This study compares the fertility effects of inducing ovulation using the GnRH analogue, dephereline, versus natural GnRH at the end of a 5-day progesterone(P4)-based protocol for fixed-time ...artificial insemination (FTAI) in in heat-stressed and non-heat stressed lactating dairy cows. Cows were given GnRH (GnRH group, n = 369) or dephereline (DEPH group, n = 379) and were inseminated 14–20 h later. Dephereline treatment increased corpus luteum (CL) size on Day 7 post-AI compared with GnRH (P < .0001) while a one-mm increase in CL size was found to give rise to a 1.1-fold increase in the pregnancy rate at FTAI (P = .001). Based on odds ratios, the interaction between treatment and heat stress had a significant effect on the ovulation failure rate (P < .01). This meant that relative to non-heat-stressed GnRH-treated cows, ovulation failure was 2.9 times more likely in heat-stressed GnRH-treated cows (P = .001), 0.3 times less likely in non-heat-stressed DEPH-treated cows (P = .04) and was similar in heat-stressed DEPH-treated cows. Further, non-heat-stressed DEPH-treated cows were more likely to conceive by a factor of 1.6 than the remaining cows (P = .03). Finally, GnRH-treated multiparous cows were 9.9 times more likely to suffer pregnancy loss than the remaining cows (P = .03). Our results indicate that, compared to treatment with GnRH, dephereline reduced the risk of ovulation failure and consequently increased the pregnancy rate under heat stress conditions. In multiparous cows, dephereline treatment also reduced the negative age effect on pregnancy maintenance.
•Compared to GnRH, dephereline favored ovulation under heat stress conditions.•Dephereline returned a higher pregnancy rate in heat-stressed cows than GnRH.•Dephereline treatment reduced the pregnancy loss rate in multiparous cows.
This study sought to examine the impact of the presence of two co-dominant (ovulatory) follicles at the time of artificial insemination (AI) on the ovulatory response to GnRH given in a fixed-time AI ...protocol. The study population comprised 622 lactating dairy cows: 306 (49.2%) with a single follicle, 198 (31.8%) with two bilateral follicles (one follicle per ovary) and 118 (19%) with two unilateral follicles (same ovary). Based on odds ratios, cows with two bilateral or unilateral follicles were less likely (by factors of 0.09 and 0.11, respectively) to undergo ovulation failure compared with cows with one follicle (P = 0.01 and P = 0.02, respectively); the likelihood of ovulation failure decreased 0.75 times with every one-mm increase in follicle diameter for cows with a single follicle, whereas individual follicle diameter was not related to ovulation failure in cows with two bilateral follicles (P = 0.001). The likelihood of double ovulation decreased 0.7 times with every one-mm diameter difference between the larger and smaller follicle for all cows with two follicles (P = 0.001), whereas cows with two unilateral follicles showed a higher (P < 0.05) double ovulation rate than cows with two bilateral follicles. In 116 (58.6%) of the cows with two bilateral follicles, only the larger follicle ovulated in 59.5% cows, whereas only the smaller one ovulated in the remaining 40.5% cows. In these cows, a one-mm size difference between the larger and the smaller follicle gave rise to a 1.12-fold increase in the ovulation failure rate for the larger follicles (P = 0.0001). Cows with two bilateral follicles were more likely (by a factor of 1.5) to conceive than cows with one follicle (P = 0.001). Significant right-left differences were not found in cows with two bilateral follicles, whereas the right ovary was more active than the left in the remaining cows. Our results indicate that cows with two co-dominant follicles at AI show different ovulation patterns to those with one dominant follicle. A higher rate of ovulation failure was observed among cows with one follicle than cows with two follicles, whereas the conception rate was higher for cows with two bilateral follicles than for the remaining cows. In cows with two follicles, double ovulations along with ovulation of the smaller follicle were related to the least size difference between the larger and smaller follicle.
•The likelihood of ovulation increased with increasing follicle diameter in cows with one follicle.•Individual follicle size was not related to ovulation failure in cows with one ovulatory follicle in each ovary.•Cows with two unilateral follicles showed a greater likelihood of double ovulation than cows with two bilateral follicles.•The conception rate was higher for cows with two bilateral follicles compared with two unilateral or a single follicle.
Reducing feeding costs in suckler beef herds to improve economic returns could have detrimental impacts on fertility. This study sought to determine whether maternal nutrient restriction during early ...pregnancy affects interferon-stimulated gene (ISG) expression in peripheral blood mononuclear cells during the peri-implantation period in two beef cattle breeds. Relationships were also examined between subnutrition and pregnancy failure defined according to ISG fold changes on Days 18 and 21 and to plasma pregnancy specific protein B (PSPB) concentrations on Day 28 post-artificial insemination (AI). Pirenaica or Parda de Montaña dams were assigned to a control (n = 23) or subnutrition (n = 30) group, receiving 100% or 65% of their estimated nutritional requirements from Day 1 to 82 post-AI, respectively. Treatment did not affect ISG expression or fertility. According to ISG fold changes (chi-square P = .023) or PSPB levels (chi-square P = .04) recorded in the subnutrition group, late embryo loss was more likely than in controls. Positive correlation was detected between Day 28 PSPB concentrations and both Day 18 MX1, MX2 and ISG15 expression, and Day 21 OAS1 expression. OAS1 and MX1 fold changes were found to be the best variables to discriminate pregnancy status. Our findings indicate that maternal nutrient restriction during the first third of pregnancy does not impair embryo signalling yet may increase the risk of pregnancy failure.
