Detection of nonpregnant cows and potential embryo losses by color Doppler ultrasound and interferon-stimulated gene expression in grazing dairy cows.

Many studies have been conducted to estimate pregnancy losses between 19 and 34 d after artificial insemination (AI) in dairy cows managed under confinement-based systems, but few studies have examined embryo mortality during this interval in dairy cows managed under gazing systems. The objectives of this prospective cohort study were (1) to assess the diagnostic value of the corpus luteum (CL) blood perfusion (BP) evaluation by Doppler ultrasound (US) to detect nonpregnant cows at 19 to 20 d post-AI, and (2) to assess the rate of potential embryo mortality between 19 to 34 d post-AI. The CL-BP of all cows included in the study (n = 131) was examined on farm by power and color mode of Doppler US and later using an image processing software by a second evaluator. The endometrium thickness and echotexture were evaluated by B-mode US at the same visit to assess if the nonpregnancy diagnosis could be improved at 19 to 20 d post-AI by this additional diagnostic tool. Blood samples were obtained at 19 to 20 d post-AI for progesterone (P4) measurement by chemiluminescence and to determine the mRNA expression of ISG by real-time PCR. Pregnancy diagnosis based on embryo visualization was performed at 33 to 34 d post-AI by US B-mode. In parallel interpretation, ISG15 and MX2 mRNA expression in leukocytes [sensitivity (Se), 100%] were regarded as suitable biomarkers for early pregnancy and were selected for molecular characterization of pregnancy at 19 to 20 d post-AI. At 19 to 20 d post-AI, 61.1% of the cows had positive CL-BP by Doppler US (Se, 98.0%), 62.7% had ISG mRNA expression in leukocytes over the cutoff point (Se, 95.7%), and 50.8% were positive, based on the combination of ISG mRNA expression, CL-BP by Doppler US, and P4 concentration (Se, 100%), and were considered as possible pregnant. At 33 to 34 d, the pregnancy rate was 37.4% diagnosed by the B-mode US. Based on the expression of the selected biomarkers in cows with active CL, we found that 28.1% of the cows could have potentially lost their pregnancy between 19 and 34 d post-AI. The Doppler US color mode showed similar accuracy and a higher negative predictive value than the genes selected as biomarkers. The additional B-mode ultrasound evaluation of the uterine stratum vasculare and the endometrium thickness improved the diagnostic accuracy. Therefore, assessing the CL-BP by Doppler US allowed early detection of nonpregnant cows at 19 to 20 d post-AI. The combination of early CL-BP by Doppler US (d 19 to 20) with early embryo detection by B-mode US (d 33-34) could be used to facilitate earlier rebreeding of dairy cows.

Heat stress and sperm production in the domestic cat.

This study aimed to assess 1) the effect of high environmental temperatures on sperm production and 2) the effectiveness of a temperature-humidity index (THI) to predict the degree of thermal stress in a cat model. Semen collection was performed by electroejaculation for 18 mo in 20 tomcats maintained under controlled photoperiod. Still, temperature and humidity were not experimentally manipulated to describe the effect of natural climate conditions on seminal samples. Ejaculates (n = 512) were then grouped according to temperature records of the sampling day and compared by temperature and THI index. Significant lower sperm parameters and increase sperm tail abnormalities were observed during warm environments (temperature and THI). Concentration and total sperm count were the most affected parameters. Environmental temperatures of 28.5 °C with 54% relative humidity (THI = 77.07) and 27.9 °C with 66% humidity (THI = 77.84) were upper thresholds of moderate thermal stress. Moreover, days with relative humidity near 90% led to severe thermal stress with temperatures as low as 26.6 °C (THI = 78.88). The current study demonstrates the detrimental effect of high environmental temperatures on sperm quality in the domestic cat. This effect is observed at lower temperatures when high relative humidity is present. In this sense, the THI was a reliable predictor of the magnitude of thermal stress experienced by cats. Thus, cats from reproductive programs should be maintained under controlled photoperiod cycles with temperatures around 20 °C and humidity around 70% to avoid semen detrimental effects.

Associations of somatic cell count with milk yield and reproductive performance in grazing dairy cows.

Poor udder health status can have a detrimental effect on milk yield and reproductive performance, leading to reductions in the dairy farm profit. The objective of this retrospective longitudinal study was to assess the associations of somatic cell count (SCC) with daily milk yield and reproductive performance. A database with 1,930,376 lactations from 867 Argentinean grazing dairy herds records collected for 14 years was used. The association of the evolution of SCC (healthy vs. new case vs. cured vs. chronic; with 150,000 SCC/mL as threshold) and of the severity of SCC [mild (150,000--400,000 SCC/mL) vs. moderate (400,000-1,000,000 SCC/mL) vs. severe (>1,000,000 SCC/mL)] with the odds for conception were estimated. Finally, the associations of the linear score of SCC (LS-SCC) with daily milk yield were estimated depending on parity and milk production quartile. The odds ratios (CI 95%) for conception at first service were 0.921 (0.902-0.941), 0.866 (0.848-0.884), and 0.842 (0.826-0.859) for the new case, cured, and chronic cows compared with healthy cows, respectively. Also, the odds ratios (CI 95%) for conception were 0.902 (0.881-0.925), 0.837 (0.808-0.866) and 0.709 (0.683-0.736) for mild, moderate and severe cases compared with healthy cows, respectively. An increase of one point of LS-SCC was associated with decreases of 0.349, 0.539, and 0.676 kg in daily milk yield for first-, second-, and third-lactation cows, respectively. In conclusion, SCC is negatively associated with the risk for conception and with daily milk yield in grazing dairy cows. This negative relationship with conception is higher when SCC increase occurs after the service date and it is influenced by severity of mastitis, and in the case of milk yield, the negative association is influenced by parity, milk production quartile, and severity of mastitis.

