Transient reductions in milk fat synthesis and their association with the ruminal and metabolic profile in dairy cows fed high-starch, low-fat diets.

Sub-acute ruminal acidosis (SARA) is sometimes observed along with reduced milk fat synthesis. Inconsistent responses may be explained by dietary fat levels. Twelve ruminally cannulated cows were used in a Latin square design investigating the timing of metabolic and milk fat changes during Induction and Recovery from SARA by altering starch levels in low-fat diets. Treatments were (1) SARA Induction, (2) Recovery and (3) Control. Sub-acute ruminal acidosis was induced by feeding a diet containing 29.4% starch, 24.0% NDF and 2.8% fatty acids (FAs), whereas the Recovery and Control diets contained 19.9% starch, 31.0% NDF and 2.6% FA. Relative to Control, DM intake (DMI) and milk yield were higher in SARA from days 14 to 21 and from days 10 to 21, respectively (P < 0.05). Milk fat content was reduced from days 3 to 14 in SARA (P < 0.05) compared with Control, while greater protein and lactose contents were observed from days 14 to 21 and 3 to 21, respectively (P < 0.05). Milk fat yield was reduced by SARA on day 3 (P < 0.05), whereas both protein and lactose yields were higher on days 14 and 21 (P < 0.05). The ruminal acetate-to-propionate ratio was lower, and the concentrations of propionate and lactate were higher in the SARA treatment compared with Control on day 21 (P < 0.05). Plasma insulin increased during SARA, whereas plasma non-esterified fatty acids and milk ß-hydroxybutyrate decreased (P < 0.05). Similarly to fat yield, the yield of milk preformed FA (>16C) was lower on day 3 (P < 0.05) and tended to be lower on day 7 in SARA cows (P < 0.10), whereas yield of de novo FA (<16C) was higher on day 21 (P < 0.01) in the SARA group relative to Control. The t10- to t11-18:1 ratio increased during the SARA Induction period (P < 0.05), but the concentration of t10-18:1 remained below 0.5% of milk fat, and t10,c12 conjugated linoleic acid remained below detection levels. Odd-chain FA increased, whereas branched-chain FA was reduced during SARA Induction from days 3 to 21 (P < 0.05). Sub-acute ruminal acidosis reduced milk fat synthesis transiently. Such reduction was not associated with ruminal biohydrogenation intermediates but rather with a transient reduction in supply of preformed FA. Subsequent rescue of milk fat synthesis may be associated with higher availability of substrates due to increased DMI during SARA.

Regulation of glucose and protein metabolism in growing steers by long-chain n-3 fatty acids in muscle membrane phospholipids is dose-dependent.

A previous study showed that long-chain n-3 polyunsaturated fatty acids (LCn-3PUFA; >18 carbons n-3) exert an anabolic effect on protein metabolism through the upregulation of insulin sensitivity and activation of the insulin signaling pathway. This study further delineates for the first time whether the anabolic effect of LCn-3PUFA on metabolism is dose responsive. Six steers were used to test three graded amounts of menhaden oil rich in LCn-3PUFA (0%, 2% and 4%; enteral infusions) according to a double 3 × 3 Latin square design. Treatment comparisons were made using iso-energetic substitutions of control oil for menhaden oil and using 6-week experimental periods. The LCn-3PUFA in muscle total membrane phospholipids increased from 8%, 14% to 20% as dietary menhaden oil increased. Feeding graded amounts of menhaden oil linearly decreased plasma insulin concentration (49, 35 and 25 µU/ml, P = 0.01). The insulin-stimulated amino acid disposal rates as assessed using hyperinsulinemic-euglycemic-euaminoacidemic clamps (20, 40 and 80 mU/kg per h) were linearly increased by the incremental administrations of menhaden oil from 169, 238 to 375 µmol/kg per h (P = 0.005) during the 40 mU/kg per h clamp, and from 295, 360 and 590 µmol/kg per h (P = 0.02) during the 80 mU/kg per h clamp. Glucose disposal rate responded according to a quadratic relationship with the incremental menhaden oil amounts (P < 0.05). A regression analysis showed that 47% of the amino acid disposal rates elicited during the hyperinsulinemic clamp was related to muscle membrane LCn-3PUFA content (P = 0.003). These results show for the first time that both protein and glucose metabolism respond in a dose-dependent manner to menhaden oil and to muscle membrane LCn-3PUFA.

