Multiple congenital musculoskeletal abnormalities in a mule foal: A case report.

Musculoskeletal abnormalities in neonate equids represent a common condition, which includes angular limb deformities, defective carpal/tarsal bone ossification, contracted limb and mandibular/maxillary prognathism. The present case report described the presentation and surgical management of multiple musculoskeletal abnormalities in a mule foal. A newborn mule foal was presented for several musculoskeletal abnormalities, such as angular deviation from the sagittal plane of both carpal joints, hind limb ligament laxity, and severe mandibular prognathism. Surgical management of mandibular prognathism was then treated through the application of a tension orthodontic wire. Postoperatively, there was a significant improvement in the correction of mandibular malocclusion and no further correction was needed. Management of other anomalies was mainly conservative, with stall rest and exercise limitations, with a considerable improvement in the first month of life. Thus, jaw malformations might be observed also in mule foals, and might be associated with multiple congenital abnormalities. Early recognition, appropriate management, and surgical treatment were essential.

Tetanus prophylaxis in horses: guidelines for New Zealand and Australia based on a critical appraisal of the evidence.

Horses are exquisitely sensitive to tetanus neurotoxin and are exposed to the risk of infection with Clostridium tetani throughout life. The vaccine against tetanus is highly effective at preventing disease, whereas tetanus in unvaccinated populations is associated with high mortality rates. Current guidelines in New Zealand and Australia for the available vaccine contain contradictions and limitations surrounding the optimal tetanus immunisation protocols for both adult horses and foals. This review critically evaluates the scientific literature on tetanus prophylaxis in horses within the context of equine practice and available products in New Zealand and Australia. The review was conducted by a panel of industry and specialist veterinarians to obtain agreement on nine equine tetanus prophylaxis guidelines for practising veterinarians. The primary protocol for tetanus toxoid (TT) immunisation consists of a three-dose series IM for all horses ≥ 6 months of age, and a four-dose series IM is proposed if commencing vaccination in foals between 3 and 6 months of age. Tetanus prophylaxis in foals < 3 months of age relies on passive immunity strategies. Following the completion of the primary protocol, a TT booster dose IM should be administered within 5 years, and every 5 years thereafter. When followed, these protocols should provide adequate protection against tetanus in horses. Additional tetanus prophylaxis guidelines are provided for veterinarians attending a horse experiencing a known "risk event" (e.g. wound, hoof abscess, surgery, umbilical infection). When a correctly vaccinated horse experiences a risk event, pre-existing immunity provides protection against tetanus. When an unvaccinated horse or one with unknown vaccination status, or a foal born to an unvaccinated dam, experiences a risk event, TT IM and tetanus antitoxin (TAT) 1,500 IU SC should be administered simultaneously at separate sites, and the TT primary immunisation protocol should subsequently be completed for the horse's respective age. In previously immunised pregnant broodmares, a TT booster dose administered 4-8 weeks prior to parturition optimises the transfer of passive immunity against tetanus to the newborn foal via colostrum; provided that post-natal IgG concentration in serum is > 800 mg/dL (8 g/L), such foals should be passively protected against tetanus up to 6 months of age. Survivors of clinical tetanus must still receive the primary protocol for vaccination against tetanus. In summary, all horses in New Zealand and Australia should be vaccinated against tetanus with protection maintained throughout life via TT booster doses, facilitated by accurate medical record keeping and client education.

Alloimmune Conditions in the Neonatal Foal.

Alloimmune disorders occur in foals when pregnant mares produce antibodies against antigens on the foal's cells or tissues, and concentrate them within colostrum. Once foals nurse and absorb colostral antibodies, they can develop hematologic or cutaneous manifestations that can occur individually or in combination. These include neonatal isoerythrolysis, a hemolytic anemia directed against factors on the foal's erythrocytes, alloimmune thrombocytopenia when the antibodies are directed against platelet antigens, alloimmune neutropenia when they are directed against neutrophil antigens, and a combination of suspected alloimmune ulcerative dermatitis, neutropenia and thrombocytopenia. Foals can also develop neutrophilic dermatitis which is suspected to be alloimmune.

Monitoring of inflammatory blood biomarkers in foals with Rhodococcus Equi pneumonia during antimicrobial treatment.

