Development and evaluation of an injectable slow-release progesterone formulation for estrus synchronization in ewes out of the breeding season.

This study was aimed at developing a type of slow-release progesterone micro-particles useable in a single intramuscular injection for estrus synchronization in non-breeding season ewes. A total of 66 ewes were randomly assigned into four groups: CIDR (n = 16): exposed to intravaginal CIDR for 12 days, and three experimental groups, i.e., T100 (n = 16), T150 (n = 17) and T200 (n = 17), receiving a single intramuscular injection of 100, 150 and 200 mg slow-release progesterone, respectively. Blood sampling was performed on all ewes at five different times, and the ELISA method measured progesterone levels. No significant differences were observed in progesterone levels among the groups in each sampling time. More than 90% of ewes in the CIDR, T100 and T150 groups and all those in T200 showed estrus behaviour, and the rate was not significantly different between groups. The difference in the mean interval from progesterone treatment to estrus was also insignificant. The parturition rate declined by increasing the dose of injected progesterone; although it was similar in CIDR and T100 groups, it decreased significantly in T150 and T200 . Since our injectable progesterone formulation was successful in the induction and synchronization of estrus in ewes out of the breeding season, it can be applied as an alternative to the conventional progesterone containing intravaginal devices.

The effect of GnRH administration/insemination time on follicular growth rate, ovulation intervals, and conception rate of Nili Ravi buffalo heifers in 7 -day-CIDR Co-synch.

The present study aimed to compare two different insemination times (72 vs 84 h) associated with an ovulation induction (GnRH) in a 7-day CIDR Co-synch to improve the conception rate of Nili Ravi buffalo heifers. Forty Nili Ravi buffalo heifers were randomly separated into two treatments based on artificial insemination (AI) timing (72 vs 84 h). All heifers were subjected to controlled internal drug release (CIDR), containing 1.38 g of progesterone for 7 days. On CIDR removal, both treatments received 150 µg of prostaglandin intramuscularly. In 7-day CIDR Co-synch (n = 20), animals were injected 100 µg of GnRH administration intramuscularly and inseminated concurrently at 72 h after CIDR removal. The remaining half (n = 20) were injected and inseminated concurrently at 84 h of CIDR removal. Pregnancy diagnosis was performed on day 40 of timed artificial insemination (TAI) with ultrasound. The follicular growth rate between 72 h after PGF2α/CIDR removal to pre-ovulatory follicle in 7-day CIDR Co-synch was more (0.102 ± 0.005 mm vs 0.079 ± 0.003 mm; P = 0.01) at 84 than 72 h. The interval from GnRH administration/TAI to ovulation was high (26.8 ± 1.64 h vs. 15.1 ± 1.25 h, P = 0.01) in 72 than 84 h. Conception rates were considerably higher in buffalo heifers inseminated at 84 h (65%) than 72 h (25%) in 7-day CIDR Co-synch protocol. In conclusion that in Nili Ravi buffalo heifers, GnRH administration/TAI after 84 h of CIDR removal allows greater follicular growth rate and shortens interval from AI to ovulation compared to the GnRH administration/TAI after 72 h of CIDR removal in 7-day CIDR-Co-synch protocol.

Dynamics of PfEMP1 Antibody Profile From Birth to 12 Months of Age in Beninese Infants.

BACKGROUND: Plasmodium falciparum-infected erythrocytes bind to specific endothelial cell receptors via members of the PfEMP1 family exported onto the erythrocyte surface. These interactions are mediated by different types of cysteine-rich interdomain region (CIDR) domains found in the N-terminal region of all PfEMP1. CIDRα1 domains bind endothelial protein C receptor (EPCR), CIDRα2-6 domains bind CD36, whereas the receptor specificity of CIDRß/γ/δ domains is unknown. METHODS: In this study, we investigated the level of immunoglobulin (Ig)G targeting the different types of PfEMP1 CIDR during the first year of life. We used plasma collected longitudinally from children of pregnant women who had been followed closely through pregnancy. RESULTS: Antibodies to CIDRα1 domains were more frequent in cord blood compared with antibodies to CIDRα2-6 domains. Higher IgG levels to EPCR-binding CIDRα1 variants positively correlated with the timing of first infections. Antibodies to all PfEMP1 types declined at similar rates to the point of disappearance over the first 6 months of life. At 12 months, children had acquired antibody to all types of CIDR domains, mostly in children with documented P falciparum infections. CONCLUSIONS: These observations agree with the notion that the timing and phenotype of first P falciparum infections in life are influenced by the immune status of the mother.

