F. Cairoli

Strategies for proteomics with incompletely characterized genomes: The proteome of Bos taurus serum

A reference map for Bos taurus serum was obtained using proteomic tools: 21 proteins, plus several serum albumin fragments, have been identified in 47 spots. One of the major acute‐phase reactants, haptoglobin, was also detected in a pathological serum. A number of technical problems had to be solved. (i) Spot resolution in two‐dimensional electrophoresis (2‐DE) is not easily optimized, as several proteins have similar molecular mass; different polyacrylamide concentration gradients were used for the analysis of various size ranges. (ii) Identification of proteins through mass spectrometry (MS) procedures is also difficult as the genome of Bos taurus is incompletely characterized. The program FASTS proved particularly useful, since it allows simultaneous searching of several unordered sequence fragments, which may be individually too short to provide a statistically valid match using BLAST.

Endocrine responses induced in anestrous goats by the administration of different hormones after a fluorogestone acetate treatment

Abstract This study was designed to investigate the endocrinological variations induced in anestrous goats by means of different hormonal stimulations. Twenty goats were divided into four groups and, after treatment for 21 days with fluorogestone acetate (FGA) in vaginal sponges, were treated as follows: (1) vehicle; (2) 500 LU. pregnant mare serum gonadotropin (PMSG) 48 h before sponge removal (s.r.); (3) 500 LU. PMSG 48 h before s.r. and 1 μg gonadotropin-releasing hormone (GnRH) every 3 h for 8 times beginning 3 h before s.r. and (4) an ampoule of human menopausal gonadotropin (HMG) equivalent to 300 LU. luteinizing hormone (LH)-like and 300 LU. follicle-stimulating hormone (FSH)-like activity at s.r. Progesterone, estradiol 17β (E2), LH, FSH and prolactin (PRL) plasma variations were analyzed by validated radioimmunoassays. The stimulation of anestrous goats with FGA alone was inadequate to induce either behavioural estrus or variations in the endocrine pattern. All the other treatments (PMSG, PMSG+ GnRH, HMG) induced an increased in estradiol 17β concentration; the highest E2 levels were induced by PMSG + GnRH treatment. The E2 peaks were followed by LH and FSH surges, which occurred at different times depending on the treatments: the LH peak was significantly (P

Agreement between ultrasonographic classification of the CL and plasma progesterone concentration in dairy cows

We evaluated the agreement between ultrasonographic characteristics of the corpus luteum (CL) and plasma progesterone (P4) concentration in dairy cows. In Phase I of the study, the ovaries of 8 cows were ultrasonographically examined, and P4 was analyzed daily from estrus (Day 0) to Day 4, then at Day 7 and Day 10, and again daily from Day 17 to the onset of next estrus. In Phase 2, the ovaries of 157 randomly selected Friesian cows were examined once by ultrasonography, and blood samples collected concurrently were analyzed for plasma P4. On the basis of the P4 values, the function of CLs was classified as follows: 1) non-secretory CL when plasma P4 was lower than 1 ng/mL (n=41); 2) evolving CL when plasma P4 was between 1 and 4 ng/mL (n=55); and 3) mid-cycle CL when plasma P4 was more than 4 ng/mL (n=61). On the basis of ultrasonographic examination, 3 additional groups were established (absence of CL, evolving CL, midcycle CL). Ultrasonographic characteristics and size of Day 3 to 4 CLs and their respective plasma P4 concentrations were not distinguishable from those of CLs observed 3 to 4 d before the subsequent estrus. The degree of agreement between the two classification was 72%. The data indicate that the functional classification of CLs is difficult to determine based on ultrasonography alone.

