pH Depression Research Articles

Cattle are more motivated for a high-concentrate diet than Sudan grass hay, despite low reticulorumen pH.

Subacute ruminal acidosis (SARA) is characterized by chronic low ruminal pH, and occurs for feedlot cattle fed high-concentrate diets. Forages slow digestion and reduce acid production. We aimed to assess how motivated finishing cattle are to access forage (Sudan grass hay, SG) via their willingness to interact with an electrified barrier. Reticulorumen pH was measured to relate the results to digestive health. Twenty-eight animals fed a high-concentrate ration ad libitum had access to 4L of one of two treatments (n = 14/treatment) fed 1×/d behind a barrier: 1) SG or 2) an additional offering of the normal ration (total mixed ration [TMR]). To access their treatment, the steer voluntarily pushed his muzzle against an electrified barrier. The electrical current was increased exponentially every 24h (0, 156, 312, 625, 1,250, 2,500, 5,000 µA) until the animal ceased accessing it. Visits to the treatment were recorded continuously 24h/d and reticulorumen pH was measured every 10min. Time with a reticulorumen pH below 5.8 was 348 ± 101 and 280 ± 76 min/24 h for SG and TMR animals, respectively; these durations meet the criterion for SARA. However, animals with access to SG were less likely to advance to the next current than TMR animals (P < 0.01) and were approximately 3× less willing to interact with higher currents than TMR (mean maximum current touched: 469 ± 169 and 1,380 ± 254 μA, respectively, mean ± SE, P = 0.01). Lower motivation to access SG was further demonstrated through fewer visits to the SG (2.4 ± 0.4 vs. 5.3 ± 0.6 #/d, P < 0.01), and less SG consumed than TMR (32.0 ± 0.1 vs. 74.0 ± 0.0 %/d, P < 0.01, measured as % due to weight differences of SG and TMR). Overall, finishing cattle valued the TMR more than SG, likely because of differences in the quantity offered, palatability, and familiarity. When rumen health was considered, SG animals visited more often (r = 0.5, P = 0.09) and showed fewer failed attempts (r = -0.5, P = 0.06) to access forage as the severity and duration of pH depression below 5.6, for example, increased. No measures of treatment use were related to pH depression for TMR animals (P ≥ 0.31). These findings provide evidence that cattle are motivated for Sudan grass hay when experiencing chronic low reticulorumen pH. However, they also contribute to the mixed evidence about the motivation for forage in this life stage, because, overall TMR was valued more highly than SG. Despite widespread pH depression, TMR cattle contrafreeloaded for additional concentration, demonstrating unexpectedly high motivation for this resource.

Challenges in the Definition and Measurement of Subacute Ruminal Acidosis in Holstein Dairy Cows: A Review

Subacute ruminal acidosis can be defined as a depression of rumen pH, which affects animal health and production. Although researchers have tried to find a solution for this disorder, it is a prevalent problem that causes considerable losses in commercial dairy cow production. This review aims to reveal critical points in current knowledge about subacute ruminal acidosis and suggest solutions for future research. The first challenging issue in subacute ruminal acidosis is the diagnosis method of this disorder, which requires appropriate statistical evaluation and modeling. In addition, biological factors should be considered to define subacute ruminal acidosis since some roles have recently been observed for different CO2 species in the rumen as a direct cause of the events. These CO2 species are sometimes more accurate than rumen pH in explaining the decrease in feed intake, milk yield, milk fat percentage, and inflammation responses. In the future, the measuring of the CO2 species in the rumen may be a replacement for pH measurement or become a factor that can greatly explain ruminal acidosis. Compared to basic methods, another challenging point is the reliability of rumen pH measurements as well as the accuracy of newly developed sensors. The reticular pH with current boluses could be measured by monitoring cows on-farm or a large number of animals in research. In conclusion, a thorough definition and precise application of new measurement devices can reveal some unknown factors for subacute ruminal acidosis in dairy cows.

