Relationship of foetal number and parity in Barbari goats to plasma profile of caprine pregnancy-associated glycoprotein (caPAG) during gestation and the early postpartum period.

This study was conducted to characterise pregnancy-associated glycoprotein (caPAG) in peripheral plasma during gestation and postpartum periods of nulliparous and multiparous does with one or two foetuses using a caPAG specific two-step sandwich ELISA system. Earliest time-points for detection of pregnancy and foetal number with appropriate cut-off values were identified. Plasma samples from 15 pregnant (multiparous: n = 8; nulliparous: n = 7; during pregnancy and postpartum period) and six non-pregnant (during oestrous cycle) goats were collected and analysed. Mean caPAG concentration was greater than the threshold for pregnancy detection (S-N = 0.40) on d22, peaked on d45 and remained unchanged until parturition. From d45 until parturition, caPAG concentration in multiparous does with two foetuses was 1.4 to 1.8 fold greater (P < 0.001) than those with one foetus. For the ELISA, 0.83 (S-N) was the most appropriate cut-off to differentiate does with two from those with a single foetus with an overall sensitivity and accuracy of 88.9% and 84.7%, respectively. Circulating caPAG concentration in multiparous goats was greater (P < 0.05) compared with nulliparous goats during the early pregnancy and postpartum periods. After parturition, caPAG concentrations markedly decreased and were basal within 14 days postpartum. In conclusion, using the caPAG specific ELISA, results indicated there were unique gestational and postpartum profiles for caPAG concentrations that are affected by number of foetuses and parity of the doe. The marked decrease in concentration of caPAG following parturition indicates there would not be compromising of the detection of subsequent pregnancies in goats using this technique.

Levels of neutral detergent fiber in diets with forage palm for dairy goats / Fibra em detergente neutro em dietas com palma forrageira para cabras leiteiras

This study aimed to evaluate the amount of fiber in the diet that is able to promote better use of nutrients in the diet of dairy goats when forage palm is the main source of roughage. Fifteen Alpine dairy goats were distributed in a 5 × 5 triple Latin square containing five treatments and five periods, and each treatment contained. We tested five diets composed of different contents of neutral detergent fiber from Tifton hay (NDFf) in the dry matter of rations containing forage palm: D1: 11.98%; D2: 18.31%; D3: 23.68%; D4: 28.76% and D5: 32.12% NDFf. The intake of dry matter, crude protein, neutral detergent fiber, non-fiber carbohydrates and ether extract decreased linearly with the increase in NDFf content. There was a linear increase of milk production and milk protein with increasing dietary NDFf. The fat, dry extract, protein, pH and acidity of the evaluated milk were not affected by increasing the levels of NDFf in the diet. In situations where dairy goats are fed forage palm and other types of forage are limited, 11.98% NDF from forage other than palm can be used as an effective source of NDF.(AU)

Objetivou-se, com este trabalho, avaliar a quantidade de fibra na dieta que é capaz de promover melhor uso de nutrientes na dieta de cabras leiteiras quando a palma forrageira é a principal fonte de forragem. Quinze cabras leiteiras alpinas foram distribuídas em um quadrado latino triplo 5 × 5 contendo cinco tratamentos e cinco períodos. Testaram-se cinco dietas compostas por diferentes teores de fibra em detergente neutro de feno de tifton (FDNf) na matéria seca de rações contendo palma forrageira: D1: 11,98%; D2: 18,31%; D3: 23,68%; D4: 28,76% e D5: 32,12% de FDNf. A ingestão de matéria seca, proteína bruta, fibra em detergente neutro, carboidratos não fibrosos e extrato etéreo diminuiu linearmente com o aumento do conteúdo de FDNf. Houve um aumento linear da produção de leite e da proteína do leite com o aumento da FDNf dietética. A gordura, a lactose, o extrato seco, o pH e a acidez do leite não foram afetados pelo aumento dos níveis de FDNf na dieta. Em situações em que as cabras leiteiras são alimentadas com palma forrageira e outros tipos de forragem são limitados, 11,98% de FDN de forragem diferente da palma podem ser usados como fonte efetiva de fibra.(AU)

Pregnancy-associated glycoproteins (PAGs) concentrations in water buffaloes (Bubalus bubalis) during gestation and the postpartum period.