•Maternal subnutrition in suckler beef cattle does not impair conceptus signalling.•Nutrient restriction in early pregnancy may increase the risk of pregnancy failure.•OAS1 and MX1 fold changes were the best variables to predict pregnancy status.
In the present study, IFN-γ (Th1), IL-17A (Th17) and IL-4 (Th2) concentrations in response to concanavalin (ConA) and Neospora caninum antigen (Nc-1) stimulation were determined in cultures of cells ...from control uninfected (n=4), naturally N. caninum-infected (n=3) and experimentally N. caninum-infected (n=6) pregnant dams and their foetuses. Experimental animals were infected at 110days of gestation and euthanized 6weeks post-infection. In culture supernatants from the dams, significantly higher IFN-γ and IL-4 levels were found in the experimentally-infected animals compared to the control or naturally-infected dams. However, among the experimentally-infected dams no significant differences in IFN-γ production were observed regardless of the incidence of live or aborted/dead foetuses, though spleen cultures of dams carrying live foetuses showed the highest levels of IFN-γ. IL-17A production was very low and occasional in the dams infected with N. caninum and did not seem to be a major regulator of IFN-γ production in this model. Experimentally infected dams with live foetuses showed higher IL-4 levels and accordingly IFN-γ/IL-4 ratios were significantly lower than ratios recorded for cows with aborted/dead foetuses. In the infected foetuses of these dams, only spleen cultures showed high levels of IFN-γ and IL-4 after Nc-1 antigen and ConA stimulation, respectively. No IL-17A was detected in the foetuses. As conclusion, although we could not clearly relate a protective immune response against N. caninum abortion only to IFN-γ levels in cell cultures, our results highlight the important role of an inverse IFN-γ/IL-4 balance in conferring protection against abortion induced by this parasite.
•IFN-γ, IL-17A and IL-4 production in pregnant dams experimentally infected with Neospora caninum and in their foetuses.•Cytokine production compared in naturally infected and N. caninum experimentally infected dams, and in control pregnant dams.•Protective immunity against abortion could not definitively linked to IFN-γ levels in cell culture supernatants.•An important role was observed of an inverse IFN-γ/IL-4 balance in protecting against N. caninum abortion.
Contents
This study examines gene expression patterns in dairy heifers experimentally infected with N. caninum during on Day 110 of pregnancy with live foetuses at euthanasia, 42 days later. The ...study population was constituted of four non‐infected controls and three infected dams. Gene expression was determined on gamma interferon (IFNγ), (Th1 pro‐inflammatory cytokine), interleukin‐4 (IL4) (Th2 pro‐gestation cytokine) or interleukin‐10 (IL10) (T regulatory cytokine) and the serine peptidase inhibitor SERPINA14 in intercaruncular, placental, uterine lymph node (UTLN) and luteal tissue samples. Intercaruncular SERPINA14 expression was negatively correlated with IFNγ expression in cotyledon samples and with IL4 expression in UTLN. No relationships were detected between cytokine gene expression at the foetal–maternal interface and SERPINA14 expression in the luteal samples. Our findings suggest that gene expression of the uterine serpin SERPINA14 correlates negatively with the expression of Th1 and Th2 cytokines at the foetal–maternal interface but not in the corpus luteum.
This study compares the fertility effects of inducing ovulation using human chorionic gonadotropin (hCG) versus gonadotropin releasing hormone (GnRH) at the end of a 5-day progesterone(P4)-based ...protocol for fixed-time artificial insemination (FTAI) in heat-stressed and non-heat stressed anestrous lactating dairy cows. Heat stress (HS) was defined as an environmental temperature-humidity index (THI) > 72. A P4 intravaginal device (CIDR) was fitted for five days and GnRH administered upon CIDR insertion and a double dose (24 h apart) of prostaglandin F2α upon CIDR removal. Cows then received either GnRH (GnRH group; n = 506), or hCG (hCG group; n = 493) 36 h after CIDR removal and were inseminated 50–56 h after CIDR removal. Ovulation failure was investigated in a subset of 425 cows: 223 and 202 receiving GnRH and hCG, respectively. Based on odds ratios, the interaction between treatment and HS had a significant effect on the ovulation failure rate (P = 0.01). This meant that compared to the rates recorded in non-heat-stressed, GnRH-treated cows (13%), ovulation failure in heat-stressed GnRH-treated cows (25.3%) was more likely by a factor of 2.3 (P = 0.04), in non-heat-stressed hCG-treated cows (2.3%) was less likely by a factor of 0.16 (P = 0.02) and was not significantly different in heat-stressed hCG-treated cows (7%). Interactions between treatment and HS and between treatment and parity had a significant effect on the pregnancy rate (P = 0.0001 and P = 0.001, respectively). The treatment-HS interaction determined that compared to the rates recorded in non-heat-stressed, GnRH-treated cows (30.5%), pregnancy in heat-stressed GnRH-treated cows (17.6%) was less likely by a factor of 0.48 than the remaining cows (P = 0.001), whereas because of the treatment-parity interaction, compared to the rates recorded in primiparous, GnRH-treated cows (31.4%), pregnancy in GnRH-treated multiparous cows (18.9%) was less likely to conceive by a factor of 0.51 than the remaining cows (P = 0.002). No significant effects of treatment on the rates of pregnancy loss or twin pregnancy were identified by binary logistic regression. In conclusion, hCG treatment given at the end of a 5-day P4-based protocol for FTAI improved ovulation and pregnancy rates in anestrous cows under conditions of HS and also had a beneficial impact on the pregnancy rate in anestrous multiparous cows throughout the year.
