Passive transfer of naturally acquired specific immunity against West Nile Virus to foals in a semi-feral pony herd

Horses naturally exposed to West Nile Virus (WNV) or vaccinated against WNV develop humoral immunity thought to be protective against development of clinical disease in exposed or infected animals. No reports evaluate the efficacy of passive transfer of naturally acquired specific WNV humoral immunity from dam to foal. The purpose of this study was to investigate passive transfer of naturally acquired immunity to WNV to foals born in a herd of semi-feral ponies, not vaccinated against WNV, in an endemic area, with many dams having seroconverted because of natural exposure. Microwell serum neutralization titers against WNV were determined in all mares and foals. Serum IgG concentration was determined in foals by serial radial immunodiffusion. Differences in IgG concentration between seropositive and seronegative foals were examined by means of the Mann-Whitney U-test. Linear regression was used to evaluate the association between mare and foal titers. Seventeen mare-foal pairs were studied; 1 foal had inadequate IgG concentration. IgG concentration was not different between seronegative and seropositive foals (P = .24). Mare and foal titers were significantly correlated in foals with adequate passive transfer of immunity (Spearman's rho = .84; P < .001); >90% of the foal's titer was explained by the mare's titer (R2 = 0.91; P < .001). Passive transfer of specific immunity to WNV is present in pony foals with adequate passive transfer of immunity born to seroconverted mares.     

Duration of maternally derived antibodies against equine influenza in newborn foals

Serum antibody concentrations against influenza A-equi-1 virus and A-equi-2 virus were measured in a group of 18 foals from birth to 4 months of age. More than 50% of the foals were seronegative to A-equi-1 virus infection by 4 weeks of age, with titers of less than or equal to 1:16. For A-equi-2 virus, more than 50% of the foals were seronegative by 2 weeks of age, with titers of less than or equal to 1:8. Passively derived antibodies against influenza A-equi-1 virus and A-equi-2 virus in foals obtained from recently vaccinated mares and from mares not vaccinated within 6 months before foaling were low in titer. The duration of passively derived antibodies was also short-lived.     

Immunologic Profiles of Peripheral Blood Leukocytes and Serum Immunoglobulin G Concentrations in Perinatal Mares and Neonatal Foals (Heavy Draft Horse)

The purpose of this study was to examine sequential changes in the immunologic parameters of perinatal mares and neonatal foals of the heavy draft horse. Blood samples were collected from clinically healthy pregnant mares and their newborn foals every week from 1 month before the expected foaling date, and 1 hour, 1 day (24-48 hours), and 1, 2, 3, and 4 weeks after foaling. Peripheral blood samples were used to examine total leukocyte counts (n = 20), differential leukocyte counts (n = 20), lymphocyte subpopulations (n = 13), lymphocyte responses to mitogens (n = 10), neutrophil phagocytic function (n = 12), and serum immunoglobulin G (IgG) concentrations (n = 10). In perinatal mares, remarkable changes observed included increased neutrophils, decreased lymphocytes, decreased CD4⁺ T lymphocytes, and decreased lymphocyte responses to mitogens at delivery. These changes were speculated to be the result of physical stress associated with delivery. In neonatal foals, increase in the phagocytic function of neutrophils, and increase in serum IgG concentration after suckling colostrum and increase of lymphocytes accompanied by physiologic growth were observed. Compared to dams, foals showed lower phagocytic function of neutrophils before suckling and fewer lymphocytes and lower lymphocyte responses to mitogens within 1 day after birth. This study revealed immunologic dynamics in perinatal mares and neonatal foals. Immunologic functions are suppressed in foaling mares and are immature in neonatal foals, especially before colostral intake. We expect these data will be useful for further studies in the field of clinical immunology, and preventive medicine.     

Maternal antibodies against equine influenza virus in foals and their interference with vaccination.