For the first time in literature this study describes the pregnancy-associated glycoprotein (PAG) profile of buffalo cows during gestation and the post-partum period using antiserum raised against PAG-molecules purified from buffalo placenta (AS#860). Ninety-eight buffalo cows, belonging to a buffalo herd subjected to a synchronization and artificial insemination (AI) program, were enrolled in this study. Blood samples were taken on days 0 (AI), 23, 25, 28, 30 and then biweekly until the end of pregnancy. Pregnancy was confirmed by ultrasonography on days 28 and 45, and by rectal palpation from day 60 onwards. Blood samples were suspended for the non-pregnant cows on day 45, while the blood of 20 buffaloes that had calved was tested every five days from the day of calving until day 50 post-calving. A cut-off value of 1.0 ng/mL was used in order to discriminate between pregnant and non-pregnant buffaloes. We used Linear Mixed models after Log(x+1) transformation to analyse the PAG concentrations. Fifty-two buffalo cows had become pregnant out of 98 synchronized (53%) and 46 remained non-pregnant (47%) as shown by ultrasonography and the PAG analysis. Significant differences (P < 0.001) in PAG concentrations were observed between the pregnant and non-pregnant buffaloes from day 23 as the PAG of the non-pregnant cows was always close to zero. Conversely, the PAG of the pregnant cows increased progressively from day AI until day 105 post-insemination and then stabilized until the end of pregnancy. Regarding pregnancy diagnosis, the sensitivity of PAG-RIA 860 system (ability of the test to correctly identify pregnant buffalo) ranged from 23% on day 23-98% on day 28 post AI; the specificity (ability to correctly identify non-pregnant buffaloes) was 100% throughout the sampling period. PAG progressively decreased from parturition to day 25 post-partum; from day 30 post-partum, the concentrations fell below 1 ng/mL and were close to 0 on the last day of observation (50 d post-partum). In conclusion, our results showed that RIA-860 is highly accurate for diagnosing pregnancy in buffaloes starting from day 28 of gestation. Furthermore, the rapid disappearance of PAG concentration after calving means that a cut-off limit in post-partum for detecting a new pregnancy is not required.

Crosstalk between uterine serpin (SERPINA14) and pregnancy-associated glycoproteins at the fetal-maternal interface in pregnant dairy heifers experimentally infected with Neospora caninum.

Infection with Neospora caninum is the leading cause of abortion in cattle. In cows naturally infected with N caninum, plasma concentrations of pregnancy-associated glycoproteins (PAG) 1 and 2 indicate fetal-placental well-being, whereas an excess of progesterone in the second trimester of gestation has been related to high abortion rate. The immunosuppressive action of progesterone on the uterus during gestation has been attributed in part to the uterine serpins (SERPINA14). This study examines expression patterns of the genes SERPINA14, PAG, and PAG2 at the fetal-maternal interface in dairy heifers experimentally infected with N caninum during the second trimester of pregnancy, when most abortions takes place in natural conditions. Irrespective of infection, expression of SERPINA14 was higher, and expression of PAG1 and PAG2 lower, for intercaruncular endometrium than for caruncles or cotyledons. Cotyledonary tissues showed the highest expression of both PAG genes but lowest expression of SERPINA14. The expression of SERPINA14 was significantly higher in intercaruncular endometrium of control dams than for infected animals, pointing to potential disruption of modulation of maternal immune function during infection. Dramatically reduced SERPINA14 was particularly apparent in infected dams with aborted fetuses. There was also a negative association between N caninum antibody titers with SERPINA14 and PAG expression in infected animals, further suggesting that N caninum infection downregulates the uterine immunosuppressive function of SERPINA14.