Immunization with virus-like particles conjugated to CIDRα1 domain of Plasmodium falciparum erythrocyte membrane protein 1 induces inhibitory antibodies.

BACKGROUND: During the erythrocytic cycle, Plasmodium falciparum malaria parasites express P. falciparum Erythrocyte Membrane Protein 1 (PfEMP1) that anchor the infected erythrocytes (IE) to the vascular lining of the host. The CIDRα1 domain of PfEMP1 is responsible for binding host endothelial protein C receptor (EPCR), and increasing evidence support that this interaction triggers severe malaria, accounting for the majority of malaria-related deaths. In high transmission regions, children develop immunity to severe malaria after the first few infections. This immunity is believed to be mediated by antibodies targeting and inhibiting PfEMP1, causing infected erythrocytes to circulate and be cleared in the spleen. The development of immunity to malaria coincides with acquisition of broad antibody reactivity across the CIDRα1 protein family. Altogether, this identifies CIDRα1 as an important vaccine target. However, the antigenic diversity of the CIDRα1 domain family is a challenge for vaccine development. METHODS: Immune responses in mice vaccinated with Virus-Like Particles (VLP) presenting CIDRα1 antigens were investigated. Antibody reactivity was tested to a panel of recombinant CIDRα1 domains, and the antibodies ability to inhibit EPCR binding by the recombinant CIDRα1 domains was tested in Luminex-based multiplex assays. RESULTS: VLP-presented CIDRα1.4 antigens induced a rapid and strong IgG response capable of inhibiting EPCR-binding of multiple CIDRα1 domains mainly within the group A CIDRα1.4-7 subgroups. CONCLUSIONS: The study observations mirror those from previous CIDRα1 vaccine studies using other vaccine constructs and platforms. This suggests that broad CIDRα1 antibody reactivity may be achieved through vaccination with a limited number of CIDRα1 variants. In addition, this study suggest that this may be achieved through vaccination with a human compatible VLP vaccine platform.

Relationship between plasma concentrations of IGF-I and clinical endometritis, and response to progesterone synchrony in dairy cows during early lactation.

The first aim of this study was to evaluate the relationship between the concentrations of plasma insulin-like growth factor-I (IGF-I) in the transition period and the incidence of clinical endometritis postpartum. The second aim was to evaluate the relationship between the concentration of plasma IGF-I in the transition period and the estrous synchrony response and pregnancy rate to a controlled internal drug release (CIDR) protocol. A total of 402 dairy cows, 250 multiparous and 152 primiparous, were enrolled. A blood sample was taken from all cows 2 and 1 wk precalving and wk 1, 3, 4, and 5 postcalving for subsequent analysis of IGF-I. Vaginal discharge score as a measurement of uterine health was recorded in wk 3, 4, and 5 postcalving. Estrous cycles of all cows were synchronized on ≥37 d in milk (mean = 59; range 37-93) using an 8-d CIDR protocol. All cows were scanned at 32 to 35 d after insemination to confirm pregnancy. Data were analyzed using Proc Mixed and Proc Logistic models in SAS (version 9.3; SAS Institute Inc., Cary, NC). Plasma IGF-I concentrations were significantly greater in the primiparous cows than in the multiparous cows, both before and after calving, and were therefore analyzed separately. In multiparous cows plasma IGF-I concentrations postcalving were significantly lower among cows with uterine infection than in those without infection, and multiparous cows with a plasma IGF-I concentration less than 40 ng/mL 1 wk after calving were 3 times more likely to suffer from a uterine infection 4 to 5 weeks postcalving (odds ratio 2.8; 95% confidence interval 1.0-7.6). Plasma IGF-I concentrations were significantly greater after calving in the 121 primiparous cows that exhibited estrus post-CIDR protocol (mean 125 ng/mL) than in the 25 primiparous cows that did not respond (mean 95 ng/mL). Multiparous cows with greater plasma IGF-I concentrations postcalving had a significantly greater chance of conceiving (mean plasma IGF-I concentration of conceiving cows = 86 ng/mL; nonconceiving = 70 ng/mL). These results show an association between greater concentrations of plasma IGF-I postcalving and response to CIDR protocol in primiparous animals (mean plasma IGF-I concentration of responders = 116 ng/mL; nonresponders = 95 ng/mL). In contrast no association was detected between concentrations of plasma IGF-I in relation to the response to the CIDR protocol in multiparous cows. In conclusion, changes in circulating concentrations of plasma IGF-I early postpartum may help predict reproductive status of dairy cows.

