Liver transcriptome response to periparturient hormonal and metabolic changes depends on the postpartum occurrence of subacute ruminal acidosis in Holstein cows.

We investigated changes in rumen fermentation, peripheral blood metabolites and hormones, and hepatic transcriptomic dynamics in Holstein cows with and those without subacute ruminal acidosis (SARA) during the periparturient period. Sixteen multiparous Holstein cows were categorized in the SARA (n = 8) or non-SARA (n = 8) groups depending on whether they developed SARA during the 2 wk after parturition. Reticulo-ruminal pH was measured continuously throughout the study. Rumen fluid, blood, and liver tissue samples were collected at 3 wk prepartum and 2 and 6 wk postpartum, with an additional blood sample collected at 0 and 4 wk postpartum. The 1-h mean pH was depressed postpartum in both groups, whereas depression was more severe in the SARA group simultaneously with significantly longer duration of time (for pH <5.6 and 5.8). Significant expression of differentially expressed genes in liver tissue (DEGs; false discovery rate corrected P < 0.1) were identified only in the non-SARA group and were further analyzed by Ingenuity Pathway Analysis software. Among the top expressed DEGs, the hepatic genes encoding lipid and cholesterol secretion (APOA1, APOA4, and G0S2) and gluconeogenesis (PC, G6PC, and PCK1) were upregulated postpartum. In silico analysis revealed the significant postpartum activation of upstream regulators, such as INSR, PPARG, and PPARGC1A. These results suggested that hepatic transcriptomic responsiveness to postpartum metabolic load and hormones were likely discouraged in cows with SARA when compared with the significant activation of genes and signaling pathways for adequate metabolic adaption to postpartum high-grain diet feeding in Holstein cows without SARA.

Forestomach developmental failure in an 11-month-old Japanese Black steer with severely retarded growth and chronic ruminal tympany.

This study reports findings from the pathological examination of the forestomach of an 11-month-old Japanese Black steer with severely retarded growth (41% of expected weight) and chronic ruminal tympany. The ruminal papillae were weakly formed (0.3-0.5 cm long) and unevenly distributed. The cellulae and cristae reticuli were underdeveloped; the cristae were 0.4-0.7 cm in height and milky white. The keratinized layer in the stratified squamous epithelium was thickened. Ruminal pH was 5.25, and ruminal volatile fatty acid concentration was 11.7 mM. The steer's severely retarded growth was considered to be caused by malnutrition due to developmental and functional failure of the forestomach.

Distinct responses in feed sorting, chewing behavior, and ruminal acidosis risk between primiparous and multiparous Simmental cows fed diets differing in forage and starch levels.

During early lactation, both primiparous (PP) and multiparous (MP) cows are commonly fed diets rich in starch and low in forages to support their high energy requirements. Yet, the PP cows experience this dietary challenge for the first time, which might result in higher odds for them to develop rumen and systemic health disorders. The primary objective of this study was to evaluate the effect of decreasing the amount of forages in the diet on chewing and sorting behaviors and rumen and systemic health variables in PP and MP dairy cows. Twenty-four lactating Simmental cows [8 PP, average dry matter intake (DMI) of 19.1 ± 1.1 kg/d; 16 MP, average DMI of 22.5 ± 1.1 kg/d] with a body weight of 737 ± 90 kg and 50 ± 22 days in milk were used in this study. Cows were first fed a total mixed ration with 60% forage and 40% concentrate [on a dry matter (DM) basis] considered marginal in forages for 2 wk. Then, cows were switched to a diet low in forages with 40% forage and 60% concentrate (on a DM basis) for 4 wk. Reticular pH was measured continuously with wireless pH-sensors inserted into the reticulum to calculate the subacute ruminal acidosis (SARA) index. Chewing activity was measured with noseband-sensor halters, and feed sorting was measured weekly. Blood samples were collected weekly and analyzed for metabolic and inflammation markers. Switching PP and MP cows from a marginal to low-forage diet decreased the time spent eating and ruminating per kilogram of DM. Primiparous cows chewed longer per kilogram of DMI than MP cows. Also, the PP cows sorted more pronounced for longer particles and against fine particles than MP cows did. Despite higher rumination activity per kilogram of DMI and the adaptive sorting behavior, the PP cows spent on average 4.6 h/d longer below a pH of 5.8 and had a higher SARA index (i.e., area pH <5.8/DMI) than MP cows, especially during the first week of the low-forage diet (9.5 vs. 4.8). The concentration of liver enzymes increased with the low-forage diet, which was especially pronounced in the PP cows. In conclusion, this study demonstrated greater susceptibility of PP cows to SARA and liver damage than MP cows fed the same diets. Although PP cows demonstrated greater chewing and ruminating activity per kilogram of DMI, as well as adapted sorting behavior in favor of large particles during the low-forage high-starch feeding, they developed more severe signs of SARA. This suggests higher forage fiber requirements for PP cows and the need for improved feeding strategies to mitigate rumen fermentation disorders during early lactation in these cows.