Synthesis of Polyaluminum Chloride Coagulant from Waste Aluminum Foil and Utilization in Petroleum Wastewater Treatment

This work investigates the potential synthesis of cost-effective polyaluminum chloride (PACl) coagulant from waste household aluminum foil and utilization for treating petroleum wastewater (PWW), especially dissolved organic compounds (DOC, like octanol–water mixture) and nonsettleable suspended (NSS-kaolin) mineral particles. Based on the Standard Practice for Coagulation–Flocculation Jar Test, the efficiency of PACl for DOC and NSS removal was evaluated in relation to the effects of the operational parameters. The results demonstrated that the as-prepared PACl has an amorphous morphology with a Keggin-type e-Al13 molecular structure {Na[AlO4(OH)24(H2O)]·xH2O and good thermal stability up to 278 °C. PACl coagulant also exhibited a higher efficiency for NSS removal than DOC by around 1.5- to 1.9-fold under broad pH (5–7), while a higher acidic/alkaline pH disrupts the sweep floc formation. An increased PACl dosage (over 25 mg/L) also caused a decrease in the coagulation efficiency by 11.7% due to Al species’ transformation and pH depression (from 6.8 to 4.9) via increased PACl hydrolysis. With a fast rotating speed of 280 rpm for 2 min, the minimum dose of PACl (10–25 mg/L) can maximize the removal efficiency of NSS (~98%) and DOC (~69%) at pH 6.5 ± 0.5 and 35 °C after 30 min of settling time. Treating actual saline PWW samples (salinity up to 187.7 g/L) also verified the high efficacy of PACl coagulation performance in reducing the turbidity and dissolved hydrocarbons by more than 75.5% and 67.7%, respectively. These findings verify the techno-economic feasibility of the as-prepared PACl coagulant in treating PWW treatment at different salinity levels.

Invited review: Effect of subacute ruminal acidosis on gut health of dairy cows.

Subacute ruminal acidosis (SARA) is assumed to be a common disease in high-yielding dairy cows. Despite this, the epidemiological evidence is limited by the lack of survey data. The prevalence of SARA has mainly been determined by measuring the pH of ruminal fluid collected using rumenocentesis. This may not be sufficiently accurate, because the symptoms of SARA are not solely due to ruminal pH depression, and ruminal pH varies among sites in the rumen, throughout a 24-h period, and among days. The impact of SARA has mainly been studied by conducting SARA challenges in cows, sheep, and goats based on a combination of feed restriction and high-grain feeding. The methodologies of these challenges vary considerably among studies. Variations include differences in the duration and amount of grain feeding, type of grain, amount and duration of feed restriction, number of experimental cows, and sensitivity of cows to SARA challenges. Grain-based SARA challenges affect gut health. These effects include depressing the pH in, and increasing the toxin content of, digesta. They also include altering the taxonomic composition of microbiota, reducing the functionality of the epithelia throughout the gastrointestinal tract (GIT), and a moderate inflammatory response. The effects on the epithelia include a reduction in its barrier function. Effects on microbiota include reductions in their richness and diversity, which may reduce their functionality and reflect dysbiosis. Changes in the taxonomic composition of gut microbiota throughout the GIT are evident at the phylum level, but less evident and more variable at the genus level. Effects at the phylum level include an increase in the Firmicutes to Bacteroidetes ratio. More studies on the effects of a SARA challenge on the functionality of gut microbiota are needed. The inflammatory response resulting from grain-based SARA challenges is innate and moderate and mainly consists of an acute phase response. This response is likely a combination of systemic inflammation and inflammation of the epithelia of the GIT. The systemic inflammation is assumed to be caused by translocation of immunogenic compounds, including bacterial endotoxins and bioamines, through the epithelia into the interior circulation. This translocation is increased by the increase in concentrations of toxins in digesta and a reduction of the barrier function of epithelia. Severe SARA can cause rumenitis, but moderate SARA may activate an immune response in the epithelia of the GIT. Cows grazing highly fermentable pastures with high sugar contents can also have a low ruminal pH indicative of SARA. This is not accompanied by an inflammatory response but may affect milk production and gut microbiota. Grain-based SARA affects several aspects of gut health, but SARA resulting from grazing high-digestible pastures and insufficient coarse fiber less so. We need to determine which method for inducing SARA is the most representative of on-farm conditions.