Effect of the side of ovulation on the uterine morphometry, blood flow, progesterone, oestradiol and nitric oxide during spontaneous and induced oestrus in lactating dairy cows.

This study investigates the changes in endometrial thickness between ipsilateral and contralateral uterine horns, diameter, blood flow area and hormonal concentrations in cows during natural and induced oestrus. In the induced group, six cows received a controlled internal device insert for 7 days. In the normal and induced oestrous group (n = 12), Doppler was performed day after day from day 5 till day 15. Oestradiol, progesterone and nitric oxide metabolites (NOMs) were measured. Results showed that dorsal, ventral and cross-section diameter of the ipsilateral horn increased during the induced oestrous group to reach a maximum on day 2 than during normal one. The total coloured area in both horns in a normal and induced group was also affected (p < .001), as the total coloured area in the ipsilateral horns, increased in the induced group, reach a maximum on day 2 after ovulation, while in normal it reaches a maximum on day of ovulation. Oestradiol increased during the induced cycle than during the normal one, but progesterone increased during days after ovulation reaches a higher value on day 10 in both groups. Nitric oxide showed two peaks, one on the day 3 and another peak on day 5. Conclusion: The ipsilateral uterine horn different diameters and thickness increased more rapidly in the induced oestrous group than the natural oestrus group. A complete analysis of variations in P4 and E2 concentrations and endometrial thickness suggested that decreases in P4 concentrations were related to an increase in endometrial thickness and that increases in E2 concentrations enhanced these endometrial changes.

Acquisition of Antibodies Against Endothelial Protein C Receptor-Binding Domains of Plasmodium falciparum Erythrocyte Membrane Protein 1 in Children with Severe Malaria.

BACKGROUND: Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) mediates parasite sequestration in postcapillary venules in P. falciparum malaria. PfEMP1 types can be classified based on their cysteine-rich interdomain region (CIDR) domains. Antibodies to different PfEMP1 types develop gradually after repeated infections as children age, and antibodies to specific CIDR types may confer protection. METHODS: Levels of immunoglobulin G to 35 recombinant CIDR domains were measured by means of Luminex assay in acute-stage (baseline) and convalescent-stage plasma samples from Papua New Guinean children with severe or uncomplicated malaria and in healthy age-matched community controls. RESULTS: At baseline, antibody levels were similar across the 3 groups. After infection, children with severe malaria had higher antibody levels than those with uncomplicated malaria against the endothelial protein C receptor (EPCR) binding CIDRα1 domains, and this difference was largely confined to older children. Antibodies to EPCR-binding domains increased from presentation to follow-up in severe malaria, but not in uncomplicated malaria. CONCLUSIONS: The acquisition of antibodies against EPCR-binding CIDRα1 domains of PfEMP1 after a severe malaria episode suggest that EPCR-binding PfEMP1 may have a role in the pathogenesis of severe malaria in Papua New Guinea.

Ovarian follicular changes and hemodynamics in Egyptian buffaloes under CIDR-PGF2α and Ovsynch-CIDR estrus synchronization treatments.