Rhodococcus equi (R. equi), a gram-positive facultative intracellular pathogen, is a common cause of pneumonia in foals and represents a major cause of disease and death. The aim of the present study was to investigate the time-depended changes in White Blood Cells (WBC), basophils (Baso), neutrophils (Neu), lymphocytes (Lymf), monocytes (Mon), eosinophils (Eos), platelet (PLT) counts, fibrinogen (Fbg) concentration, interferon (IFN-α, IFN-γ) and interleukins (IL-2 and IL-10) in foals with clinical R. equi pneumonia. The main treatment was with azithromycin-rifampicin for 14 days. Blood was sampled prior to, 7 and 14 days after starting therapy. Treatment was associated with significantly decreased counts of WBC, (25.6 ± 6.7 and 14.2 ± 2,7 × 103/ml), Neu (18.6 ±6.2 and 10.7 ± 3.1 × 103/ml), Mon (1.5 ± 0.5 and 0.9 ± 0.2 × 103/ml) and Fbg (539 ± 124 and 287 ± 26 g/dl) between day 0 and day 14. IL-2 and IL-10 concentrations were significantly increased (P = 0.028, P = 0.013, respectively) after treatment, whereas IFN-α and IFN-γ concentrations were not. The diagnostic potentials of INF-α, INF-γ, IL-2 and IL-10 per se seems not very high, however, the study suggests that the activity change of selected interleukins in the course of the disease may be associated with amelioration. We concluded that patterns of serum concentration changes of INF-α, INF-γ, IL-2 and IL-10 may help in the study of the innate immune response in foals during infection and treatment of R. equi pneumonia.

Case report: Findings in ovaries development from an aborted equine fetus.

An aborted female foal was submitted for necropsy. During the gross examination, the ovaries were pale, grayish, and enlarged (6 × 5 cm), with a well-developed vascular structure surrounding the external surface; the cut surface of the ovaries showed a brownish parenchyma with white follicular areas mainly localized in the peripheral region. The ovaries were fixed for histological investigations. The histological evaluation of the ovaries showed polygonal-shaped cells with abundant cytoplasm and round or oval nuclei, arranged in cords of single cells. The tissue architecture was characterized by the presence of lobular-like tissues with a central vein. The tissue mimicking hepatocytes was delimited by a mature fibrous tissue and was surrounded by the normal ovarian tissue characterized by germinal epithelium and primordial follicular structures. Based on the histological findings, a diagnosis of bilateral ovarian hamartoma was carried out initially. For a better characterization of the ovarian tissue, the expression of tissue-specific (liver and ovary) markers was investigated using immunohistochemistry. Following the immunohistochemical analysis, the hamartoma diagnosis was excluded. The ovaries exhibited unique characteristics different from those of adult horse ovaries as well as unique morphological features different from other mammalian species. This case report enhances our understanding of ovaries at a later stage of pregnancy and unveils unique characteristics of horse ovaries development, avoiding misdiagnosis with pathological findings, hamartomas, or neoplasia.

Occurrence of ultrasonographic assessed placental abnormalities, treatments, pregnancy outcome, and subsequent fertility on a large warmblood stud farm: A retrospective field study.

Little is known about the incidence and outcome of high-risk pregnancies in equine practice and clinical studies on spontaneous occurring placentitis cases and treatments are missing. Therefore, the aims of this retrospective field study were to (1) describe the incidence and severity of ultrasonographic assessed placental abnormalities (UPA) in 4,192 pregnancies on a large commercial warmblood stud farm in 2017 - 2019 and (2) characterize these UPA cases and their pregnancy outcome. UPA severity (Placental abnormality score (PSc) 1-3; low to high), nine treatment regimens (TM1-9) used in UPA cases and treatment duration as well as subsequent fertility were analyzed in the group of UPA mares. The proportion of pregnancies affected by UPA was 4.2 % (n=177/4192). Placental abnormality severity was scored as PSc1 (51.4 %), PSc2 (32.8 %) and PSc3 (15.8 %). The generalized mixed model revealed PSc was affected by mare age and mare status (own pregnancy (OP) or embryo transfer recipient (ER)) (P=0.035) with ER mares having increased PSc compared with mares having their own pregnancy. Abortion occurred in 17/177 (9.6 %) UPA pregnancies. Overall, at the end of the next season, 61.1 % of UPA mares were pregnant, 32.0 % barren, and 6.9 % open (n=175). Pregnancy was established in 62/91(68.1 %) of mares with PSc1, 31/58 (53.4 %) with PSc2 and 14/26 (53.8 %) with PSc3. Most pregnancies were achieved in the first 81/107 (75.7 %) or second 18/107 (16.8 %) inseminated cycle. In conclusion, early detection and treatment of ultrasonographic assessed placental abnormalities can save high-risk pregnancies in > 90 % of cases with a satisfying subsequent fertility.