356 Anomalies of Carbohydrate Digestion and Metabolism by Ruminants

Abstract Carbohydrate, derived either from plant cell walls (neutral detergent fiber; NDF) from fresh or conserved forages consumed by cows and growing cattle or from starch of grains and concentrates in commercial feedlot diets, provides most of the dietary energy for ruminant growth and production. The ruminants’ ability to harvest forage from remote, inaccessible locations, to scavenge energy from NDF of feeds and byproducts of grain largely indigestible by non-ruminants, and to employ non-protein N to form protein-rich, nutrient-dense foods for humans are key ecological advantages over non-ruminants. Outlined below are some topics needing research attention. First, feed composition tables invariably indicate that the digested energy (DE) of carbohydrates is 4.4 kilocalories per gram, yet the gross energy (GE) of purified carbohydrate polymers (starch; cellulose, glycogen) averages 4.18; monomers (hexoses, pentoses) average 3.74. Having DE exceed GE violates the first Law of Thermodynamics! Might this 5 percent discrepancy reflect underestimations of fecal lignin or of protein digestibility due to metabolic fecal N? Favoring feeds rich in carbohydrate, this discrepancy confounds formulation of least-cost diets. Secondly, site of digestion of carbohydrate alters its metabolizable energy (ME). Fermentation, though required to yield ME from NDF, generates methane and heat. In contrast, starches and sugars that escape ruminal fermentation, if digested in the small intestine, circumvent these losses increasing their relative ME. Unfortunately, when ruminally fermented, starch, either directly or via pH depression, reduces NDF fermentation; might ruminal NDF digestion be increased by feeding NDF plus buffer versus concentrate diet components asynchronously? Third, carbohydrate metabolites lost in urine (lactate, pentoses, hippuric acid) need further quantification. Finally, ruminally-formed metabolites (methylglyoxal, methane) adversely affect animal health and energetics. Ruminal modification (site- or time-targeted enzymes or inhibitors; altering the ruminal microbiome with pre- or probiotics, inoculants, nutraceuticals) should help enhance ruminant productivity and sustainability.

Ruminal pH and its relationship with dry matter intake, growth rate, and feed conversion ratio in commercial Australian feedlot cattle fed for 148 days.

Measurement of ruminal pH throughout a 148-day feeding period in cattle fed commercial diets and to relate this to feed intake, growth rate and feed conversion ratio. Factors contributing to variation in rumen pH, including meal frequency, duration and weight, and, total daily intake, were also evaluated. Forty-eight cattle were randomly allocated to two pens and 12 randomly selected from each pen had rumen pH monitoring boli inserted. Ruminal pH was measured every 10 min and feed intake was measured continually. The cattle were fed a commercial feedlot diet for 148 days and weighed into and out of the feedlot to measure growth rate and to calculate feed conversion ratio. Cattle from both pens were registered to collect individual feed intake data using the GrowSafe® feed management system. Mean ruminal pH decreased with days on feed. Mean daily dry matter intake was the major contributor to greater average daily gain and lower ruminal pH. Lower mean ruminal pH was associated with greater average daily gain and lower feed conversion ratio, where it remained above the threshold of 5.6. There was no association between ruminal pH and average daily gain or feed conversion ratio for mean ruminal pH below 5.6. Ruminal acidosis can occur at any time during the feeding period, and the risk could be greater as days on feed increase. Feedlot production outcomes are not improved by ruminal pH depression below the threshold of 5.6.

Preweaning to postweaning rumen papillae structural growth, ruminal fermentation characteristics, and acute-phase proteins in calves

This study evaluated pre- to postweaning ruminal structural development, fermentation characteristics, and acute-phase protein levels in calves with a high milk replacer (MR) feeding rate prior to weaning. Six ruminally cannulated Holstein bull calves were fed MR (150 g/L) at 15% of body weight (BW) in 2 equal volumes daily. Volumes were adjusted weekly based on BW. Calves were weaned using a 1-step weaning method, with MR decreased by 50% at the end of wk 5 and full weaning at the end of wk 6. Calf starter, chopped straw, and water were offered ad libitum. Intake was recorded daily, and BW was recorded weekly. From wk 5 to 12, ruminal pH was continuously measured using a ruminal pH bolus. Ruminal fluid was collected weekly from wk 5 to 12 for measurement of short-chain fatty acid concentrations and quantification of total bacteria and protozoa. Rumen papillae were obtained at wk 5, 6, 7, 8, and 12 for histological analysis. Serum amyloid A and lipopolysaccharide-binding protein were measured weekly. Data were analyzed using GLIMMIX procedure of SAS (SAS Institute Inc., Cary, NC), with week as a fixed effect and calf as a random effect. During the weaning step-down, starter intake was 3-fold higher and continued to increase until wk 12. Body weight increased from birth to wk 12; however, BW did not change during wk 6, 7, and 8, possibly due to low metabolizable energy intake caused by the weaning strategy. Preweaning ruminal pH was below 5.8 for approximately 936.3 ± 125.99 min/d, implying ruminal acidosis. Furthermore, ruminal pH below 5.8 reached a peak at wk 8 with 1,203.9 ± 227.65 min/d below pH 5.8 and slowly decreased to 388.1 ± 189.82 min/d below pH 5.8 at wk 12. Papillae surface area, length, and width increased during wk 12 compared with wk 5. Corneum thickness increased by week, whereas spinosum/basale thickness only increased during wk 8 compared with wk 5. The acute-phase protein concentration was highest at wk 1 and then decreased and remained constant until wk 12. In conclusion, even before step-down weaning, calves experienced ruminal acidosis despite low starter intake. Further, the observed prolonged ruminal pH depression suggests that dietary rumen adaptation after weaning can take several weeks in calves with a high MR feeding rate preweaning. The prolonged depressed ruminal pH did not affect acute-phase proteins and this finding, along with the other results, suggests that rumen epithelium barrier integrity is not compromised during weaning.

