K. Gary Magdesian

Mutations in MITF and PAX3 cause "splashed white" and other white spotting phenotypes in horses.

During fetal development neural-crest-derived melanoblasts migrate across the entire body surface and differentiate into melanocytes, the pigment-producing cells. Alterations in this precisely regulated process can lead to white spotting patterns. White spotting patterns in horses are a complex trait with a large phenotypic variance ranging from minimal white markings up to completely white horses. The “splashed white” pattern is primarily characterized by an extremely large blaze, often accompanied by extended white markings at the distal limbs and blue eyes. Some, but not all, splashed white horses are deaf. We analyzed a Quarter Horse family segregating for the splashed white coat color. Genome-wide linkage analysis in 31 horses gave a positive LOD score of 1.6 in a region on chromosome 6 containing the PAX3 gene. However, the linkage data were not in agreement with a monogenic inheritance of a single fully penetrant mutation. We sequenced the PAX3 gene and identified a missense mutation in some, but not all, splashed white Quarter Horses. Genome-wide association analysis indicated a potential second signal near MITF. We therefore sequenced the MITF gene and found a 10 bp insertion in the melanocyte-specific promoter. The MITF promoter variant was present in some splashed white Quarter Horses from the studied family, but also in splashed white horses from other horse breeds. Finally, we identified two additional non-synonymous mutations in the MITF gene in unrelated horses with white spotting phenotypes. Thus, several independent mutations in MITF and PAX3 together with known variants in the EDNRB and KIT genes explain a large proportion of horses with the more extreme white spotting phenotypes.

Clinical and biochemical abnormalities in endurance horses eliminated from competition for medical complications and requiring emergency medical treatment: 30 cases (2005–2006)

OBJECTIVE To describe the clinical and clinicopathologic abnormalities in endurance horses eliminated from competition and requiring emergency medical treatment. DESIGN Retrospective study over a 2-year period (2005-2006). Ten horses that successfully completed the ride in 2006 were included for comparison. SETTING Temporary equine emergency field hospital. ANIMALS All horses (n=30) that were removed from endurance competition and treated for a metabolic abnormality were studied. INTERVENTIONS Horses were treated with IV fluids and analgesics. Monitoring included lab work (PCV, total protein, and electrolytes) and serial physical examinations. Statistical analysis included descriptive statistics and parametric and nonparametric comparisons (ANOVA, Friedmans test, and Kruskal-Wallis) where appropriate. MEASUREMENTS AND MAIN RESULTS The clinical diagnoses identified included colic, esophageal obstruction, poor cardiovascular recovery, myopathy, and synchronous diaphragmatic flutter. As a group, these sick horses had lower plasma chloride and potassium and higher total plasma protein concentrations as compared with 10 healthy horses that successfully completed the ride (P<0.05, <0.01, and <0.05 for chloride, potassium, and total protein, respectively). Horses with colic had a lower PCV as compared with horses with poor recovery and those with synchronous diaphragmatic flutter (P<0.05). All horses, including colics, were treated medically and discharged to owners. CONCLUSIONS Based on the results of this study, the prognosis for horses requiring emergency veterinary treatment after being removed from endurance competition (for metabolic reasons) appears to be good if horses are withdrawn from competition under the same criteria outlined in this study. Biochemical abnormalities tend to be mild and do not necessarily aid in delineating sick horses from successfully completing horses. None of the horses with gastrointestinal disease required abdominal surgery.

Neonatal Foal Diarrhea

Diarrhea is a significant cause of morbidity and mortality in the neonatal foal. Numerous noninfectious and infectious agents are responsible for enterocolitis and enteritis. This article provides an overview of the differential diagnoses for neonatal diarrhea and general and specific guidelines for therapy.

Use of quantitative real-time PCR for the detection of Salmonella spp. in fecal samples from horses at a veterinary teaching hospital.

A quantitative real-time (RT)-PCR assay was developed to detect Salmonella spp. in the feces of 911 equine species admitted to a veterinary hospital. Fresh feces and feces enriched for 24h in selenite broth were assessed by conventional culture and by RT-PCR targeting the Salmonella invA gene. The detection limit for the RT-PCR assay was 3 and 10 organisms, respectively, when spiked samples were purified from selenite broth and feces. The analytical specificity was 100% based on the detection of a panel of 40 salmonella serotypes from five serogroups and the lack of cross-reactivity with non-related micro-organisms. Although Salmonella spp. were not cultured from fresh feces, the organism was cultured from 6/911 (0.6%) of broth-enriched samples. The bacterial load in enriched samples varied from 3 to 861,037 salmonella invA gene copies/μL DNA. The RT-PCR assay had an overall relative accuracy of 98%, a relative sensitivity of 100% and a relative specificity of 98%, when compared to conventional culture. The judicious use of such a RT-PCR method has the potential to reduce the risk of nosocomial infections such as salmonellosis through the provision of highly accurate and rapid pathogen detection.