Ultrasonographic appearance of tissue is a better indicator of CL function than CL diameter measurement in dairy cows

In this study, the ovaries of 99 randomly selected Friesian cows were examined by ultrasonography measuring the diameter and evaluating the appearance of corpora lutea (CLs) in order to assess the most reliable method for their functional classification. Concurrently, blood samples were taken and analyzed for plasma progesterone (P4) concentration. On the basis of the ultrasonographic measurement of the diameter of the CL, three groups were established: (A) CL not detected (n = 30), (B) CL psi < 20 mm (n = 22), and (C) CL psi > or = 20mm (n = 47). On the basis of the ultrasonographic appearance, three different groups were established: (A) CL not detected (n = 30), (B) evolving CL (n = 25), and (C) mid-cycle CL (n = 44). On the basis of the P4 values, CLs were functionally classified in the following three groups: (A) CL not detected when plasma P4 was lower than 1 ng/ml (n = 27), (B) evolving CL when plasma P4 was between 1 and 4 ng/ml inclusive (n = 29), and (C) mid-cycle CL when plasma P4 was more than 4 ng/ml (n = 43). The degree of agreement between plasma P4 concentrations and either ultrasonographic classification (diameter or appearance) was highly significant (P < 0.001). However, the results of the present study suggest that for the evaluation of functional classification of the CL in cows ultrasonographic appearance is more reliable than the evaluation of the diameter.

Hair cortisol level as a retrospective marker of hypothalamic-pituitary-adrenal axis activity in horse foals.

Stimulation of the hypothalamic-pituitary-adrenal (HPA) axis and elevated cortisol concentrations in fetal plasma are associated with foal maturity, viability and adaptation to independent life. However, non-invasive measurement of cortisol in hair samples has not yet been validated in horses. The current study developed a radioimmunoassay to analyse cortisol in horse hair and was used to measure cortisol hair concentration at birth and at 30 and 60 days of age as a retrospective study of HPA axis activity. Cortisol was detectable in the hair of foals from birth until 2 months, but decreased with time and varied greatly between individuals. Analysis of hair cortisol could be useful for non-invasive retrospective studies of HPA axis activity in perinatal horse.

The use of oxytocin for the reduction of cow placental retention, and subsequent endometritis

Oxytocin was administered to reduce incidence of retained placenta and uterine infections that could delay subsequent conception. Three hundred and fifty multiparous Friesian cows, each with spontaneous delivery of a single calf were divided randomly into two groups. Some (n = 175) were injected with 30 IU of oxytocin immediately after delivery and again 2-4 h later, while the remainder formed an untreated control group. The placental retention 24 h after parturition was 24.6% and 10.9% in control and treated animals respectively (P < 0.01). Endometritis occurred in 51.6% of the animals following placental retention as compared to 10.4% of those with normal expulsion of the fetal membranes (P < 0.001). A comparison of reproductive indices showed a statistically significant improvement of fertility in treated cows with the average interval from calving to conception being reduced from 124.4 d to 93.7 d (P < 0.0001).

Relationship between pharmacological induction of estrous and/or ovulation and twin pregnancy in the Thoroughbred mares

The aim of this study was to evaluate the possible relationship of pharmacological induction of estrous and/or ovulation with the occurrence of twin pregnancies in Thoroughbred mares. Out of 680 mares, 356 received one of the following treatments during the estrous cycle in which they became pregnant: injection of 0.5mg of cloprostenol at the ultrasonographic detection of a CL (n=86); injection of 5000 IU human chorionic gonadotropin (hCG) immediately before mating (n=221); injection of 0.5mg of cloprostenol at the ultrasonographic detection of a CL plus injection of 5000 IU hCG immediately before mating on cloprostenol-induced estrous (n=49). The other 324 mares, not treated for induction of estrous or ovulation in the estrous cycle resulting in pregnancy, were used as control group. The occurrence of twin and single pregnancies in treated and control mares underlines that the percentage of twin pregnancy in treated mares (16.6%) was statistically significantly higher (P<0.0001; odds ratio, OR=2.87) than the percentage of twinning in the control group (6.5%). Comparison of the occurrence of twins between treatments revealed a statistically significant difference between mares treated with hCG alone compared to animals given prostaglandin F2alpha (PGF2alpha) plus hCG. The results show a statistically significant difference for each treatment compared to controls, with the least difference (P<0.05; OR=2.18) for the comparison between hCG treatment group and controls, a significance of P<0.01; OR=3.05 for the comparison between PGF2alpha treatment and controls, and a highly statistically significant difference (P<0.0001; OR=6.37) for the comparison between PGF2alpha plus hCG-treated animals and controls.