Variations in mammary protein metabolism during the natural filling of the udder with milk over a 12-h period between two milkings: leucine kinetics.

To define the temporal variations of whole body and mammary leucine kinetics over a 12-h period between two milkings, we used two groups of four Holstein cows, one in their second and the other in their third or fourth lactation. Cows were infused with L-[1-13C]leucine during the 12-h interval between two milkings. Blood was sampled every 30 min during that period from arterial and mammary sources. Timeafter milking did not affect whole body irreversible loss rate of leucine but affected whole body leucine oxidation, which broadly followed variations in arterial plasma leucine concentration. Similarly, mammary leucine irreversible loss rate and leucine used for protein synthesis were not affected by time after milking. Leucine oxidation by the mammary gland was, however, affected by time after milking. It increased by 15% from the first 2-h period to the following 4-h period and then decreased by 13% over the following 2-h period. A 21% increase in leucine oxidation was observed from 8 to 10 h after milking, and then it decreased by 26% over the last 2-h period. Protein degradation expressed as percentage of mammary leucine flux followed a similar temporal pattern. Leucine used for protein synthesis by the mammary gland was unaltered over time after milking, suggesting that the increased availability of leucine resulting from mammary protein breakdown would increase intracellular concentrations of leucine, which would have favored its catabolism. Overall, these results confirm the high metabolic activity of the mammary gland, as protein synthesis by the mammary gland averaged 43% of whole body protein synthesis.

Variations in mammary metabolism during the natural filling of the udder with milk over a 12-h period between two milkings.

Two groups of four Holstein cows, one in their second and the other in their third or fourth lactation, were used to study temporal variations of mammary metabolism over a 12-h period between two milkings. Blood samples were collected every 30 min from an artery and a mammary vein during a 12-h interval between two milkings. Isoleucine, leucine, lysine, methionine, and phenylalanine mammary net fluxes varied or tended to change over time after milking with a similar pattern between whole blood and plasma. For these amino acids, whole blood and plasma net fluxes reached their maximum over the first 8 h after milking. Simultaneously, respiratory quotients decreased linearly and varied from 2.31 to 2.01 during the first 8 h of the period, suggesting active mammary lipogenesis. From 8 to 12 h after milking, mammary amino acid net fluxes decreased, while mammary oxygen uptake tended to increase with a concomitant decrease in the respiratory quotient reaching 1.84 to 1.40. These findings suggest that, beginning 8 h after milking, mammary uptake of amino acids starts to decrease and catabolic processes appear promoted; this phenomenon could help to explain the increase in milk production reported in the literature with increased milking frequency.

Penicillin concentrations in serum, milk, and urine following intramuscular and subcutaneous administration of increasing doses of procaine penicillin G in lactating dairy cows.