Transcriptomic Changes in the Rumen Epithelium of Cattle after the Induction of Acidosis.

The transition from normal forage to a highly fermentable diet to achieve rapid weight gain in the cattle industry can induce ruminal acidosis. The molecular host mechanisms that occur in acidosis are largely unknown. Therefore, the histology and transcriptome profiling of rumen epithelium was investigated in normal and acidosis animals to understand the molecular mechanisms involved in the disease. The rumen epithelial transcriptome from acidosis (n=3) and control (n=3) Holstein steers was obtained using RNA-sequencing. The mean values of clean reads were 70,975,460&plusmn;984,046 and 71,142,189&plusmn;834,526 in normal and acidosis samples, respectively. In total, 1,074 differentially expressed genes were identified in the two groups (P&lt;0.05), of which 624 and 450 genes were up- and down-regulated in the acidosis samples, respectively. Functional analysis indicated that the majority of the up-regulated genes had a function in filament organization, positive regulation of epithelial and muscle fiber concentration, biomineral tissue development, negative regulation of fat cell differential, regulation of ion transmembrane transport, regulation of cell adhesion and butyrate, as well as short-chain fatty acid absorption that was metabolized as an energy source. Functional analysis of the down-regulated genes revealed effects in immune response, positive regulation of T-cell migration, regulation of metabolic processes, and localization. Furthermore, the results showed a differential expression of genes involved in the Map Kinase and Toll-like receptor signaling pathways. The IL1B, CXCL5, IL36A, and IL36B were significantly down-regulated in acidosis rumen tissue samples. The results suggest that rapid shifts to rich fermentable carbohydrates diets cause an increase in the concentration of ruminal volatile fatty acids, tissue damage, and significant changes in transcriptome profiles of rumen epithelial.

Genome-wide identification and characterization of novel long non-coding RNA in Ruminal tissue affected with sub-acute Ruminal acidosis from Holstein cattle.

Sub-acute ruminal acidosis is a type of metabolic disorder in which affected cattle show a considerable depression of rumen pH. This leads to a dramatic decline in productivity and consequent loss of income for many dairy farms. The objective of the present study is to identify and characterize novel long non-coding RNAs (lncRNAs) in Holstein cattle affected by sub-acute ruminal acidosis. Two replicates from six animals were sequenced that bioinformatically analyzed. Results showed 6679 novel lncRNAs among which 12 intergenic lncRNAs showed differential expression (p value ≤0.05). GO and KEGG analysis revealed that calcium signaling and G protein couple-receptor pathways may be involved in regulating metabolic processes during sub-acute ruminal acidosis. Furthermore, other biological processes including transmembrane transport, adult behavior, neuroactive ligand-receptor interaction, GABAergic synapse, cholinergic synapse were significantly enriched. The present data suggest that these differentially expressed lncRNAs may play regulatory roles in modulating biological processes associated with sub-acute ruminal acidosis in cattle rumen.

Physiological effects of starter-induced ruminal acidosis in calves before, during, and after weaning.

The objectives were to nutritionally induce or blunt ruminal acidosis in young calves and to compare indicators of rumen and systemic health. Ten bull calves (n = 5/diet) were ruminally cannulated at 3 wk of age and received milk replacer and 1 of 2 calf starter diets that were designed to cause (AC; pelleted, 42.7% starch, 15.1% neutral detergent fiber, 57.8% nonfiber carbohydrates) or blunt (BL; texturized, 35.3% starch, 25.3% neutral detergent fiber, 48.1% nonfiber carbohydrates) ruminal acidosis. Mean birth weight was 38.7 ± 1.3 kg. Body weight and calf starter intake were measured weekly. Rumen contents were sampled at -8, -4, 0, 2, 4, 8, 12, and 24 h relative to starter feeding during wk 6, 8, 10, 12, 14, and 16 of age. Blood was collected from the jugular vein during the same weeks for complete blood cell count, blood pH, and partial pressures of oxygen and carbon dioxide. Rate of starter consumption was assessed during wk 16. Marker systems were used to estimate liquid passage and volatile fatty acid absorption rates. Calves were slaughtered at 17 wk, and rumen tissue was collected and assessed for papillae length, width, and degree of tissue degradation. Mean ruminal pH ± standard error was 5.37 ± 0.24 and 5.63 ± 0.24 for AC and BL calves, respectively. Lowest pH values were observed the week after weaning. Total ruminal volatile fatty acid concentrations were 131.5 and 124.8 ± 2.4 mM in AC and BL calves, respectively, and increased with age and time after feeding. Dry matter intake was lower in AC calves at wk 4 and remained lower through wk 16. Rate of starter consumption was also lower in AC calves at wk 16. Body weight also was also lower for AC calves from wk 5 through 16. Blood hemoglobin and hematocrit were lower in AC calves, but other blood characteristics were not different. Rumen volume increased with age and tended to be greater in BL calves. Passage rate and papillae length and width were not different between diets, but AC calves experienced a greater degree of tissue degradation. Ruminal acidosis symptoms in calves appear similar to those in adult cattle, and the etiology of the disease seems to follow similar mechanisms. It is clear from this study that symptoms can be moderated by diet, but further research is needed to determine whether symptoms can be nutritionally prevented or whether calves that experience ruminal acidosis are more susceptible to the disease as adults.

