Use of prostaglandin E2 to ripen the cervix of the mare prior to induction of parturition

Eleven light-breed pregnant mares (335 to 347 d gestaton) were used to evaluate the use of prostaglandin E2 as a cervical ripening agent prior to induction of parturition during the months of April and May. Six hours prior to induction, each mare's cervix was examined per vagina for softness and dilation. Each mare was then assigned to 1 of 2 treatment groups: Group PGE mares (n = 7) received 2.0 to 2.5 mg prostaglandin E2 deposited intracervically; Group SAL mares (n = 4) received 0.5 mL of sterile NaCl deposited intracervically. Six hours later, the mares were readied for parturition by wrapping the tail, scrubbing and rinsing the perineum and udder, and examining the cervix as previously described. Each mare was then administered 15 U, i.v. oxytocin at 15-min intervals until the chorioallantois ruptured. Intervals from initial oxytocin injection until rupture of the chorioallantois, from initial oxytocin injection until delivery of the foal, from delivery of the foal until the foal stood unassisted, and from delivery of the foal until the foal suckled were recorded. Mean cervical dilation immediately prior to induction of parturition tended to be greater in Group PGE mares (3.9 +/- 1.7 cm) than in Group SAL mares (1.9 +/- 1.9 cm; P = 0.10). Mean change in cervical dilation over the 6-h period prior to induction (3.4 +/- 1.9 cm vs 1.5 +/- 2.1 cm), mean number of injections of oxytocin required until the chorioallantois ruptured (1.9 +/- 0.7 vs 2.5 +/- 1.0), and mean intervals from initial injection of oxytocin to rupture of the chorioallantois (20 +/- 10 min vs 28 +/- 19 min) and delivery of the foal (28 +/- 7 min vs 34 +/- 22 min) were not different between Group PGE and SAL mares, respectively (P > 0.10). The proportion of foals that stood within 1 h of birth also did not differ between Group PGE foals (6/7; 86%) and Group SAL foals (3/4; 75%; Chi-square = 0.17; P > 0.10). The proportion of foals that nursed within 2 h of birth was higher in Group PGE foals (6/7; 86%) than in Group SAL foals (1/4; 25%; Chi-square = 4.02; P < 0.05). Premature separation requiring manual rupture of the chorioallantois at the vulvar labia occurred in 1 Group PGE mare (cervical dilation of 1.5 cm at time of induction) and 1 Group SAL mare (cervix closed and firm at time of induction). Foals born from the 2 mares with premature placental separation had the longest intervals from initial oxytocin injection to delivery, delivery to ability to stand unassisted, and delivery to suckling within their respective treatment groups. In summary, it appears that cervical ripening prior to induction of parturition favors shorter deliveries and foal vigor. Intracervical administration of prostaglandin E2 may prove useful for ripening the cervix of the mare prior to induction of parturition. Further studies are indicated to determine optimal dosage and method of administration of prostaglandin E2.     

Concentrations of altrenogest in plasma of mares and foals and in allantoic and amniotic fluid at parturition

Treatment with the progestin altrenogest is widely used in pregnant mares. The fact that foals born from healthy mares treated with altrenogest until term suffered from neonatal problems raises the question of direct effects of altrenogest on vital functions in the neonate. We have therefore investigated altrenogest concentrations in maternal and neonatal blood plasma and in fetal fluids. Pregnant mares were treated with altrenogest orally once daily (0,088 mg/kg bodyweight, n = 7) or left untreated (n = 8) from 280 d of gestation until foaling. Altrenogest concentration was determined in plasma of the mares, their foals and in amniotic and allantoic fluid. The concentration of altrenogest in plasma from treated mares (2.6 ± 1.0 ng/mL) was significantly lower than in plasma from their foals immediately after birth (5.6 ± 1.9 ng/mL; p < 0.05), but was significantly higher than in their fetal fluids (amniotic fluid: 0.4 ± 0.1 ng/mL; p < 0.05; allantoic fluid: 3.0 ± 1.5 ng/mL). Altrenogest was undetectable in maternal and fetal plasma and fetal fluids of control pregnancies at all times. Altrenogest concentration in plasma of foals from treated mares was strongly correlated to the altrenogest concentration in plasma of their dams (r = 0.938, p < 0.001) and in amniotic (r = 0.886, p < 0.001) and allantoic fluid (r = 0.562, p < 0.05). A significant decrease in altrenogest concentration between the time periods 0-15 min, 30-120 min, and 180-360 min after parturition was seen in the plasma from foals born to altrenogest-treated mares. In conclusion, our data demonstrate that altrenogest reaches the equine fetus at high concentrations.     