This study aimed at investigating the efficacy of two protocols of estrous synchronization on follicular changes and hemodynamics. Pluriparous Egyptian buffaloes (n = 36) were synchronized either with controlled internal drug release (CIDR)-PGF2α (7-days CIDR insert with PGF2α injected on the 6th day; n = 18) or Ovsynch-CIDR (Ovsynch protocol concurrent with 7-days CIDR insert; n = 18). Blood sampling and ovarian ultrasound examinations (Grayscale, color and power Doppler modes) were conducted on the Day of CIDR removal, estrus, and luteal phase. Mean follicle diameter (MFD), first (1st-LF) and second (2nd-LF) largest follicle diameters, and E2 levels significantly increased in the CIDR-PGF2α group at CIDR withdrawal. Ovsynch-CIDR markedly fortified higher follicle population, MFD, and 1st-LF diameter at estrus and corpus luteum (CL) volume at the luteal phase in concomitant with increases (P < 0.05) in E2 (at estrus) and P4 (at luteal phase). At CIDR removal, the blue pixels in the dominant follicle (DF) were higher (1.5 times; P = 0.054) in the Ovsynch-CIDR than in the CIDR-PGF2α. At estrus, total blood flow (TBF) and power Doppler pixels (PDP) to DF(s) were noticeably higher (seven and 1.6 times; respectively) in the Ovsynch-CIDR than in CIDR-PGF2α (5906 ± 237 vs. 830 ± 60 pixels, P < 0.01 and 5479 ± 322 vs. 3377 ± 19 pixels, P < 0.05; respectively). At the luteal phase, TBF and PDP to the CL increased in the Ovsynch-CIDR group than in the CIDR-PGF2α group (11060 ± 965 vs. 7963 ± 480 pixels, 1.4 times, P = 0.05 and 18900 ± 1350 vs. 13220 ± 568 pixels, 1.1 times, P = 0.005; respectively). In conclusion, based on the improvement in synchronized follicular activity and hemodynamics, the Ovsynch-CIDR regimen is persuaded in Egyptian buffaloes.

Onset of oestrus and periovulatory events in sheep exposed to 5 and 14 days of CIDR treatment with and without eCG.

The present study supports that 5-day short-term CIDR treatments without administration of eCG are equally effective for inducing oestrus behaviour, preovulatory LH discharge and ovulation in sheep than classical protocols based on 14-day treatments plus eCG at CIDR withdrawal. However, the implementation of a 5-day protocol without eCG for fixed-time artificial insemination would be adapted to a later timing of ovulation (p < .05).

Ovarian hormones and antioxidant biomarkers in dromedary camels synchronized with new and re-used controlled intravaginal drug release (CIDR)/GPG (Ovsynch) program during breeding season.

This study aimed to investigate the effect of CIDR, re-used wCIDR, and Ovsynch protocols for the synchronization of follicular waves on ovarian hormones, oxidative stress, and antioxidant biomarkers during the breeding season. Dromedary camels (N = 18) were divided into three equal groups. The first group received CIDR. The second group received previously used wCIDR after thorough cleaning and disinfection. The third group was subjected to GPG protocol. Progesterone (P4), estradiol (E2) l, total antioxidant capacity (TAC), catalase (CAT), lipid peroxide product (MDA), nitric oxide (NO), and glutathione reduced (GSH) were measured. Days during CIDR affected P4 (P = 0.0001), E2 (P = 0.047), TAC (P = 0.01), NO (P = 0.028), and GSH (P = 0.005). Days during re-used wCIDR effected P4, TAC, CAT, NO, GSH, and MDA (P ≤ 0.001). Days during GPG effected P4, E2, TAC, GSH (P = 0.0001), MDA (P = 0.036), and NO (P = 0.02). CIDR-treated camels had high P4 (P = 0.0001), E2 (P = 0.0001), TAC (P = 0.012), and NO (P = 0.0001), with low GSH (P = 0.001), and MDA (P = 0.003). Exogenous progesterone improved ovarian hormones and the antioxidant capacity and minimized the oxidative stress than the GPG treatment and is recommended for future reproductive management of camels.

Immunization with Recombinant Plasmodium falciparum Erythrocyte Membrane Protein 1 CIDRα1 Domains Induces Domain Subtype Inhibitory Antibodies.

