Genetics and physiology of leptin in periparturient dairy cows

In dairy cattle, the increase in milk yield has been accompanied by a more negative energy balance (EB) during early lactation and a decrease in fertility. As the hormone leptin is involved in regulation of nutritional status and reproductive function this hormone is an interesting protein to investigate during the periparturient period in dairy cattle. This study was performed to get insight into the function of leptin during the periparturient period and to perform an association study between polymorphisms in the bovine leptin gene and leptin receptor gene and fertility as well as production traits. Leptin concentrations in the periparturient cow undergo remarkable changes; leptin concentrations were high during late pregnancy and declined to a nadir at parturition. Genetic analysis of the leptin gene indicated that a combination of three polymorphisms located at a 135 bp region of the leptin promoter explained most of the variance in prepartum leptin concentrations. The two extreme genotype combinations could be used to investigate the function of leptin concentrations in pregnant cows. A polymorphism located on intron 2 of the leptin gene explained a significant part of the variation in milk yield. On the promoter region of the leptin gene an SNP was detected that was associated with first postpartum luteal activity (FPLA). This SNP could be a candidate marker for fertility in dairy cows. Another SNP on the leptin promoter was associated with energy balance and dry matter intake (DMI) where a higher dry matter intake occurred together with a higher energy balance. Two genotype combinations of the aforementioned three associated SNPs were defined which had a good milk yield together with a good energy balance and fertility. Calculations of an economical value per trait have to validate if one of these genotype combinations would be a possible candidate to be used in selection.     

环境酸度对紫花苜蓿早期生长和生理的影响

采用沙培的方式研究酸性土壤中土壤酸度对紫花苜蓿生理和生长的影响。结果表明,pH值在5～6的弱酸性环境为苜蓿的最适生长环境,这一酸度范围内幼苗生长正常或受到促进,如单株叶片数高于对照(pH=7)处理23.6%～27.2%、单叶面积高出对照20.87%～25.50%,叶绿素含量高出对照31.2%～27.9%,差异均达显著水平(P<0.05)。而出苗率、株高、根瘤数、根瘤等级、根长和地上、地下生物量亦高于对照;而pH值低于4的环境酸度为低适生长环境,可使苜蓿幼苗受到严重胁迫,出苗率、株高、单叶面积、单株叶片数、根瘤数、根长和地上、地下生物量均低于其他处理。在不同土壤酸度下,质膜透性、叶绿素含量、光化学效率(Fv/Fo)等均存着显著差异(P<0.05),pH＜4的各项生理指标均明显低于pH≥5的处理,其中地上生物量和根系活力分别不足对照处理的55.6%和61.1%(P<0.05)。     

Relation between cervical mucus sperm toxicity and pregnancy in cows]

Cervical secretion was collected from 99 dairy cows of the Black Pied Lowland breed at an age of 24 to 109 months with a minimum performance of 4000 kg of milk annually in spontaneous oestrus 42 days post partum at the minimum. It was demonstrated by the radioimmunological determination of progesterone in skim milk that from all the breeding cows (n = 99) the oestral secretion was collected during an optimum insemination time (x = 0.19 ng.ml-1 progesterone). Of the statistical set of 99 cows, 55 (i.e. 56.57%) got in calf and 43 (i.e. 43.43%) remained barren. As found, the conception results were not influenced by the rank of the insemination after parturition and the conception capacity of the cows was not influenced by the length of time from parturition to the respective inseminations. It was demonstrated in our investigation that the conception capacity of the cows increases as the time of sperm survival prolongs in the cervical secretion in vitro. The highest conception capacity (80%) was recorded in the cows whose cervical secretion was free of spermiotoxic action. In such cases the motility of the spermatozoa in the oestral mucus was maintained at a temperature of 38 degrees C for longer than 300 minutes. On the other hand, it was demonstrated that the conception capacity of the cows declines with shortening time of sperm survival in the cervical mucus.     

The physiology of natural parturition in cows]

In the year 1990 free parturitions of 339 cows were followed on three farms; these parturitions were compared with the last parturitions of these cows when the tender's assistance was available. The spontaneous free parturitions were followed in the open air in litter boxes which were located outside the barns where the groups of about twenty highly pregnant cows and heifers were housed for two months before their parturitions all the year round irrespective of the weather conditions. In addition to direct continuous observations, a host of zooveterinary data filed for these groups of animals were evaluated, concerning particularly those animals which could not be directly followed for the time reasons. Through direct observations the time schedule of the given parturition was evaluated and recorded above all, including all changes in the behaviour of the cow itself, and also behavioural changes in the group of animals or the behaviour of particular groups of animals toward each other. Applying the same criteria, the behaviour of born calves toward the mother, behaviour of calves toward each other and their behaviour toward other breeding cows was evaluated. The method of disinterested observation was used, this means the principle of noninterference with the most actions observed, with the exception of such situations that could result in the calf death. Another principle was to minimize through these observations all negative effects that could influence the behaviour of individual animals and also the behaviour of the whole group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between certain physical properties of cervical mucus and fertility in cows