Treatment of acute rumen lactic acidosis with intravenous hypertonic sodium chloride or bicarbonate solutions followed by intraruminal water.

The effectiveness of 7% hypertonic saline solution (HSS) and 6% hypertonic bicarbonate solution (HBS) followed by intraruminal water administration for the treatment of acute rumen lactic acidosis (ARLA) has not been evaluated yet. The hypothesis of the study is that treatment with HBS causes a faster correction of imbalances and clinical recovery than treatment with HSS. ARLA was induced in six healthy mature female sheep with sucrose twice. The sheep received both treatment regimens in a cross over design. After 18 h of induction, rumen lavage was performed and IV infusion of 7% HSS (4 mL/kg BW) or 6% HBS (6.7 mL/kg BW) was done over 5 min, followed by intraruminal administration of water (8% BW). The solutions provided 4.8 mmol/kg BW of sodium. Physical and laboratory tests were performed for 168 h. Both treatments increased plasma volume. After HSS treatment, pH and HCO3- returned to baseline values at 144 h and BE at 168 h. With HBS treatment, pH returned to baseline at 24 h, and HCO3- and BE at 48 h. In both treatments, the ruminal motility and the maximum hay intake were restored at 96 and 120 h, respectively. In conclusion, administration of HSS or HBS followed by intraruminal water corrects dehydration and mild-to-moderate metabolic acidosis in sheep with ARLA. Treatment with HBS promoted a faster correction of metabolic acidosis and could be indicated for the most severe cases.

Review: Metabolic challenges in lactating dairy cows and their assessment via established and novel indicators in milk.

The increasing lactational performance of dairy cows over the last few decades is closely related to higher nutritional requirements. The decrease in dry matter intake during the peripartal period results in a considerable mobilisation of body tissues (mainly fat reserves and muscle mass) to compensate for the prevailing lack of energy and nutrients. Despite the activation of adaptive mechanisms to mobilise nutrients from body tissues for maintenance and milk production, the increased metabolic load is still a risk factor for animal health. The prevalence of production diseases, particularly subclinical ketosis is high in the early lactation period. Increased ß-hydroxybutyrate (BHB) concentrations further depress gluconeogenesis, feed intake and the immune system. Despite a variety of adaptation responses to nutrient and energy deficit that exists among dairy cows, an early and non-invasive detection of developing metabolic disorders in milk samples would be useful. The frequent and regular milking process of dairy cows creates the ability to obtain samples at any stage of lactation. Routine identification of biomarkers accurately characterising the physiological status of an animal is crucial for decisive strategies. The present overview recapitulates established markers measured in milk that are associated with metabolic health of dairy cows. Specifically, measurements of milk fat, protein, lactose and urea concentrations are evaluated. Changes in the ratio of milk fat to protein may indicate an increased risk for rumen acidosis and ketosis. The costly determination of individual fatty acids in milk creates barriers for grouping of fatty acids into saturated, mono- and polyunsaturated fatty acids. Novel approaches include the potential of mid-IR (MIR) based predictions of BHB and acetone in milk, although the latter are not directly measured, but only estimated via indirect associations of concomitantly altered milk composition during (sub)clinical ketosis. Although MIR-based ketone body concentrations in milk are not suitable to monitor the metabolic status of the individual cow, they provide an estimate of the overall herd or specific groups of animals earlier in a particular stage of lactation. Management decisions can be made earlier and animal health status improved by adjusting diet composition.

[Feed refusal in fattening bulls because of maize silage contamination by Jimson weed (Datura stramonium)]. / Futterverweigerung bei Mastbullen durch eine Kontamination der Maissilage mit Gewöhnlichem Stechapfel (Datura stramonium).

