Diagnostic performance of the BHBCheck ß-hydroxybutyrate meter for hyperketonaemia in Indian cows and buffaloes.

The objective of the study was to evaluate the diagnostic performance of the electronic handheld BHBCheck meter (BHM) (PortaCheck, Inc., USA) to determine whole blood, plasma and serum ß-hydroxybutyrate (BHB) against serum BHB determined using reference laboratory method of Randox D-3 Hydroxybutyrate Ranbut assay (RSM) in Indian dairy cows and buffaloes. Blood samples were collected by puncturing coccygeal vessels for determining whole blood, serum and plasma BHB using BHM and serum BHB using RSM from 217 cows (Gir breed; median 42 DIM and 3rd lactation) and 223 buffaloes (non-descript; median 39 DIM and 3rd lactation) from nearby herds. The Pearson's correlation between whole blood (0.988; 0.987), plasma (0.985; 0.983) and serum (0.985; 0.983) BHB determined using the BHM and serum BHB determined with the RSM in Indian cows and buffaloes, respectively, were significant. Bland-Altman plot demonstrated an excellent agreement between whole blood, plasma and serum BHB determined with BHM, against the serum BHB determined with RSM in Indian cows and buffaloes, respectively. For hyperketonaemia with reference serum BHB cut-off values ≥ 1.2 and 1.4 mmol/L determined with RSM, it recorded optimized BHB thresholds, sensitivity and specificity for whole blood (≥ 0.9 to 1.0 mmol/L; 91 to 95% and 88 to 98%), plasma (≥ 0.9 to 1.0 mmol/L; 91 and 100%) and serum (≥ 0.9 to 1.0 mmol/L; 92 to 100% and 85 to 94%) with BHM in cows and buffaloes, respectively. In conclusion, BHB determined with BHM demonstrated an excellent correlation, agreement and test characteristics with BHB determined with RSM and hence can accurately determine whole blood, plasma and serum BHB in cows and buffaloes.

Body temperature in early postpartum dairy cows.

A strategy widely adopted in the modern dairy industry is the introduction of postpartum health monitoring programs by trained farm personnel. Within these fresh cow protocols, various parameters (e.g., rectal temperature, attitude, milk production, uterine discharge, ketones) are evaluated during the first 5 to 14 days in milk (DIMs) to diagnose relevant diseases. It is well documented that 14% to 66% of healthy cows exhibit at least one temperature of 39.5 °C or greater within the first 10 DIM. Although widely adopted, data on diagnostic performance of body temperature (BT) measurement to diagnose infectious diseases (e.g., metritis, mastitis) are lacking. Therefore, the objective of this study was to identify possible factors associated with BT in postpartum dairy cows. A study was conducted on a commercial dairy farm including 251 cows. In a total of 217 cows, a vaginal temperature logger was inserted from DIM 2 to 10, whereas 34 cows did not receive a temperature logger as control. Temperature loggers measured vaginal temperature every 10 minutes. Rectal temperature was measured twice daily in all cows. On DIM 2, 5, and 10, cows underwent a clinical examination. Body temperature was influenced by various parameters. Primiparous cows had 0.2 °C higher BT than multiparous cows. Multiparous cows that calved during June and July had higher BT than those that calved in May. In primiparous cows, this effect was only evident from DIM 7 to 10. Furthermore, abnormal calving conditions (i.e., assisted calving, dead calf, retained placenta, twins) affected BT in cows. This effect was more pronounced in multiparous cows. Abnormal vaginal discharge did increase BT in primiparous and multiparous cows. Primiparous cows suffering from hyperketonemia (beta-hydroxybutyrat ≥ 1.4 mmol/L) had higher BT than those not affected. In multiparous cows, there was no association between hyperketonemia and BT. The results of this study clearly demonstrate that BT is influenced by various parameters in dairy cows. Therefore, these parameters have to be considered when interpreting measurements of BT in dairy cows. This information helps to explain the high incidence of type I and II errors when measuring BT and clearly illustrates that measures of BT should not be used as a single criterion to decide whether or not to provide antibiotic treatment to dairy cows. However, research-based test characteristics of other parameters (e.g., vaginal discharge) alone or in combination with BT are still lacking.