Effect of cat seminal plasma and purified llama ovulation-inducing factor (ß-NGF) on ovarian function in queens.

The present study aimed to determine the effect of cat seminal plasma and purified llama ovulation-inducing factor (ß-NGF) on ovarian activity in queens. Queens (n = 6) were used for all the treatments in a crossover design with an interval time between treatments of three interestrus intervals. Forty-eight hours after the detection of an estrus vaginal cytology, queens were given cat seminal plasma (subcutaneous or intramuscular), purified llama ovulation-inducing factor (15 or 35 µg), hCG (75 UI), saline, or were mated with a male. A total of 192 estrous cycles were observed. Estrus length and serum estradiol concentration were 6 ± 1 days (range 2-10 d) and 38 pg/mL (range 10-75 pg/mL), respectively. Queens mated and given hCG showed higher serum progesterone concentration and longer interestrus interval (47 ± 5 d) than that of controls (10 ± 3 d). Sixty-seven percent of queens (4/6) treated with subcutaneous cat seminal plasma, and 17% of those treated with purified llama ß-NGF showed high serum progesterone concentrations along with prolonged interestrus. However, intramuscular administration of cat seminal plasma produced interestrus intervals similar to controls (15 ± 5 d) and basal serum progesterone concentration (<0.50 ng/mL). This study demonstrates that the subcutaneous administration of cat seminal plasma induced ovulation in queens. Therefore, molecules present in cat seminal plasma, contribute to the induction of ovulation in queens. Identifying those molecules will improve the knowledge of queen's reproductive physiology. Also, it could offer a physiologic alternative to induce ovulation in queens when reproductive biotechnologies are used.

Uterine endometrial cytology, biopsy, bacteriology, and serum C-reactive protein in clinically healthy diestrus bitches.

The aim of this work was to assess the agreement between endometrial cytology and uterine biopsy for the diagnosis of endometritis (END), the bacterial populations isolated from the vagina and uterus of bitches having END, and the measurement of C- reactive protein as a diagnostic tool for diagnosis of END in clinically healthy bitches. Fifty privately-owned intact, clinically healthy bitches, in diestrus, without a history of progestin administration, aged between 8 months and 6 years old and weighing between 5 and 28 kg were used in this study. Bitches were included in a program for breeding control at a municipal pet public shelter. Before ovariohysterectomy (OVX) samples for vaginal cytology and bacteriology, and blood samples were taken. After OVX endometrial cytology, bacteriological samples and biopsy were collected. Histologic examination was performed to confirm the uterine condition. Blood samples were centrifuged and stored at -20 °C until progesterone (P4) and C-reactive protein was measured. Samples for bacteriological culture were collected, and swabs were placed into Stuart's transport medium and transported to the laboratory. On histopathologic examination, the most common observation was END (27/50), followed by normal endometrium (NE; 18/50), cystic endometrial hyperplasia (CEH; 2/50), atrophy (2/50) and fibrosis with degeneration of the endometrial glands (1/50). Low degree of agreement was observed between results obtained by endometrial cytology samples and results obtained by biopsy in endometrial diagnosis (Kappa Coefficient: -0.19). In vaginal samples, ß-hemolytic Streptococcus, Staphylococcus spp., Escherichia coli, Proteus spp., Corynebacterium spp., and Klebsiella pneumoniae were the bacteria most often found. In uterine samples, only four samples from END showed bacterial grow. C-reactive protein frequency was higher in END (6/23, 23%) vs NE (0/16, 0%; Van der Waerden P-value = 0,0302). Our results support the hypothesis that END is a frequent finding in uterine biopsy and could be associated with subfertility and infertility in the bitch. A low degree of agreement was observed between the diagnostic results from the uterine biopsy and endometrial cytology. Bacteriology would not be recommended as a diagnostic tool because no bacteria highly associated with uterine diseases were isolated from bitches with END. Finally, the usefulness of C-reactive protein concentration as a marker for END in bitches could not be conclusively demonstrated.

Effect of milk production on reproductive performance in dairy herds.

The objective of the present study was to assess the relationship between individual cow milk yield and fertility, accounting for the contextual effect of the herd. A data set including 657,968 lactations from 677 dairy herds in Argentina from 2001 to 2012 was used. The odds of pregnancy by 100 d in milk (DIM) were assessed by a multilevel logistic model (with cow as the first and herd as the second hierarchical level), and time to pregnancy was assessed by a proportional hazards regression model. Multilevel logistic models included the fixed effects of milk yield by 80 DIM, parity, year, and calving season at cow level and quartiles of herd milk yield by 80 DIM as a contextual effect. The proportional hazards model included the effect of daily cow-level milk yield as time-dependent variable, with milk yield at herd level as the stratification variable. Cows producing 1 standard deviation over the mean milk yield of their herd had 1.3 percentage point lower pregnancy by 100 DIM (from 31.4 to 30.1%; odds ratio = 0.942) when in herds in the top quartile of milk yield, whereas they increased 0.5 percentage points (from 27.9 to 28.4%) when in herds in the lowest quartile of milk yield. Only 4% of the observed variation in pregnancy by 100 DIM was explained by the random effect of the herd. Similarly, cows producing 1 standard deviation (8 kg/d) greater than the herd mean daily milk had 1.3% lower hazard of pregnancy (hazard ratio = 0.987) at 63 DIM in herds in the top quartile of milk yield, whereas they had 14.8% higher hazard (hazard ratio = 1.148) in herds in the lowest quartile of milk yield. The magnitude of the negative association between the cow's daily milk yield and the hazard of pregnancy increased with DIM. In conclusion, the relationship between milk yield and reproductive performance is statistically significant, but the effect size is practically small and is modulated by herd production level.