Eight healthy, non-pregnant, crossbred Holstein dairy cows (557-682 kg) within their first 3 months of lactation (13-21.5 kg of milk/day) were used. Cows were kept in tie stalls for the whole experiment. The 8 cows were randomly assigned to 2 (IM and SC) 4 x 4 balanced Latin square design experiments. Doses of procaine penicillin G (PPG) (300000 IU/mL) in each square were 7000, 14000, 21000 and 28000 IU/kg and were injected IM or SC once daily for 5 consecutive days. Volumes of PPG per site of injection never exceeded 20 mL. Blood was collected to determine the Cmax, Tmax, and AUC; urine and milk were also taken to measure the persistence of PPG in these fluids. Results show that serum Cmax and Tmax were only slightly affected by increasing the doses or the route of administration, whereas the AUC was linearly increased in relation to the dose injected in both modes of injection. In the urine, Cmax varied from 160 to 388 IU/mL and Tmax from 72-120 h during 5 consecutive days of PPG injection. A dose effect in Cmax was observed only for the IM route of administration and no variation (P > 0.05) was found between the IM and SC routes. Milk Cmax concentrations were only increased by the dose regimen in the IM group. At doses of 21000 and 28000 IU/kg, the IM group had a higher (P > 0.05) Cmax when compared with the SC groups. Milk PPG residues were not detectable over 96 h following the last IM injection, independently of the dose injected. However milk PPG residues were detected for up to 132 h following the last SC injection. These results show that when PPG is injected IM once daily in volumes not exceeding 20 mL/site at doses as high as 28000 IU/kg, the withdrawal period should be at least 96 h. Therefore, in the present model, there was no advantage to inject PPG by SC route to improve PPG kinetic parameters as the AUC, Cmax, or Tmax.

[Determination of the potassium balances in diary cows and the examination of daily and lactation period-associated variations. / Evaluation des équilibres potassiques chez la vache laitière et étude de ses variations journalières et selon le stade de production.

The objectives of this study were to evaluate a method of determining the internal and external potassium balances in dairy cows and to determine daily and lactation period-associated variations of these balances. Ingested potassium, milk potassium concentration (LK) and potassium urinary fractional clearance (FeK) defined the potassium external balance. Serum (SK) and red blood cell (CIEK) potassium concentrations defined the potassium internal balance. Whole blood, urine, and milk were collected at different times of the day from 19 cows, grouped in 5 lactation periods. Significant differences were observed for LK between lactation periods. Significant variations occurred during the day for CIEK, LK, and FeK. Thus, in order to monitor potassium balances during a time period, samples should be collected at the same time each day. This method of determining both potassium balances simultaneously could be useful in the study of the bovine hypokalemia syndrome.

Etiology, forms, and prognosis of gastrointestinal dysfunction resembling vagal indigestion occurring after surgical correction of right abomasal displacement.

The objectives of this study were to determine the etiology and types of vagal indigestion (VI) occurring after right displacement of the abomasum or abomasal volvulus (RDA/AV), and the prognosis for each type. Data of cows presented for RDA/AV from a retrospective (n = 288) and a prospective (n = 132) study were used. Vagal indigestion occurred in 39 and 22 cows in each study, respectively. A necropsy was performed in 29 cases. Gastric compartment dilation compatible with VI type III or IV occurred in 23 cases. An abnormal gastric wall was detected in 22 cases. Peritonitis was present in 18 cows. Vagal nerve lesions were present in 5 out of 13 cases studied. Clinical, hematological, and necropsy results suggested a classification of VI with respect to presence or absence of peritonitis. Gastric wall damage, peritonitis and vagal nerve lesions appear important in the etiology. Considering peritonitis occurrence, antimicrobial therapy appears necessary in the treatment of RDA/AV.

External pudic venous reflux into the mammary vein in lactating dairy cows.

A representative blood sample from the mammary vein depends on the functional integrity of the valves in the external pudic vein (EPV). To determine if the EPV valves maintain blood flow into the inguinal direction during the second and subsequent lactations, we used eight lactating cows catheterized in the EPV, the lateral branch of the cranial mammary vein (MV), and the external pudic artery (EPA). The averaged daily milk yields were 25.0 +/- 1.8 kg in cows in second lactation and 31.5 +/- 2.9 kg in older cows. The relative time taken by a pulse dose of p-amino hippuric acid (PAH) injected into the EPV, to reach the EPA and the MV, was measured in a first trial. In a second trial, we assessed the extent of alteration of the mammary PAH blood concentration with blood originating from other tissues using a continuous infusion of PAH into the EPA simultaneously with blocking or not any EPV backflux. From the first experiment, the PAH injected into the EPV appeared first in the EPA and then in the MV in cows in second lactation, suggesting that blood flow was towards the inguinal region. But in a third-lactation cow, the order of appearance was reversed. In parallel, the occlusion trial demonstrated that the concentration of PAH in the MV was diluted by 14 to 39% with blood draining nonmammary tissues only in cows in third or fourth lactation. This resulting reversed flow from the EPV towards the MV would have a detrimental impact on conclusions of mammary gland metabolism studies conducted with cows in their third lactation or higher.