Anti-lipopolysaccharide antibody mitigates ruminal lipopolysaccharide release without acute-phase inflammation or liver transcriptomic responses in Holstein bulls.

Anti-lipopolysaccharide (LPS) antibody administration has the potential benefits of neutralizing and consequently controlling rumen-derived LPS during subacute ruminal acidosis. Four Holstein bulls were used in this crossover study with a 2-week wash-out period. Anti-LPS antibody (0 or 4 g) was administered once daily for 14 days. Significantly lower ruminal LPS and higher 1-h mean ruminal pH were identified in the 4 g group. However, blood metabolites, acute-phase proteins, cytokines, and hepatic transcriptomes were not different between the two groups. Therefore, anti-LPS antibody administration mitigated ruminal LPS release and pH depression without accompanying responses in acute-phase inflammation or hepatic transcriptomic expression.

Anti-lipopolysaccharide antibody administration mitigates ruminal lipopolysaccharide release and depression of ruminal pH during subacute ruminal acidosis challenge in Holstein bull cattle.

The effects of anti-lipopolysaccharide (LPS) antibody on rumen fermentation and LPS activity were investigated during subacute ruminal acidosis (SARA) challenge. Eleven Holstein cattle (164 ± 14 kg) were used in a 3 × 3 Latin square design. Cattle were fed a roughage diet on days −11 to −1 (pre-challenge) and day 2 (post-challenge), and a high-grain diet on days 0 and 1 (SARA challenge). For 14 days, 0-, 2-, or 4-g of anti-LPS antibody was administered once daily through a rumen fistula. Ruminal pH was measured continuously, and rumen fluid and blood samples were collected on days −1, 0, 1, and 2. Significantly lower ruminal LPS activity on day 1 was observed in the 2- and 4-g groups than those in the 0-g group. In addition, significantly higher 1-hr mean ruminal pH on SARA challenge period (days 0 and 1) was identified in the 4-g group than in the 0-g group. However, rumen fermentation measurements (total volatile fatty acid [VFA], VFA components, NH3-N and lactic acid) and peripheral blood metabolites (glucose, free fatty acid, beta-hydroxybutyrate, total cholesterol, blood urea nitrogen, aspartate aminotransferase and gamma-glutamyl transferase) were not different among the groups during the experimental periods. Therefore, anti-LPS antibody administration mitigates LPS release and pH depression without the depression of rumen fermentation and peripheral blood metabolites during SARA challenge in Holstein cattle.

Effects of starch concentration of close-up diets on rumen pH and plasma metabolite responses of dairy cows to grain challenges after calving

The objective of this study was to evaluate the effects of starch concentration of close-up diets on plasma concentrations of energy metabolites and rumen pH of dairy cows after calving. Eighteen multiparous Holstein dairy cows (mean parity = 2.78; mean body weight = 708 kg; mean body condition score = 3.08) fitted with ruminal cannulas were assigned to treatment balanced for parity, body condition score, and expected calving date. Cows were enrolled in the study at d 28 ± 3 before the expected calving date and fed either a low-starch (LS; 14.0% starch) or high-starch (HS; 26.1% starch) diet until parturition. All cows were fed a common diet after calving (25.1% starch). A grain challenge was performed on d 7 ± 2 and 21 ± 2 after calving by dosing 6.35 kg (dry matter basis) of finely ground barley and wheat grain (1:1) into the rumen via cannula. Feeding the HS diet before calving increased the duration (369 vs. 49 min/d) and area of pH below 5.8 (85.1 vs. 5.2 pH × min/d) during d -10 to -8. In addition, even though all cows were fed a common diet after calving, HS cows tended to have longer duration (177 vs. 76 min/6 h) and greater area of pH below 5.8 (67.8 vs. 20.3 pH × min/6 h) during a grain challenge on d 7. Plasma concentration of insulin tended to be greater in cows fed the HS diet (1.40 vs. 1.09 ng/mL), whereas plasma free fatty acid concentration was not different between treatments during the grain challenge on d 7. During the grain challenge on d 21, neither rumen pH nor blood metabolites were different between the HS and LS cows. These findings suggested that feeding an HS diet during the close-up period does not mitigate rumen pH depression but may exacerbate it after calving compared with feeding an LS diet.