Tumor necrosis factor-α is transferred to equine neonates via colostrum but is not associated with their health status.

We followed the hypothesis that equine neonates with reduced transfer of tumor necrosis factor-α (TNFα) are at increased risk of neonatal infection. We investigated TNFα concentrations in colostrum of healthy mares and blood of their neonates in a non-hospitalized population of Warmblood mares where delivery, neonatal adaptation and health was closely monitored by veterinarians. Concentration of TNFα and IgG was determined in colostrum respective milk and in neonatal blood collected immediately after delivery and 18 h thereafter in 97 foals that were assigned to groups failure of passive transfer (FPT; n = 31) and control (CON; n = 66) based on serum IgG concentration at 18 h of age. Foal health was assessed repeatedly during the first 24 h of life. Statistical analysis was done with p < 0.05 indicating significance. There were no significant differences between foal groups FPT and CON regarding age and parity of dams, gestation length (FPT 343 ± 10, CON 340 ± 8 days) and foal sex. Concentrations of TNFα in colostrum at birth and in foals at 18 h varied but did not differ between groups (colostrum FPT 6.1 ± 9.1, CON 9.9 ± 31.5 ng/ml; foal FPT 2.3 ± 5.9, CON 2.4 ± 5.3 ng/ml; n.s.). There was an increase in the mean serum TNFα concentration until 18 h in foals (n.s. between groups). Results of the present study confirm previous findings of TNFα transfer from the mare to the neonate via colostrum but do not suggest that transfer of TNFα via colostrum is important for protection of the neonate against infectious diseases.

Evaluating smartphone-based optical readouts for immunoassays in human and veterinary healthcare: A comparative study.

Recent advances have significantly enhanced the use of smartphone devices for medical diagnostics. This study uses high-resolution cameras in mobile devices to capture and process bioassay images, enabling the quantification of diverse biomarkers across a range of diagnostic tests conducted on 96-well microplates. The study evaluates the effectiveness of this technology through protein quantification techniques and immunoassays that generate colorimetric responses at specific wavelengths. It includes the assessment of bicinchoninic acid and Bradford protein quantification methods, alongside a conventional immunoassay for detecting mare antibodies in colostrum to monitor foal immunodeficiencies. Further application involves the readout of magneto-actuated immunoassays aimed at quantifying bacteria. The results obtained from benchtop spectrophotometry at 595, 562, and 450 nm are compared with those acquired using a smartphone, which identified color intensities in shades of blue, purple, and yellow. This comparison yields promising correlations for the samples tested, suggesting a high degree of accuracy in the smartphone capability to analyze bioassay outcomes. The analysis via smartphone is facilitated by a specific app, which processes the images captured by the phone camera to quantify color intensities corresponding to different biomarker concentrations. Detection limits of 12.3 and 22.8 µg mL-1 for the bicinchoninic acid assay and 36.7 and 45.4 µg mL-1 for the Bradford are obtained for protein quantification using the spectrophotometer and the smartphone app, respectively. For mare's antibodies in colostrum, the values are 1.14 and 1.72 ng mL-1, while the detection of E. coli is performed at 2.0 x 104 and 2.9 × 104 CFU mL-1, respectively. This approach offers further advantages, including wide availability, cost-effectiveness, portability, compared to traditional and expensive benchtop instruments.

Follicular Dynamics and Pregnancy Rates during Foal Heat in Colombian Paso Fino Mares Bred under Permanent Grazing.

