Complete caecal bypass without ileal transection for caecal impaction in horses: seven clinical cases (1997-2007).

OBJECTIVES: To report the clinical outcome in seven horses following use of a newly described surgical technique for treating caecal impaction. METHODS: The medical records of seven horses with caecal impaction treated surgically using a stapling technique to create a complete caecal bypass without ileal transection were reviewed. Data were obtained from the records and through telephone interviews with case-associated personnel. RESULTS: The mean age was 10 years (range 2-22 years) and duration of colic ranged from 24 h to 2 weeks. Five horses had type II motility dysfunction and the remaining two had type I. Mean surgical time was 185 min (range 146-245 min) and the horses were hospitalised for a mean of 12.4 days (range 9-22 days); 71% (5/7) were discharged from hospital and all five were alive 60 days from the surgery date. One horse was lost to follow-up. The four (66.7%) available remaining horses were alive ≥ 1 year (long-term survivors). CONCLUSIONS: Complete caecal bypass without ileal transection for clinical cases of caecal impaction had comparable outcomes to complete bypass with ileal transection. The technique is easy to perform, has the potential to reduce surgical time, compared with traditional bypass techniques, and may reduce the risk of intraoperative abdominal contamination. It is recommended for use in clinical cases in which caecal bypass is desirable.

Surgical repair of coxofemoral luxation in a horse.

A 4-year-old castrated male Miniature Horse was evaluated because of severe right hind limb lameness of 5 days' duration. The diagnosis of craniodorsal luxation of the right coxofemoral joint was made by physical examination and radiographic imaging. Closed reduction was attempted but was unsuccessful. Surgical reduction was successfully performed, using toggle pin, synthetic capsular reconstruction, and trochanteric transposition techniques. No postoperative complications were observed. Follow-up 26 months after surgery revealed no recurrence of the luxation and no evidence of lameness. These surgical techniques are used successfully for repair of coxofemoral luxations in small animals. To our knowledge, there has been no report of these techniques attempted in horses. These surgical techniques may have merit for the treatment of coxofemoral luxations in small equine patients.

Prevalence and prognostic importance of hypomagnesemia and hypocalcemia in horses that have colic surgery.

OBJECTIVE: To determine the prevalence of hypomagnesemia and hypocalcemia in horses with surgical colic. ANIMALS: 35 horses with surgically managed colic. PROCEDURE: Serum concentrations of total magnesium (tMg2+) and calcium (tCa2+), as well as ionized magnesium (iMg2+) and calcium (iCa2+) were analyzed before surgery and 1, 3, 5, and 7 days following surgery. A lead-II ECG and pertinent clinical data were also obtained at each time. RESULTS: Preoperative serum tMg2+ and iMg2+ concentrations were below the reference range in 6 (17%) and 19 (54%) horses, respectively. Serum concentrations of tCa2+ and iCa2+ were less than the reference range in 20 (57%) and 30 (86%) horses before surgery. Horses with strangulating lesions of the gastrointestinal tract had significantly lower preoperative serum concentrations of iMg2+ and iCa2+, as well as a higher heart rate than horses with nonstrangulating lesions. Horses that developed postoperative ileus had significantly lower serum concentrations of iMg2+ after surgery. Serum concentrations of magnesium and calcium (total and ionized) correlated significantly with the PR, QRS, QT, and corrected QT (QTc) intervals. Horses that were euthanatized at the time of surgery (n = 7) had significantly lower preoperative serum concentrations of iMg2+, compared with horses that survived. Neither serum magnesium nor calcium concentrations were predictors of hospitalization time or survival. CONCLUSIONS AND CLINICAL RELEVANCE: Hypomagnesemia and hypocalcemia were common during the perioperative period, particularly in horses with strangulating intestinal lesions and ileus. Serum concentrations of tMg2+ and tCa2+ were less sensitive than iMg2+ and iCa2+ in detecting horses with hypomagnesemia and hypocalcemia.

Treatments resulting in pregnancy in nonovulating, hormone-treated oocyte recipient mares.

Synchronization of follicle growth between oocyte donor and recipient mares is difficult. To avoid this, recipient mares in a clinical program were used during a period of low follicular activity, and were treated with estrogen before transfer and progesterone after transfer. Five pregnancies were established after oocyte transfer to nonovulating, hormone-treated recipient mares. One pregnancy was lost before 30 d gestation, and the other 4 foals were carried to term. One foal died at birth. Establishment and maintenance of pregnancy in these mares indicates that nonovulating, hormone-treated mares may offer an alternative to cyclic recipients in oocyte transfer programs.

Birth of a foal after oocyte transfer to a nonovulating, hormone-treated recipient mare.

