Subclinical (SCK) and clinical (CK) ketosis are metabolic disorders responsible for big losses in dairy production. Although Fourier-transform mid-infrared spectrometry (FTIR) to predict ketosis in cows exposed to great metabolic stress was studied extensively, little is known about its suitability in predicting hyperketonemia using individual samples, e.g. in small dairy herds or when only few animals are at risk of ketosis. The objective of the present research was to determine the applicability of milk metabolites predicted by FTIR spectrometry in the individual screening for ketosis. In experiment 1, blood and milk samples were taken every two weeks after calving from Holstein (n = 80), Brown Swiss (n = 72) and Swiss Fleckvieh (n = 58) cows. In experiment 2, cows diagnosed with CK (n = 474) and 420 samples with blood β-hydroxybutyrate [BHB] <1.0 mmol/l were used to investigate if CK could be detected by FTIR-predicted BHB and acetone from a preceding milk control. In experiment 3, correlations between data from an in farm automatic milk analyser and FTIR-predicted BHB and acetone from the monthly milk controls were evaluated. Hyperketonemia occurred in majority during the first eight weeks of lactation. Correlations between blood BHB and FTIR-predicted BHB and acetone were low (r = 0.37 and 0.12, respectively, P < 0.0001), as well as the percentage of true positive values (11.9 and 16.6%, respectively). No association of FTIR predicted ketone bodies with the interval of milk sampling relative to CK diagnosis was found. Data obtained from the automatic milk analyser were moderately correlated with the same day FTIR-predicted BHB analysis (r = 0.61). In conclusion, the low correlations with blood BHB and the small number of true positive samples discourage the use of milk mid-infrared spectrometry analyses as the only method to predict hyperketonemia at the individual cow level.

The purpose of the present review is to describe how human physiology at very low carbohydrate intakes relates to the criteria for nutritional essentiality. Although we did not limit ourselves to one particular type or function of carbohydrates, we did primarily focus on glucose utilisation as that function was used to determine the recommended daily allowance. In the general population, the human body is able to endogenously synthesise carbohydrates, and does not show signs of deficiency in the absence of dietary carbohydrates. However, in certain genetic defects, such as glycogen storage disease type I, absence of dietary carbohydrates causes abnormalities that are resolved with dietary supplementation of carbohydrates. Therefore, dietary carbohydrates may be defined as conditionally essential nutrients because they are nutrients that are not required in the diet for the general population but are required for specific subpopulations. Ketosis may be considered a physiological normal state due to its occurrence in infants in addition to at very low carbohydrate intakes. Although sources of dietary carbohydrates can provide beneficial micronutrients, no signs of micronutrient deficiencies have been reported in clinical trials of low-carbohydrate ketogenic diets. Nonetheless, more research is needed on how micronutrient requirements can change depending on the dietary and metabolic context. More research is also needed on the role of dietary fibre during a low-carbohydrate ketogenic diet as the beneficial effects of dietary fibre were determined on a standard diet and several studies have shown beneficial effects of decreasing non-digestible carbohydrates.

The experiments reported in this research communication aimed to compare the serum nonesterified fatty acid (NEFA) composition in ketotic cows and healthy cows during the perinatal period. NEFAs play significant roles in etiology and pathology of ketosis. We hypothesized that ketotic cows will display a different serum NEFA composition compared to healthy controls, and fatty acid related indicators for ketosis prediction can be screened. Pre-partum healthy cows were recruited, and blood samples were collected on −7, 3, 7, 14 and 21 d postpartum. Cows were further divided into a healthy control group (C group, n = 6) and a ketosis group (K group, n = 6) if blood β-hydroxybutyric acid levels exceeded 1.2 mm during the experiment. NEFA composition was then analyzed by means of Gas Chromatography-Mass Spectrometer (GC-MS). Only C12 : 0% was significantly higher in C group than K group on 7 d pre-partum (P < 0.05), when the cows were not diagnosed with ketosis. Five fatty acids displayed statistical differences in composition between C and K group (P < 0.05), namely C12 : 0, C16 : 0, C17 : 0, C18 : 1n9 and C22 : 1n9. Saturates%, unsaturates%, mono-unsaturates% and saturates/unsaturates were also different between C and K group (P < 0.05). Of note, C18 : 1n9/C12 : 0 and C18 : 1n9/C22 : 1n9 in K group were significantly higher than those in controls on 7 d pre-partum (P < 0.05). It is suggested that the ratios show potential as indicators for prediction of ketosis.