Juvenile idiopathic Epilepsy in Egyptian Arabian foals: 22 cases (1985-2005)

BACKGROUND Epilepsy is relatively uncommon in horses compared with other species and limited information is available. HYPOTHESIS The objectives of the study were to describe the age of onset, clinical signs, clinicopathologic data, electroeticephalographic findings, treatment, and outcome, including long-term prognosis in Arabian foals with idiopathic epilepsy. ANIMALS Twenty-two foals were included in the study. MATERIALS AND METHODS Medical records from 1985 to 2005 were reviewed. RESULTS The age of onset of affected foals ranged from 2 days to 6 months. Seizures were characterized by generalized tonic and clonic motor activity, staring, and loss of consciousness. The most common postictal signs were transient blindness and abnormal mental status. The interictal neurologic examination was otherwise normal. Clinicopathologic data and imaging diagnostics were normal except in 4 foals that developed complications. Electroencephalography revealed epileptiform activity in 9 of 13 foals. Foals were treated with benzodiazepines for the short-term; whereas phenobarbital was used over the long-term. Potassium bromide was added in 3 foals in which seizures were multiple, frequent, and difficult to control. The long-term prognosis was favorable with cessation of seizures by 1 year of age. The most common complication was head trauma. The most common concurrent disease was pneumonia. CONCLUSIONS AND CLINICAL IMPORTANCE Juvenile idiopathic epilepsy of Egyptian Arabian foals has an early clinical onset but appears to be self-limiting. Information obtained from this study strongly suggests a heritable condition that merits further investigation.

Parenteral nutrition in neonatal foals: clinical description, complications and outcome in 53 foals (1995-2005).

This retrospective study describes the use of and complications associated with parenteral nutrition (PN) administration to 53 equine neonates at the University of California Veterinary Medical Teaching Hospital. Medical records were examined and information obtained on signalment, physical examination, clinical diagnosis, outcome, total hospitalization time, insulin administration, microbiology culture results, other complications (i.e. thrombophlebitis) and necropsy findings. Complete blood count and serum biochemistry analytes, venous blood gas, serum electrolyte and glucose concentrations, and blood lactate concentration results were compared before and during PN administration in all foals. Seventeen foals (32%) developed hypertriglyceridemia (>200mg/dL). Triglyceride concentrations >200mg/dL were significantly (P=0.049) associated with non-survival. Forty-seven foals (89%) developed hyperglycemia (blood glucose >120mg/dL) and eight (15%) developed catheter-related complications (thrombosis or local sepsis). Packed cell volume, total protein, creatinine, blood urea nitrogen, and sorbitol dehydrogenase concentrations decreased while foals were on PN, while serum chloride concentration increased. This study highlighted that hypertriglyceridemia during the acute phase of neonatal illness may be detrimental to outcome, and that the safety of lipid-containing solutions in foals warrants further study.

Successful Treatment and Polymerase Chain Reaction (PCR) Confirmation of Tyzzer's Disease in a Foal and Clinical and Pathologic Characteristics of 6 Additional Foals (1986–2005)

BACKGROUND Tyzzers disease is a rapidly progressive and highly fatal hepatitis of foals caused by Clostridium piliforme. Survival of a confirmed case has not been reported previously. HYPOTHESIS Successful therapy of C. piliforme infection in foals is possible. Polymerase chain reaction (PCR) can be used to diagnose Tyzzers disease antemortem or postmortem. ANIMALS Seven foals were included in the study. METHODS Retrospective study was made to evaluate the clinical and pathologic characteristics of foals with Tyzzers disease. Medical records of the Veterinary Medical Teaching Hospital at University of California Davis were reviewed. Foals <3 months old were included in the study if typical clinical signs were present and histologic examination identified multifocal coagulative necrosis and hepatitis with intracytoplasmic filamentous bacilli, consistent with C. piliforme. A real-time TaqMan assay was developed to detect C. piliforme gene sequences in liver tissue from affected foals. RESULTS Median survival time from onset of disease in nonsurviving foals was 30 hours (mean 34.5 +/- 20.1; range, 16-62 hours). Common clinical findings included lethargy, recumbency, seizures, and fever. Laboratory findings included metabolic acidosis, hypoglycemia and increased activity of hepatobiliary enzymes. Treatment consisted of IV fluids, antimicrobial and antiinflammatory drugs, and parenteral nutrition. One filly survived, whereas 6 died. Postmortem examination of the 6 foals that died disclosed hepatomegaly with multifocal necrosis. Liver tissue from 4 foals was positive for C. piliforme gene sequences using PCR. CONCLUSIONS AND CLINICAL IMPORTANCE Although the mortality rate of Tyzzers disease is high, successful outcome is possible if intensive care is initiated promptly. PCR can be used for early and specific diagnosis.