Mutual grooming and nearest-neighbor relationships among foals of Equus caballus

A 3-year study was carried out on the developmental behavior of foals from birth to 24 weeks of age and the behavior of mares living with foals. Mutual-grooming partners of foals were primarily other foals. The peak frequency of mutual grooming occurred during Weeks 9–12, when fillies mutual-groomed 1.6 times h⁻¹ and colts mutual-groomed 0.9 times h⁻¹. Fillies mutual-groomed more frequently than colts (P < 0.025). Fillies mutual-groomed randomly with colts and other fillies (P < 0.05), whereas colts mutual-groomed almost exclusively with fillies (P = 0.03). At all ages studied, if a foal's nearest neighbor was not its mother, it was more likely to be another foal than would be expected if the foal was associating randomly with non-mother ponies. Fillies were more likely than expected to have a filly rather than a colt as their nearest neighbor (P = 0.01). Thus, during their first few months of life, the foals studied exhibited patterns of behavior which were consistent with the development of the usual social milieu of unmanaged adults, in which several mares form a cohesive herd with one or more stallions associating with them.     

Amniotic fluid and blood lactate concentrations in mares and foals in the early postpartum period

Amniotic fluid (AF) lactate concentration and time-dependent changes in blood lactate concentration in mares after parturition have never been evaluated. In this study, the venous blood lactate concentration of mares and foals during the first 72 h of the postpartum period was assessed, and the concentration of lactate in the AF collected during delivery and the utility of its measurement for evaluating the foal's health were investigated. This prospective observational study was carried out on mares attended at delivery. They were divided into mares delivering healthy (Group 1) and sick (Group 2) foals. The following samples were collected: AF and umbilical blood at delivery, mare's and foal's jugular blood every 12 hours from parturition until 72 h postpartum (T0-T72). Sixty-two mares were enrolled in Group 1 and 19 in Group 2. In Group 2, the survival rate was 68.4%. The median blood lactate of the foals at T0 was 3.60 mmol/L in Group 1 and 5.05 mmol/L in Group 2. The monitoring of the blood lactate concentration showed a significant time-dependent decrease from T24 in the foals (P < 0.01) and from T12 in the mares (P < 0.01). Lactate concentration over time was significantly different between healthy and sick foals (P < 0.01) but not between mares with normal and dystocic delivery (P = 0.08). A significant difference (P = 0.04) was detected as regards AF lactate concentration between Group 1 (median 14.99 mmol/L) and Group 2 (median 12.61 mmol/L). For the first time, AF lactate concentration was evaluated during parturition, and significantly higher levels were found in mares delivering healthy foals. This was an unexpected and very interesting result which warrants further investigation involving a larger number of mares. Additional studies are needed before either mare's blood or AF lactate concentration can be used in a clinical setting.     

Absence of effects from immunocontraception on seasonal birth patterns and foal survival among barrier island wild horses

Despite a large body of safety data, concern exists that porcine zonae pellucidae (PZP) immunocontraception--used to manage wild horse populations--may cause out-of-season births with resulting foal mortality. Our study at Assateague, Maryland indicated the effects of immunocontraception on season of birth and foal survival between 1990 and 2002 on wild horses from Assateague Island. Among 91 mares never treated, 69 (75.8%) of foals were born in April, May, and June (in season). Among 77 treated mares, 50 (64.9%) were born in season. Of 29 mares foaling within 1 year after treatment (contraceptive failures), 20 (68.9%) were born in season. Of 48 mares treated for greater than 2 years then withdrawn from treatment, 30 (62.5%) of 48 foals were born in season. There were no significant differences (p <.05) between either treatment group or untreated mares. Survival did not differ significantly among foals born in or out of season or among foals born to treated or untreated mares. Data indicate a lack of effect of PZP contraception on season of birth or foal survival on barrier island habitats.     

