 times higher compared to CG cheeses. Moreover, a greater presence of aldehyde compounds and alcohols were observed in the cheeses of experimental group. Conclusion: The present study pointed out that EL supplementation may improve the chemical and physical qualities of goat milk and cheeses.

Evaluating fermentation quality, in vitro digestibility and aerobic stability of a total mixed ration ensiled with different additives on Tibet plateau

  • Dong, Zhihao;Wang, Siran;Zhao, Jie;Li, Junfeng;Liu, Qinhua;Bao, Yuhong;Shao, Tao
    • Animal Bioscience
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    • v.34 no.2
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    • pp.223-232
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    • 2021
  • Objective: To investigate the improvement in utilization efficiency of total mixed ration (TMR) on Tibetan plateau, TMR were ensiled with different additives. Methods: A total of 150 experimental silos were prepared in a completely randomized design to evaluate the six treatments: i) control (without additive), ii) Lactobacillus buchneri (L. buchneri), iii) acetic acid, iv) propionic acid, v) 1,2-propanediol; and vi) 1-propanol. After 90 days of ensiling, silos were opened for fermentation quality and in vitro analysis, and then subjected to an aerobic stability test for 14 days. Results: Treating with L. buchneri, acetic acid, 1,2-propanediol and 1-propanol decreased propionic acid contents and yeast number, whereas increased (p<0.05) pH, acetic acid and ethanol contents in the fermented TMR. Despite increased dry matter (DM) loss in the TMRs treated with 1,2-propanediol and 1-pronanol, additives did not affect (p>0.05) all in vitro parameters including gas production at 24 h (GP24), GP rate constant, potential GP, in vitro DM digestibility and in vitro neutral detergent fibre digestibility. All additives improved the aerobic stability of ensiled TMR to different extents. Specially, aerobic stability of the ensiled TMR were substantially improved by L. buchneri, acetic acid, 1,2-propanediol, and 1-propanol, indicated by stable pH and lactic acid content during the aerobic stability test. Conclusion: L. buchneri, acetic acid, 1,2-propanediol, and 1-propanol had no adverse effect on in vitro digestibility, while ensiling TMR with the additives produced more acetic acid and ethanol, subsequently resulting in improvement of aerobic stability. There is a potential for some fermentation boosting additives to enhance aerobic stability of fermented TMR on Tibetan plateau.

Evaluation of liquid and powdered forms of polyclonal antibody preparation against Streptococcus bovis and Fusobacterium necrophorum in cattle adapted or not adapted to highly fermentable carbohydrate diets

  • Cassiano, Eduardo Cuellar Orlandi;Perna, Flavio Junior;Barros, Tarley Araujo;Marino, Carolina Tobias;Pacheco, Rodrigo Dias Lauritano;Ferreira, Fernanda Altieri;Millen, Danilo Domingues;Martins, Mauricio Furlan;Pugine, Silvana Marina Piccoli;de Melo, Mariza Pires;Beauchemin, Karen Ann;Meyer, Paula Marques;Arrigoni, Mario de Beni;Rodrigues, Paulo Henrique Mazza
    • Animal Bioscience
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    • v.34 no.1
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    • pp.74-84
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    • 2021
  • Objective: Feed additives that modify rumen fermentation can be used to prevent metabolic disturbances such as acidosis and optimize beef cattle production. The study evaluated the effects of liquid and powdered forms of polyclonal antibody preparation (PAP) against Streptococcus bovis and Fusobacterium necrophorum on rumen fermentation parameters in ruminally cannulated non-lactating dairy cows that were adapted or unadapted to a high concentrate diet. Methods: A double 3×3 Latin square design was used with three PAP treatments (control, powdered, and liquid PAP) and two adaptation protocols (adapted, unadapted; applied to the square). Adapted animals were transitioned for 2 weeks from an all-forage to an 80% concentrate diet, while unadapted animals were switched abruptly. Results: Interactions between sampling time and adaptation were observed; 12 h after feeding, the adapted group had lower ruminal pH and greater total short chain fatty acid concentrations than the unadapted group, while the opposite was observed after 24 h. Acetate:propionate ratio, molar proportion of butyrate and ammonia nitrogen concentration were generally greater in adapted than unadapted cattle up to 36 h after feeding. Adaptation promoted 3.5 times the number of Entodinium protozoa but copy numbers of Streptococcus bovis and Fibrobacter succinogens genes in rumen fluid were not affected. However, neither liquid nor powdered forms of PAP altered rumen acidosis variables in adapted or unadapted animals. Conclusion: Adaptation of cattle to highly fermentable carbohydrate diets promoted a more stable ruminal environment, but PAP was not effective in this study in which no animal experienced acute or sub-acute rumen acidosis.