Foals that were born to mares vaccinated twice a year against influenza had moderate to high haemagglutination-inhibition antibody titers at 24 hours after birth. The foals were vaccinated at six and ten weeks of age, and again at three to five months after birth. Four months after the third vaccination no antibodies against A/H7N7 and A/H3N8 influenza viruses were detected in these foals. Thus, maternal antibodies probably prevented the development of antibodies against equine influenza virus after vaccination. Foals born to the same mares one year later were monitored to determine the rate of decline of maternal antibodies against influenza viruses. Antibody titers of the foals shortly after birth were similar to those of the mares at foaling. The antibodies persisted for three to six months, and their biological half-life was estimated to be approximately 38 days. Two vaccinations of foals against influenza after the maternal antibodies had virtually disappeared resulted in an antibody response in most, but still not all, foals. These findings suggest that foals should not be vaccinated against influenza until maternal antibodies have disappeared.     

Induction of parturition in mares: effect on passive transfer of immunity to foals

Parturition was induced in 11 mares, using a synthetic prostaglandin. Eight mares, not treated, were used as controls. There was no significant difference between the serum immunoglobulin G (IgG) concentrations of the treated and control mares. The concentration of IgG in the colostrum of treated mares compared favorably with that reported for naturally foaling mares. Four foals from treated mares died or were euthanatized because of weakness during the 1st 24 hours after birth. The mean IgG concentration in the surviving foals from treated mares at 24 to 36 hours of age was 1,561 mg/100 ml, which was significantly (P less than 0.01) lower than the mean concentration of 2,731 mg/100 ml in foals from control mares. The mean serum IgG concentration in foals from control mares was significantly (P less than 0.01) greater than that of their dams, whereas the mean serum IgG concentration of the foals from treated mares was significantly (P less than 0.01) lower than that of their dams.     

Vaccination of pregnant ponies against equine rhinopneumonitis

Bovine herpesvirus 1247 (one dose) was given subcutaneously to five pregnant pony mares between 227 and 319 days of their gestations. There were no adverse clinical reactions, and the virus was not recovered from nasal swabs collected during a 2-week period after vaccination. Four ponies foaled full-term, live, healthy foals. The foal of the fifth mare (No. 1) was found dead, but on the basis of the pathologic and virologic examinations, the virus was not considered to be the cause of the death. At 3 weeks after vaccination, the pregnant pony mares had a 13- to 250-fold increase in serum antibody titer to equine herpesvirus-1. A virulent-virus challenge exposure of all pony mares at 208 days after vaccination resulted in antibody titers greater than those just before this exposure. Virus was recovered from nasal swabs from vaccinated mares only on postexposure day 1, whereas the one control (nonvaccinated) pony shed virus for at least 3 days after challenge exposure. The immunogenic and the nonabortifacient characteristics of the herpesvirus 1247 in pregnant pony mares indicate that it may be useful to vaccinate horses against equine herpesvirus-1.     

Immune potency of lyophilized, killed vaccine for equine viral arteritis and its protection against abortion in pregnant mares

Immune potency test was conducted in horses by inoculating a killed vaccine for equine viral arteritis (EVA) which had been freeze-dried and contained aluminum hydroxide adjuvant. Serum neutralizing (SN) antibody to equine arteritis virus (EAV) was detected at maximal titers of 1:80 to 1:640, 1 to 2 weeks after 2-dose vaccination of 6 female horses. However, 6 pregnant mares inoculated with the vaccine which had been kept in storage for 1 year at 4°C produced much higher titers ranging from 1:320 to 1:1280. A maximal mean titer of 1:199.5 occurred in the 1st and 2nd week after 2-dose inoculation with the nonpreserved vaccine, whereas a maximal mean titer of 1:794.3 occurred in the 2nd week using the preserved vaccine. The horses showed no systemic or local adverse reactions clinically or hematologically after vaccination. Four of the 6 vaccinated pregnant mares were exposed to the Bucyrus strain of EAV but resisted challenge exposure, while 3 nonvaccinated control pregnant mares revealed acute EVA causing abortion and death. Isolation of EAV was positive from the body tissues of the aborted and dead fetuses and their dams, but was negative from the vaccinated mares. No significant rise of SN antibody titers was detected in the vaccinated mares following challenge exposure, suggesting that the vaccine can protect against EAV infection in pregnant mares and prevent abortion or death.     