No studies have evaluated the peripartum follicular dynamics resulting in foal heat under tropical environments. We aimed to assess retrospectively the peripartum follicular dynamics in Colombian Paso Fino mares that were inseminated at the foal heat, becoming pregnant or not. Records including follicular dynamics of pregnant mares prepartum and from foaling until foal heat ovulation were assessed in Colombian Paso Fino mares (CPF, n = 24) bred under permanent grazing in a tropical herd in Colombia. The number of ovarian follicles >10 mm before foaling and the largest follicle (F1) growth rate (mm/day) from foaling until the F1 reached the largest diameter (pre-ovulatory size) at the foal heat were assessed. Mares were inseminated at foal heat with 20 mL of semen (at least 500 million live spermatozoa) with >75% motility and 80% viability from a stallion of proven fertility. Ovulation was confirmed the day after follicles had reached the largest diameter. Quantitative data from follicular growth, the day at ovulation, from mares that became pregnant (PM) or not (NPM) at 16 days post-insemination were compared by one-way ANOVA, repeated measures ANOVA (follicle growth rate data) or Chi-square test (edema and cytology scores data). Epidemiological data, gestation length, and the number of follicles on third prepartum days did not significantly differ between PM and NPM (p > 0.05). Seventy-one percent of mares (17/24) got pregnant. Ovulatory follicles grew faster in the NPM group (n = 7), which ovulated between the seventh and ninth postpartum days, compared to PM (n = 17), which ovulated between the 11th and 13th postpartum days. Pre-ovulatory follicle diameter in PM (48.57 ± 0.8 mm) was significantly larger than in NPM (42.99 ± 1.0 mm) (p < 0.05). In addition, the PM edema score (2.93 ± 0.32 mm) on ovulation day was significantly lower (p < 0.05) than NPM (4.47 ± 0.05 mm). First postpartum ovulation occurred at 12.6 ± 0.3 and 8.5 ± 0.4 days (p < 0.05) in PM and NPM, respectively. Colombian Paso Fino mares bred under permanent grazing under tropical rainforest conditions with no foaling or postpartum complications showed a 71% gestation rate when inseminated at foal heat when ovulation occurs between the second and third postpartum week.

Trimethoprim-sulfonamide: a valid antimicrobial treatment in foals?

Trimethoprim-sulfonamide (TMPS) combinations are widely used to treat a range of infectious diseases in horses, but some equine practitioners are reluctant to use them for treatment of both neonatal and older foals. Considering the emergence of increased antimicrobial resistance, the use of protected antimicrobials commonly prescribed to foals should be avoided and alternative first-line therapy considered, where appropriate. This review examines the characteristics and pharmacokinetics of TMPS and its suitability for treatment of foals. Data regarding dosage and route of administration are reported on the basis of recent publications in foals. The review intends to share significant information about the common infections that are most likely responsive to TMPS treatment in foals and, as such, where TMPS might be considered a suitable first-line therapeutic option.

Changes in aspects of hoof and distal limb conformation in foals by radiographic evaluation.

This study investigated age-related radiographic changes in the distal parts of the forelimbs by radiographic evaluation and identified the radiographic changes associated with diseases specific to foals. The hoof angle (HA), distal phalanx angle (P3A), distal phalanx palmer angle (P3PA), distal interphalangeal joint angle (DIPJA), and metacarpophalangeal joint angle (MPJA) on lateromedial radiographs of forelimbs were measured on the day after birth (Day 1); at 1, 2, 4, 6, and 8 weeks of age; and then at monthly intervals until 12 months of age. HA and P3A significantly increased from 1 day to 4 weeks and 4 weeks to 3 months of age. The P3PA increased dramatically from 1 day to 1 week, 1 week to 2 weeks, and 2 weeks to 8 weeks of age, and then decreased after 3 months of age. DIPJA significantly decreased from 1 day to 2 weeks of age before increasing from 3 to 5 months of age. MPJA increased with age until 4 weeks, slightly decreased from 2 to 4 months of age, and then gradually decreased from 4 to 6 months of age. The findings indicate that foals' forelimbs typically show flexion of the fetlock and a broken backward hoof-pastern axis just after birth, an upright fetlock until 4 months of age, and a change to a mature conformation after 6 months of age. Physiological variants were correlated with the occurrence of common foal diseases during the radiographic evaluation periods.

Comparing microbiotas of foals and their mares' milk in the first two weeks after birth.