Prevalence of subclinical ketosis and relationships with postpartum diseases in European dairy cows.

Subclinical ketosis (SCK) is defined as concentrations of ß-hydroxybutyrate (BHBA) ≥ 1.2 to 1.4 mmol/L and it is considered a gateway condition for other metabolic and infectious disorders such as metritis, mastitis, clinical ketosis, and displaced abomasum. Reported prevalence rates range from 6.9 to 43% in the first 2 mo of lactation. However, there is a dearth of information on prevalence rates considering the diversity of European dairy farms. The objectives of this study were to (1) determine prevalence of SCK, (2) identify thresholds of BHBA, and (3) study their relationships with postpartum metritis, clinical ketosis, displaced abomasum, lameness, and mastitis in European dairy farms. From May to October 2011, a convenience sample of 528 dairy herds from Croatia, Germany, Hungary, Italy, Poland, Portugal, Serbia, Slovenia, Spain, and Turkey was studied. ß-Hydroxybutyrate levels were measured in 5,884 cows with a handheld meter within 2 to 15 d in milk (DIM). On average, 11 cows were enrolled per farm and relevant information (e.g., DIM, postpartum diseases, herd size) was recorded. Using receiver operator characteristic curve analyses, blood BHBA thresholds were determined for the occurrence of metritis, mastitis, clinical ketosis, displaced abomasum, and lameness. Multivariate binary logistic regression models were built for each disease, considering cow as the experimental unit and herd as a random effect. Overall prevalence of SCK (i.e., blood BHBA ≥ 1.2 mmol/L) within 10 countries was 21.8%, ranging from 11.2 to 36.6%. Cows with SCK had 1.5, 9.5, and 5.0 times greater odds of developing metritis, clinical ketosis, and displaced abomasum, respectively. Multivariate binary logistic regression models demonstrated that cows with blood BHBA levels of ≥ 1.4, ≥ 1.1 and ≥ 1.7 mmol/L during 2 to 15 DIM had 1.7, 10.5, and 6.9 times greater odds of developing metritis, clinical ketosis, and displaced abomasum, respectively, compared with cows with lower BHBA blood levels. Interestingly, a postpartum blood BHBA threshold ≥ 1.1 mmol/L increased the odds for lameness in dairy cows 1.8 (95% confidence interval: 1.3 to 2.5) times. Overall, prevalence of SCK was high between 2 to 15 DIM and SCK increased the odds of metritis, clinical ketosis, lameness, and displaced abomasum in European dairy herds.

Technical note: evaluation of data loggers for measuring lying behavior in dairy calves.

Lying behavior might indicate how the animal interacts with its environment and is an important indicator of cow and calf comfort. Measuring behavior can be time consuming; therefore, behavioral recording with the help of loggers has become common. Recently, the Hobo Pendant G data logger (Onset Computer Corp., Bourne, MA) was validated for measuring lying behavior in cows but no work to date has validated this logger for measuring lying behavior in calves. The objective of this study was to test the accuracy of the Hobo Pendant G data logger for measuring total lying time and frequency of lying bouts in dairy calves. In 2 experiments (experiment 1: thirty-seven 2-h observation periods; experiment 2: nineteen 24-h observation periods), we tested the effect of 2 different recording intervals, the effect of attachment to different legs, and the effect of removing short, potentially erroneous readings. We found an excellent relationship when comparing the 30-s and 60-s recording intervals. For total lying time and bout frequency, the highest correlation was found when the logger was attached to the hind legs and recording was conducted with a 60-s sampling interval. In experiment 2, average total lying time was 1,077 ± 54 min/24 h (18.0 ± 0.9h/24h), with an average frequency of 19.4 ± 4.5 bouts per day. Predictability, sensitivity, and specificity for experiment 2 were >97% using the 60-s recording interval and removing single readings of lying or standing from the data set compared with direct observation as reference. The data logger accurately measured total lying time and bout frequency when the sampling interval was ≤ 60 s and short readings of lying and standing up to 1 min were converted into the preceding behavior. The best results were achieved by attaching the logger to the right hind leg.

Effect of heat stress on body temperature in healthy early postpartum dairy cows.