Plasmodium falciparum malaria pathogenesis is tied to the sequestration of parasites in the microvasculature. Parasite sequestration leading to severe malaria is mediated by P. falciparum erythrocyte membrane protein 1 (PfEMP1) binding to endothelial protein C receptor (EPCR) via its CIDRα1 domains. CIDRα1 domains are targets of naturally acquired immunity, and a vaccine eliciting antibodies inhibiting the EPCR binding of CIDRα1 could potentially prevent disease and death from malaria. CIDRα1 domains have diversified in sequence to escape immune recognition but preserved structure to maintain EPCR binding. The EPCR-binding CIDRα1 domains separate into six major sequence types predicted to form a conserved structure in which only the amino acids essential for EPCR binding are highly conserved. Here, we investigated whether antibodies elicited by vaccination with single or multiple recombinant CIDRα1 domains are able to bind and inhibit diverse CIDRα1 domains. We found that EPCR binding-inhibitory antibodies to CIDRα1 variants closely related to those used for vaccination are readily elicited, whereas antibodies binding distant CIDRα1 variants are sporadically generated and are rarely inhibitory. Despite this, sequence similarity correlated poorly with the ability of induced antibodies to inhibit across diverse variants, and no continuous sequence regions of importance for cross-inhibitory antibodies could be identified. This suggested that epitopes of cross-variant inhibitory antibodies were predominantly conformational. Vaccination with immunogens engineered to focus immune responses to specific epitopes or an optimal choice of multiple CIDRα1 variants may improve elicitation of broadly reactive and inhibitory antibody responses.

Efficacy of using previously used controlled internal drug release (CIDR) insert on the reproductive performance, hormone profiles and economic measures of sheep.

This study was conducted using 120 multiparous Awassi ewes during the breeding season to compare the effects of using previously used controlled internal drug release (CIDR) on the hormone profiles, reproductive performance and economic measures of ewes. Ewes were randomized to receive one of five previously used CIDR (previously used for 6, 12, 18, 24 or 30 days) or the new CIDR as a control for 6 days (CIDR6, CIDR12, CIDR18, CIDR24, CIDR30, and CIDR0 [control], respectively). Blood samples were collected on four occasions, at the time of CIDR insertion, after 3 days of insertion, and at the time of withdrawal and insemination. Serum estradiol (E2) and progesterone (P4) concentrations were measured. Timed insemination was performed 48 hr post-CIDR withdrawal. Pregnancy was diagnosed by ultrasonography 23 days after insemination and confirmed on day 35. The heat detection rate was significantly (p < 0.05) higher in the CIDR0 and CIDR6 groups than in the CIDR18 and CIDR30 groups. The total pregnancy rate and fecundity were significantly (p < 0.05) higher in the CIDR6 group than in other groups. P4 level was significantly (p < 0.05) higher in the CIDR0 group than in the CIDR30 group at the time of removal. At each time point, the E2 level was significantly (p < 0.05) higher in the CIDR6 group than at the other groups. The total variable cost, total cost, return and net profit were higher in the CIDR6 and CIDR0 groups than in the other groups. In conclusion, although previously used CIDRs are efficient at synchronizing oestrus in ewes, the duration of previously usage significantly affected the reproductive parameters and economic profit. CIDRs previously used for 6 days and new CIDRs provided the highest fertility and fecundity rates, besides return and net profit. Economically, it is not advisable to use CIDRs that previously used for 12 days or more.

Effectiveness of controlled internal drug release device treatment to alleviate reproductive seasonality in anestrus lactating or dry Barki and Rahmani ewes during non-breeding season.

This study aimed to evaluate the effectiveness of hormonal treatments on ovarian activity and reproductive performance in Barki and Rahmani ewes during non-breeding season. Forty-eight multiparous ewes, 24 Barki and 24 Rahmani ewes were divided into two groups, 12 lactating and 12 dry ewes for each breed. Controlled internal drug release (CIDR) device was inserted in all ewes for 14 days in conjunction with intramuscular 500 IU equine chronic gonadotrophin (eCG) at day of CIDR removal. Data were analysed using PROC MIXED of SAS for repeated measures. Breed, physiological status and days were used as fixed effects and individual ewes as random effects. Barki ewes recorded higher (p < .05) total number of follicles, number of large follicles, serum estradiol concentration and estradiol: progesterone (E2 :P4 ) ratio compared to Rahmani ewes. Lactating ewes recorded higher (p < .05) number of small follicles and lower concentration of total antioxidant capacity (TAC) compared to dry ewes. Number and diameter of large follicles recorded the highest (p < .05) values accompanied with disappearance of corpora lutea at day of mating. Serum progesterone concentration recorded lower (p < .05) value at day of mating and the highest (p < .05) value at day 35 after mating. CIDR-eCG protocol induced 100% oestrous behaviour in both breeds, but Rahmani ewes recorded longer (p < .05) oestrous duration compared to Barki. Conception failure was higher (p < .05) in Barki compared to Rahmani ewes. In conclusion, CIDR-eCG protocol was more potent in improving ovarian activity in Barki compared to Rahmani ewes, but this protocol seems to induce hormonal imbalance in Barki ewes that resulted in increasing conception failure compared to Rahmani ewes.