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy-Associated Glycoproteins 1 and 2 in Foetal Fluids.

Plasma concentrations of PAG-1 are used for pregnancy diagnosis and as a marker of placental/foetal well-being, while those of PAG-2 may be an indicator of abortion risk in Neospora caninum-infected cows. Studies have shown that N. caninum infection modifies PAG-1 and PAG-2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG-1, PAG-2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG-2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non-infected foetuses. In infected foetuses, significantly higher concentrations of PAG-1 (p < 0.001) and PAG-2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non-infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG-1 and PAG-2 levels and pH values than fluids from non-infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.

Experimental Neospora caninum infection modifies trophoblast cell populations and plasma pregnancy-associated glycoprotein 1 and 2 dynamics in pregnant dairy heifers.

Neospora caninum is an obligate intracellular protozoan that causes abortion in cattle worldwide. Plasma concentrations of pregnancy-associated glycoprotein 1 (PAG-1), produced by binucleate trophoblast cells, are used for pregnancy diagnosis and as a marker of foetal-placental well-being, while PAG-2, produced by both mono- and binucleate trophoblast cells, may serve as an indicator of abortion risk. In prior work, natural N. caninum infection was found to modify plasma PAG-1 and PAG-2 patterns. The present study examines PAG-1 and PAG-2 dynamics and trophoblast cell populations following experimental infection with N. caninum. The study population was comprised of 17 N. caninum seronegative Holstein-Friesian heifers. On day 110 of gestation, 6 heifers were inoculated intravenously with 10(7) taquyzoites of N. canimum. Plasma samples for PAG-1 and PAG-2 determinations were collected on days 0, 7, 14, 21 and 42 post infection. During the study course, pregnancy was normally expressed in all controls while three infected heifers lost their foetuses. All heifers were euthanised on day 42 post infection and placentome samples from the 14 non-aborting heifers were collected to examine trophoblast cell populations. Plasma PAG-1 and PAG-2 concentrations in non-infected heifers increased until the day of euthanasia while non-aborting infected heifers showed a temporary fall in PAG-1 (P<0.004) and PAG-2 (P<0.002) concentrations from 7 to 14 days post infection (dpi). The two dams aborting at 14 and 21 dpi and a third dam with a mummified foetus upon euthanasia showed dramatic PAG-1 and PAG-2 reductions from 14 dpi to undetectable levels upon euthanasia. A stereological study of placentomes revealed significantly higher relative proportions of mono- (P=0.035) and binucleate (P=0.029) trophoblast cells at 42 dpi in non-infected heifers than infected non-aborting heifers. According to our findings, following experimental N. caninum infection on day 110 of gestation, non-aborting heifers showed a brief reversible drop in plasma PAG-1 and 2 concentrations two weeks later and reduced proportions of bi- and mono-nucleate trophoblast cells 42 days after infection. In aborting dams, dramatically reduced PAG levels were related to severe placental damage and a non-viable pregnancy.

Milk prolactin response and quarter milk yield after experimental infection with coagulase-negative staphylococci in dairy heifers.