BACKGROUND: The mare-foal relationship is essential for the well-being and growth of a foal. Mare's milk provides a foal with nutrients, protective immunity, and microbes. Within the first two weeks of life, there is a risk for a foal to suffer from diarrhea, particularly "foal heat diarrhea" which happens at about the time of a mare's estrus cycle but is more likely due to transitions in the microbiota in the foal's gastrointestinal (GI) tract. We hypothesized that this GI microbiota transition could be caused by changes in lysozyme and microbial populations in the mare's milk. To test this hypothesis, fifteen mare-foal pairs were followed in the first 15 days post-foaling. Every other day milk was collected from mares and rectal swabs were collected from foals. Lysozyme activity in the mare's milk was measured using a fluorescence assay. Microbial DNA was isolated from the milk and swabs and the V4 domain of 16 S rRNA genes were PCR amplified and sequenced using Illumina MiSeq technology. Microbial populations were analyzed using DADA2 and phyloseq within R. RESULTS: Mare's milk lysozyme activity peaked for samples at Day 1 and levels dropped to 72.5% of Day 1 activity by Day 15; however, microbial populations in the mare's milk did not vary significantly over the two weeks. Furthermore, levels of microbial diversity found in foal rectal swabs were initially similar to microbial diversity seen in mare's milk; however, over the first fifteen days, diversity increased for the foal rectal swab microbiota and swab microbial populations differed from milk microbes. A transition occurred shifting from microbes from the phylum Proteobacteria early in rectal swabs to those primarily from the phyla Firmicutes and Bacteroidota after the first few days post-foaling. These phyla contained several families and genera of microbes that promote utilization of milk components in healthy gut transition. Microbial abundance levels correlated more with days post-parturition than with lysozyme activity and mare's milk microbial populations. CONCLUSIONS: The findings suggest that much of the microbial populations responsible for the transition of the foal's gut comes from sources outside of mare's milk species and levels of lysozyme activity.

The impact of age on vitamin D receptor expression, vitamin D metabolism and cytokine production in ex vivo Rhodococcus equi infection of equine alveolar macrophages.

Rhodococcus equi (R. equi), a pneumonia-causing intracellular bacterium, results in significant morbidity and mortality in young foals, while healthy adult horses rarely develop disease. Survival and replication within alveolar macrophages (AMφ) are the hallmarks of R. equi's pathogenicity. The vitamin D receptor (VDR) and its ligand, the active vitamin D metabolite 1,25(OH)2D, are important in immune responses to intracellular bacteria. The vitamin D/VDR pathway regulates the downstream production of cytokines in infected human AMφ. The immunomodulatory role of the vitamin D/VDR pathway in equine leukocytes is unknown. The objective of the current study was to determine the impact of R. equi infection and age on synthesis of 1,25(OH)2D, VDR expression, and cytokine production in an ex vivo model of R. equi infection in equine AMφ. AMφ were collected from ten healthy foals at 2-, 4- and 8-weeks old and from nine healthy adult horses once via bronchoalveolar lavage. AMφ were mock infected (CONTROL) or infected with a virulent laboratory strain of R. equi for 7 days (INFECTED). VDR expression was determined via RT-qPCR from cell lysates. 1,25(OH)2D and cytokines were measured in cell supernatant by immunoassays. VDR expression was impacted by age (P = 0.001) with higher expression in AMφ from 8-week-old foals than from 2-week-old foals and adults. There was no significant effect of infection in foal AMφ, but in adults, relative VDR expression was significantly lower in INFECTED AMφ compared to CONTROL AMφ (P = 0.002). There was no effect of age or infection on 1,25(OH)2D concentration (P > 0.37). Mean TNFα production was significantly higher from INFECTED compared to CONTROL AMφ from 4- and 8-week-old foals and adults (P < 0.005). Mean IFNγ production was significantly higher from AMφ from foals at 8-weeks-old compared to 2-weeks-old (P = 0.013) and higher from INFECTED AMφ than from CONTROL AMφ in foals at 4-weeks-old and in adults (P < 0.027). The proportion of samples producing IL-1ß and IL-10 was also significantly higher from INFECTED compared to CONTROL AMφ isolated from 4-week-old foals (P < 0.008). Similarly, in adult samples, IL-17 was produced from a greater proportion of INFECTED compared to CONTROL samples (P = 0.031). These data document age-associated changes in VDR expression and cytokine production in equine AMφ in response to R. equi infection. This preliminary investigation supports the need for further research to fully elucidate if the vitamin D pathway has an immunomodulatory role in the horse.

Serum activities of complement 1q and antibodies to the virulence-associated protein A are lower in foals that develop rhodococcal pneumonia.