A nonovulating, hormone-treated mare was used successfully as an oocyte recipient. The mare's ovarian activity was suppressed using progesterone and estrogen treatment. This treatment was stopped, then estrogen was administered for 3 d prior to the transfer. An oocyte was recovered from the follicle of a donor mare and was transferred via flank laparotomy into the recipient's oviduct. The recipient mare was inseminated 7 h before transfer. The recipient was treated with intramuscular progesterone from the day after transfer until 47 d after transfer, and then with oral altrenogest until 150 d gestation. A normal colt was born at 321 d gestation, and was shown by DNA analysis to be the progeny of the donor mare. This is the first report of fertilization and embryo development to term after transfer of oocytes to a nonovulating mare, and, to our knowledge, the first of its kind in any domestic species.

Reliability of using results of abdominal fluid analysis to determine treatment and predict lesion type and outcome for horses with colic: 218 cases (1991-1994).

OBJECTIVE: To assess clinical utility of abdominal fluid analysis in predicting outcome, lesion type, and whether medical or surgical treatment is indicated for horses with colic. DESIGN: Retrospective study. ANIMALS: 218 horses > 1 year old. PROCEDURE: Horses were classified on the basis of age, site of lesion, lesion type (nonstrangulating vs strangulating), type of treatment (medical vs surgical), and outcome (survival vs nonsurvival). Sensitivity and specificity of using age and results of abdominal fluid analysis, individually and in combination, to predict lesion type, type of treatment, and outcome were determined. RESULTS: Most single variables evaluated were found to have low sensitivity, specificity, and predictive value for determining lesion type, whether medical or surgical treatment was indicated, and outcome. When evaluated in series, abdominal fluid color and specific gravity had a high positive predictive value for lesion type, and patient age and abdominal fluid color had a high positive predictive value for outcome. CLINICAL IMPLICATIONS: Results of abdominal fluid analysis cannot be used alone to accurately predict lesion type, whether medical or surgical treatment is needed, or outcome for horses with colic.

Immunoprecipitation and virus neutralization assays demonstrate qualitative differences between protective antibody responses to inactivated hepatitis A vaccine and passive immunization with immune globulin.

Antibodies to hepatitis A virus (anti-HAV) were measured in children from two separate vaccine trials (n = 70) 4 weeks after a dose of inactivated hepatitis A vaccine (VAQTA). The geometric mean titers (GMTs) of anti-HAV were 49.3 and 45.2 mIU/mL by immunoassay, while reciprocal GMTs of neutralizing anti-HAV were 6.5 and 15.0 by an 80% radioimmunofocus inhibition test (RIFIT) and 55.6 and 92.0 by antigen reduction assay (HAVARNA). The GMT of antibody detected by radioimmunoprecipitation (RIPA) was > or =401. These data establish serologic correlates of protection against disease and show that RIPA is most sensitive for detection of early vaccine-induced antibody. Sera collected from adults (n = 20) 7 days after administration of immune globulin contained similar antibody levels by immunoassay (45.1 mIU/mL) and slightly higher GMTs of neutralizing antibody (27.5 by RIFIT and 146 by HAVARNA) but negligible precipitating antibody (GMT, 5.6). These results are best explained by differences in the affinity of antibodies for virus following active versus passive immunization.

Safety, tolerability, and immunogenicity of two regimens of Oka/Merck varicella vaccine (Varivax) in healthy adolescents and adults. Oka/Merck Varicella Vaccine Study Group.

A multicenter clinical trial was conducted among 757 healthy adolescents and adults, 13-54 years, to compare two regimens of Oka/Merck varicella vaccine with respect to safety, tolerability, and immunogenicity. Participants were randomized to receive two injections of vaccine either four or eight weeks apart and were followed for clinical reactions and serologic response. The two vaccine regimens were equally well tolerated. The seroconversion rates (gpELISA) four weeks after injection 1 and 2 were 72 and 99%, respectively, for those who received vaccine four weeks apart and 78 and 99%, respectively, for those who received vaccine eight weeks apart. The differences in seroconversion rates were not statistically significant. However, delaying the second dose to eight weeks resulted in a higher antibody titer one month after the second injection. Administration of a two-dose regimen of varicella vaccine to susceptible adolescents and adults is well tolerated and highly immunogenic.

Combined use of complement and anti-immunoglobulin in an enhanced neutralization assay for antibodies to varicella-zoster virus.

An enhanced neutralization assay was developed to permit the sensitive, specific, and reproducible measurement of antibodies to varicella-zoster virus (VZV). Optimal neutralization was achieved using a combination of guinea pig complement (C') and rabbit anti-human IgG. This provided 625-, 160- and 13- to 64-fold increases in dilution endpoints of human post-zoster serum, varicella-zoster immune globulin and representative sera from recipients of live attenuated varicella vaccine, respectively, above those measured in the absence of C' and anti-IgG. The specificity of the assay was shown by the absorption of serum neutralization capacity with VZV-specific antigen and the lack of concordance between antibody titers to VZV with those to either herpes simplex virus type-2 or cytomegalovirus. The antibody status of recipients of live attenuated varicella vaccine was established from the amount of neutralizing activity produced at a single optimal serum dilution.