Diet selection and performance of horses grazing on different heathland types

The number of horses in northern Spanish mountains has increased in recent decades, but little is known about their grazing behaviour, performance and potential for foal meat production. This research aimed to study the diet selection, liveweight (LW) changes and parasitic status of dry and lactating mares, and foals’ LW gains, grazing on heathlands with different botanical composition. The experimental design consisted of three vegetation types: dominated by heather (Ericaceae) species (H), dominated by gorse (Ulex gallii; G) and co-dominated by gorse and heath-grasses (G-G), with four replicates per treatment (12 paddocks of 1.2 ha). The study lasted three grazing seasons (2010–12). Each year, 24 crossbred mature mares (310±52 kg LW) were used, managing one lactating mare with her foal plus one non-lactating mare per paddock from May to late summer or early autumn. In the case of H paddocks, animals had to be removed before (late August to early September) because of apparent loss of body condition. Animals were periodically weighed. Mares’ diet composition was estimated using alkane markers, analysing the discrepancies in alkane concentrations between dietary plant components and faeces. Faecal samples were also analysed for gastrointestinal nematodes ova. Chemical composition of the main plant components (i.e. heather, gorse and grasses) revealed a low nutritive value, averaging 79, 115 and 113 g CP/kg dry matter (DM), respectively, that could restrict livestock performance. Mares initially selected gorse and grasses (0.47 and 0.40, respectively, in 2010), increasing heather consumption over time (from 0.13 in 2010 to 0.29 in 2012) as gorse availability decreased. The performance of both mares and foals was lower in H compared with G and G-G paddocks (−216 v. 347 g/day for mares, P<0.01; 278 v. 576 g/day for foals, P<0.05), whereas LW changes were more favourable in dry mares than in lactating ones (241 v. 78 g/day; P<0.05). Small strongyle (Cyathostominae) egg counts in mares’ faeces increased across the grazing season with no differences between treatments. These results indicate that grazing by horses on gorse- and grass-gorse-dominated shrublands could be sustainable at least during part of the year (4 to 6 months). However, heather-dominated heathlands are not able to meet the nutritional needs of horses even for a short time (2 to 4 months). Nevertheless, the low nutritive quality of these vegetation communities, especially in autumn, requires animal access to other pastures with a higher nutritive value, or supplementary feeding, to enhance foals’ growth and maintain sustainable grazing systems with productive herds.     

Activation of foal neutrophils at different ages by CpG oligodeoxynucleotides and Rhodococcus equi

Toll-like receptor 9 (TLR9) activation stimulates protective immune responses against intracellular pathogens by phagocytes, including neutrophils. This study examined TLR9-mediated neutrophil activation in neonatal foals. Unmethylated CpGs, ligands for TLR9, were used to stimulate equine neutrophils, either purified or in contact with other peripheral blood leukocytes. Rhodococcus equi was used as another stimulus in parallel. TLR9 mRNA was constitutively expressed at a similar level in purified equine neutrophils across different ages from birth to adulthood, and expression was not affected by either CpG or R. equi. Purified foal neutrophils were directly sensitive to CpG stimulation, reflected by enhanced reactive oxygen species generation following fMLP stimulation, and by expressing significantly (P < 0.05) greater mRNA of IFN-γ, IL-8, IL-12p35, and significantly (P < 0.05) decreased TNF-α mRNA. In comparison, purified foal neutrophils stimulated by R. equi showed significantly (P < 0.05) increased mRNA production of IL-6, IL-8, IL-23p19, and TNF-α. Neutrophils co-cultured with other leukocytes expressed a distinct profile of cytokine mRNA than purified neutrophils in response to CpG stimulation, whereas the profile was very similar following R. equi stimulation irrespective of neutrophil purity. When co-cultured with other leukocytes, foal neutrophils were significantly (P < 0.05) activated at birth by B-class CpGs and produced IL-6, IL-8, IL-12p40, and IL-23p19 at similar magnitudes to those at 2 months of age. In foal neutrophils at birth, R. equi significantly (P < 0.05) induced all cytokines stimulated by CpGs (except IL-12p40), as well as TNF-α. Our results indicate that foal neutrophils were sensitive to CpG or R. equi activation as early as at birth, and that B-class CpGs enhanced foal neutrophil functions in vitro.     