OBJECTIVE: To determine the effects of transfusion of Rhodococcus equi hyperimmune plasma (REHIP) on serum concentrations of complement component 1q (C1q) and to examine the association of serum C1q and anti-rhodococcal antibodies of newborn foals with subsequent development of rhodococcal pneumonia. ANIMALS/SAMPLES: Foals (n = 205) from 2 Thoroughbred breeding farms in New York transfused with REHIP between January 1, 2022, and December 1, 2022. PROCEDURES: Blood was collected immediately before transfusion with REHIP and again from the contralateral vein immediately after transfusion. Foals were followed through weaning for clinical and ultrasonographic evidence of rhodococcal pneumonia. Serum samples were tested by ELISA for concentrations of C1q and for activity of IgG1 and IgG4/7 recognizing the virulence-associated protein A (VapA) of R equi. Logistic regression analysis was used to determine the association between rhodococcal pneumonia and levels of C1q and anti-VapA IgG1 and IgG4/7. RESULTS: REHIP significantly decreased C1q concentrations immediately after transfusion. Accounting for effects of farm and birth month, estimated odds of pneumonia were 2.1-fold (P = .0330) higher for foals with pretransfusion C1q concentrations less than or equal to the population median and 3.3-fold (P = .0051) higher for foals with posttransfusion IgG1 activity in the lower quartile. CLINICAL RELEVANCE: Both C1q and IgG appear to contribute to protection against R equi, and IgG1 appears to be especially important. Increasing IgG1 concentrations targeting rhodococcal proteins in REHIP or serum of foals might improve protection against R equi foal pneumonia.

Could assisted reproductive techniques affect equine fetal membranes and neonatal outcome?

Embryo transfer (ET) and intracytoplasmic sperm injection (ICSI) are widely used in equine species, but their effects on fetal adnexa and neonates have not been investigated yet. The aim of this study was to retrospectively evaluate whether pregnancies obtained by ET or ICSI could be associated with the presence of macroscopic alterations of fetal membranes (FM) and umbilical cord (UC) and if the use of these techniques could influence neonatal outcome. Sixty-six light breed mares hospitalized at the Veterinary Teaching Hospital, University of Bologna, for attending delivery were included in the study. Mares were divided into Artificial Insemination (AI; 32/66 mares, 48 %), Embryo Transfer (ET; 12/66 mares, 18.2 %) and Intracytoplasmic Sperm Injection (ICSI; 22/66 mares, 33 %) groups. All the medical reports of mares and their foals were reviewed and data about mare, pregnancy, foaling, fetal membranes, umbilical cord and foal were recorded. The occurrence of dystocia resulted statistically different between AI group and ICSI group (p = 0.0066), and between AI group and ET group (p = 0.044). Macroscopic examination of FM revealed alterations in 30/66 mares (46 %): 8/32 in AI (25 %), 7/12 in ET (58 %) and 15/22 in ICSI (68 %) with significant lower incidence in AI compared to ET (p = 0.04) and ICSI (p = 0.002) groups. Alterations reported were chorionic villi hypoplasia, chorioallantois edema, allantois cysts, necrotic areas and greenish-grey concretions. Total length of UC resulted significantly shorter in ICSI group (49 ± 9 cm; p < 0.03) compared to AI (60 ± 17 cm) and ET (59 ± 15 cm). However, there were no differences in the incidence of foals' diseases at birth and in foals' survival among groups (p > 0.05). The results demonstrate that transfer of in vivo or in vitro produced embryos may lead to alterations of placental development, as observed in other species, without being associated with a higher incidence of neonatal morbidity and mortality. Further studies about trophoblast development, FM histological evaluation, and placental gene expression should be carried out to clarify the mechanisms underlying the placental alterations.

Intramuscular but not nebulized administration of a mRNA vaccine against Rhodococcus equi stimulated humoral immune responses in neonatal foals.