A sudden decrease in feed intake (within 24 hours) without an impaired general condition occurred on a bull-rearing and -fattening farm in western Germany in December 2017. One day later, some of the bulls displayed signs of ruminal tympany (free gas bloat) and the reduction in feed intake became more severe. The maize silage was recognized as the cause of the observed symptoms, because clinical signs first appeared after taking the silage from a different localization within the bunker silo. Maize in this particular section was harvested from a separate area previously affected by wind damage (numerous bent maize plants), where no crop protection (herbicide application) was performed. Fruits and seeds of Jimson weed were identified in the incriminated maize silage samples. Following replacement of the contaminated parts of the maize silage with non-contaminated material and treatment of the clinically affected animals (anti-tympanic medication plus live yeast preparation), the feed intake returned to the normal level and the affected animals recovered within 6 days. The cause of the symptoms was probably the anti-nutritive properties of Datura stramonium that affect the motor function of the gastrointestinal tract.

Metabolic and stress responses in dairy cows fed a concentrate-rich diet and submitted to intramammary lipopolysaccharide challenge.

Feeding dairy cows diets rich in grain often leads to subacute rumen acidosis (SARA), which might affect their responsiveness to immunogenic stimuli such as exogenous lipopolysaccharide (LPS), and can lead to metabolic alterations. The main objective of this study was to investigate if SARA affects the stress and metabolic health responses resulting from an intramammary LPS challenge. Before the intramammary LPS challenge, the SARA cows showed higher blood glucose and a tendency for higher lactate and aspartate aminotransferase as well as a trend toward lower ß-hydroxybutyrate (BHBA) and γ-glutamyltransferase compared with control cows. After the LPS challenge, the serum cortisol concentration markedly increased and the calcium concentration decreased both in SARA and control cows. In SARA-LPS cows, however, the lactate concentration increased due to the LPS infusion, whereas it remained unchanged in the control cows. A lower serum BHBA concentration was found in SARA-LPS compared with control-LPS cows. Higher non-esterified fatty acid concentrations were found in control-LPS cows shortly before the LPS challenge compared with SARA cows, challenged or not with LPS, whereas it did not differ from SARA-LPS cows thereafter. In conclusion, the results suggest that intramammary LPS challenge induced stress and lowered calcium concentration in all dairy cows, whereby this challenge showed lower BHBA and higher lactate levels in cows with SARA conditions.

Epigenetic mechanisms contribute to decrease stearoyl-CoA desaturase 1 expression in the liver of dairy cows after prolonged feeding of high-concentrate diet.

Subacute ruminal acidosis (SARA) of dairy cattle is a widely occurring but not very overt metabolic disorder thought to impair milk composition. The enzyme stearoyl-CoA desaturase 1 (SCD1) is rate-limiting for the formation of Δ-9 unsaturated fatty acids and thus crucially involved in controlling lipid metabolism in the liver. It is known that SCD1 expression is downregulated during SARA, but the underlying molecular mechanisms are unknown. To study these mechanisms, we enrolled 12 healthy multiparous mid-lactation Holstein cows into a diet-induced SARA experiment. Six cows were fed a high-concentrate diet for 18 weeks (60% content of high-concentrate to 40% forage; HC group), whereas the others received a low-concentrate diet ad libitum (40% high-concentrate content to 60% forage; LC group). Sustained low ruminal pH values (pH 5.6 maintained for 4 h/d) and reduced milk yield performance (2.07 kg/d less than LC cows) verified that SARA had been induced in the HC group. Results showed a significantly decreased concentrations of cis-9 monounsaturated long-chain fatty acids in plasma collected from hepatic but not portal veins. This was matched by reduced SCD1 mRNA and protein concentrations in HC livers. The expression levels of genes related to lipid formation (DGAT1 and PLIN2) were downregulated during SARA, whereas those of catabolic genes (CPT1A, CPT2, and ACOX1) and some inflammatory genes were upregulated. Expression of SCD1 was downregulated through reduced transcription and abundance of the transcription factor sterol regulatory element-binding protein 1 (SREBP1c).This effect was augmented by local chromatin tightening and DNA methylation at and around the SREBP1c binding site in the SCD1 promoter. Chromatin immunoprecipitation assays confirmed that SARA reduced SREBP1c binding at the SCD1 promoter; hence, epigenetic mechanisms are involved in regulating the expression of genes related to long-chain fatty acid modification, partially through downregulation of both SCD1 and SREBP1c in the liver. Our results suggest that in addition to inflammatory genes, SCD1 is also involved in SARA-induced epigenetic regulation and its associated metabolic changes. This knowledge might help to provide a target for intervening against the detrimental metabolic effects of SARA.