Coagulase-negative staphylococci (CNS) are the most common bacteria involved in subclinical mastitis in dairy cows. Remarkably, CNS-infected dairy heifers produce more milk than uninfected heifers. Because the lactation hormone prolactin (PRL) is also involved in mammary gland immunity, we investigated the milk PRL response and the mammary quarter milk yield following experimental CNS challenge. Eight healthy Holstein-Friesian heifers in mid-lactation were experimentally infected using a split-udder design with 3 different CNS strains: one Staphylococcus fleurettii (from sawdust bedding) and 2 Staphylococcus chromogenes strains (one isolate from a teat apex, the other isolate from a chronic intramammary infection). Three mammary quarters per heifer were simultaneously inoculated with 1.0×10(6) cfu, whereas the remaining mammary quarter was infused with sterile phosphate-buffered saline, serving as a control. An existing radioimmunoassay was modified, validated, and used to measure PRL frozen-thawed milk at various time points until 78h after challenge. The mean milk PRL level tended to be higher in the CNS-challenged mammary quarters compared with the control mammary quarters (7.56 and 6.85ng/mL, respectively). The increase in PRL over time was significantly greater in the CNS-challenged mammary quarters than in the control mammary quarters. However, no difference was found in the PRL response when comparing each individual CNS strain with the control mammary quarters. The mean mammary quarter milk yield tended to be lower in the CNS-infected mammary quarters than in the control mammary quarters (1.73 and 1.98kg per milking, respectively). The greatest milk loss occurred in the mammary quarters challenged with the intramammary strain of S. chromogenes. Future observational studies are needed to elucidate the relation between PRL, the milk yield, and the inflammatory condition, or infection status, of the mammary gland.

Pregnancy-associated glycoprotein (PAG) concentration in plasma and milk samples for early pregnancy diagnosis in Lacaune dairy sheep.

In the present study, four RIA systems (RIA-1 to -4) based on two antisera raised against ovine pregnancy-associated glycoproteins (ovPAGs), combined with an ovine or a bovine PAG tracer were used to measure PAG concentrations in plasma and milk samples of dairy ewes. Blood and milk samples were collected on different days of gestation: 0, 18, 20, 22, 25, 28, 32, 42, and 49. From day 20 onward, the PAG in plasma could be detected in all pregnant ewes using the four RIA systems. By using milk, except for RIA-1, the other systems showed a sensitivity of 100% from day 28 of gestation onward. In plasma, PAG concentrations were higher in multiple than in single pregnancies, while no clear relationship was observed in milk. In conclusion, milk is a good alternative to plasma for early pregnancy diagnosis in sheep from day 28 to day 42.

Effects of crossbreeding on endocrine patterns determined in pregnant beef/dairy cows naturally infected with Neospora caninum.

Neospora caninum is an intracellular coccidian parasite causing abortion worldwide in dairy cattle. Studies have shown that N caninum infection modifies endocrine patterns and that beef cows or cows with a crossbreed pregnancy, especially for a greater maternal-paternal genetic distance, show a reduced risk of abortion when naturally infected compared with pure-breed Holstein Friesian (HF) pregnancies. This study examined the effects of crossbreeding on plasma progesterone (P4), pregnancy-associated glycoprotein (PAG)-1 and -2, and prolactin patterns produced during gestation in N caninum-infected beef/dairy cows. We analyzed 74 pregnancies in nonaborting cows carrying a single fetus established in 26 lactating HF cows bred with Holstein bulls (H-H), 13 HF cows bred with Limousin bulls (H-L), 12 HF cows used as recipients of transferred in vivo-produced frozen-thawed Rubia Gallega (beef breed; RG) embryos (H-RG-ET), and 23 RG suckling cows bred with RG bulls (RG-RG). Of the cows, 29 (39%) were seropositive for N caninum. Blood samples for hormone and placental protein determinations were collected on Days 30, 60, 90, 120, 150, and 180 of gestation. Procedures of repeated measures analysis of variance throughout gestation revealed higher PAG-1 levels in the H-L and H-RG-ET groups compared with H-H and RG-RG. Higher prolactin levels were detected in RG-RG than in the remaining groups. N caninum seropositivity and fetal genotype had a significant effect on PAG-2 concentrations, such that highest levels of PAG-2 occurred in RG-RG seropositive cows and lowest in H-H seropositive cows, whereas Neospora-seropositive cows showed lower P4 concentrations than their seronegative partners. In conclusion, chronic N caninum infection modifies endocrine patterns of PAG-2 and P4 during pregnancy, whereas PAG-1 and prolactin concentrations are affected by breed and fetal genotype, irrespective of Neospora infection status.

Correlation between acute and chronic 24-hour blood pressure response to resistance training in adult women.