The effect of post-mating hCG or progesterone administration on reproductive performance of Afshari × Booroola-Merino crossbred ewes.

To investigate the efficiency of hCG/CIDR after breeding to increase the reproductive performance, 35 synchronized ewes were mated with fertile rams and were assigned to three treatment groups. Ewes in hCG group (n = 12) received 400 IU hCG on day 11 post-mating, and ewes in CIDR group (n = 11) received CIDR from day 7 until day 19 post-mating. Ewes in the control group (n = 12) did not receive any treatment. Blood samples were collected on days 7, 12, 17, and 22 post-mating. Plasma P4 concentrations were higher on days 12 and 17 post-mating in hCG- and CIDR-treated groups (P < 0.05). However, the concentrations of P4 on day 22 post-mating in hCG and control groups were higher than that in CIDR group (P < 0.05). Ewes in hCG group produced more quadruplets (P < 0.05) and triplets, and as a result, they had a larger litter size (P < 0.05). The lamb mortality rate by weaning in hCG group (3.6%) was less than that in control (11.8%; P < 0.05) and CIDR (9.1%; P > 0.05) groups. Post-mating administration of hCG or CIDR did not affect the lamb birth weight in single and quadruplet births (P > 0.05), but the birth weight of twin lambs was higher in the hCG and CIDR groups (P < 0.05). Weaning weights of twin lambs were higher in the hCG and CIDR groups (P < 0.05). In conclusion, hCG/CIDR administration post-mating increased the maternal P4 concentrations and enhanced reproductive performance.

Incorporation of estradiol benzoate to CIDR protocol improves the reproductive responses in crossbred dairy heifers.

The present study was designed to determine the effect of estradiol benzoate (EB) on reproductive response following a controlled internal drug release (CIDR) protocol in crossbred (Sahiwal × Friesian) dairy heifers. In the first trial, a total of 100 crossbred dairy heifers were treated with CIDR protocol for 7 days and injected with the PGF2α on day 6. After 24 h of CIDR removal, one group (EB = 50) was injected with estradiol benzoate whereas the other (control = 50) remained untreated. Estrus intensity and response were recorded visually and ovulation rate was recorded by ultrasonography. All heifers were artificially inseminated at 48 and 60 h following CIDR removal. Heifers were scanned for pregnancy within days 30-40 of artificial insemination (AI). In the second trial, two subgroups of heifers were included to observe the estrus and ovulatory events. The results of the first trial revealed that estrus response was achieved 100% in both the treatment groups. Estrus intensity (2.9 ± 0.1 vs. 2.0 ± 0.7) and ovulation rate (100 vs. 88%) differed significantly (P < 0.05) between the EB and control groups. However, a tendency for higher pregnancy per AI was observed (54 vs. 36%; P = 0.07) in EB than that in control groups. The results of the second trial revealed that a significantly (P < 0.05) shorter estrus and earlier ovulatory events were observed in EB-treated heifers. It is concluded that the incorporation of estradiol benzoate to the CIDR protocol is helpful to improve the estrus signs and enhance the ovulation and the pregnancy per AI in crossbred dairy heifers.

Neonatal milk supplementation in lambs has persistent effects on growth and metabolic function that differ by sex and gestational age.