Diurnal changes in the haemoglobin level, red blood cell number and mean corpuscular haemoglobin in foals during the first 13 weeks of life and in their lactating mothers

1. In six foals and their mothers (of Standard breed) diurnal changes in the haemoglobin level, red blood cell number and mean corpuscular haemoglobin during the first 13 weeks of foal life were studied. 2. Studies begun when a foal reached 7 days of age and were repeated every two weeks till 13 weeks of foal life. Blood was taken every 6 hr in foals and every 4 hr in mares. 3. No diurnal rhythmicity in parameters studied either in foals or in mares was found. 4. Decrease of haemoglobin level in lactating mares (14.0-12.5 g/100 ml) without changes in red blood cell number were observed. Mean corpuscular haemoglobin changed distinctly from one study day to another. 5. In foals were stated: significantly higher haemoglobin level and erythrocyte number and lower mean corpuscular haemoglobin than in mares.     

Development of diurnal rhythm in some metabolic parameters in foals

1. The development of diurnal rhythm activity of FDPA, AspAT and A1AT and in levels of cortisol, T3 and T4 was observed in the blood serum of six foals. 2. The studies began when a foal was 7 days old and were repeated every month until foals reached 1 year of age. Blood samples were taken every 4 hr for one day each month. 3. As a control group four barren mares were used, kept and examined in the same conditions. 4. In mature mares, diurnal rhythms in activity of A1AT (acrophase at 2200 hr), AspAt (2400 hr) and cortisol (0630 hr) but in T3 only in summer months (acrophase at 0100 hr) were observed. 5. During the first 6 months of foal life, significantly higher mean levels of FDPA, A1AT, T3 and T4 than in control mares were found. 6. The cortisol level in foals was half as much as that of mature mares throughout the year. 7. In foals the diurnal rhythm in A1AT activity occurred in the 5th month and in AspAt--in the 12th month (acrophase at 2400 hr), but in cortisol levels it was developed already in the second month of foal life (acrophase at 0830 hr).     

Spatial relations between mares and foals of the Welsh pony (Equus caballus)

Welsh pony mares and foals (Equus caballus) were usually found to be within 1 or 5 m of each other during the first week of the foal's life and gradually spent more time at greater distances as the foals became older. There was an overall levelling of the trend during the 9th–15th weeks of life of the foal, followed by a second period of change during weeks 16–24. Through weeks 21–24, mares and foals spent at least half of their time within 5 m of each other. Proximity was primarily due to foal activity except during foal recumbency. During the first 8 weeks of the foal's life, a mare remained close by when it was recumbent, either by grazing in a circle around it or by standing upright beside it. Mares and foals were most likely to be close together when they were resting upright with the other ponies in the herd and most likely to be far apart when the foal was playing. Similarities in patterns of spatial relationship between the foals of a given mare were demonstrated. There was no difference between colts and filies in the development of independence.     

Efficacy of domperidone gel in an induced model of fescue toxicosis in periparturient mares

The objective was to evaluate the efficacy of domperidone in the prevention of reproductive complications of fescue toxicosis in periparturient mares. Pregnant mares at ≤310 days of gestation were fed ≥200 μg ergovaline per kg diet daily in endophyte-infected fescue hay and seed, starting ≥30 days before their expected foaling date (EFD: 340 days after breeding). Thirty-five mares were randomized to a treatment group to receive either domperidone gel (n = 20, 1.1 mg/kg, PO, once daily) or placebo (n = 15). Treatment was initiated 10 to 15 days before the EFD and continued for 5 days after foaling. “Treatment success” was defined as foaling within 14 days of the EFD, adequate mammary development on the day of foaling, and adequate lactation for 5 days postpartum. Twenty-seven mares were included in the effectiveness analysis. More mares in the domperidone group (12/13, P < 0.0001) were treatment successes than in the control group (1/14). Gestation length was shorter (P = 0.0011), and lactation at foaling (P = 0.0011) was better for the domperidone-group mares. Foals from two control mares were born dead and four others died or were euthanized within a few days after birth, compared with one foal death (an autolyzed twin) from a domperidone-treated mare. Plasma IgG concentrations were evaluated in 24 foals. Failure of passive transfer of immunoglobulins (IgG <800 mg/dL) occurred in 13/16 (81%) foals of domperidone-group mares and 7/8 (88%) foals of control mares. In conclusion, the reproductive complications of fescue toxicosis in periparturient mares induced by a fescue seed/hay model were prevented by treatment with domperidone.     