Maternal antibody passively transferred interferes with rabies vaccination in hamsters

Transference and interference of maternal immunity to offspring after rabies vaccination were studied in hamsters. Females were vaccinated or not before mating and offspring were vaccinated at the age of 7, 14, 21 and 30 days. Other pups were maintained as controls. Thirty days after vaccination pups were challenged intracerebrally with CVS virus. Mouse neutralization tests were used to verify antibody titers. Mortality of 97.0, 76.9, 60.9 and 24.0% was observed in pups vaccinated at 7, 14, 21 and 30 days respectively, born from vaccinated dams, while in pups from non-vaccinated dams, mortality was 51.4, 28.6, 8.7 and 0.0%. Statistically significant associations were found between mortality and age at vaccination, by simple linear regression with y=-3.1169x + 120.8 (p = 0.008; r2=0.98) for litters vaccinated and born from vaccinated dams and y=-2.2541x + 62.7495 (p = 0.03; r2=0.93) for pups vaccinated and born from non-vaccinated dams. Immunological response to vaccination in pups born from vaccinated mothers was delayed 11 days, when compared to that observed in pups of non-vaccinated mothers.     

Absorption of bovine colostral immunoglobulins G and M in newborn foals

The uptake of colostral IgG and IgM, their serum half-lives, and the rates of endogenous synthesis of IgG and IgM were evaluated in 6 newborn foals fed bovine colostrum (principals) and 6 foals allowed to suckle their dams (controls). The principal foals were fed 400 ml of bovine colostrum (IgG, 10,000 mg/dl and IgM, 200 mg/dl) at 2-hour intervals, from 2 to 20 hours after foaling (total dose, 4 L). Serum IgG and IgM concentrations were determined by single radial immunodiffusion from birth to 98 days of age. At foaling, principal foals had no detectable serum equine IgG, but 1 control foal had serum equine IgG of 185 mg/dl. After ingestion of colostrum, there was no significant difference in the maximal serum bovine IgG concentration (range, 1,350 to 3,300 mg/dl) in the principal foals, and maximal serum equine IgG concentration in the control foals (range, 500 to 6,000 mg/dl). The calculated biological bovine and equine IgG half-life in the principal and control groups was 9.4 and 26 days, respectively. Endogenous IgG synthesis was first detected in 1 principal foal at 3 days of age, but was detected first between 28 and 42 days in the other principal foals. Starting on day 56 there was no significant difference in serum equine IgG concentration between groups. At foaling, foals in both groups had low equine IgM concentrations. In the control foals, there was marked individual variation in the increases in equine IgM concentration (range, 5 to 73 mg/dl) after ingestion of colostrum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Investigation of antigen specific lymphocyte responses in healthy horses vaccinated with an inactivated West Nile virus vaccine

West Nile virus (WNV) is a single-stranded, enveloped RNA virus capable of causing encephalitic disease in horses. Unvaccinated horses are at risk for developing WNV disease in endemic geographic regions. Effective vaccination reduces disease frequency and diminishes disease severity in vaccinated individuals that become infected with WNV. Recent data indicate CD4+ lymphocytes are required for effective protection against disease; in particular, cross talk between CD4+ and CD8+ lymphocytes must be functional. The objective of this project was to investigate immune responses in horses throughout a series of three vaccinations using a commercial inactivated vaccine under natural conditions. Immune responses to vaccination were determined by neutralizing antibody titers with plaque reduction neutralization test (PRNT), IgM titer (capture ELISA), WNV specific antibody Ig subclass responses, WNV lymphocyte proliferative responses and intracellular cytokine expression. Horses were vaccinated with a series of three vaccines at 3-week intervals using an inactivated product. An initial measure of immune activation following vaccination was determined by evaluating changes in lymphocyte cytokine expression. Interferon (IFN) gamma and interleukin (IL)-4 expressing CD4+ lymphocytes significantly increased 14 days following initial vaccination compared to unvaccinated horses (P<0.05). IFN-gamma expressing CD8+ lymphocytes also increased and remained elevated for 110 days. Antigen specific lymphocyte proliferative responses were significantly increased up to 90 days following the third vaccination (P<0.05). As expected, vaccinated horses produced increased neutralizing antibody based on PRNT data and WNV antigen-specific Ig subclass responses compared with unvaccinated horses (P<0.05). Our data indicate that WNV vaccination with an inactivated product effectively induced an antigen-specific antibody responses, as well as CD4+ and CD8+ lymphocyte activation.     