Rattlesnake envenomation in horses: 58 cases (1992-2009)

OBJECTIVE To characterize signalment, clinical and laboratory findings, treatment, and outcome in horses with rattlesnake envenomation in northern California. DESIGN Retrospective case series. ANIMALS 58 client-owned horses evaluated for rattlesnake envenomation at 2 referral hospitals from 1992 to 2009. PROCEDURES Records of horses with rattlesnake envenomation were reviewed, and data concerning signalment, clinical and laboratory findings, treatment, and outcome were collected. In addition, a rattlesnake-bite severity score (RBSS) was assigned to each horse. Variables were compared between horses that survived and those that did not. RESULTS The overall mortality rate was 9%. Nine horses received antivenin; no complications were reported and none of the 9 died. The most common laboratory findings associated with severity of envenomation were thrombocytopenia, hypoproteinemia, hyperlactatemia, and a high RBSS. CONCLUSIONS AND CLINICAL RELEVANCE Most horses in this study had a good prognosis after being bitten by rattlesnakes. Laboratory and clinical examination findings may be useful for identifying horses with a poorer prognosis. Treatment with antivenin may be beneficial and warrants further evaluation.

Evaluation of a rapid agglutination method for detection of equine red cell surface antigens (Ca and Aa) as part of pretransfusion testing

BACKGROUND Blood typing before transfusion minimizes the risk of transfusion reactions and prevents immunization of the recipient against incompatible RBC antigens. The major RBC antigens that warrant identification before packed RBC or whole blood transfusions in horses are Ca and Aa. Standard blood-typing protocols are time-consuming (2.5-3.0 hours) and impractical in emergency settings. OBJECTIVES The purpose of this study was to determine whether equine RBCs could be typed for Ca and Aa antigens using sera from horses with RBC antibodies in a modified rapid (15 minute) blood-typing protocol. METHODS Serum was obtained from a horse with anti-Ca antibodies and from another horse with anti-Aa antibodies. The presence of agglutinating antibodies was confirmed with antibody screening. Venous blood samples, collected in citrate-phosphate-dextrose, were obtained from 21 horses of various breeds. Samples were blood typed in the Veterinary Medical Teaching Hospital Hematology Laboratory using standard methodology. Washed RBCs from each of the 21 horses were incubated individually with anti-Ca and anti-Aa sera at dilutions of 1:4, 1:8, and 1:16 for 15 and 30 minutes at room temperature and 37 degrees C. RESULTS Of the 21 horses, 13 were identified as Aa+/Ca+, four were Aa+/Ca-, two were Aa-/Ca+, and two were Aa-/Ca-. All 17 Aa-positive horses had a positive agglutination reaction at all dilutions of anti-Aa serum, incubation times, and temperatures, while all Aa-negative horses were negative. Each Ca-positive horse had a positive agglutination reaction at all incubation time points and temperatures up to the 1:16 dilution of the anti-Ca serum. All Ca-negative horses were negative at all times, temperatures, and dilutions of anti-Ca serum. Use of the modified protocol on 26 hospitalized horses resulted in accurate typing, based on complete antibody screens. CONCLUSIONS These results support the hypothesis that equine RBCs can be blood typed using a rapid (15 minute) protocol, at room temperature, for the presence of Ca and Aa antigens using equine-derived antisera. This technique may be beneficial for pretransfusion testing of equine patients in an emergency setting.

Determination of body water compartments in neonatal foals by use of indicator dilution techniques and multifrequency bioelectrical impedance analysis

OBJECTIVE To determine values for total body water (TBW), extracellular fluid volume (ECFV), intracellular fluid volume (ICFV), and plasma volume (PV) in healthy neonatal (< 24 hours old) foals and to create a multifrequency bioelectrical impedance analysis (MF-BIA) model for use in neonatal foals. ANIMALS 7 healthy neonatal foals. PROCEDURES Deuterium oxide (0.4 g/kg, IV), sodium bromide (30 mg/kg, IV), and Evans blue dye (1 mg/kg, IV) were administered to each foal. Plasma samples were obtained following an equilibration period, and the TBW, ECFV, ICFV, and PV were calculated for each foal. An MF-BIA model was created by use of morphometric measurements from each foal. RESULTS Mean ± SD values were obtained for TBW (0.744 ± 0.024 L/kg), ICFV (0.381 ± 0.018 L/kg), ECFV (0.363 ± 0.014 L/kg), and PV (0.096 ± 0.015 L/kg). The 95% limits of agreement between the MF-BIA and indicator dilution techniques were within ± 2 L for TBW and ECFV. CONCLUSIONS AND CLINICAL RELEVANCE Fluid volumes in neonatal foals were found to be substantially larger than fluid volumes in adult horses. Multifrequency bioelectrical impedance analysis may be a useful technique for predicting TBW, ICFV, and ECFV in neonatal foals.