Invited review: Practical feeding management recommendations to mitigate the risk of subacute ruminal acidosis in dairy cattle.

Rumen health is of vital importance in ensuring healthy and efficient dairy cattle production. Current feeding programs for cattle recommend concentrate-rich diets to meet the high nutritional needs of cows during lactation and enhance cost-efficiency. These diets, however, can impair rumen health. The term "subacute ruminal acidosis" (SARA) is often used as a synonym for poor rumen health. In this review, we first describe the physiological demands of cattle for dietary physically effective fiber. We also provide background information on the importance of enhancing salivary secretions and short-chain fatty acid absorption across the stratified squamous epithelium of the rumen; thus, preventing the disruption of the ruminal acid-base balance, a process that paves the way for acidification of the rumen. On-farm evaluation of dietary fiber adequacy is challenging for both nutritionists and veterinarians; therefore, this review provides practical recommendations on how to evaluate the physical effectiveness of the diet based on differences in particle size distribution, fiber content, and the type of concentrate fed, both when the latter is part of total mixed ration and when it is supplemented in partial mixed rations. Besides considering the absolute amount of physically effective fiber and starch types in the diet, we highlight the role of several feeding management factors that affect rumen health and should be considered to control and mitigate SARA. Most importantly, transitional feeding to ensure gradual adaptation of the ruminal epithelium and microbiota; monitoring and careful management of particle size distribution; controlling feed sorting, meal size, and meal frequency; and paying special attention to primiparous cows are some of the feeding management tools that can help in sustaining rumen health in high-producing dairy herds. Supplementation of feed additives including yeast products, phytogenic compounds, and buffers may help attenuate SARA, especially during stress periods when the risk of a deficiency of physically effective fiber in the diet is high, such as during early lactation. However, the usage of feed additives cannot fully compensate for suboptimal feeding management.

Key role of short-chain fatty acids in epithelial barrier failure during ruminal acidosis.

Subacute ruminal acidosis is induced by high concentrations of short-chain fatty acids (SCFA, mainly acetate, propionate, and butyrate) that release protons to decrease the pH of the ruminal digesta. This low pH, in turn, is thought to damage epithelial barrier function. The present study applied a model of simulated ruminal acidosis ex vivo to investigate if SCFA directly contribute to epithelial barrier failure beyond their role as proton donors. Epithelial tissues from the rumen of slaughtered sheep were mounted in Ussing chambers and incubated under 3 different conditions. Two groups were incubated in the absence of SCFA at mucosal pH 6.1 (control) and pH 5.1, respectively, for 7 h. A third group was first incubated in a mucosal solution containing 100 mM SCFA at pH 5.1 for 2 h and, thereafter, in a mucosal solution without SCFA at pH 6.1 for the remaining 5 h. Transepithelial conductance (Gt), short-circuit current (Isc), and fluorescein fluxes were determined. After 7 h of incubation, the expression levels of claudin-1, claudin-4, claudin-7, and occludin were measured by quantitative reverse-transcription PCR and Western blot. Furthermore, the local distribution of these tight junction (TJ) proteins was examined by confocal laser scanning microscopy. A 7-h incubation at pH 5.1 in the absence of SCFA did not influence either Gt or fluorescein flux rates of ruminal tissues ex vivo compared with the control. In contrast, incubation at pH 5.1 with SCFA for only 2 h induced increases in Gt and fluorescein flux rates that continued even after tissues were returned back to pH 6.1. Expression analysis showed that pH 5.1 without SCFA for 7 h induced no changes in mRNA expression of claudin-1, claudin-4, claudin-7, and occludin and a selective decrease in protein expression of only claudin-4 compared with the control. However, a 2-h incubation at pH 5.1 in the presence of SCFA decreased the mRNA-expression of claudin-7, as well as the protein expression of claudin-4, claudin-7, and occludin. The decreased expression of these TJ proteins in the group incubated with SCFA was also evident in immunohistochemistry. Immunohistochemistry additionally evidenced a considerable retraction of all tested TJ proteins out of the TJ in that group. We conclude that a low mucosal pH of 5.1 is tolerated well by ruminal epithelia for several hours. However, a low pH in combination with SCFA induces damage to the TJ and disturbs barrier function, which is not immediately reversible upon the removal of the acidotic insult.

Towards practical application of sensors for monitoring animal health: the effect of post-calving health problems on rumination duration, activity and milk yield.