Histologic and ultrastructural changes after large-colon torsion, with and without use of a specific platelet-activating factor antagonist (WEB 2086), in ponies.

The role of platelet-activating factor (PAF) in mediating the colonic damage that develops after large-colon torsion was studied in 14 ponies. Morphologic changes in areas of the ascending colon and selected abdominal and thoracic viscera after 1 hour of large-colon torsion and 3 to 5 hours of reperfusion were determined, as well as the protective effects of systemic administration of a specific PAF antagonist (WEB 2086). Ponies were selected then allocated at random and in equal numbers to 2 groups that received 1 of 2 treatments prior to induction of large-colon torsion: group 1--control (saline solution), and group 2--WEB 2086 (3 mg/kg of body weight loading dose and 3 mg/kg/h for the remainder of the study). In each pony, full-thickness tissue specimens from the gastrointestinal tract--cecum, pelvic flexure, left and right ventral colon, and right dorsal colon--heart, left lung, liver, left adrenal gland, spleen, and right kidney were collected and histologically evaluated. Edema, mucosal necrosis, and neutrophil infiltration in colonic sections were graded from 0 (normal) to 3 (most severe changes). Sections of liver and lung from 3 ponies in each group, and colon from 1 pony in each group, also were examined by transmission electron microscopy to determine the presence of ultrastructural alterations. Ischemia and reperfusion induced marked changes in all sections of colon in all ponies: moderate to severe submucosal edema, moderate necrosis of the superficial epithelium and lamina propria, and necrosis of the mucosal crypt epithelium. Extravascular neutrophil accumulation was evident in all sections of colon and cecum, but not in other tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Development of the formalin-inactivated hepatitis A vaccine, VAQTA from the live attenuated virus strain CR326F.

The development of the formalin-inactivated hepatitis A vaccine, VAQTA, culminates nearly two decades of the basic science studies of VAQTA in hepatitis A virology at the MRL. The master seed virus for production of VAQTA is derived from the F'(P18) variant of the strain CR326F which has been studied in human clinical trials and shown to the highly attenuated. The antigen is highly purified to make possible the consistency and thoroughness of its inactivation by formalin. Phase I clinical studies of VAQTA were initiated in 1989 and have progressed since that time to the recent Phase III clinical trials which demonstrated efficacy of a single dose of the vaccine in preventing clinical hepatitis A disease in pediatric populations in Monroe, NY.

The effects of freeze-drying on the potency and stability of live varicella virus vaccine.

Investigations into methods for improving the potency and stability of live varicella-zoster virus (Oka strain) vaccines have included the use of different lyophilization procedures which yielded products with different moisture levels. Three procedures were used: an 8-hour controlled-vacuum (0.47 mBars) procedure, a 14-hour controlled-vacuum (0.14 mBars) procedure, and a 48-hour high-vacuum (less than 0.07 mBars) procedure. Samples were stored for 24 months at -24 degrees C, -15 degrees C (in a frost-free freezer), and 4 degrees C. Potency was determined by a plaque assay in MRC-5 cells; moisture content was measured by the Karl Fisher method. Moisture content was 6 to 8 percent for the product made using the 8-hour procedure, 2 to 7 percent for the 14-hour procedure, and 0.5 to 1.5 percent for the 48-hour procedure. In addition to higher moisture, the 8-hour procedure resulted in a higher initial potency, indicating a lower loss during lyophilization, and better stability than did the 14- and 48-hour procedures. Although the initial potency from the 14-hour procedure was not statistically different from that for the 48-hour procedure, the product made with the 14-hour procedure did have better stability characteristics than that made with the 48-hour procedure.

Modified cases of chickenpox after varicella vaccination: correlation of protection with antibody response.

Four thousand forty-two healthy children and adolescents, ages 12 months to 17 years, were vaccinated with a single dose of live attenuated varicella vaccine (VARIVAX; Merck Sharp and Dohme Research Laboratories) containing approximately 1000 to 1625 plaque-forming units/dose during clinical trials conducted from 1987 to 1989. Clinical follow-up of vaccinees revealed that 2.1 and 2.4% of vaccinees developed modified cases of varicella in the first and second years, respectively, after vaccination. Most of those who developed varicella postvaccination had an attenuated illness, characterized by fewer lesions and a lower incidence of fever (greater than or equal to 100 degrees F, oral) than after natural infection. The likelihood of developing varicella postvaccination decreased (P less than 0.0001) as the 6-week postvaccination glycoprotein-based enzyme-linked immunosorbent assay titer increased. In addition the number of lesions in these cases tended to decrease (P = 0.07 for Year 1 and P = 0.02 for Year 2) as the 6-week glycoprotein-based enzyme-linked immunosorbent assay titer increased. Thus the 6-week postvaccination glycoprotein-based enzyme-linked immunosorbent assay titer can be used as a surrogate marker for protection from natural disease.