Time From Neuraxial Anesthesia Placement to Delivery is Inversely Proportional to Umbilical Arterial Cord pH at Scheduled Cesarean Delivery

(Am J Obst Gynecol. 2019;220:389.e1–389.e9) While the maternal risks of cesarean delivery are well documented in the literature, less is known about the risks to the fetus. Neuraxial block-related hypotension and maternal obesity have been found to contribute to uterine hypoperfusion and umbilical arterial pH depression. Examining the time between spinal anesthesia placement and delivery could provide insight into these risks to the fetus awaiting cesarean delivery. The aim of this study was to evaluate neonatal umbilical arterial pH in relation to predelivery time intervals during scheduled cesarean delivery.

Combinations of non-invasive indicators to detect dairy cows submitted to high-starch-diet challenge

High-starch diets (HSDs) fed to high-producing ruminants are often responsible for rumen dysfunction and could impair animal health and production. Feeding HSDs are often characterized by transient rumen pH depression, accurate monitoring of which requires costly or invasive methods. Numerous clinical signs can be followed to monitor such diet changes but no specific indicator is able to make a statement at animal level on-farm. The aim of this pilot study was to assess a combination of non-invasive indicators in dairy cows able to monitor a HSD in experimental conditions. A longitudinal study was conducted in 11 primiparous dairy cows fed with two different diets during three successive periods: a 4-week control period (P1) with a low-starch diet (LSD; 13% starch), a 4-week period with an HSD (P2, 35% starch) and a 3-week recovery period (P3) again with the LSD. Animal behaviour was monitored throughout the experiment, and faeces, urine, saliva, milk and blood were sampled simultaneously in each animal at least once a week for analysis. A total of 136 variables were screened by successive statistical approaches including: partial least squares-discriminant analysis, multivariate analysis and mixed-effect models. Finally, 16 indicators were selected as the most representative of a HSD challenge. A generalized linear mixed model analysis was applied to highlight parsimonious combinations of indicators able to identify animals under our experimental conditions. Eighteen models were established and the combination of milk urea nitrogen, blood bicarbonate and feed intake was the best to detect the different periods of the challenge with both 100% of specificity and sensitivity. Other indicators such as the number of drinking acts, fat:protein ratio in milk, urine, and faecal pH, were the most frequently used in the proposed models. Finally, the established models highlight the necessity for animals to have more than 1 week of recovery diet to return to their initial control state after a HSD challenge. This pilot study demonstrates the interest of using combinations of non-invasive indicators to monitor feed changes from a LSD to a HSD to dairy cows in order to improve prevention of rumen dysfunction on-farm. However, the adjustment and robustness of the proposed combinations of indicators need to be challenged using a greater number of animals as well as different acidogenic conditions before being applied on-farm.

Feeding Diets Moderate in Physically Effective Fibre Alters Eating and Feed Sorting Patterns without Improving Ruminal pH, but Impaired Liver Health in Dairy Cows.

Simple SummaryThis study tested the hypothesis that the content of physically effective fibre in the diet modifies eating and feed sorting patterns of cows, which is reflected in rumen and systemic health variables. We observed that switching cows from a diet high to a diet moderate in physically effective fibre (17.8%; 28% starch on dry matter basis) in an attempt to stabilize rumen pH variations altered eating, sorting, and chewing behaviour. However, this attempt did not prevent ruminal pH drop and the impairment of liver health variables. Therefore, our study emphasizes the importance of increasing the level of physically effective fibre for particles >8 mm beyond 17%–18% of the diet when (barley grain-based) starch content is at a 28% level.The main challenge in dairy cattle feeding is to find a balance between the energy and physically effective fibre (peNDF), required to maintain rumen health. In an attempt to regulate the balance between energy intake and rumen buffering, we hypothesized that the content of peNDF in the diet modifies eating and feed sorting patterns of the cows. Sixteen lactating Simmental cows were switched from a diet high in peNDF, with which they were fed for one week, to a diet moderate in peNDF for four weeks. Data showed that during the moderate peNDF feeding the cows increased sorting for medium-sized particles and avoided both long and very fine particles. In addition, cows decreased their eating time per meal, but increased the number of meals per day, obviously attempting to decrease the amount of fermentable substrate per time unit while maintaining high levels of nutrient/energy intake. Although these changes during the moderate peNDF feeding went along with a lower diurnal variation of ruminal pH, feeding of the latter diet did not prevent ruminal pH drop and increased the level of all liver enzymes, indicating liver tissue damage. In conclusion, the altered eating, chewing, and sorting behaviour of the cows during the moderate peNDF feeding could not alleviate the deficiency in peNDF, which resulted in ruminal pH depression and impairment of liver health variables.