The presence of anti-Yersinia pseudotuberculosis immunoglobulins in equine serum

The research was conducted on clinically healthy mares (n = 40) and foals (n = 78) during Y. pseudotuberculosis associated enzootics. The animals were divided into groups: I to IV--mares, IA to IVA--their offsprings, IB to IVB--foals which mothers were not treated with any medicaments. The animals in group I, IA and IB were injected with PBS; in group II, IIA and IIB--with Y. pseudotuberculosis strain-based vaccine, in group III, IIIA and IIIB--with P. acnes strain-based immunostimulator; in group IV, IVA and IVB--with P. acnes strain-based immunostimulator and (5 days after the immunostimulator injection) Y. pseudotuberculosis strain-based vaccine. The presence of antibodies was determined by means of ELISA. The study revealed anti-Yersinia pseudotuberculosis IgG only in 19 mares before, and in 25 mares and 26 foals 3 weeks after vaccination. The mean extinction 3 weeks after vaccination amounted to: II-0.489, IV-2.578, iiA-0.572, IVA-0.974, IIB-0.312, iVB-0.418. The cut-off extinction value was 0.154. The presence of anti-Yersinia pseudotuberculosis IgG before vaccination in the sera of clinically healthy mares may suggest that Y. pseudotuberculosis infection occurs definitely more often than is expected. Vaccination preceded by immunostimulation appeared to be the most efficient method of treatment against yersiniosis.     

Safety of multiple,submucosal inoculations of a live attenuated strangles vaccine in pregnant mares

A live attenuated vaccine against Streptococcus equi was administered submucosally in the upper lip to 224 pregnant and healthy mares to evaluate its safety. After a primary immunisation the mares were inoculated every 3 months until foaling. As control group, 206 mares of the same breeding farm were administered the solvent of the vaccine submucosally. None of the 430 mares presented any clinical evidence of strangles and neither local nor systemic reactions to vaccination were noticed. There was no association between abortion and vaccination. Furthermore no case of S. equi infection was revealed at post mortem examinations of aborted foals.     

Immunologic responses to West Nile virus in vaccinated and clinically affected horses

OBJECTIVE: To compare neutralizing antibody response between horses vaccinated against West Nile virus (WNV) and horses that survived naturally occurring infection. DESIGN: Cross-sectional observational study. ANIMALS: 187 horses vaccinated with a killed WNV vaccine and 37 horses with confirmed clinical WNV infection. PROCEDURE: Serum was collected from vaccinated horses prior to and 4 to 6 weeks after completion of an initial vaccination series (2 doses) and 5 to 7 months later. Serum was collected from affected horses 4 to 6 weeks after laboratory diagnosis of infection and 5 to 7 months after the first sample was obtained. The IgM capture ELISA, plaque reduction neutralization test (PRNT), and microtiter virus neutralization test were used. RESULTS: All affected horses had PRNT titers > or = 1:100 at 4 to 6 weeks after onset of disease, and 90% (18/20) maintained this titer for 5 to 7 months. After the second vaccination, 67% of vaccinated horses had PRNT titers > or = 1:100 and 14% had titers < 1:10. Five to 7 months later, 33% (28/84) of vaccinated horses had PRNT titers > or = 1:100, whereas 29% (24/84) had titers < 1:10. Vaccinated and clinically affected horses' end point titers had decreased by 5 to 7 months after vaccination. CONCLUSIONS AND CLINICAL RELEVANCE: A portion of horses vaccinated against WNV may respond poorly. Vaccination every 6 months may be indicated in certain horses and in areas of high vector activity. Other preventative methods such as mosquito control are warranted to prevent WNV infection in horses.     