Clinical response and immunomodulation following experimental challenge of calves with type 2 noncytopathogenic bovine viral diarrhea virus.

Eight calves between 16 and 18 weeks of age that were seronegative to bovine viral diarrhea virus (BVDV), bovine leucosis virus and bovine immunodeficiency-like virus were infected (day 0) intranasally with the type 2 noncytopathogenic Canadian 24515 field isolate of BVDV in order to evaluate the effect of BVDV infection on certain clinical, hematological and immunological parameters. All virus-exposed animals developed fever and showed a significant (P < 0.05, 0.01 or 0.001) drop in the number of circulating leucocytes (neutrophils, lymphocytes and monocytes) by day 3 or 5 post-exposure (PE), which continued to the end of the experiment at day 12 PE. BVDV was consistently isolated from the peripheral blood buffy coat cells from day 5 PE, and also from selected tissues (spleen, thymus, mesenteric and submaxillary lymph nodes, small intestine, lungs and thyroid gland) that were collected at the time of euthanasia of the animals at day 12 PE. Diminished significant (P < 0.05) percentages of peripheral blood mononuclear cells (PBMCs) expressing at their surface either B7 and MHC II molecules were observed in virus-exposed calves at days 7, 10 and/or 12 PE, when compared to virus-nonexposed control calves (n = 5). However, no changes in the percentages of PBMCs expressing either B4 or MHC I molecules were observed throughout the experiment. Finally, a significant (P < 0.05 or 0.01) enhanced phagocytic capability of the PBMCs, as analyzed by flow cytometry, was observed in virus-exposed animals at days 3, 5, 7, 10 and 12 PE, when compared to control calves. These results demonstrated the virulence of the 24515 isolate of BVDV in 4 to 4.5 month-old calves, and suggest that type 2 BVDV infection in calves is associated with dysregulation of certain immunological functions.

Description of 14 cases of bovine hypokalaemia syndrome.

The records of 14 cases of bovine hypokalaemia observed between 1983 and 1996 were reviewed. The most common history included a protracted, often infectious, disease. All age groups were represented. Although previously reported as a risk factor, isoflupredone acetate had not been administered to five of the cases. The following clinical signs were recorded in 10 cases: abnormal position of the head and neck, severe weakness, rumen hypomotility or atony, abnormal faeces, anorexia and tachycardia. Cardiac dysrhythmia was observed in six cases. Acid-base imbalance (alkalosis in 10 cases), hyperglycaemia and increased activities of aspartate aminotransferase and creatine kinase were associated with hypokalaemia ranging from 1.35 to 2.49 mmol/litre. Treatments included symptomatic treatment, supportive care and potassium chloride given intravenously and orally at an average total daily dose of 42 g/100 kg bodyweight (26 g by mouth and 16 g intravenously) for an average of five days. Eleven cases recovered after an average of three days.

Prevalence and geographical distribution of Giardia spp. and Cryptosporidium spp. in dairy farms in Québec.

Giardiasis and cryptosporidiosis are parasitic infections of humans, domestic animals, and wildlife. In cattle, direct transmission through the contamination of barns and/or pasture appears to be the principal mode of infection. In Canada there is only one study reported in the literature on the prevalence of giardiasis and one study on the prevalence of cryptosporidiosis. These studies were done in Alberta and Manitoba, respectively. The purpose of this study was to determine the prevalence of Giardia spp. and Cryptosporidium spp. infections in dairy farms in Québec. Calves were sampled from 505 dairy farms. We are reporting that 45.7% of the farms were found to be positive for Giardia spp. and 88.7% were infected with Cryptosporidium spp.