Subacute ruminal acidosis suppressed the expression of MCT1 in rumen of cows.

Subacute ruminal acidosis (SARA) is characterized by the depression of ruminal pH and an increase in the concentrations of short-chain fatty acids (SCFAs) and lipopolysaccharide (LPS) in the rumen of cows. The onset of SARA was linked to the accumulation of SCFAs. However, the mechanism of SCFAs transport is unknown. The proton-linked monocarboxylate transporter (MCT1) plays a vital role in the transportation of SCFAs. The goal of this study was to elucidate the distribution of MCT1 along the gastrointestinal tract of calves and adult cows; the expression change of MCT1 in SARA cows and the effect of ruminal pH, SCFAs, and LPS on MCT1 expression in rumen epithelial cells in vitro. The results indicated the presence of MCT1 along the gastrointestinal tract of calves and adult cows, most abundantly expressed in the rumen. Importantly, the expression of MCT1 was decreased in the rumen epithelium of SARA cows, and the expression of MCT1 was restored in the SARA treatment group. In vitro, LPS, low rumen fluid pH, high concentrations of SCFAs (90 mM acetate, 40 mM propionate, and 30 mM butyrate), and high concentrations of acetate, propionate, and butyrate, respectively, inhibited the expression of MCT1 in rumen epithelial cells. Taken together, these results indicated that LPS, low ruminal pH, and high concentrations of SCFAs decreased the expression of MCT1, further aggravating the accumulation of SCFAs in the rumen by decreasing the absorption of SCFAs.

Changes in ruminal and reticular pH and bacterial communities in Holstein cattle fed a high-grain diet

BackgroundSubacute ruminal acidosis (SARA) is characterized by a ruminal pH depression, and microbiota can also be affected by a higher acidity and/or dietary changes. Previous studies have revealed similar patterns in pH reduction in the rumen and reticulum, whereas changes in reticular pH and bacterial community following a high-grain diet are not fully understood. The aim of this study was to investigate the changes in reticular pH and bacterial community structure following a high-grain diet simultaneously with those in the rumen.ResultsSARA was diagnosed when ruminal and reticular pH remained under 5.6 for 350 ± 14 and 312 ± 24 min/d, respectively, on the last day of the CON period. During the CON period, lower proportion of acetic acid and higher proportion of butyric acid were observed compared with the HAY period. The proportions of acetic acid and propionic acid were lower and higher, respectively, in the rumen compared with the reticulum. From 454 pyrosequencing analysis, the relative abundance of several genera differed significantly between the two periods and the two locations. During the HAY period, higher relative abundances of Prevotella, Eubacterium, Oscillibacter, and Succiniclasticum and lower relative abundances of Ruminococcus, Clostridium, and Olsenella were identified compared with the CON period. Furthermore, the relative abundance of Eubacterium was lower in the rumen compared with the reticulum. Bacterial diversity indices were significantly different between the HAY and CON periods, being higher in the HAY period. The quantitative real-time PCR showed that the copy numbers of several cellulolytic bacteria (Fibrobacter succinogenes and Ruminococcus albus) were higher during the HAY period.ConclusionA high-grain diet showed similar impacts on the pH, fermentation, and bacterial community structure in the rumen and reticulum. During the CON period, ruminal and reticular pH decreased following the high-grain challenge, and lower bacterial diversity and changes in the bacterial composition, similarity, and bacterial copy numbers were observed due to a higher acidity and dietary changes compared with the HAY period. These changes may influence the fermentative ability of the rumen and reticulum.