The perinatal environment has a major influence on long-term health and disease risk. Preterm birth alters early-life environment and is associated with altered metabolic function in adulthood. Whether preterm birth per se or the early nutritional interventions used to support growth in preterm infants underpins this association is unknown. Lambs born preterm, following dexamethasone induction of labour, or spontaneously at term were randomised to receive nutrient supplementation, analogous to the milk fortifier used clinically or water as a control for the first 2 weeks after birth. Thereafter, nutrition was not different between groups. Growth was monitored, and the glucose-insulin axis function was assessed in juvenile (4 months) and adult life (14 months). Early nutrition influenced adult metabolic function and body composition to a greater extent than preterm birth. In supplemented females, arginine-stimulated insulin secretion was increased in preterm but reduced in term-born juveniles compared with controls (repeated-measures ANOVA P<0·01). In supplemented preterm males, adult weight, ponderal index (PI) and fasting insulin concentrations were elevated compared with preterm controls (weight, 75 (sem 3) v. 69 (sem 2) kg; PI, 48·0 (sem 2·1) v. 43·7 (sem 1·7) kg/m3; fasting insulin, 0·19 (sem 0·02) v. 0·10 (sem 0·02) ng/ml). Conversely, supplemented term-born males had reduced adult weight, PI and fasting insulin concentrations compared with term-born controls (weight, 64 (sem 2) v. 70 (sem 2) kg; PI, 44·4 (sem 1·8) v. 48·2 (sem 1·7) kg/m3; fasting insulin, 0·09 (sem 0·02) v. 0·14 (sem 0·02) ng/ml; all group×supplement interactions P<0·05). Adult metabolic health may reflect both gestational age at birth and early nutrition. Human studies are urgently needed to investigate the adult sex-specific health implications of neonatal nutritional strategies.

Meta-analysis of progesterone supplementation during timed artificial insemination programs in dairy cows.

A systematic review of the literature was performed with the objective to evaluate the effects of progesterone supplementation using a single intravaginal insert during timed artificial insemination (AI) programs on fertility in lactating dairy cows. A total of 25 randomized controlled studies including 8,285 supplemented cows and 8,398 untreated controls were included in the meta-analysis. Information regarding the presence of corpus luteum (CL) at the initiation of the synchronization protocol was available for 6,883 supplemented cows and 6,879 untreated controls in 21 experiments. Studies were classified based on service number (first AI vs. resynchronized AI), use of presynchronization (yes vs. no), and insemination of cows in estrus during the synchronization protocol (inseminated in estrus and timed AI vs. timed AI only). Reproductive outcomes of interest were pregnancy per AI (P/AI) measured on d 32 (27 to 42) and 60 (41 to 71) after AI, and pregnancy loss between d 32 and 60 of gestation. Random effects meta-analyses were conducted and treatment effect was summarized into a pooled risk ratio with the Knapp-Hartung modification (RRK+H). The effect of moderator variables was assessed using meta-regression analyses. Progesterone supplementation increased the risk of pregnancy on d 32 [RRK+H = 1.08; 95% confidence interval (CI) = 1.02-1.14] and 60 after AI (RRK+H = 1.10; 95% CI = 1.03-1.17). The benefit of progesterone supplementation was observed mainly in cows lacking a CL at the initiation of the timed AI program (d 60: RRK+H = 1.18; 95% CI = 1.07-1.30) rather than those with CL (d 60: RRK+H = 1.06; 95% CI = 0.99-1.12). Progesterone supplementation benefited P/AI in studies in which all cows were inseminated at timed AI (d 60: RRK+H = 1.20; 95% CI = 1.10-1.29), but not in studies in which cows could be inseminated in estrus during the timed AI program (d 60: RRK+H = 1.04; 95% CI = 0.92-1.16). Progesterone supplementation tended to reduce the risk of pregnancy loss (RRK+H = 0.84; 95% CI = 0.67-1.00). Service number and presynchronization did not influence the effect of progesterone supplementation on fertility. In summary, progesterone supplementation using a single intravaginal insert during the timed AI program increased P/AI mostly in cows without CL and reduced the risk of pregnancy loss in lactating dairy cows. Insemination of cows in estrus during the synchronization protocol eliminated the benefit of supplemental progesterone on P/AI.

Preovulatory follicle diameter, growth rate and time of ovulation during induced oestrus using a CIDR® in trypanotolerant female Bos taurus N'Dama cattle.