Aggression and social spacing in light horse (Equus caballus) mares and foals

Aggression and social spacing were studied in 14 light horse mares and their foals living at pasture. Focal samples were collected on each mare-foal dyad for 6 to 10.5 h from 2 months of foal age until weaning at approximately 4 months of age. Observations on foals continued until approximately 6 months of age for 7.5 to 10.5 h per foal. Every 2 min the identities of all individuals within 5 m were recorded. All occurrences of agonistic behavior, and the participants, were recorded during the focal samples. In addition, during feeding of supplemental grain, all occurrences of agonistic behavior by all subjects were recorded. Significant correlations were found between mare rank and the rank of foals both prior to and after weaning. Before weaning, the rank of the foal was significantly correlated with birth order. No significant correlation between birth order and foal rank was found for the post-weaning hierarchy. An animal's gender had no significant effect on foal rank or the choice of preferred associate. Both prior to and after weaning, foals associated preferentially with the foal of their dam's most preferred associate. In addition, significant positive correlations were found between rank of mares and foals and the rate at which they directed aggression to other herd members.     

Endogenous transplacental transmission of Neospora hughesi in naturally infected horses

Over a 2-yr study period, we investigated possible endogenous transplacental transmission of Neospora hughesi in 74 mare and foal pairs following the diagnosis of neuronal neosporosis in a weanling foal. Presuckle and postsuckle serum of each foal, serum and colostrum of each periparturient mare, and serum of each mare and foal pair, collected at 3-mo intervals thereafter, were tested for N. hughesi using an indirect fluorescent antibody test (IFAT). Furthermore, whole blood and colostrum samples and placentae were tested for the presence of N. hughesi by real-time PCR. The mares' seroprevalence at foaling based on IFAT (titer ≥ 160) was 52 and 6% in 2006 and 2007, respectively. Colostral antibodies against N. hughesi were detected in 96 and 11% of the mares in the 2-yr study. With the exception of 3 foals, all remaining foals were born seronegative to N. hughesi. Passive transfer of colostral antibodies to N. hughesi was documented in 15 foals. Three foals born from 2 different mares had presuckle antibodies at a titer ranging from 2,560 to 20,480. All 3 foals were born healthy. Two foals were born to the same dam that also gave birth to the weanling diagnosed with neuronal neosporosis in 2005. The third foal was born to a second mare with no previous foaling history at the farm. Seroconversion was documented in 10 foals and 9 mares over the 2-yr study. All blood and colostrum samples tested PCR negative for N. hughesi. Only 1 placenta collected in 2007 from the mare with the 2 congenitally infected foals tested PCR positive for N. hughesi. In conclusion, N. hughesi persisted in this population via endogenous transplacental infection.     

Specific Immune Response of Mares and their Newborn Foals to Actinobacillus spp. Present in the Oral Cavity

Oral swab samples, serum and colostrum was taken from 15 mares and 14 of their foals, within 24 h of birth. The presence of antibody against Actinobacillus spp. isolated from the oral cavity was investigated using agar gel immunodiffusion. Antibodies against 48 out of the 77 Actinobacillus isolates from all horses in the study were present in the respective sera of 13 mares and 9 foals. In 11 mother-foal pairs, the antibody content of the foal serum was similar to that of the mare, and in 9 cases this was reflected in the antibody content of colostrum from the mare. The results indicate that an immune response to Actinobacillus spp. colonising the oral cavity is present in many adult horses and that this immune response can be transferred from mother to foal via colostrum.     