The object of the present investigation was to determine several physical properties of cervical mucus in spontaneous oestrus cows in relation to fertility. Because, it is very difficult to determine the exact time of the beginning of oestrus in cows, the aim of the present study was to investigate if physical properties of cervical mucus at the time of artificial insemination (AI) are related to conception or not. A total of 93 cows of Friesian breed were used. The animals exhibited spontaneous oestrus, without being submitted to any hormonal treatment. Samples of cervical mucus were collected 5-30 min before AI and pH, viscosity, spinnbarkeit (spinability), crystallization and penetration were measured. Pregnancy diagnosis was performed by rectal palpation 3 months later. The results obtained from cows that conceived (44 animals), compared to those obtained from cows that did not conceive (49 animals), were the following: a) viscosity had been significantly lower (p < 0.05), b) crystallization had been significantly higher (p < 0.05), and c) pH, spinnbarkeit and penetration of spermatozoa into cervical mucus did not differ. In conclusion, the best time for AI is when viscosity is below 20 mm H2O and crystallization is above 3. Viscosity and crystallization could be related to ovulation time, but this needs further investigation.     

Effect of norgestomet treatment on plasminogen activator activity in the cervical mucus and the endometrium in dairy cows

The aim of this study was to determine the effect of norgestomet treatment, in the absence or the presence of a functional corpus luteum (CL), on plasminogen activators activity (PAA) in the cervical mucus and the endometrium in dairy cows. Eleven days after oestrus (Day 0 = oestrus), 38 cows were randomly assigned to one untreated control group (n = 9) and three treatment groups (S(1), S(2) and S(3)). Animals of S(1) group (n = 9) received an implantation of norgestomet on the outer surface of the ear for 8 days, simultaneous injection of oestradiol valerate 5 mg and norgestomet 3 mg, i.m., and on Day 19 an injection of ECG 500 IU, i.m. Animals of S(2) group (n = 11) received the treatment of S(1) group, plus an administration of PGF(2)alpha on Day 10 for the regression of CL. Animals of S(3) group received the treatment of S(2) group, plus two additional norgestomet implants inserted on Day 16 for 36 h. Both types of plasminogen activators [the tissue-type (t-PA) and the urokinase-type (u-PA)] were detected in the cervical mucus and the endometrium of the cows. Plasminogen activators activity in the cervical mucus was higher in control group than in S(1), S(2) and S(3) groups (P < 0.001). In contrast, endometrial PAA did not differ among groups (P > 0.05). Oestradiol-17beta concentrations on Day 21 were higher in S(2) group than in control group (P < 0.01) and S(3) group (P < 0.05). Progesterone concentrations did not differ among groups (P > 0.05). Oestradiol-17beta concentrations could positively affect cervical mucus PAA in control group (P < 0.1), but not in other groups (P > 0.05). These findings suggest that control of estrous cycle by norgestomet administration, in dairy cows, exerts a suppressive effect on plasminogen activators synthesis and/or secretion in the cervical mucus, regardless of the absence or the presence of the CL. On the contrary, endometrial PAA is not affected by norgestomet treatment.     

Association between physical properties of cervical mucus and ovulation rate in superovulated cows

Possible associations between certain physical properties of cervical mucus (CM) and ovulation rate were studied in 21 superovulated Holstein cows. In CM samples collected at the beginning of estrus (0 h) and in 4 h intervals for the following 24 h, the pH, the spinnbarkeit (spinability), and the crystallization value were measured. Blood samples, collected at the same time points with CM samples, were assessed for progesterone and estradiol concentrations. At 48 h the number of ovulated follicles was counted by transrectal ultrasonography and the animals were allotted into 2 groups according to the occurrence of at least one (group A, n = 16) or no (group B, n = 5) ovulations. The pH was lower (P < 0.05) at 8 h (7.00 + 0.24) in group A compared with group B (7.55 + 0.12). In group A, spinnbarkeit was significantly lower at 0 h and 20 h, and higher at 8 h and 16 h compared with group B (0 h: 2.50 + 0.82 versus 6.95 + 0.41; 20 h: 3.00 + 1.89 versus 5.38 + 0.94; 8 h: 7.00 + 0.87 versus 2.75 + 0.43; 16 h: 7.00 + 1.41 versus 4.30 + 0.71, for groups A versus B, respectively). Crystallization was significantly lower at 4 h (2.00 + 0.63) and 20 h (1.50 + 0.82) in group A compared with group B (3.13 + 0.32 at 4 h and 3.00 + 0.41 at 20 h). Progesterone at all time points, and estradiol at 16 h, 20 h, and 24 h were lower (P < 0.05) in group A than in group B. The pH, crystallization, estradiol, and progesterone differed (P < 0.05) within one group, while sbk differed within both groups.Our results imply that during the periovulatory period, steadily low progesterone concentrations trigger alterations of certain CM characteristics, while extremely high estradiol concentration could prevent the occurrence of these alterations.     