The aim of this study was to assess the dose (300 to 600 IU) effects of equine chorionic gonadotropin (eCG) on the preovulatory follicle diameter, growth rate and time of ovulation characterized by echography. The eCG was injected at the end (D0) of the 7-day treatment with a controlled internal device release (CIDR®) and a PGF2α being injected 2 days before the removal of the CIDR® (d-2). The 120 N'Dama female were distributed into five experimental groups. The control group (n = 26) was treated with physiological saline at the removal of the CIDR®, while the animals in the four treated groups received, respectively, 300 IU (n = 25), 400 IU (n = 24), 500 IU (n = 22) and 600 IU (n = 23) of eCG. The diameter of the preovulatory follicle was significantly higher (P < 0.05) in the animals treated with 300 IU (10.1 ± 1.4 mm) than in untreated animals (9.3 ± 1.2 mm). Follicle growth rate was significantly (P < 0.05) higher in treated animals (1.0 ± 0.4 mm/day) than in the control group (0.9 ± 0.4 mm/day). The average interval between the time of eCG injection and ovulation was similar in the non-treated (83.7 ± 14.4 h) and treated animals (79.7 ± 11.9). Treated animals showed a significant increase in the percentage of ovulation (94.7 % compared to 73.1 %) (P < 0.01). Use of eCG contributed towards synchronising the time of ovulation between 72 to 96 h, which would facilitate the use of systematic insemination.

Inhibition of storage pathology in prenatal CLN5-deficient sheep neural cultures by lentiviral gene therapy.

The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are inherited neurodegenerative lysosomal storage diseases caused by mutations in several different genes. Mutations in CLN5 cause a variant late-infantile human disease and some cases of juvenile and adult clinical disease. NCLs also occur in animals, and a flock of New Zealand Borderdale sheep with a CLN5 splice-site mutation has been developed for model studies. Dissociated mixed neural cells from CLN5-deficient foetal sheep brains contained no obvious storage bodies at plating but these accumulated rapidly in culture, mainly in microglial cells and also in neurons and astrocytes. Accumulation was very obvious after a week, as monitored by fluorescent microscopy and immunostaining for subunit c of mitochondrial ATP synthase. Photography at intervals revealed the dynamic nature of the cultures and a flow of storage bodies between cells, specifically the phagocytosis of storage-body containing cells by microglia and incorporation of the storage bodies into the host cells. No storage was observed in cultured control cells. Transduction of cell cultures with a lentiviral vector expressing a C-terminal Myc tagged CLN5 resulted in secretion of post-translationally glycosylated and processed CLN5. Transduction of CLN5-deficient cultures with this construct rapidly reversed storage body accumulation, to less than half in only six days. These results show that storage body accumulation is reversible with enzyme correction and support the use of these cultures for testing of therapeutics prior to whole animal studies.

Comparison of synchronisation and fertility after different modifications of the ovsynch protocol in cyclic dairy cows.

The objectives of this study were to evaluate the effects of (1) double doses of PGF2α administration or (2) an exogenous progesterone (CIDR) applied concurrently with, or (3) the day after, first GnRH of Ovsynch (GnRH-1), on synchronisation and fertility during the Ovsynch protocol. All cows (n = 378) received the Ovsynch protocol (GnRH-7d-PGF2α-56h-GnRH-18h-TAI). The 'OVS' group (n = 105) received only the Ovsynch protocol. The 'OVS-PGF' group (n = 118, GnRH-7d-PGF2α-12h-PGF2α-44h-GnRH-18h-TAI) received an extra dose of PGF2α 12 h later on Day 7. The 'OVS-7CIDR' group (n = 78, GnRH+CIDR-7d-PGF2α-56h-GnRH-18h-TAI) received a CIDR for 7 days between GnRH-1 and PGF2α. In the 'OVS-6CIDR' group (n = 77, GnRH-24h-CIDR-6d-PGF2α-56h-GnRH-18h-TAI), CIDR was applied one day after GnRH-1 and removed 6 days later. When all cows were evaluated, the responses to GnRH-2 were higher (P = 0.005) in cows that responded to GnRH-1 (95.4%) compared to the cows that did not respond (87.6%). The pregnancy rates at 31 and 62 days for each group were 48.6% and 42.9% in the OVS, 54.2% and 52.5% in the OVS-PGF, 52.6% and 48.7% in the OVS-7CIDR, and 55.8% and 49.3% in the OVS-6CIDR groups. Thus, none of the three different treatments has an effect on increasing the out-comes of the Ovsynch protocol in cyclic lactating dairy cows.