Nutritional ketosis, induced via either the classical ketogenic diet or the use of emulsified medium-chain triglycerides, is an established treatment for pharmaceutical resistant epilepsy in children and more recently in adults. In addition, the use of oral ketogenic compounds, fractionated coconut oil, very low carbohydrate intake, or ketone monoester supplementation has been reported to be potentially helpful in mild cognitive impairment, Parkinson’s disease, schizophrenia, bipolar disorder, and autistic spectrum disorder. In these and other neurodegenerative and neuroprogressive disorders, there are detrimental effects of oxidative stress, mitochondrial dysfunction, and neuroinflammation on neuronal function. However, they also adversely impact on neurone–glia interactions, disrupting the role of microglia and astrocytes in central nervous system (CNS) homeostasis. Astrocytes are the main site of CNS fatty acid oxidation; the resulting ketone bodies constitute an important source of oxidative fuel for neurones in an environment of glucose restriction. Importantly, the lactate shuttle between astrocytes and neurones is dependent on glycogenolysis and glycolysis, resulting from the fact that the astrocytic filopodia responsible for lactate release are too narrow to accommodate mitochondria. The entry into the CNS of ketone bodies and fatty acids, as a result of nutritional ketosis, has effects on the astrocytic glutamate–glutamine cycle, glutamate synthase activity, and on the function of vesicular glutamate transporters, EAAT, Na+, K+-ATPase, Kir4.1, aquaporin-4, Cx34 and KATP channels, as well as on astrogliosis. These mechanisms are detailed and it is suggested that they would tend to mitigate the changes seen in many neurodegenerative and neuroprogressive disorders. Hence, it is hypothesized that nutritional ketosis may have therapeutic applications in such disorders.

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 β-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.

Dairy cows with ketosis display severe oxidative stress as well as high blood concentrations of non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHB). Cytochrome P4502E1 (CYP2E1) plays an important role in the induction of oxidative stress. The aim of this study was to investigate CYP2E1 expression and activity in the liver of clinically ketotic cows (in vivo) and the effects of NEFA and BHB on CYP2E1 expression and activity in hepatocytes (in vitro). Dairy cows with clinical ketosis exhibited a low blood concentration of glucose but high concentrations of NEFA and BHB. Hepatic mRNA, protein expression, and activity of CYP2E1 were significantly higher in cows with clinical ketosis than in control cows. In vitro, both NEFA and BHB treatment markedly up-regulated the mRNA and protein expressions as well as activity of CYP2E1 in cow hepatocytes. Taken together, these results indicate that high levels of NEFA and BHB significantly up-regulate the expression and activity of hepatic CYP2E1, and may be influential in the induction of oxidative stress in cows with clinical ketosis.