Measurement of body temperature is the most common method for an early diagnosis of sick cows in fresh cow protocols currently used on dairy farms. Thresholds for fever range from 39.4 °C to 39.7 °C. Several studies attempted to describe normal temperature ranges for healthy dairy cows in the early puerperium. However, the definition of a healthy cow is variable within these studies. It is challenging to determine normal temperature ranges for healthy cows because body temperature is usually included in the definition. Therefore, the objectives of this study were to identify factors that influence body temperature in healthy dairy cows early postpartum and to determine normal temperature ranges for healthy cows that calved in a moderate (temperature humidity index: 59.8 ± 3.8) and a hot period (temperature humidity index: 74.1 ± 4.4), respectively, excluding body temperature from the definition of the health status. Furthermore, the prevalence of fever was calculated for both periods separately. A subset of 17 (moderate period) and 15 cows (hot period) were used for analysis. To ensure their uterine health only cows with a serum haptoglobin concentration ≤ 1.1 g/L were included in the analysis. Therefore, body temperature could be excluded from the definition. A vaginal temperature logger that measured vaginal temperature every 10 min was inserted from Day 2 to 10 after parturition. Additionally rectal temperature was measured twice daily. Day in milk (2 to 10), period (moderate and hot), and time of day had an effect on rectal and vaginal temperature. The prevalence of fever (≥ 39.5 °C) was 7.4% and 28.1% for rectal temperature in the moderate and hot period, respectively. For vaginal temperature (07.00 to 11.00 h) it was 10% and 33%, respectively, considering the same threshold and period. This study demonstrates that body temperature in the early puerperium is influenced by several factors (day in milk, climate, time of day). Therefore, these factors have to be considered when interpreting body temperature measures to identify sick cows. Furthermore, the prevalence of fever considering different thresholds is higher during hot than moderate periods. However, even in a moderate period healthy cows can exhibit a body temperature that is considered as fever. This fact clearly illustrates that fever alone should not be considered the decision criterion whether a cow is allocated to an antibiotic treatment, although it is the most important one that is objectively measurable.

Endogenous and exogenous progesterone influence body temperature in dairy cows.

Three experiments were conducted to determine the effect of endogenous progesterone (P4) on body temperature comparing lactating, pregnant with lactating, nonpregnant cows, and to study the effect of exogenous P4 administered via a controlled internal drug release (CIDR) insert on body temperature in lactating dairy cows. Body temperature was measured vaginally and rectally using temperature loggers and a digital thermometer, respectively. In experiment 1, 10 cyclic lactating cows (3 primiparous, 7 multiparous) and 10 lactating, pregnant cows (3 primiparous, 7 multiparous) were included. Vaginal temperatures and serum P4 concentrations were greater in pregnant cows (vaginal: 0.3±0.01°C; P4: 5.5±0.4 ng/mL) compared with nonpregnant cows. In experiment 2, estrous cycles of 14 postpartum healthy, cyclic, lactating cows (10 primiparous, 4 multiparous) were synchronized, and cows were assigned randomly to 1 of 2 treatments (CIDR-P4 or CIDR-blank). A temperature logger was inserted 1 d after ovulation using a P4-free CIDR (CIDR-blank) and a CIDR containing 1.38g of P4 (CIDR-P4) in the control (n=7) and the P4-treated group (n=7), respectively. On d 3 after P4 treatment, vaginal temperature was 0.3±0.03°C greater compared with that on d 1 and d 5. In experiment 3, 9 cyclic multiparous lactating cows were enrolled 1±1 d after confirmed ovulation and a temperature logger inserted. Two days later, a CIDR-P4 was inserted on top of the CIDR-blank. On d 5±1 and d 7±1, respectively, the CIDR-P4 and CIDR-blank with the temperature logger were removed. During the CIDR-P4 treatment (48h), vaginal temperature was 0.2±0.05°C and 0.1±0.05°C greater than during the pre- and post-treatment periods (48h), respectively. Serum P4 concentration peaked during CIDR-P4 treatment (2.2±0.8 ng/mL) and was greater than during the pre-treatment period (0.2±0.2 ng/mL) for 48h. An increase in vaginal temperature could be due to endogenous and exogenous P4. However, a correlation between serum P4 concentrations and body temperature did not exist. Further investigations are warranted to better understand the pathways of the thermogenic effect of P4 on body temperature.