The purpose of the present study was to correlate the acute and chronic decrease in blood pressure (BP) following resistance training (RT). 13 normotensive women (18-49 years) completed an acute whole body RT session with 3 sets of 10 repetitions at 60% 1RM and then 8 weeks of RT as follows: 3/week, 3 sets of 8-12 repetitions maximum. Systolic (SBP) and diastolic BP (DBP) were measured up to 60 min and 24 h following RT (acute and chronic). The greatest acute decrease of SBP (108.5±7.0 mmHg) and DBP (71.5±6.4 mmHg) values over the 60-min period were reduced compared to pre-exercise (117.3±11.7 and 79.3±8.2 mmHg, respectively; p<0.05). The chronic effect on resting BP was observed only for those presenting acute post-exercise hypotension (PEH). The change in both SBP and DBP following acute RT was correlated with the chronic change in resting SBP and DBP (r>0.5; p≤0.05). The change in 24 h BP after acute RT was correlated with the chronic reduction in SBP (r=0.74) and DBP (r=0.80). The magnitude of PEH is a promising candidate for the prediction of individual BP-related training efficacy.

Gamma interferon production and plasma concentrations of pregnancy-associated glycoproteins 1 and 2 in gestating dairy cows naturally infected with Neospora caninum.

Gamma interferon (IFN-γ) production and cross-breed pregnancy have been attributed a role in protecting dairy cows infected with Neospora caninum against abortion. Plasma levels of pregnancy-associated glycoproteins-1 (PAG-1) are a marker of placental/foetal well-being and of PAG-2 is an abortion risk indicator in chronically N. caninum-infected animals. The present study examines, in cross-breed pregnancies, interactions between IFN-γ production and levels of PAG-1 and PAG-2 in non-aborting naturally Neospora-infected dairy cows. Data were obtained from 60 pregnant Holstein-Friesian cows: 44 Neospora-seropositive and 16 Neospora-seronegative; 12 became pregnant using Holstein-Friesian semen and 48 using Limousin semen. Blood samples were collected on Days 40, 90, 120, 150, 180 and 210 of gestation. Gamma interferon was only detected in the plasma of nine of the 44 Neospora-seropositive cows, all of them became pregnant using Limousin semen. Through GLM procedures, in cows inseminated with Limousin semen and Neospora-seropositive cows showing no IFN-γ production, PAG-1 concentrations were high and increased throughout gestation compared to the levels detected in cows inseminated with Holstein-Friesian semen and Neospora-seropositive cows producing IFN-γ, respectively. In Neospora-seronegative cows and in Neospora-seropositive cows showing no IFN-γ production, significantly increased PAG-2 concentrations were observed on gestation Day 120. Our findings indicate that IFN-γ production correlates negatively and the production of antibodies against N. caninum is uncorrelated with plasma PAG concentrations during gestation in Neospora-infected dairy cows. Accordingly, IFN-γ production could be linked to the transplacental migration of tachyzoites, which may cause a reduction in PAG levels.

Plasma concentrations of pregnancy-associated glycoproteins measured using anti-bovine PAG-2 antibodies on day 120 of gestation predict abortion in dairy cows naturally infected with Neospora caninum.

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy-associated glycoprotein-2 (PAG-2) concentrations throughout pregnancy and (ii) whether plasma PAG-2 concentrations could predict abortion in N. caninum-infected cows. The study was performed on a commercial Holstein-Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non-aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post-insemination) in non-aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG-1 and PAG-2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG-2 concentrations throughout gestation (between-subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora-seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG-2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG-2 concentrations and the risk of abortion in Neospora-infected dairy cows. Thus, plasma PAG concentrations measured using anti-boPAG-2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals.

The inseminating bull and plasma pregnancy-associated glycoprotein (PAG) levels were related to peripheral leukocyte counts during the late pregnancy/early postpartum period in high-producing dairy cows.