High-yielding cows may suffer from negative energy balance during early lactation, which can lead to ketosis and delayed ability of returning to cyclicity after calving. Fast recovery after calving is essential when breeding for improved fertility. Traditionally used fertility traits, such as the interval from calving to first insemination (CFI), have low heritabilities and are highly influenced by management decisions. Herd Navigator™ management program samples and analyses milk progesterone and β-hydroxybutyrate (BHB) automatically during milking. In this study, the genetic parameters of endocrine fertility traits (measured from milk progesterone) and hyperketonemia (measured from milk BHB) in early lactation were evaluated and compared with traditional fertility traits (CFI, interval from calving to the last insemination and interval from first to last insemination) and the milk yield in red dairy cattle herds in Finland. Data included observations from 14 farms from 2014 to 2017. Data were analyzed with linear animal models using DMU software and analyses were done for first parity cows. Heritability estimates for traditional fertility traits were low and varied between 0.03 and 0.07. Estimated heritabilities for endocrine fertility traits (interval from calving to the first heat (CFH) and commencement of luteal activity (C-LA)) were higher than for traditional fertility traits (0.19 to 0.33). Five slightly different hyperketonemia traits divided into two or three classes were studied. Linear model heritability estimates for hyperketonemia traits were low, however, when the threshold model was used for binary traits the estimates became slightly higher (0.07 to 0.15). Genetic correlation between CFH and C-LA for first parity cows was high (0.97) as expected since traits are quite similar. Moderate genetic correlations (0.47 to 0.52) were found between the endocrine fertility traits and early lactation milk yield. Results suggest that the data on endocrine fertility traits measured by automatic systems is a promising tool for improving fertility, specifically when more data is available. For hyperketonemia traits, dividing values into three classes instead of two seemed to work better. Based on the current study and previous studies, where higher heritabilities have been found for milk BHB traits than for clinical ketosis, milk BHB traits are a promising indicator trait for resistance to ketosis and should be studied more. It is important that this kind of data from automatic devices is made available to recording and breeding organizations in the future.

High concentrations of non-esterified fatty acid (NEFA) and β-hydroxybutyrate (BHBA) in cows’ blood caused by ketosis are associated with inflammatory states. We hypothesised that ketosis in postparturient dairy cows would result in altered levels on inflammation-related proteins not only in plasma but also in the milk fat globule membranes (MFGM). Thirty cows were selected from a dairy farm in Heilongjiang, China. Inflammatory milk fat globule membrane proteins were detected using ELISA kits, and a fully automatic biochemical analyser was used to measure the concentrations of BHBA, NEFA, glucose (GLU) and triglyceride (TG) in plasma. MFGM protein from milk of ketotic cows contained significantly different concentrations of acute-phase response proteins (complement C3 (C3), prothrombin (F2), alpha-1-acid glycoprotein (ORM1), inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4), alpha-2-HS-glycoprotein (AHSG), complement C9 (C9), complement regulatory protein variant 4 (CD46)) in comparison with milk from non-ketotic cows. Blood concentrations of C3, complement C9 (C9), tumour necrosis factor α (TNFα), MFGM C3, monocyte differentiation antigen CD14 (CD14) and ORM1 levels were correlated with energy balance. ITIH4 and CD46 increased, and AHSG and ORM1 decreased before the onset of ketosis. These biomarkers offer potential as predictors and monitors of ketosis in at-risk cows.

In this Research Paper we hypothesised that the temporary insulin resistance seen during the transition period in dairy cows may cause significant differences in the efficacy of PG at different sampling periods and that in some cases this effect will be dose dependent. Eighty four sampling sets were generated by studying 7 multiparous Holstein cows repeatedly at 4 sampling periods of 3 d length (dry cows: days 40, 39 and 38 antepartum; close up cows: days 10, 9 and 8 antepartum; fresh cows: days 3, 4 and 5 post-partum; lactating cows: days 38, 39 and 40 post-partum). On each of these days 3 h after morning feeding propylene glycol was drenched in different dosages of 100, 300 or 500 ml once per day (cross over study). The different doses were applied in an alternating order (Latin square). Blood samples were taken before, every 30 min up to 4 h, after 6 and 12 h after PG application. Following parameters have been measured: insulin, non-esterified fatty acids (NEFA), betahydroxybutyrate (BHB), bilirubin, cholesterol, potassium, aspartate aminotransferase (AST) and glutamate dehydrogenase (GLDH). Revised Quantitative Insulin Sensitivity Check Index (RQUICKI) was calculated. It was found that glucose, insulin, NEFA, BHB, bilirubin and potassium concentrations were influenced differently by the three defined dosages of propylene glycol at four different sampling periods. Whereas RQUICKI, cholesterol, AST and GLDH did not differ between the sampling periods and treatments. The major results of the study are that the effect of PG is dose-dependent and that the effect of PG is depending on the time of application according to calving. It can be concluded that in fresh cows higher dosages are necessary to provoke similar effects in comparison to dry, close up and lactating cows. Although the study did not compare to topdressing of PG from the results it is reasonable to believe that bolus application of a specific PG volume is necessary to provoke the effect.