Are mule pregnancies really longer than equine pregnancies? Comparison between mule and equine pregnancies

In equine management, it is important to predict the approximate foaling date of mares to monitor parturition and allow early identification and intervention of problems during the perinatal period. There are no studies comparing accurate gestational length (GL) when mares are carrying mule foals and no controlled comparisons between GL of mares pregnant with equine or mule foals. Therefore, the objectives of this study were to compare GL of mares pregnant with equine or mule foals and establish normal reference values for GL of mares pregnant with male and female mules. Gestational length of Mangalarga Paulista breed mares pregnant with equine (n = 54) or with mule (n = 54) foals during the breeding seasons of 2007 to 2016 was evaluated. The mean GL was 347.2 ± 1.4 days (range of 326–368 days) and 341.1 ± 1.6 days (range of 307–360 days) for equine and mule pregnancies, respectively. The normal GL reference for mule pregnancies was 316.9–365.3 days. Therefore, GL of equine pregnancies was longer than of mule pregnancies. Gestational length was not different when pregnancies resulted in females or males within each group. This study established an important reference value for normal GL of mule pregnancies, which can be used by practitioners to estimate and predict foaling dates more accurately.     

Plasma prolactin concentrations in mares and their neonates after oxytocin induction of parturition

Studies were undertaken to investigate the effects of oxytocin induction on prolactin release in term (Group II) and preterm (Group III) mares and to compare these effects to spontaneously foaling mares (Group I). Since physiological concentrations of prolactin in blood have not been measured in the neonatal foal, experiments were designed to monitor prolactin in the cord artery and jugular blood of the foals from all groups of mares. Although prolactin levels varied in term mares (Group I and II) during the last 11 days of pregnancy, an increase was observed between Day -6 and Day 0 (2.7 and 11.9 ng/ml respectively; P less than 0.1). The average concentration of prolactin over the last 4 days (Days -3 to 0) had increased by 40% when compared to the average concentration on Days -6, -5, and -4. These findings indicate a rising trend which appears to occur concomitantly with changes in concentrations of 2 mammary components tested, sodium and potassium. Prolactin concentrations did not significantly increase in term mares after oxytocin treatment or in spontaneously foaling mares. However, the preterm induced mares had higher prolactin concentrations during the first stage of labor (19.3 +/- 7.2 ng/ml) than prior to treatment with oxytocin (4.7 +/- 2.0 ng/ml; P less than 0.01). Levels of prolactin in all groups significantly declined by 20-min post-placental expulsion. For the first 30 min after birth, prolactin concentrations in foals from oxytocin-induced mares appeared to be 2-fold higher than those from spontaneously foaling mares. Thereafter, prolactin values declined to baseline values by 48 hrs. When comparing cord arterial plasma with cord venous plasma in each group, prolactin concentrations were similar. However, the average prolactin levels in both the cord artery and vein appeared higher (ave: 1.1 ng/ml) in Group II and III than in Group I (less than 0.5 ng/ml). From these results, the authors suggest that 1) prolactin may have a role in regulating mammary secretory products in mares just prior to parturition; 2) oxytocin may increase prolactin secretion in preterm induced mares; 3) oxytocin induction may have a short term effect to increase circulatory prolactin concentrations in neonates in utero regardless whether their dams were treated preterm or term.     

Epidemiological studies of equine herpesvirus 1 (EHV-1) in Thoroughbred foals: a review of studies conducted in the Hunter Valley of New South Wales between 1995 and 1997.

Sero-epidemiological studies conducted between 1995 and 1997 on two large Thoroughbred stud farms in the Hunter Valley of NSW showed clear evidence of EHV-1 infection in foals as young as 30 days of age. Similarly, serological evidence suggested that these foals were infected with EHV-1 from their dams or from other lactating mares in the group, with subsequent foal to foal spread of infection prior to weaning. These studies also provided evidence of EHV-1 infection of foals at and subsequent to weaning, with foal to foal spread of EHV-1 amongst the weanlings. These data indicated that the mare and foal population was a reservoir of EHV-1, from which new cases of infection propagated through the foal population both before and after weaning. The results of these studies support the long standing management practices of separating pregnant mares from other groups of horses to reduce the incidence of EHV-1 abortion. Also, these results have important implications for currently recommended vaccination regimens, as the efficacy of vaccination in already latently infected horses is unknown.     