It has been established that the immunologic and endocrine status of the peripartum dairy cow determines the animal's subsequent productive and reproductive performance. Thus, at parturition reduced immune functions of peripheral blood polymorphonuclear cells (PMN) has been observed after a peak in pregnancy-associated glycoproteins (PAGs), and, more recently, the inseminating bull was linked to plasma levels of bovine PAGs in pregnant Holstein-Friesian dairy cows. The present study sought to determine whether changes in leukocyte counts during the peripartum period, indicative of the animal's immune status, could be related to the inseminating bull and to PAG levels. Ninety-six clinically healthy, single pregnant cows in a commercial dairy herd were selected. Four samples were collected before parturition (on gestation Days 220-226, 234-240, 248-254, and 262-268) and two samples after parturition (on Days 14-21, and 28-34 postpartum) to analyze total and differential blood cell counts. Based on GLM analysis procedures of variance for repeated measures, the inseminating bull was found to affect counts of total leukocytes and lymphocytes (P < 0.001; between-subject effects) throughout the peripartum period. In addition, cows with high plasma PAG levels (> 900 ng/ml) on Day 262-268 of gestation had higher numbers of total leukocytes and neutrophils throughout the peripartum (P < 0.001; between-subject effects). Young animals (≤ 1 lactation) had higher total leukocyte and lymphocyte counts than older cows (2 or more lactations) throughout the study period. These results reveal a clear relationship between the inseminating bull or plasma PAG levels and peripheral leukocyte counts during the peripartum period in dairy cows.

Pregnancy-associated glycoprotein, chymosin and pepsinogen immunoreactivity of proteins extracted from fetal gastric tissue in bovine species.

The objective of this work was to investigate the expression of gastric aspartic proteinases in fundic and pyloric mucosa removed from bovine fetuses. For this purpose, fractions issued from classical biochemical protocols were analyzed by proteolytic method, by PAG-RIA and by Western blot with the use of antisera raised against both pepsinogens and PAG. A strong reaction of proteins extracted from the fundic mucosa collected at the beginning of pregnancy was revealed with both anti-bPAG-I and anti-bPAG-II antisera, suggesting the expression of pepsinogen F in bovine species. Concerning pyloric mucosa, the analysis by Western blot highlighted a very strong immunoreaction with the anti-bovine chymosin serum. Amino-terminal sequencing allowed to identify bovine fetuin and albumin in fundic extracts, chymosin in the pyloric mucosa extracts, as well as some unknown proteins in both mucosa. Despite no N-terminal microsequence corresponding to the hypothetical pepsinogen F could be identified, it cannot be excluded that an existing bovine pepsinogen F-like molecule could be degraded during the purification procedure or that co-purified proteins could be responsible for masking its N-terminal microsequence.

Acute cardiorespiratory and metabolic responses during resistance exercise in the lactate threshold intensity.

The aims were both to determine lactate and ventilatory threshold during incremental resistance training and to analyze the acute cardiorespiratory and metabolic responses during constant-load resistance exercise at lactate threshold (LT) intensity. Ten healthy men performed 2 protocols on leg press machine. The incremental test was performed to determine the lactate and ventilatory thresholds through an algorithmic adjustment method. After 48 h, a constant-load exercise at LT intensity was executed. The intensity of LT and ventilatory threshold was 27.1±3.7 and 30.3±7.9% of 1RM, respectively (P=0.142). During the constant-load resistance exercise, no significant variation was observed between set 9 and set 15 for blood lactate concentration (3.3±0.9 and 4.1±1.4 mmol x L(-1), respectively. P=0.166) and BORG scale (11.5±2.9 and 13.0±3.5, respectively, P=0.783). No significant variation was observed between set 6 and set 15 for minute ventilation (19.4±4.9 and 22.4±5.5 L x min(-1), respectively, P=0.091) and between S3 and S15 for VO2 (0.77±0.18 and 0.83±0.16 L x min(-1), respectively, P=1.0). Constant-load resistance exercise at LT intensity corresponds to a steady state of ventilatory, cardio-metabolic parameters and ratings of perceived exertion.