Enhancing soluble phosphate concentration in sludge liquor by pressurised anaerobic digestion

Recovery of phosphate from wastewater is challenging, with one of the best opportunities being recovery from sludge anaerobic digestion liquor, as struvite. However, this is limited by the proportion of total phosphorous which is soluble, due to in-digester metal ion precipitation. High-pressure anaerobic digestion may enable enhanced phosphate solubility (and hence recovery potential), without the use of added acid, due to an increased liquid phase CO2 concentration. This was tested at 2, 4, and 6 bar absolute (bara) vs a 1 bara control reactor, fed with activated sludge. Increased pressure significantly (p = 0.0008), increased the fraction of phosphate that was soluble, ranging from 52% at 1 bara, to 75% at 6 bara. Model based analysis indicated that the main reason for increased solubility was pH depression (down to 6.4 at 6 bara), rather than changes in ion pairing (with carbonates) or increases in ionic activity. However, biological performance was adversely impacted, with a substantial loss in VS and COD destruction (on the order of 5%–10% absolute). No organic acid accumulation was observed. Bacterial and archaeal communities were significantly impacted (p∼0.0003–0.0005), with a shift to specific organisms, including Bacteroidales Rikenellaceae within the bacteria, and a Deep Sea Euryarchaeotal Group at 2 bara, and Methanocellaceae within the archaea at 4 and 6 bara. The work indicates that high-pressure operation is a technically viable option to improve phosphate recovery, and produce a high-methane biogas product, but that the loss of overall conversion needs to be further addressed, possibly through two-stage digestion.

Cystic Fibrosis Transmembrane Conductance Regulator Regulates Pannexin 1 Channel Opening for Acidosis‐induced ATP Release from Cardiomyocytes

ATP is released in the heart under a variety of conditions, including hypoxia, ischaemia and catecholamine stimulation. Under physiological conditions, extracellular ATP, and its breakdown product, adenosine, contribute to local vasodilation and regulation of coronary blood flow. We investigated the mechanism of ATP release in cardiomyocytes isolated from adult rat heart.Treatment with lactic acid produced pH depression and enhanced ATP release from cardiomyocytes. On the other hand, simulated ischaemia (metabolic inhibition plus anoxia) lowered ATP release. The acidosis‐induced ATP release was abolished by the specific CFTR inhibitor, CFTRinh‐172, CFTR pore blocker, GlyH‐101, or CFTR siRNA, suggesting that CFTR was involved. Forskolin and IBMX, agents that activate CFTR by elevating the intracellular cAMP, also increased ATP release, confirming the role of CFTR. However, blockade of Pannexin1 channels by Fast Green FCF, Brilliant Blue FCF, or pannexin1 siRNA, attenuated the ATP release during acidosis or forskolin treatment, and further lowered the ATP release during simulated ischaemia, suggesting that Pannexin1 functions as the ATP release channel. Immunofluorescence imaging suggested that CFTR and Pannexin1 were not co‐localized. Inhibition of either the Na+/H+ exchanger (NHE) with amiloride or the Na+/Ca2+ exchanger (NCX) with SN6 or KB‐R7943 abolished acidosis‐induced ATP release. FAK inhibitor 14, FAK autophosphorylation inhibitor, can attenuate the forskolin or lactic‐acid‐stimulated ATP release. Furthermore, forskolin or lactic‐acid‐stimulated ATP release was abolished by bicarbonate free medium or caspase inhibitor. Thus, we have demonstrated that the acidosis‐induced ATP release was involved in calcium induced CFTR open. The cyclic AMP, which activates mitochondrial PKA, is generated within mitochondria by the bicarbonate‐regulated soluble adenylyl cyclase (sAC). Hence, we propose that the CFTR Cl channel opens during acidosis, allowing the movement of bicarbonate into the cell, which activate the HCO3‐sAC‐cAMP‐PKA (mito‐sAC) signaling cascade and indirectly modulates Pannexin1 gating by activation of caspase.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Use of solid phosphorus fractionation data to evaluate phosphorus release from waste activated sludge

Waste activated sludge (WAS) can become an important source of phosphorus (P). P speciation was examined under anaerobic conditions, with different pH (4, 6 and 8) and temperatures (10, 20 and 35 °C). Aqueous P was measured and an extraction protocol was used to find three solid phosphorus fractions. A pH of 4 and a temperature of 35 °C gave a maximum of 51% of total P solubilized in 22 days with 50% of total P solubilized in 7 days. Batch tests indicate that little pH depression is needed to release non-apatite inorganic P (including microbial polyphosphate), while a pH of 4 rather than 6 will release more apatite inorganic P, and that organic P is relatively more difficult to release from WAS. Fractionation analysis of P in WAS can aid in design of more efficient methods for P recovery from WAS.

Relationship between pH of ruminal fluid during subacute ruminal acidosis and physiological response of the Polish Holstein-Friesian dairy cows.