Three sources of sensory data: cow's individual rumination duration, activity and milk yield were evaluated as possible indicators for clinical diagnosis, focusing on post-calving health problems such as ketosis and metritis. Data were collected from a computerised dairy-management system on a commercial dairy farm with Israeli Holstein cows. In the analysis, 300 healthy and 403 sick multiparous cows were studied during the first 3 weeks after calving. A mixed model with repeated measurements was used to compare healthy cows with sick cows. In the period from 5 d before diagnosis and treatment to 2 d after it, rumination duration and activity were lower in the sick cows compared to healthy cows. The milk yield of sick cows was lower than that of the healthy cows during a period lasting from 5 d before until 5 d after the day of diagnosis and treatment. Differences in the milk yield of sick cows compared with healthy cows became greater from 5 to 1 d before diagnosis and treatment. The greatest significant differences occurred 3 d before diagnosis for rumination duration and 1 d before diagnosis for activity and milk yield. These results indicate that a model can be developed to automatically detect post-calving health problems including ketosis and metritis, based on rumination duration, activity and milk yield.

Rumination time and reticuloruminal temperature as possible predictors of dystocia in dairy cows.

The objectives of this study were to explore changes of rumination time and reticuloruminal pH and temperature of dairy cows and heifers (means ± standard deviation; age = 5.8 ± 1.9; parity = 2.7 ± 1.4; body condition score = 3.2 ± 0.2) with eutocic (EUT, n = 10) and dystocic calving (DYS, n = 8). The recording period lasted from 3 d before calving until 7 d in milk. For the comparison of rumination time and reticuloruminal characteristics between groups, time to return to baseline (the time interval required to return to baseline from the delivery of the calf) and area under the curve (AUC; both for prepartum and postpartum periods) were calculated for each parameter. Rumination time decreased from baseline 28 h before calving both for EUT and DYS cows; after 20 h before calving, it decreased to 32.4 ± 2.3 and 13.2 ± 2.0 min/4 h between 8 and 4 h before delivery in EUT and DYS cows, respectively, and then it decreased below 10 and 5 min during the last 4 h before calving. Until 12 h after delivery, rumination time reached 42.6 ± 2.7 and 51.0 ± 3.1 min/4 h in DYS and EUT dams, respectively; however, AUC and time to return to baseline suggested lower rumination activity in DYS cows than in EUT dams for the 168-h postpartum observational period. Reticuloruminal pH decreased from baseline 56 h before calving both for EUT and DYS cows, but did not differ between groups before delivery. Reticuloruminal pH showed a decreasing tendency and clear diurnal variation after calving for both EUT and DYS cows, with slightly higher AUC values in DYS cows. In DYS cows, reticuloruminal temperature decreased from baseline 32 h before calving by 0.23 ± 0.02°C, whereas in EUT cows such a decrease was found only 20 h before delivery (0.48 ± 0.05°C). The AUC of reticuloruminal temperature calculated for the prepartum period was greater in EUT cows than in DYS cows. During the first 4 h after calving, reticuloruminal temperature decreased from 39.68 ± 0.09 to 38.96 ± 0.10°C and from 39.80 ± 0.06 to 38.81 ± 0.08°C in EUT and DYS cows, respectively, and reached baseline levels after 35.4 ± 3.4 and 37.8 ± 4.2 h after calving in EUT and DYS cows, respectively. Based on our results, continuous monitoring of changes in rumination time and reticuloruminal temperature seems to be promising in the early detection of cows with a higher risk of dystocia. Depressed rumination activity of DYS cows after calving highlights the importance of the postpartum monitoring of cows experiencing difficulties at calving. The effect of dystocia on postpartum reticuloruminal pH was not pronounced.

Technical note: Preliminary evaluation of an automated indwelling rumen temperature bolus measurement system to detect pyrexia in preweaned dairy calves.

Indwelling rumen temperature bolus (RTB) systems have the potential to offer a convenient and timely method of detecting pyrexia, indicative of active infectious disease. The aim of this pilot study was to evaluate the utility of using RTB systems in preweaned dairy calves. First, an in vitro study was performed to evaluate the accuracy of the RTB in its immediate environment. Thirteen RTB were immersed in a hot water bath (WB). Variably collected RTB temperatures were then matched to WB temperatures, which varied from 36 to 41°C, with 1h spent at each temperature. Second, an in vivo study was performed to evaluate the ability of the RTB to predict a rectal thermometer (RT) temperature. Ten healthy heifer calves less than 1wk of age were administered an RTB. Rectal thermometer and matched RTB temperatures were taken hourly, over a 6-h period, 1 day per week during wk 1, 3, 5, 7, and 9 of age. During each 6-h observation period, calves were offered both water and milk feedings and temperatures recorded every 15min for 1h thereafter. For both studies, the relationship between RTB and one of WB (in vitro) or RT temperature (in vivo) was described by calculating a concordance correlation coefficient (CCC) and by use of a multivariable linear regression model with repeated measures. For the in vivo study, the model also controlled for week and breed. Diagnostic test characteristics were calculated for the ability of individual RTB measures to detect pyrexia (RT ≥39.5°C). For the in vitro study, the association between the RTB and a known temperature was strong (CCC=0.95), but the RTB measures underestimated the temperature of the water bath by 0.43±0.08°C. For the in vivo study, the association between RT and the RTB temperature measurement in a calf was weaker (CCC=0.29); the average RTB temperature was 0.33±0.06°C lower than the RT temperature. The sensitivity (29%) and positive predictive value (17%) of using individual RTB measures to detect a fever was low. The results of this pilot study suggest that an individual RTB measurement may not be a good diagnostic test to detect pyrexia in calves.