Anti-abortion and Fertility Vaccine Potential of Defined Double Deletion (ΔaroAΔhtrA) Mutant (S30) of Salmonella Abortusequi in Equids

The present study on defined double deletion (ΔaroAΔhtrA) mutant (S30) of Salmonella enterica serovar Abortusequi evaluated it for safety, immunogenicity, and efficacy as a vaccine candidate in equids. The candidate strain was found safe in equids (foals, male and female horses and donkeys, and pregnant and nonpregnant mares) and induced good humoral and cell-mediated immunity on administration through oral route. The strain was not excreted in feces of vaccinated animals. The vaccine candidate administered orally (1 × 10¹¹ cfu per animal) protected mares even after 180 days of inoculation against abortion on challenge with wild-type S. Abortusequi strain whereas in nonvaccinated control, all mares aborted. The vaccination in infertile mares resulted in regaining of fertility in 67%–80% thoroughbred mares at two different breeding farms. Further, the humoral immunity was transferred to foals from vaccinated mothers through colostrum, but no placental transfer was evident. Thus, the vaccine under study may be recommended for use in equids to control S. Abortusequi infection–associated abortions and also to enhance fertility of temporarily infertile mares in endemic areas.     

Changes in liver copper concentration of thoroughbred foals from birth to 160 days of age and the effect of prenatal copper supplementation of their dams

OBJECTIVES: To monitor the change in liver copper concentration of Thoroughbred foals from birth to 160 days of age and to determine the effects of supplementation by two injections of copper edetate given to dams in late gestation on the liver copper concentration of their foals at birth. PROCEDURE: Ten mares pregnant to the same stallion were randomised into two groups on the basis of age, liver copper concentration and expected foaling date. The treatment group mares were given 100 mg and 250 mg copper edetate intramuscularly during the ninth and tenth months of gestation respectively. Foals had liver biopsies taken weekly in the first month of life, then monthly for four months. Foals were euthanased at 160 days of age; liver samples were taken and the copper concentrations were determined. RESULTS: Two distinct patterns of age dependent decline in liver copper concentration were evident. The mean (+/- SD) liver copper concentration of the foals was high at birth (374 +/- 130 mg/kg DM), and for seven it declined to adult values by 160 days of age (21 +/- 6 mg/kg DM). In three foals the decline was at a slower rate than in the other seven and at 160 days of age the mean concentration was 162 +/- 32 mg/kg DM. Repeated measures analysis showed significant differences between each biopsy (P < 0.01) and between 'normal' and 'accumulator' foals (P < 0.002). Copper injections given to mares in late pregnancy had no effect on the liver copper concentration of foals at birth. CONCLUSIONS: The significance of the two patterns of age dependant decline in liver copper concentration is unknown. Parenteral copper supplementation of the dam in late gestation had no effect on the liver copper concentration of the foal at birth.     

Serum antibodies in mares and foals to Actinobacillus equuli whole cells, outer membrane proteins, and Aqx toxin

Actinobacillus equuli is carried in the alimentary tract of mares and can cause severe septicemia of neonatal foals. A hemolytic subspecies, A. equuli subsp. haemolyticus, and a non-hemolytic subspecies, A. equuli subsp. equuli, have been identified. Hemolytic strains produce the RTX toxin Aqx. The purpose of this study was to demonstrate sequentially in two sets of mare-foal pairs antibodies to A. equuli whole bacterial cells, outer membrane proteins, and recombinant Aqx and to compare the transfer of antibodies to these antigens between mares and their foals. Two mare/foal sets of sera were evaluated. Cohort A consisted of 18 mare-foal pairs obtained in the spring of 2005. Cohort B consisted of 10 mare-foal pairs obtained in the spring of 2006. For both sets, mare and foal sera were obtained immediately after foaling and prior to nursing (time 0) as well as at 12 and 24h and daily thereafter for 7 days. For Cohort B, sera were also obtained 30 days after birth. At parturition all mares had detectable antibodies to A. equuli whole cells and outer membranes; however, of those mares, two in Cohort A had undetectable antibodies to Aqx and their foals likewise had undetectable anti-Aqx antibodies. Antibodies against whole cells, outer membrane proteins, and Aqx were readily transferred from mares to foals. In most cases, there were significant correlations (p<0.05) between antibodies against whole cells, outer membrane proteins, and Aqx in mares' sera at the time of parturition and foal sera 24 after birth. Antibodies against the three antigen preparations had declined insignificantly (p>0.05) by day 30.