Morphological changes in the jejunum of calves naturally infected with Giardia spp. and Cryptosporidium spp.

Giardiosis and cryptosporidiosis are frequently diagnosed in calves at the large animal clinic of the veterinary school. Few studies have been reported in the literature regarding pathogenesis of these two intestinal protozoa. The aims of this study were to follow the histological changes in the villi and crypts and the changes in the number of intraepithelial lymphocytes in the jejunum of naturally infected calves during the acute phase of infection. For this purpose, 29 calves aged between 7 and 10 days were bought at a local auction. The animals were housed in individual pens to avoid cross-contamination. Fecal samples were examined microscopically for the presence of Giardia cysts and Cryptosporidium oocysts, three times per week for a period of 45 days. Six calves did not pass any cysts or oocysts and were used as controls. Fifteen calves passed Giardia cysts only, five passed both cysts and oocysts, and three passed oocysts only. The villus to crypt ratio index was 1.76 in the control group and 1.08 in the Giardia-infected group. In the Cryptosporidium-infected calves, the ratio was 1.18 and calves infected with both parasites had an index of 1.37. The number of intraepithelial lymphocytes per millimeter of jejunum tissue was 21 in the control group. This number was doubled in the calves infected with Giardia, but was slightly lower in the other infected groups. All of the infected calves had intermittent diarrhea and mucus was seen in many fecal samples.

Evolution of insulin-like growth factor-1, prostaglandin E2, and mitogenic activity of bovine mammary primary lymph during the dry period and lactogenesis.

Four pregnant cows near the end of lactation were fitted with a catheter in a lymph duct afferent to the supramammary lymph node. Cows were dried off 3 d after surgery, and samples of lymph were collected daily from the day of surgery until 4 d postpartum. Samples of blood and mammary secretions were taken before and after drying off and at parturition. Concentrations of most metabolites were lower in lymph than in serum. Concentrations of IGF-I and prostaglandin E2 were not affected at drying off but decreased and increased, respectively, at parturition. All IGF-binding proteins that were present in serum were also present in lymph fluid, but the binding activity was lower. Mitogenic activities of lymph samples taken at various physiological stages were determined on mammary epithelial (MAC-T) and fibroblast cell lines. Lymph was mitogenic, but mitogenic activity was not related to physiological stages. The correlation was high between mitogenic activity of lymph on MAC-T cells and the content of prostaglandin E2 in lymph. Supplementation of lymph with additional prostaglandin E2 increased mitogenic activity, and neutralization of lymph by antibodies reduced mitogenic activity. Basal medium conditioned by the epithelial cell line contained 100 to 250 pg/ml of immunoassayable prostaglandin E2.

[Cesarians on dairy cows: 159 cases]. / Césarienne chez la vache laitière: 159 cas.

A retrospective study on 159 cesareans on dairy cows referred between 1985 and 1989 to the veterinary teaching hospital is reported. The objective of the study was to evaluate the probability and the factors associated with survival and subsequent calving. The risk of dying for the cows was 24% following surgery. The risk of dying differed if the cow had an emphysematous, dead, or live calf; it was 63%, 21%, and 14%, respectively. Of all the cows returned to the farm and for which there was a breeding recorded, 64% calved again. But only 28% of all the cows returned home and, therefore, only 17% of all the referred cases calved again. We used multivariate analysis to obtain a predictive model on prognosis following surgery. The prognosis was only influenced by the calf condition at admission.

Long-term effects of human growth hormone-releasing hormone and photoperiod on hormone release and puberty in dairy heifers.