Chemical contamination of the cervical mucus in cows in relation to pregnancy and age factors]

Applying the method of absorption atom spectrometry (AAS), the contamination of cervical mucus by chemical elements (Cd, Pb, Hg, Cr, Cu, Zn) was demonstrated in the oestrus period of cows in different ecological agricultural regions of the North Moravian region. The results of observation revealed only a statistically significant difference (P less than 0.05) in Cd contents in the test groups; test group mean = 0.015 micrograms.g-1 Cd, control group mean = 0.006 micrograms.g-1. Cr findings in cervical mucus are of priority importance, the same applies to Cu findings. Zinc was found to be an element influencing negatively the conception of cows. The Zn values in cervical mucus of nonpregnant cows were demonstrated to be significantly higher (P less than 0.01) (conception--group = 0.841 micrograms.g-1, conception + group = 0.219 micrograms.g-1 Zn. So called sum of chemical elements: Cd + Pb + Hg + Cr + Cu + Zn was proposed and evaluated as a picture of the total contamination of cervical mucus. This characteristic was also influenced by the Zn findings in the group of pregnant and nonpregnant cows. Zn: Cd antagonism in cervical secretions of barren and pregnant cows was statistically highly significant (P less than 0.01). Chemical elements in cervical mucus were determined for the first time in a digested sample of cervical secretion. The correlation coefficients used for the cows (n = 99) from a contaminated region showed that the content of chemical elements in cervical secretion was not influenced by the age of cows. The content of chemical elements in cervical secretion was not increasing with the age.     

Effects of Saccharomyces cerevisiae on ruminal pH and microbial fermentation in dairy cows: Yeast supplementation on rumen fermentation

An experiment was conducted with eight ruminally-cannulated cows using a crossover design with 2 periods to determine the effects of yeast supplementation on rumen fermentation. Holstein dairy cows in late lactation were either supplemented with 0.5 g/hd/d of Saccharomyces cerevisiae, an active dry yeast (CNCM-1077, Levucell SC20 (r) SC, Lallemand Animal Nutrition) or not supplemented (control). A basal diet consisting of 60% forage and 40% concentrate (DM basis) was fed once daily to both groups of cows throughout the entire experiment. Ruminal pH was measured continuously every 22 min using a pH probe that was placed in the ventral rumen sac for 6 consecutive days. Volatile fatty acid and ammonia N concentrations in the rumen were measured on days 5 or 6 of the 12-d period for each cow and DM intake was monitored throughout the experiment. Data were analyzed using a mixed-effects model with repeated measures. There were no differences in dry matter intake between treatments. Mean ruminal pH was greater (P < 0.05) when yeast was supplemented (6.53 ± 0.07) compared with the control (6.32 ± 0.07). Average maximum and minimum ruminal pH were also greater (P < 0.05) when yeast was supplemented (7.01 ± 0.09 and 5.97 ± 0.08, respectively) compared with the control (6.80 ± 0.09 and 5.69 ± 0.09, respectively). Time spent under the subacute acidosis threshold, pH less than 5.6, was lower (P < 0.05) with yeast supplementation compared with control cows. No difference was observed for ruminal ammonia N concentrations (mean = 14.0 ± 1.2 mg/dL) between treatments. Total VFA concentration (mM) in the rumen tended to be lower (P = 0.10) in the yeast-supplemented cows (107.3 ± 6.35) than in the control cows (122.4 ± 6.35), which could be related to the greater pH observed with yeast supplementation. Supplementing dairy cows with active dry yeast in the current experiment increased the mean, minimum and maximum ruminal pH; decreased time spent in subacute rumen acidosis, and tended to decrease total VFA concentration in the rumen compared with control cows.     

Effects of forage source and particle size on chewing activity,ruminal pH,and saliva secretion in lactating Holstein cows

The present study investigated the effects of dietary forage source (quality) and particle size on chewing activity, saliva secretion, and ruminal pH. Twelve multiparous lactating Holstein cows, four of which were ruminally cannulated, were used in a replicated 4 × 4 Latin square experimental design with a 2 × 2 factorial arrangement of treatments. Cows fed wild‐rye hay diets had longer daily eating times than cows fed oaten hay diets. Treatments had no effect on ruminating time; therefore, resting time varied inversely to eating time. Neither the rate nor the amount of saliva secretion while eating, ruminating, or resting was affected by diet, resulting in similar total daily saliva secretions across treatments (231 L/day). Total volatile fatty acids (VFAs) in the ruminal fluid from animals fed oaten hay diets were higher than those from animals fed wild‐rye hay diets; further, VFAs increased with decreasing forage particle size (FPS). Consistent with elevated VFA concentrations, reducing FPS and including oaten hay in the diet decreased mean ruminal pH and increased the daily time of ruminal pH under 5.8. Results of this study suggest that forage source and particle size affect ruminal pH might be via variations in VFA production rather than increased salivary recycling of buffering substrates.