Timpanismo em bovinos, secundário à obstrução esofágica por citrus limon (limão siciliano) / Bloat in cattle secondary to esophageal obstruction by citrus limon (sicilian lemon)

Timpanismo ruminal consiste na distensão acentuada do rúmen e retículo, devido a incapacidade do animal em expulsar gases produzidos durante o processo fisiológico da fermentação. O timpanismo pode ser ocasionado de forma primária, por formação de espuma estável, ou secundária, devido a anormalidades funcionais e/ou físicas que interferem na eructação. Nesse trabalho, são descritos os aspectos epidemiológicos, clínicos e anatomopatológicos da ocorrência de timpanismo secundário em bovinos, decorrente da obstrução esofágica aguda por limões sicilianos. Cinco bovinos, de um lote de 210, foram afetados. Os bovinos eram suplementados com resíduo de tangerina (Citrus reticulata) no cocho e na última carga desse subproduto, havia limões sicilianos inteiros misturados ao resíduo. Os cinco animais afetados eram da raça Aberdeen Angus e tinham entre 12-24 meses de idade. Todos apresentaram sinais clínicos caracterizados principalmente por mucosas cianóticas, grave timpanismo, desconforto abdominal, acentuada dispneia e taquicardia, atonia ruminal, desidratação, decúbito e morte. O curso clínico variou entre 24 a 48 horas. Na necropsia, os cinco bovinos apresentavam grave obstrução esofágica por limões nas porções: cranial (logo após a laringe [1/5]), porção medial (entrada do tórax [1/5]) e final (próximo ao cárdia [3/5]). Nas áreas de oclusão, observou-se extensa necrose e ulceração da mucosa esofágica. O conteúdo ruminal dos bovinos estava seco e misturado com limões inteiros. No esôfago de dois animais havia linha de timpanismo. As lesões histológicas eram visualizadas principalmente no esôfago, na região da obstrução, onde se evidenciaram alterações degenerativas, necróticas e ulcerativas acentuadas no revestimento epitelial.

Ruminal bloat (acute timpany) in ruminants is a marked rumen-reticular distension which results from more gas being produced during the physiologic process of fermentation than is eliminated by eructation. This condition may present itself as either primary timpany due to the formation of stable foam or secondary timpany resulting from functional and/or physical disturbances compromising eructation. This paper describes the epidemiological, clinical, and anatomopathological aspects of acute timpany in cattle secondary to esophageal obstruction by sicilian lemons. Five out of a herd of 210 cattle were affected. Cattle were supplemented with tangerine (Citrus reticulata) residues in a trough. In the last batch of this feed there were whole sicilian lemons mixed with the tangerine residue. The five affected cattle were 12-24 month-old Aberdeen-Angus. All of the five presented clinical signs characterized mainly by cyanotic mucous membranes, severe timpany, abdominal discomfort, marked dyspnea and tachycardia, ruminal atony, dehydration, recumbence and death. Clinical course lasted from 24 to 48 hours. Necropsy findings in the five affected cattle were similar and included complete esophageal obstruction by lemons in the cranial esophagus (immediately cranial to the larynx [1/5]) medial esophagus (at the thoracic inlet [1/5]) and caudal esophagus (close to the cardia [3/5]). At the occluded sites the esophageal mucosa was necrotic and ulcerated. Ruminal content was dried and admixed with whole lemons. In the esophagus o two affected bovine a bloat line was observed. Histological lesions were observed mainly in the esophagus at the sites of obstruction and consisted of marked degenerative, necrotic and ulcerative changes in the esophageal mucosal epithelium.

Short communication: Noninvasive indicators to identify lactating dairy cows with a greater risk of subacute rumen acidosis.