•Compared to GnRH treatment, hCG led to a lower incidence of ovulation failure throughout the year.•Heat-stressed cows receiving hCG returned a higher pregnancy rate than those receiving GnRH.•Multiparous cows receiving hCG were more likely to become pregnant than their peers receiving GnRH.
This study examines the possible effects of progesterone (P4) supplementation during the time of pregnancy recognition, from Days 15 to 17 post-artificial insemination (AI), on reproductive ...performance in high-producing dairy cows. Cows in their 15th day post-AI were alternately assigned to a control, no-treatment group (C: n = 257) or treatment group (P4: n = 287) on a weekly rotational basis according to the chronologic order of their gynecologic visit. On the basis of the odds ratio, the interaction treatment × previous placenta retention had a significant effect (P = 0.02) on conception rate. Thus, cows in P4 that had not suffered a retained placenta were 1.6 times more likely to conceive 28 to 34 days post-AI than the remaining cows. In nonpregnant cows, treatment had no effect on subsequent return to estrus or AI interval and neither were any effects of treatment observed on twin pregnancy and early fetal loss rates. The results of this study demonstrate the efficacy of P4 supplementations during the time of pregnancy recognition after AI in cows without a clinical history of placenta retention.
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.
•Cytokine gene expression analyzed in N. caninum experimental infection in cattle.•Study of infected aborting and non-aborting dams, control heifers and their foetuses.•Up-regulated Th1, Th2 and Treg ...in infected dams with live foetuses and their foetuses.•Down-regulated Th2 and Treg cytokine in infected dams with aborted/non-viable foetuses.•Recovery of expression levels in dams a few weeks after an abortion occurred.
Neospora caninum is a major cause of abortion in cattle. However, it is not known why not all infected animals abort. In this study, Th1 (IFN-γ), Th2 (IL4) and T reg (IL-10) cytokine gene expression was examined by real time PCR using the TaqMan approach in all of these dams and their foetuses after experimental infection with the isolate Nc-Spain7 at 110 days of pregnancy and euthanasia 6 weeks after infection. In prior published work, foetal death was observed in three of six infected dams and transplacental infection in all the 6 infected foetuses. In the spleen of the dams, IL-4 expression was down-regulated in dams with aborted/non viable foetuses compared to both uninfected dams (controls, n=3) and infected dams with live fetuses at euthanasia. In the lymph nodes draining the placenta, up-regulated expression of IL-4 was observed in infected dams with live foetuses compared to control dams. In the placenta, infected dams with live foetuses had significantly up-regulated IFN-γ in both caruncle and cotyledon and up-regulated IL-10 in cotyledon compared to control dams. Infected live foetuses showed up-regulated expression of IFN-γ and IL-10 in foetal spleen, and showed downregulated expression of IL-4 in the thymus compared to control uninfected foetuses. Expression of any cytokine in the thymus was significantly lower compared to the levels observed in foetal spleen. The results indicate an up-regulated expression of Th1, Th2 and Treg in infected dams with live foetuses and in their foetuses. On the other hand, down-regulation of Th2 immune responses and Treg cytokines were observed in infected dams which had aborted or had non-viable foetuses at euthanasia, suggesting an immunological recovery of cytokine gene expression levels in dams a few weeks after an abortion occurred.
•Concentrations of pregnancy-associated glycoproteins 1 (PAG-1) and 2 (PAG-2) in response to experimental infection on day 110 of gestation with N. caninum in dairy heifers.•Significant lower ...relative proportions of bi- and mono-nucleate trophoblast cells was detected in non-aborting Neospora-infected heifers 42 days post infection, compared to non-infected animals.•Non-aborting Neospora-infected dairy heifers showed a brief transient reduction in PAG-1 and PAG-2 concentrations from 7 to 14 days post infection.•Infected aborting dams showed a dramatic decline in PAGs until undetectable levels were recorded on day 42 post infection.•Following infection with N. caninum, heifers suffer placental damage, though, in some animals, this damage will be overcome and pregnancy will successfully continue to term.
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 107 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.
PDF