The objective of this study was to design and validate a mathematical model to detect post-calving ketosis. The validation was conducted in four commercial dairy farms in Israel, on a total of 706 multiparous Holstein dairy cows: 203 cows clinically diagnosed with ketosis and 503 healthy cows. A logistic binary regression model was developed, where the dependent variable is categorical (healthy/diseased) and a set of explanatory variables were measured with existing commercial sensors: rumination duration, activity and milk yield of each individual cow. In a first validation step (within-farm), the model was calibrated on the database of each farm separately. Two thirds of the sick cows and an equal number of healthy cows were randomly selected for model validation. The remaining one third of the cows, which did not participate in the model validation, were used for model calibration. In order to overcome the random selection effect, this procedure was repeated 100 times. In a second (between-farms) validation step, the model was calibrated on one farm and validated on another farm. Within-farm accuracy, ranging from 74 to 79%, was higher than between-farm accuracy, ranging from 49 to 72%, in all farms. The within-farm sensitivities ranged from 78 to 90%, and specificities ranged from 71 to 74%. The between-farms sensitivities ranged from 65 to 95%. The developed model can be improved in future research, by employing other variables that can be added; or by exploring other models to achieve greater sensitivity and specificity.

A time series model was used to describe the daily milk yield of healthy cows in the first 48 days of lactation. A moving average (MA) model of order 1 on the first-order differences of the data was selected — the same chosen in a previous study by Deluyker et al. (1990). When the model was used to generate predicted daily yields in a second data set, for cows in which clinical ketosis had been diagnosed, it was found that significant deviations of actual yield below the daily forecast occurred from 3 days before the day of diagnosis. The model appeared to be transportable to healthy cows from another herd. Threshold values were defined to identify ailing animals by their deviation from predicted yield. However, the thresholds were not very sensitive, and required that a fairly high level of false positives be accepted (25% of a healthy herd over a 5-day period).

Various dairy cattle production and health characteristics were studied with a view to identify easily available and measurable factors associated with the incidence of ketosis. The analyses were carried out using data from the Langhill Dairy Cattle Research Centre. Two approaches were used to assess the relative risk to cows of getting ketosis: one using information known at the beginning of lactation and one using information collected as the lactation progressed. In both approaches analyses were carried out using different amounts of the available information to simulate differences between recording systems. In the first approach the following were found to relate significantly to the level of recorded ketotic incidents: parity; ketosis in the previous lactation; calving condition score; 305-day milk yield in the previous lactation; and the average milk protein percentage in the previous lactation. The effects of these were quantified. In the second approach, where the change in ketosis incidence rate over the weeks of lactation was investigated, the average dry-matter intake in the previous week and changes in live weight and body condition score over the previous week were found to have a significant effect on the probability of getting ketosis in the coming week. The risk assessments varied depending on the information used and a flexible approach is recommended if potential risk factors are to be successfully incorporated into decision support systems.

Acetone concentration in milk is correlated with subclinical and clinical ketosis and also with milk yield and reproductive efficiency. The purpose of this study was to assess a threshold of milk acetone at which cows tend to be adversely affected. The critical values were estimated on data from 11690 lactations where the change in daily milk yield from weeks 1 to 2 to weeks 5 to 6 of lactation was evaluated in relation to different milk acetone concentrations. Daily milk yield tended to be reduced in cows with acetone concentrations between 0·7 and 1·4 mmol/l, but responses were not entirely consistent. At concentrations >l·4 mmol/l, daily milk yield was clearly reduced. Long-term milk yield (day 0 to 100 and day 0 to 200) was about 190 kg of 40 g/kg fat-corrected milk (FCM) less in cows with acetone concentrations >l·4 vis-à-vis <0·7 mmol/l. Reproductive efficiency was also impaired in cows with acetone concentrations >l·4 mmol/l; 4·9 days longer interval from calving to first service (first parity and parity 2+ cows) and 5·7 times a greater risk of cystic ovaries (parity 1), compared with cows with milk acetone <0·7 mmol/l. However, no significant effects on long-term milk yield or on reproductive efficiency were found in cows with acetone concentrations 0·7 to 1·4 vis-à-vis <0·7 mmol/l. The results suggest that 1·4 mmol/l acetone in milk may be used as the most important critical value, as higher concentrations are detrimental to productivity. The interval 0·7 to 1·4 mmol acetone per I milk may be used as a warning class, since early lactation yield may be reduced.