Selected Metabolic and Hormonal Profiles during Maintenance of Spontaneous Ovarian Cysts in Dairy Cows

Information is lacking regarding the relationship between metabolic and hormonal profiles and the maintenance of spontaneous ovarian cyst disease in dairy cows. For this reason, the concentrations of non-esterified fatty acids (NEFA), insulin-like growth factor I (IGF-I) and cortisol (C) were investigated during the spontaneous course of ovarian cyst disease in dairy cows (n=6) between the 7th and 16th weeks post-partum (PP). The control group consisted of normally cycling cows (n=6). Blood samples were collected twice a day, and plasma was analysed using different techniques. Progesterone and 15-ketodihydro-PGF(2α) plasma profiles were investigated to confirm the ovulatory or anovulatory conditions of the cows. Cortisol plasma levels were not significantly different among sampling times within each group or between the two groups. NEFA plasma levels were significantly higher in cycling cows compared to cystic cows at the 16th week PP (p<0.01), but with rather low values, indicating by now sparse mobilization of fat stores. Insulin-like growth factor I plasma concentrations were higher in cystic cows during the 8th, 10th, 11th (p<0.01) and 16th week PP (p<0.05), indicating that the presence of ovarian cysts coincides with increased IGF-I levels. These results suggest no influence of cortisol and NEFA levels in cysts maintenance, while a possible involvement of IGF-I can be suspected not only in the pathogenesis, as already known, but also in the maintenance of spontaneous cystic ovarian disease in cattle.

Reproductive performance of dairy cows with luteal or follicular ovarian cysts after treatment with buserelin

In dairy farm management economic losses resulting from cystic ovarian degeneration are well known. In spite of this, neither the definition nor the aetiopathology of ovarian cysts are clear and agreed upon. Also the usual classification in luteal and follicular cysts, requiring ultrasound examination together with assessment of P4 to be accurate, is not very helpful in field conditions. Consequently a single treatment is often provided for both types of cysts, and since the 1970s treatments with GnRH and its analogues have been considered very useful. Nevertheless differences in recovery rates after GnRH treatment in animals with either luteal or follicular cysts are reported. Thus, the aim of this study was to evaluate recovery rate, recovery time and conception rate after treatment with buserelin (GnRH-analogue) in cows with ovarian luteal or follicular cysts. In a 5 years period, 150 cows with cysts out of a total of 990 animals, were detected and treated intravenously between 45 and 60 days PP with 20μg buserelin. No statistically significant differences were found in recovery rates and in conception rates between the two types of cysts. Comparison of recovery times showed significantly shorter recovery for cows with luteal cysts. The results emphasise the usefulness of GnRH to treat ovarian cysts regardless of their type, in relation to both recovery and conception rates. Intervals from treatment to resumption of ovarian activity were affected by the characteristics of ovarian cysts, with a faster recovery for the luteal type.

Recovery Rate and Recovery Time in Cows with Luteal and Follicular Ovarian Cysts after Treatment with Buserelin

M.C. Veronesi1*, M. Battocchio1, F. Cairoli1, A. Comin2 and M. Isola3 1Istituto di Clinica Ostetrica e Ginecologica Veterinaria, Facolta di Medicina Veterinaria, Universita di Milano, Milano; 2Dipartimento di Scienze Alimentari, Facolta di Medicina Veterinaria, Universita di Udine, Udine; 3Cattedra di Biometria e Statistica Medica, Facolta di Medicina, Universita di Udine, Udine, Italy *Correspondence: Dipartimento di Scienze Cliniche Veterinarie, Sezione di Clinica Ostetrica e Ginecologica Veterinaria, Facolta di Medicina Veterinaria, V ia Celoria, 10, 20133 Milano, Italy E-mail: maria.veronesi@unimi.it