Validity of prepartum changes in vaginal and rectal temperature to predict calving in dairy cows.

The prevalence of dystocia is high in many dairy herds and is associated with stillbirth and negative effects for the cow. An accurate predictor of calving would enable supervision of cows more precisely to a relevant time interval so that obstetrical assistance can be provided in a timely manner. This might help to decrease calf mortality rate. Evidence exists that cows exhibit a decrease in body temperature before the onset of calving. The performance of a decrease in body temperature as a test to predict the onset of calving in dairy cows has not been investigated. The objective was to investigate test criteria of a decrease in vaginal and rectal temperature as predictors of calving in dairy cows. In 3 experiments, temperature loggers (Minilog 8, Vemco Ltd., Halifax, Canada) were inserted into the vagina of cows before calving (n = 85), and rectal temperatures were measured twice daily in 55 of these cows. Vaginal temperatures were 0.2 to 0.3 °C and 0.6 to 0.7 °C lower on the day of calving compared with 24 and 48 h before calving, respectively. Rectal temperatures were 0.3 to 0.5 °C and 0.4 to 0.6 °C lower on the day of calving compared with 24 and 48 h before calving, respectively. Vaginal temperatures exhibited a diurnal rhythm during the 120 h before calving, which continued on a lower level during the 48 h preceding parturition. In the 3 experiments, a decrease in vaginal temperature of ≥ 0.3 °C over 24h could predict calving within 24h, with sensitivity ranging from 62 to 71% and specificity ranging from 81 to 87%. Similarly, a decrease in rectal temperature measured at 0730 h of ≥ 0.3 °C could predict calving within 24h, with sensitivity from 44 to 69% and specificity from 86 to 88%. Although dairy cows exhibit a distinctive decrease in vaginal and rectal temperatures commencing approximately 48 h before calving, detecting this decrease does not determine the onset of calving precisely. Nevertheless, it can provide valuable information in addition to the traditional signs (i.e., relaxation of the sacrosciatic ligament) that calving is imminent.

Body temperature around induced estrus in dairy cows.

The overall objective of this study was to study the influence of induced estrus on body temperature, comparing 5 distinct intervals around induced estrus and to determine the diurnal pattern from 4 ± 1 d before to 4 ± 1 d after induced estrus. Sixteen estrous cycles of 9 postpartum dairy cows were synchronized with 2 injections of PGF(2α), 10 d apart. After the second PGF(2α) injection on d 10, temperature loggers were inserted into the vaginal cavity for a 12 ± 1-d period. Two days later, a third dose of PGF(2α) was injected to induce estrus. After confirmation of a corpus luteum, loggers were removed on d 5 ± 1. Observation of estrus, rectal palpation, and ultrasound scanning to determine ovulation were carried out every 4 ± 1h, beginning at 12h after the third PGF(2α) injection. Blood samples from the vena coccygea mediana were collected twice daily from d 11 to 12 and every 4 ± 1h after the third PGF(2α) injection until ovulation. Vaginal temperature was recorded every 5 min and averaged to hourly means for the following 5 periods: 1) 48 h preceding the third PGF(2α) injection, 2) from the third PGF(2α) injection to first signs of estrus, 3) estrus to ovulation, 4) a 4-h interval in which ovulation occurred, and 5) a 96-h post-ovulation period. High body temperatures (39.0 ± 0.5 °C) and low progesterone (P4) concentrations (<0.5 ng/mL) were observed during estrus, whereas low body temperatures were observed from PGF(2α) injection to estrus (38.6 ± 0.3 °C) and around ovulation (38.5 ± 0.2 °C), respectively. An association between body temperature and serum P4 concentrations did not exist. However, P4 concentrations on d 11 and 12 were high (5.0 ± 1.5 ng/mL) and decreased (0.9 ± 0.2 ng/mL) after ovulation. Diurnal temperature rhythms were similar before and after estrus. Vaginal temperature before estrus (d 11 and 12) was slightly (0.1 °C) higher compared with the post-ovulation period.