The opportunistic intracellular bacterial pathogen Rhodococcus equi elicits type I interferon by engaging cytosolic DNA sensing in macrophages

Rhodococcus equi is a major cause of foal pneumonia and an opportunistic pathogen in immunocompromised humans. While alveolar macrophages constitute the primary replicative niche for R. equi, little is known about how intracellular R. equi is sensed by macrophages. Here, we discovered that in addition to previously characterized pro-inflammatory cytokines (e.g., Tnfa, Il6, Il1b), macrophages infected with R. equi induce a robust type I IFN response, including Ifnb and interferon-stimulated genes (ISGs), similar to the evolutionarily related pathogen, Mycobacterium tuberculosis. Follow up studies using a combination of mammalian and bacterial genetics demonstrated that induction of this type I IFN expression program is largely dependent on the cGAS/STING/TBK1 axis of the cytosolic DNA sensing pathway, suggesting that R. equi perturbs the phagosomal membrane and causes DNA release into the cytosol following phagocytosis. Consistent with this, we found that a population of ~12% of R. equi phagosomes recruits the galectin-3,-8 and -9 danger receptors. Interestingly, neither phagosomal damage nor induction of type I IFN require the R. equi’s virulence-associated plasmid. Importantly, R. equi infection of both mice and foals stimulates ISG expression, in organs (mice) and circulating monocytes (foals). By demonstrating that R. equi activates cytosolic DNA sensing in macrophages and elicits type I IFN responses in animal models, our work provides novel insights into how R. equi engages the innate immune system and furthers our understanding how this zoonotic pathogen causes inflammation and disease.     

Maternal care and foal social relationships in a herd of Sorraia horses: Influence of maternal rank and experience

The influence of maternal rank and experience on patterns of maternal care and social relationships of foals were investigated in a managed herd of Sorraia horses, Equus caballus. Social interactions and spatial relationships of 13 foals (seven females and six males) born to seven mares were examined from birth to 10 months of life, within the three major periods of foal development. Conflict over suckling between dam and foal was not generally affected by rank and experience, but higher-ranking mothers allowed more suckling during late lactation than lower-ranking mothers. Foals of higher-ranking mares spent more time in proximity to the mother during socialization. Maternal rank and experience did not significantly affect maternal protectiveness, foal independence from the mother or the development of affiliative relationships between foals and group members. Foals of higher-ranking mares received lower frequencies of aggression from other horses only in the first month of life. Dominance relationships among foals depended mainly on aggressiveness and were not associated with maternal rank. The large variability in maternal behaviour, the absence of a significant association between maternal rank and body condition at parturition and the stable social environment within this herd may partly account for the reported results.     

Diurnal changes in lactic and pyruvic acid levels and pH values in foals during the first 13 weeks of their life and in their lactating mothers.

1. The diurnal changes in the levels of lactic (LA) and pyruvic (PA) acids and values of pH were studied in standard-bred mares and their foals, for 13 weeks of foal life, throughout 2 years. 2. The studies began when a foal was 7 days old and were repeated every 2 weeks until foals reached 13 weeks of age. 3. Blood samples were taken every 4 hr for one day, each second week. 4. In the LA, PA levels and pH values no diurnal rhythm was stated in lactating mares during study periods. 5. In foals the diurnal rhythm in LA and PA occurred in the 3rd month of their life, with the acrophase at night hours. 6. The significant correlations between foal and their mother metabolites studied were observed. 7. The environmental factors (air temperature, humidity) influence fluctuations of the amplitude in parameters studied in the horse blood.     

Oral Administration of Electron-Beam Inactivated Rhodococcus equi Failed to Protect Foals against Intrabronchial Infection with Live,Virulent R. equi

There is currently no licensed vaccine that protects foals against Rhodococcus equi–induced pneumonia. Oral administration of live, virulent R. equi to neonatal foals has been demonstrated to protect against subsequent intrabronchial challenge with virulent R. equi. Electron beam (eBeam)-inactivated R. equi are structurally intact and have been demonstrated to be immunogenic when administered orally to neonatal foals. Thus, we investigated whether eBeam inactivated R. equi could protect foals against developing pneumonia after experimental infection with live, virulent R. equi. Foals (n = 8) were vaccinated by gavaging with eBeam-inactivated R. equi at ages 2, 7, and 14 days, or gavaged with equal volume of saline solution (n = 4), and subsequently infected intrabronchially with live, virulent R. equi at age 21 days. The proportion of vaccinated foals that developed pneumonia following challenge was similar among the vaccinated (7/8; 88%) and unvaccinated foals (3/4; 75%). This vaccination regimen did not appear to be strongly immunogenic in foals. Alternative dosing regimens or routes of administration need further investigation and may prove to be immunogenic and protective.     