Effect of restraint stress on plasma concentrations of cortisol, progesterone and pregnancy associated-glycoprotein-1 in pregnant heifers during late embryonic development.

The aim of the present study was to evaluate the effect of restraint stress, which is commonly practised in the field, on plasma concentrations of cortisol, progesterone (P4) and bovine pregnancy-associated glycoprotein-1 (boPAG-1) in pregnant heifers between Days 30 to 40 of gestation. Twelve Holstein-Friesian heifers between Days 30 (Day 0 of experiment) and 40 (Day 10 of experiment) of pregnancy in a Hungarian dairy farm were used in the present study. The heifers were exposed to an acute stressor consisting of immobilisation (restraint stress) in a crush for 2 h (Group 1, n = 6) on Day 2 (Hour 48) and for 2 × 2 h (Group 2, n = 6) on Days 2 and 3 (Hour 72) of the experiment. Transrectal ultrasonography (7.5 MHz linear-array rectal transducer) was performed daily from Day 0 to Day 10 of the experiment to detect embryonic heartbeat or the fate of the conceptus. Blood samples were withdrawn before each ultrasonographic examination. Additional blood samples were withdrawn by 1 and 2 h (at Hours 49 and 50 in Groups 1 and 2 and Hours 73 and 74 in Group 2) of the onset of applying the stressor. Plasma cortisol, P4 and boPAG-1 concentrations were measured by radioimmunoassay. Acute restraint stress significantly (P < 0.001) increased the plasma cortisol level in pregnant heifers at 1 h of the exposure to the stressor at Days 2 (48 h) and 3 (72 h) of the experiment. On the other hand, the restraint stress did not affect the concentration of P4 and boPAG-1 concentrations in both groups. In conclusion, restraint stress for 2 h during early pregnancy in heifers increased blood cortisol, but it did not affect the concentrations of P4 and boPAG-1 between Days 30 to 40 of gestation.

Clinical implications of induced twin reduction in dairy cattle.

Embryo reduction may prevent the negative effects of twinning in dairy cattle; however, the technique may carry an additional risk of pregnancy loss. The aim of this study was to evaluate the effect on pregnancy maintenance of embryo reduction by manual amnion rupture in unilateral and bilateral twin pregnant cows. A secondary objective was to examine the dynamics of endocrine factors following the treatment. On Day 35-41 of gestation 55 cows bearing two live twin embryos (28 bilateral, 27 unilateral) were randomly assigned to a twin reduction group (n = 27; cows fitted with a progesterone releasing intra-vaginal device for 21 days after manual amnion rupture) or control group (n = 28; untreated cows). Pregnancy loss before Day 90 was recorded in nine control and eleven twin reduction cows (32.1% vs 40.7%, respectively, p = 0.508). Logistic regression models indicated that laterality was the only variable significantly affecting pregnancy loss. The pregnancy loss risk was 8.7 times higher for unilateral than for bilateral twin pregnancies (59.3% vs 14.3%, respectively, P < 0.001) yet was similar in the unilateral control and unilateral twin reduction cows (62.3% vs 53.8%, respectively, P = 0.581). In contrast, four of 14 cows with bilateral twin pregnancies undergoing twin reduction lost their pregnancies while no losses were recorded in control cows with bilateral pregnancies (P = 0.049). A rise in plasma progesterone concentration was detected on the day following treatment in the twin reduction group and concentrations remained high within the first week of treatment. Plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations fell between Day 35-41 and Day 42-48, regardless of treatment. Our findings indicate that embryo reduction by manual amnion rupture did not carry an additional risk of pregnancy loss for unilateral twin pregnancies, whereas it increased the risk of pregnancy failure in bilateral twin pregnancies. However, benefits of preventing cows from delivering twins might also be considered when assessing the success of embryo reduction in bilateral twin pregnancies.

Plasma urea nitrogen in relation to pregnancy rate in dairy sheep.