There is a paucity of information on the metabolic effects of undernutrition of the ewe carrying multiple fetuses in late pregnancy. In the present study the effects of induction of ketosis from 132 d gestation in ewes carrying twin fetuses were compared with a control group. The ewes were well fed up to 132 d. Ketotic ewes showed a loss of condition score from 3·7 (SE 0·11) at 130 d gestation to 3·0 (SE 0·15) 10 d later after clinical recovery, compared with control twin-pregnant ewes (P < 0·01). The weight loss during the same time period was from 70·6 (SE 2·7) kg at 130 d to 64·2 (SE 2·7) kg at 140 d gestation. As expected, both groups lost weight and condition score in the first 28 d of lactation. Induction of ketosis caused a significant shortening of the gestation period to 142·8 (SE 0·7) d compared with 150 (SE 0·4) d in normal twin-pregnant ewes (P < 0·001). Ewes with induced ketosis recovered clinically and showed a normal feed intake by 3·4 (SE 0·07) d; three required treatment. Induction of ketosis resulted in reduction of hepatic uptake of bromosulphthalein (P < 0·01) and its biliary excretion (P < 0·05), metabolic clearance rate (P < 0·001), fractional clearance (P < 0·001) and 15 and 30 min retention compared with control twin-pregnant ewes. Most values had returned to normal by the first week of lactation. It is thought that in human pregnancy similar changes in bromosulphthalein clearance may be related to reduced binding sites for bromosulphthalein in the liver caused by increased circulating oestrogens. Induction of ketosis resulted in a significant hypoglycaemic (P < 0·01), ketotic (P < 0·001) state compared with well-fed twin-pregnant ewes. These changes could be correlated with the severity of the clinical signs, together with a significant rise in plasma urea (P < 0·001) and NH3 (P < 0·05) concentrations. Again, the return of most of these values to normal by the first week of lactation lends support to the reversibility of hepatic lesions caused by fatty infiltration of the liver. The seventy of this condition in naturally occurring cases suggests that factors other than undernutrition may be contributory, such as the general body condition of the ewe and glucose metabolism by the liver, including the conversion of propionate to glucose.

The effects of acetone concentration in milk, feeding frequency, and intake of concentrates and energy in total diet on milk yield were studied in 38624 lactations from 474 herds during 3 years. Herd-related data on feeding factors were collected once per indoor feeding period. Milk acetone concentrations higher than 0·40 mmol/l were deemed to denote hyperketonaemia, whether subclinical or clinical. The lactation curves of ketotic cows had an abnormal shape, with an inverted peak in early lactation. Most of the reduction in milk yield took place during the first 100 days of lactation with an estimated loss of proportionately 0·085 (acetone concentration ≤0·40 v. >2·00 mmol/l). The loss in 200-day milk yield was estimated to 328 kg 40 g/kg fat-corrected milk (FCM) at an overall mean of 5056 kg. If a herd's performance is monitored by lactation curves, a low peak — and hence an apparently high persistency – can indicate a ketosis problem. More frequent feeding of concentrates was correlated with a proportionately 0·033 to 0·074 higher milk yield in multiparous cows, but inconsistent between years. A more variable proportional response (−0·035 to 0·131) was observed in primiparous cows depending on interactions with, for example, breed. A greater total energy supply, including a larger amount of concentrate (maximum in lactation), was correlated with a higher milk yield, about 0·06 kg FCM per day per MJ metabolizable energy and 0·5 kg FCM per day per kg concentrate, respectively. However, increased feeding of concentrates at calving (parity 1) and 15 days after calving (multiparous cows) was correlated with decreased milk yield, by about −0–3 and −0–5 kg FCM per day per kg concentrate, respectively.