Effects of Administration of Live or Inactivated Virulent Rhodococccus equi and Age on the Fecal Microbiome of Neonatal Foals

Background

Rhodococcus equi is an important pathogen of foals. Enteral administration of live, virulent R. equi during early life has been documented to protect against subsequent intrabronchial challenge with R. equi, indicating that enteral mucosal immunization may be protective. Evidence exists that mucosal immune responses develop against both live and inactivated micro-organisms. The extent to which live or inactivated R. equi might alter the intestinal microbiome of foals is unknown. This is an important question because the intestinal microbiome of neonates of other species is known to change over time and to influence host development. To our knowledge, changes in the intestinal microbiome of foals during early life have not been reported. Thus, the purpose of this study was to determine whether age (during the first month of life) or administration of either live virulent R. equi (at a dose reported to protect foals against subsequent intrabronchial challenge, viz., 1×10¹⁰ colony forming units [CFU]) or inactivated virulent R. equi (at higher doses, viz., 2×10¹⁰ and 1×10¹¹ [CFU]) altered the fecal microbiome of foals.

Methodology/Principal Findings

Fecal swab samples from 42 healthy foals after vaccination with low-dose inactivated R. equi (n = 9), high-dose inactivated R. equi (n = 10), live R. equi (n = 6), control with cholera toxin B (CTB, n = 9), and control without CTB (n = 8) were evaluated by 454-pyrosequencing of the 16S rRNA gene and by qPCR. No impact of treatment was observed among vaccinated foals; however, marked and significant differences in microbial communities and diversity were observed between foals at 30 days of age relative to 2 days of age.

Conclusions

The results suggest age-related changes in the fecal microbial population of healthy foals do occur, however, mucosal vaccination does not result in major changes of the fecal microbiome in foals.     

Effect of corticotherapy on proteomics of endometrial fluid from mares susceptible to persistent postbreeding endometritis

The objective was to determine the effects of corticotherapy, in the presence and absence of uterine inflammation, on proteomics of endometrial fluid from mares susceptible to endometritis. In 11 mares, estrus was induced seven times with 5 mg PGF_2α given at 14-day intervals. The first estrus was a control (no treatment). During the third estrus, mares received glucocorticoid (GC) treatment (20 mg isoflupredone acetate) every 12 h, for three consecutive days. The fifth estrus was the Infected treatment (intrauterine infusion of 1 × 10⁹ colony-forming unit/mL Streptococcus equi subspecies zooepidemicus). Finally, the seventh was a combination of GC + Infected treatment (infusion of bacteria 24 h after the first GC treatment). At 12 h after the end of each treatment, uterine samples were collected and submitted to two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for protein separation and mass spectrometry. Both GC treatment and uterine lumen infection induced proteomic alterations in the endometrial fluid of susceptible mares, characterized by an increase, decrease, or both in the relative optic density and/or frequency of inflammatory acute phase proteins (APP), with major alterations occurring when corticotherapy was applied in the presence of an infectious process. Corticotherapy in the presence of infection increased α₁-antitrypsin (AAT), transthyretin (TT), and actin, but reduced immunoglobulin G, whereas intrauterine infection increased haptoglobin (Hp) and apolipoprotein A-1 (ApoA-1) and decreased transferrin (TF). Infection reduced levels of α₁-antitrypsin and transthyretin, whereas corticotherapy in the presence of infection increased their frequency. We concluded that GC influenced the immune response, not only as suppressors, but also as enhancers of local defense mechanisms, through an immunomodulatory action. Short-term corticotherapy could be beneficial for treatment of uterine infectious processes in the mare.