Forty-eight Holstein dairy heifers (98.9 kg BW; 3 mo old) were subjected for 246 d to twice-daily s.c. injections of saline (CTL) or human growth hormone-releasing hormone (GRH; 5 micrograms/kg BW) and to photoperiods of 8 h of light (L): 16 h of dark (D) or 16L:8D according to a 2 x 2 factorial arrangement of treatments. Jugular blood samples were collected from 16 heifers at 3, 4, 8, and 11 mo of age to monitor prolactin, growth hormone, and estradiol-17 beta. Plasma progesterone concentrations were monitored weekly in all heifers as an index of puberty (> 1 ng/mL). Growth hormone release was induced by GRH (P < .001) throughout the trial; area under the GH curve (AUC) averaged 1,582 vs 3,643 ng.min-1.mL-1 in CTL vs GRH heifers. However, GRH-induced GH response was less (P < .05) after the second daily injection. There was also an interaction (P = .08) between GRH, photoperiod, and days of treatment on GRH-induced GH response; AUC was greater in GRH-16L:8D than in GRH-8L:16D heifers at 3 mo but less at 8 mo of age. The PRL concentrations were similar for both photoperiods at 3 mo (36.4 vs 41.7 ng/mL) and 8 mo (16.2 vs 12.8 ng/mL) of age but were greater in 16L:8D vs 8L:16D heifers at 4 mo (18.4 vs 39.3 ng/mL) and 11 mo (26.3 vs 44.1 ng/mL) of age (photoperiod x day interaction, P < .001). Photoperiod of 16L:8D vs 8L:16D reduced (P < .01) weight at puberty in CTL heifers (251 vs 303 kg BW) and to a lesser extent in GRH-treated heifers (271 vs 284 kg BW; GRH x photoperiod interaction, P = .10). In conclusion, GH response is maintained throughout 8 mo of GRH treatment, and a 16L:8D photoperiod will reduce age and weight at puberty in heifers. Furthermore, refractoriness to photoperiod-induced PRL changes was detected.

Hormonal changes following an acute stress in control and somatostatin-immunized pigs.

Sixteen Yorkshire pigs (49 +/- 2 kg BW at 17 weeks) were immunized against somatostatin (SRIF; 4 males, 4 females) or its conjugated protein, bovine serum albumin (BSA; controls; 4 males, 4 females). Immunizations were done at 10, 12 and 14 weeks of age. Jugular vein cannulae were surgically inserted at 17 weeks of age. Five d later, half of each sex from the control and SRIF-immunized groups were stressed. The other half were subjected to the same stress 48 hr later. On both days, remaining animals were used as unstressed controls. The stress consisted of 5 min of snare restraint. Blood samples were collected from all pigs on both days at -20, -15, -10, -5, 0 (beginning of stress), 2, 6, 10, 15, 20, 30, 40, 60, 90, 120, 150, 180 and 240 min. Samples were radioimmunoassayed for cortisol, growth hormone (GH), prolactin (Prl), insulin, triiodothyronine (T3), thyroxine (T4) and insulin-like growth factor I (IGF-I). Mean antibody titers against SRIF (1:150 dilution) at 15 weeks were 0.49 +/- .09% and 54.5 +/- 4.9% for control and SRIF immunized pigs, respectively. Gender and immunization against SRIF had no effect on any of the variables measured (P greater than 0.05), except for T3 levels which were greater in females than in males (P less than 0.05). The stress by time of sampling interaction was significant (P less than 0.01) for all hormones measured. Cortisol values almost tripled within 15 min of stress, reaching concentrations above 100 ng/mL. Maximal increases were seen at 2 min for T4 (14%), at 6 min for T3 (36%), at 15 min for Prl (46%) and at 10 min for insulin (141%). An increase of 129% in GH concentration was present at 20 min in stressed pigs; however, an increase of 97% was also seen at 120 min in control pigs. Concentrations of IGF-I decreased (21%) by 60 min in the stressed pigs and remained depressed for up to 150 min. Stress associated with snare restraint, therefore, induces major changes in the concentrations of a series of hormones in growing pigs. On the other hand, immunization against SRIF did not alter any of the hormonal profiles measured. Since snare restraint is widely used to handle pigs during jugular puncture, any study of hormonal secretion in this species should be carried out under carefully controlled conditions in terms of blood sampling technique.