The aim of the study was to investigate the effect of ruminal fluid pH depression on biochemical indices of blood, urine, feces, and milk, and to determine which of them may be helpful as a marker for the diagnosis of subacute ruminal acidosis (SARA). Ruminal fluid samples were obtained by rumenocentesis from 305 cows representing 13 commercial dairy herds. The herds were selected based on percentages of cows with an assigned value of ruminal fluid pH segregated into three groups as: SARA-positive herd, if at least 25% of the ruminal fluid samples indicated a pH < 5.6; SARA-risk herd, if less than 25% of ruminal fluid samples indicated a pH < 5.6, but at least 33% showed a pH ≤ 5.8; and SARA-negative herd, if less than 25% of the ruminal fluid samples indicated a pH < 5.6, but less than 33% exhibited a pH = 5.8. Moreover, the dairy cows were divided according to the ruminal fluid pH into three groups as follows: healthy cows (HC, pH>5.80, n = 196), risk cows (RC, pH 5.8 - 5.6, n = 51), and acidotic cows (AC, pH < 5.6, n = 58). Almost 19% (58/305) of the cows were classified as acidotic (pH < 5.6) and 46.2% of the herds as SARA-positive. In the AC group, higher concentrations of insulin-like growth factor-I (IGF-I), non-esterified fatty acids (NEFA), rectal temperature and lower blood pH, compared with those of the HC group, were recorded. Moreover, in the SARA-positive herds, higher concentrations of IGF-I and the lowest blood pH, compared with SARA-negative herds, were observed. The lowering of ruminal fluid pH increased the blood IGF-I and NEFA concentrations and the rectal temperature and decreased the blood pH. These measures are indicators of the physiological changes that occur as part of the pathogenesis of the condition and may be helpful for the diagnosis of the SARA syndrome when serial measurements are conducted.

Thiamine supplementation facilitates thiamine transporter expression in the rumen epithelium and attenuates high-grain-induced inflammation in low-yielding dairy cows

An experiment was conducted to uncover the effects of increasing dietary grain levels on expression of thiamine transporters in ruminal epithelium, and to assess the protective effects of thiamine against high-grain-induced inflammation in dairy cows. Six rumen-fistulated, lactating Holstein dairy cows (627 ± 16.9 kg of body weight, 180 ± 6 d in milk; mean ± standard deviation) were randomly assigned to a replicated 3 × 3 Latin square design trial. Three treatments were control (20% dietary starch, dry matter basis), high-grain diet (HG, 33.2% dietary starch, DM basis), and HG diet supplemented with 180 mg of thiamine/kg of dry matter intake. On d 19 and 20 of each period, milk performance was measured. On d 21, ruminal pH, endotoxic lipopolysaccharide (LPS), and thiamine contents in rumen and blood, and plasma inflammatory cytokines were detected; a rumen papillae biopsy was taken on d 21 to determine the gene and protein expression of toll-like receptor 4 (TLR4) signaling pathways. The HG diet decreased ruminal pH (5.93 vs. 6.49), increased milk yield from 17.9 to 20.2 kg/d, and lowered milk fat and protein from 4.28 to 3.83%, and from 3.38 to 3.11%, respectively. The HG feeding reduced thiamine content in rumen (2.89 vs. 8.97 μg/L) and blood (11.66 vs. 17.63 μg/L), and the relative expression value of thiamine transporter-2 (0.37-fold) and mitochondrial thiamine pyrophosphate transporter (0.33-fold) was downregulated by HG feeding. The HG-fed cows exhibited higher endotoxin LPS in rumen fluid (134,380 vs. 11,815 endotoxin units/mL), and higher plasma concentrations of lipopolysaccharide binding protein and pro-inflammatory cytokines when compared with the control group. The gene and protein expression of tumor necrosis factor α (TNFα), IL1B, and IL6 in rumen epithelium increased when cows were fed the HG diet, indicating that local inflammation occurred. The depressions in ruminal pH, milk fat, and protein of HG-fed cows were reversed by thiamine supplementation. Thiamine supplementation increased thiamine contents in rumen and blood, and also upregulated the relative expression of thiamine transporters compared with the HG group. Thiamine supplementation decreased ruminal LPS (49,361 vs. 134,380 endotoxin units/mL) and attenuated the HG-induced inflammation response as indicated by a reduction in plasma IL6, and decreasing gene and protein expression of pro-inflammatory cytokines in rumen epithelium. Western bottling analysis showed that thiamine suppressed the protein expression of TLR4 and the phosphorylation of nuclear factor kappa B (NFκB) unit p65. In conclusion, HG feeding inhibits thiamine transporter expression in ruminal epithelium. Thiamine could attenuate the epithelial inflammation during high-grain feeding, and the protective effects may be due to its ability to suppress TLR4-mediated NFκB signaling pathways.