The objective of the current study was to evaluate if milk urea nitrogen (MUN) and milk fat content could be used as the noninvasive indicator to identify cows with greater or lower risk of subacute ruminal acidosis (SARA). Our hypothesis was that cows with lower MUN and milk fat content would have greater risk of SARA, whereas cows with higher MUN and milk fat content would have lower risk of SARA. In the screening study, 35 late-lactating Holstein cows (DIM=250±71.1; BW=601±45.4kg) were fed a high-grain diet containing 35% forage and 65% concentrate mix ad libitum for 21 d. Concentration of MUN ranged from 5.7 to 13.9Mg/dL among the 35 cows, and the average milk fat content was 3.5%. Then, 5 cows with highest MUN concentrations with milk fat higher than 3.5% were selected as animals that presumably have low risk of SARA, and 5 cows with lowest MUN concentrations with milk fat less than 3.5% were selected as animals that presumably have high risk of SARA. These 10 animals were ruminally cannulated during the subsequent dry period. As 1 low-risk cow was culled due to fatty liver, 9 animals (DIM=122±33.2; BW=615±49.1kg) were used in the subsequent study in the following lactation. All cows were fed a high-grain diet consisting of 35% forage and 65% concentrate mix ad libitum for 21 d. Ruminal pH was measured every 30 s for 72 h. Minimum (5.75 vs. 5.30) and mean ruminal pH (6.35 vs. 6.04) was higher for low- compared with high-risk animals. In addition, duration of rumen pH below 5.8 was shorter in low-risk animals (52.5 vs. 395min/d). These results suggested that MUN and milk fat content in late-lactating cows fed a high-grain diet may be used to identify cows that have higher or lower risk of SARA.

Effects of a selection of additives on in vitro ruminal methanogenesis and in situ and in vivo†NDF digestibility.

The effects of 18 essential oils, yeast, Quebracho tannin and Quillaja saponin on ruminal methane and gas production (GP) were studied in vitro. A lactating cow diet was incubated with rumen inoculum. Doses of the additives (mg/L) were: essential oils = 500; tannin = 67; yeast = 8.35 and 16.7; and saponin = 300. Lemongrass, estragole, eugenol, geraniol, limonen, thyme oil and thymol produced less gas (overall mean 33.8 mL/200 mg dry matter (DM)) than control (43.6 mL/200 mg DM; P < 0.001). Methane produced (mL/200 mg DM) by guaiacol (10.7), lemongrass (9.6), limonene (11.4), thyme oil (10.9) and thymol (2.1) was lower than control (12.5) (P < 0.001). Methane percentage on total GP was lower (P < 0.001) for cinnamic alcohol (25.4), guaiacol (24.5), thymol (19.7) and vanillin (26.3) than control (28.8). In a second experiment, thymol, guaiacol and yeast were added to the diet of dry fistulated cows to determine in situ neutral detergent fiber digestibility (NDFD) of six forages and in vivo dietary NDFD. Thymol and yeast decreased in situ NDFD after 24 h (33.9% and 33.5% vs. 38.1%; P = 0.008). Thymol decreased in vivo total tract NDFD (40.8% vs. 51.4%; P = 0.02). Differences in GP and methane levels were registered within classes of additives. A careful selection of additives may allow for the manipulation of ruminal fermentation.

Transabdominal ultrasonography of the ruminal mucosa as a tool to diagnose subacute ruminal acidosis in adult dairy bulls: a pilot study.

Subacute rumen acidosis (SARA) is frequently encountered in ruminants on high-concentrate rations and characterized by mild to moderate rumen pH depression. Although the measurement of rumen pH is considered as a gold standard approach to diagnose SARA, rumen fluid collection is conceived as a cumbersome and invasive procedure. In the present study, the suitability of transabdominal ultrasonography to identify structural changes of the rumen mucosa associated with SARA was explored. Five adult rumen canulated bulls previously adjusted to a roughage-based ration were switched in 10-day intervals to rations with increasing concentrate content. Seven rations with a concentrate content ranging from 5% to 95% were fed. The rumen pH and rumen mucosal thickness were measured at the end of each 10-d feeding period. The left flank was divided into 12 acoustic windows and the mean rumen mucosal thickness for each acoustic window determined for each ration. Repeated measure ANOVA and stepwise regression analyses were conducted. Increasing the dietary concentrate content resulted in thickening of rumen mucosa which was most pronounced in the upper part of the ventral rumen sac and the least pronounced in the uppermost and lowermost parts of the rumen. The intersection of a horizontal line going through the costochondral junction and a vertical line coming from the third lumbar vertebra was found to be most suitable to identify animals with rumen pH < 5.5 at 4 h post feeding. We conclude that transabdominal ultrasonography of the rumen mucosa has the potential to be a suitable, non-invasive diagnostic tool to identify adult ruminants with SARA.