Effect of human growth hormone-releasing factor and(or) thyrotropin-releasing factor on growth, carcass composition, diet digestibility, nutrient balance, and plasma constituents in dairy calves.

Sixty male dairy grain-fed calves, raised from 70 to 223 kg BW in individual crates, were used in a 2 X 2 factorial arrangement to determine the effect of administration of human growth hormone-releasing factor (1-29)NH2 (GRF) and(or) thyrotropin-releasing factor (TRF). Calves received twice-daily s.c. injections of .9% NaCl (control), GRF (5 micrograms/kg BW), TRF (1 micrograms/kg BW) or GRF (5 micrograms/kg BW) plus TRF (1 micrograms/kg GTRF). Average daily gain and days on feed were not affected by treatments, but TRF treatment increased (P less than .05) total intake of dry matter (DM) and feed conversion ratio: 3.00, 3.02, 3.08, and 3.22 kg DM/kg weight gain for control, GRF, TRF, and GTRF, respectively. During two 7-d periods, after 66 and 75 d of treatment, feces and urine were collected from 40 calves (5 per treatment per period). Treatment with GRF increased (P less than .05) digestibility of DM, nitrogen (N), and energy and tended (P less than .20) to increase N retention. At slaughter, withers height was increased (P = .05) by GRF and carcass length was increased (P less than .05) by TRF. Pituitary and liver weights were increased (P less than .05) by TRF. The combination of GRF and TRF slightly increased (P less than .10) protein content and decreased (P less than .05) fat content of the 9-10-11th rib section. After d 1, GRF treatment chronically increased (P less than .05) insulin concentrations and also increased (P less than .10) IGF-I concentrations on d 29 and 57. In summary, chronic treatment with GRF and(or) TRF did not improve growth or efficiency, although GRF increased digestibility of DM, N, and energy and the GRF plus TRF combination resulted in slightly leaner carcasses.

Dose effect of human growth hormone-releasing factor and thyrotropin-releasing factor on hormone concentrations in lactating dairy cows.

Two experiments were conducted to study the effects of growth hormone-releasing factor (GRF) and thyrotropin-releasing factor (TRF) administration on hormone concentrations in dairy cows. In the first trial, 12 cows were used on 5 consecutive days to determine the effect of four sc doses of GRF (0, 1.1, 3.3 and 10 micrograms.kg-1 BW) and three sc doses of TRF (0, 1.1 and 3.3 micrograms.kg-1 BW) combined in a factorial arrangement. GRF and TRF acted in synergy (P = .02) on serum growth hormone (GH) concentration even at the lowest dose tested and GH response to the two releasing factors was higher than the maximal response observed with each factor alone. TRF increased (P less than .01) prolactin (Prl), thyrotropin (TSH), triiodothyronine (T3) and thyroxine (T4) concentrations similarly at the 1.1 and 3.3 micrograms.kg-1 doses and GRF did not interact (P greater than .40) with TRF on the release of these hormones. In the second trial, the effect of GRF (3.3 micrograms.kg-1 BW, sc) and TRF (1.1 micrograms.kg-1 BW, sc) was tested at three stages (18, 72 and 210 days) of lactation on serum Prl and TSH concentrations. Eighteen cows (n = 6 per stage of lactation) were used in two replicates of a 3 X 3 latin square. The TRF and GRF-TRF treatments were equipotent (P greater than .05) in increasing Prl and TSH concentrations. Prl and TSH responses were similar (P greater than .40) throughout lactation. In summary, GRF at doses ranging from 1.1 to 10.0 micrograms.kg-1 and TRF at doses ranging from 1.1 to 3.3 micrograms.kg-1 act in synergy on GH release and do not interact on Prl, TSH, T3 and T4 concentrations in dairy cows. Furthermore, Prl and TSH response to TRF are not affected by stage of lactation.