The aim of this field study was to investigate the relationship of plasma urea nitrogen (PUN) with the pregnancy rate in lactating Awassi × Merino ewes. One hundred and eighty-five Awassi × Merino ewes were used in the present study. Ewes were fed a diet containing 17.4% crude protein and were milked twice a day by the milking machine. The ewes were synchronized for estrus by insertion of intravaginal sponges containing 30 mg flurogestone acetate for 14 days. At the time of sponge removal each ewe was administered eCG (600 IU). All ewes were inseminated twice with fresh semen into the external os of the cervix at 48 and 56 h after sponge removal. The day of insemination was considered as Day 0 for calculating the gestational period. Blood samples were collected from each ewe at Days 0, 18 for measurement of PUN concentrations and at Day 22 after AI for measurement of pregnancy-associated glycoprotein (PAG) by radioimmunoassay (RIA). Thirty-eight ewes (20.5%) were confirmed pregnant by PAG-RIA test at Day 22 and by ultrasonography at Day 80. The mean (±S.D.) concentration of PUN in all ewes at Day 0 was 12.7±4.6 mmol/L. There were non-significant differences in the level of PUN between pregnant and non-pregnant ewes at Days 0 (12.2±4.2 mmol/L vs. 12.8±4.7 mmol/L, respectively) and 18 (9.6±2.9 mmol/L vs. 10.4±4.0 mmol/L, respectively) after AI. Mean PUN concentrations decreased significantly from Day 0 to Day 18 after AI in both pregnant and non-pregnant ewes. By using logistic regression analysis, there was no effect of PUN concentrations on the probability of pregnancy occurrence in the studied ewes (odds ratio: 0.97; 95% confidence interval: 0.9-1.05; P=0.45). In conclusion, there was no evidence of a relationship between PUN concentration and pregnancy rate for lactating Awassi × Merino ewes in the present study because of low pregnancy rate observed.

Alternative method for quantification of alfa-amylase activity.

A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 A degrees C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing alpha-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale alpha-amylase over a wide range (2.4 to 7,500 U.mL-1) in scientific investigation as well as in teaching laboratory activities.

Neospora caninum and coxiella burnetii seropositivity are related to endocrine pattern changes during gestation in lactating dairy cows.

Q fever is a zoonotic infection caused by Coxiella burnetii that is endemic worldwide. Domestic ruminants are a source of infection for humans. Given the suggestion that the bacterium recrudesces during pregnancy in cattle, this study was designed to determine whether C. burnetii infection affects hormonal patterns, such as progesterone, cortisol, pregnancy associated glycoproteins (PAG), and prolactin during gestation in lactating cows. Possible interactions with Neospora caninum were also explored. The study was performed on 58 gestating non-aborting cows. Blood samples for hormone determinations were collected on Days 40, 90, 120, 150, 180, and 210 of gestation. For antibody determinations, blood was collected at day 40 postinsemination and postpartum. By GLM repeated measures analysis of variance, we established the effects of production and reproductive variables as well as Coxiella and Neospora seropositivity related to changes on cortisol, PAG, progesterone, and prolactin levels. Coxiella antibody levels were significantly related to cortisol, PAG, and plasma progesterone concentrations, whereas Neospora seropositivity was linked to plasma progesterone concentrations. The interaction between Coxiella and Neospora seropositivity was correlated with cortisol and plasma progesterone levels, whereas the interaction seropositivity against C. burnetii-plasma cortisol concentration was related to plasma PAG levels. Finally, an effect of lactation number only was observed on plasma prolactin. Our findings suggest that both the N. caninum and C. burnetii infection or the presence of both modify endocrine patterns throughout gestation. Cows seropositive to both, Neospora and Coxiella, showed higher plasma progesterone levels than the remaining animals examined. Seropositivity to C. burnetii was associated with placental damage and diminishing PAG levels throughout the second half of gestation, along with increased plasma cortisol levels on Day 180 of gestation.