OBJECTIVE: Design and evaluate immune responses of neonatal foals to a mRNA vaccine expressing the virulence-associated protein A (VapA) of Rhodococcus equi. ANIMALS: Cultured primary equine respiratory tract cells; Serum, bronchoalveolar lavage fluid (BALF), and peripheral blood mononuclear cells (PBMCs) from 30 healthy Quarter Horse foals. METHODS: VapA expression was evaluated by western immunoblot in cultured equine bronchial cells transfected with 4 mRNA constructs encoding VapA. The mRNA construct with greatest expression was used to immunize foals at ages 2 and 21 days in 5 groups: (1) 300 µg nebulized mRNA (n = 6); (2) 600 µg nebulized mRNA (n = 4); (3) 300 µg mRNA administered intramuscularly (IM) (n = 5); (4) 300 µg VapA IM (positive controls; n = 6); or (5) nebulized water (negative controls; n = 6). Serum, BALF, and PBMCs were collected at ages 3, 22, and 35 days and tested for relative anti-VapA IgG1, IgG4/7, and IgA activities using ELISA and cell-mediated immunity by ELISpot. RESULTS: As formulated, nebulized mRNA was not immunogenic. However, a significant increase in anti-VapA IgG4/7 activity (P < .05) was noted exclusively in foals immunized IM with VapA mRNA by age 35 days. The proportion of foals with anti-VapA IgG1 activity > 30% of positive control differed significantly (P = .0441) between negative controls (50%; 3/6), IM mRNA foals (100%; 5/5), and IM VapA (100%; 6/6) groups. Natural exposure to virulent R equi was immunogenic in some negative control foals. CLINICAL RELEVANCE: Further evaluation of the immunogenicity and efficacy of IM mRNA encoding VapA in foals is warranted.

Evaluation of a smartphone electrocardiograph in healthy foals and comparison to standard base-apex electrocardiography.

Smartphone-based technology for ECG recording has recently spread as a complementary tool for electrocardiographic screening and monitoring in adult horses and in other animal species. The present study aimed to assess the feasibility and accuracy of a smartphone-based ECG in healthy foals. This was a prospective observational study (authorization n. 45,865/2016) including 22 foals aged less than 21 days. A reference standard base-apex ECG (rECG) was acquired, and a smartphone ECG (sECG) was recorded immediately after by using a smartphone-based single lead electrocardiograph. All ECG tracings were evaluated in a blind fashion by a single board-certified cardiologist, who judged whether the tracings were acceptable for interpretation and performed ECG measurements and diagnosis. The Spearman correlation coefficient, the Cohen's k test and the Bland-Altman test were used to assess the agreement between sECG and rECG. All sECG tracings were acceptable for interpretation. All foals showed sinus rhythm on both rRCG and sECG tracings, with perfect agreement in heart rate classification (κ = 0.87; p < 0.001). No clinically relevant differences were found in the assessment of waves and intervals duration. Concerning P wave and QRS complex polarity, the percentage of agreement between rECG and sECG was 78% and 83%, respectively. About ECG tracing quality, rECG and sECG showed a substantial agreement (κ = 0.624; p < 0.001). In conclusion, the smartphone-based ECG device tested in the present study recorded good quality single-lead ECG tracings in foals, reliable for heart rate and ECG measurements, but different polarity of P waves and QRS complexes was found in some foals in comparison to rECG.

Hematology and clinical chemistry in mule foals from birth to two months of age: A preliminary study.

In horses and donkeys, age-related changes in hematological and biochemical parameters preclude the use of normal values of adults in the evaluation of foals. This study aimed to obtain data on hematological and biochemical parameters of mule foals from birth up to the second month of life and to assess age-related changes in order to determine if dedicated reference ranges are required in younger animals. Blood samples from seven healthy mule foals were obtained at birth before colostrum consumption, 24 h, 48 h of life, and then weekly until the second month of life. Results were expressed as mean and standard deviation or median, minimum, and maximum values if showing non-gaussian distribution. Kruskal-Walls and Dunn tests were used to verify the differences among sampling times. Significance was set at P < 0.05. Red blood cell count, packed cell volume and hemoglobin decreased from 24 h to one week of age. Mean corpuscular volume and mean corpuscular hemoglobin decreased over the first month. White blood cells increased from birth to seven days of life. Aspartate amino transferase increased while alkaline phosphatase decreased in the first week of life. Urea, creatinine, and lactate decreased, while glucose concentrations increased at 24 h. Ionized calcium and magnesium and total sodium and potassium showed no changes. In mule foals, several laboratory parameters may be the same or intermediate, lower or higher than in equine or donkey foals, but also compared to all other adult species. The preliminary results suggest that for mule foals, age influences hematological and biochemical parameters.