Oka/Merck varicella vaccine in healthy children: final report of a 2-year efficacy study and 7-year follow-up studies.

A large double-blind, randomized, placebo-controlled trial of live attenuated Oka/Merck varicella vaccine was conducted among healthy children, 1-14 years of age. During the first varicella season, the efficacy of the vaccine among susceptible children was 100%1. During the second varicella season, 22 children were diagnosed with varicella; 21 cases in placebo recipients and one in a vaccine recipient. The overall efficacy of the vaccine through two varicella seasons was 98%. After the code for the study was broken, the original group of vaccine recipients continued to be followed for development of varicella. The estimated proportion of vaccine recipients who remained varicella-free at the end of 7 years was 95%. The 23 cases of varicella that occurred in vaccine recipients over the 7-year period were considerably milder than natural varicella. The average number of lesions was 53, 50% of the children had non-vesicular rashes, and 14% of the children had a temperature greater than or equal to 38.9 degrees C (102 degrees F), oral. The persistence of antibody in a subset of vaccine recipients followed for 6 years was 100%.

Further evaluation of a live hepatitis A vaccine in marmosets.

Live, attenuated F' hepatitis A vaccine virus was studied in vivo in Saguinus labiatus marmosets for possible reversion to virulence, for possible establishment of persistent infection and for its capacity as a parenterally administered vaccine to induce immunity to oral infection. Serial transmission of the virus in S. labiatus, using infectious stool extracts for the second and third passages, produced no evidence of reversion of the F' vaccine virus to virulence. Monitoring for live HAV in stools over a 135-day period post-inoculation of marmosets with the F' vaccine revealed no evidence of persistent infection. Vaccinated animals were also shown to be resistant to infection on challenge by the oral route as well as by the previously demonstrated parenteral route.

Varicella vaccine (VARIVAX) in healthy children and adolescents: results from clinical trials, 1987 to 1989.

A total of 3303 healthy children and adolescents, aged 12 months to 17 years, were vaccinated with one of five production lots of a live attenuated varicella vaccine (VARIVAX) containing 1000 to 1625 plaque-forming units per dose. The vaccine was generally well tolerated. Ninety-six percent (2381/2475) of vaccinees responded to vaccination by producing antibody as measured by a glycoprotein-based enzyme-linked immunosorbent assay; 99% (569/576) of those tested maintained antibody at 1 year following vaccination. The incidence of varicella following household exposure in vaccinees was approximately 12%; household contact historically results in 87% infection. Nearly all of the vaccinees who had varicella after vaccination had a clinically modified disease.

A simple antigen-reduction assay for the measurement of neutralizing antibodies to hepatitis A virus.

A simplified hepatitis A virus (HAV) antigen-reduction neutralization assay (HAVARNA) was developed to permit the measurement of biologically active antibodies in recipients of candidate HAV vaccines. Degrees of neutralization were measured from the reduction in the amount of HAV antigen synthesized by 7-10 days after infection of MRC-5 (fetal human diploid lung) cell cultures. Sera producing a greater than or equal to 50% reduction in viral infectivity were scored as neutralizing. The assay was applied to demonstrate serum HAV neutralizing activity in 10 of 10 and 9 of 10 recipients of 10(7) and 10(6) TCID50 doses, respectively, of the Merck CR326F (F' variant) live attenuated vaccine. The dilution end points of selected sera ranged from 1:10 to 1:640. The dilution end point of the World Health Organization reference globulin no. 1 was 1:530,000 (0.2 mlU/ml of HAV antibody). The HAVARNA provided a rapid, sensitive, and reproducible means to measure neutralizing antibodies to HAV.

Antibody assays suitable for assessing immune responses to live varicella vaccine.

An enzyme-linked immunosorbent assay for antibodies to varicella-zoster virus (VZV), using purified viral glycoproteins as antigen (gpELISA), was compared with other assays for measuring vaccine-induced antibody responses. The gpELISA was more sensitive than conventional assays, proved highly specific for VZV and agreed well with an assay for neutralizing antibody activity. It was successfully applied to large-scale testing of live varicella vaccine in humans.

Effects of heparin treatment on colonic torsion-associated hemodynamic and plasma eicosanoid changes in anesthetized ponies.

Large colon torsion frequently is a fatal condition in horses. The purpose of the study reported here was to determine systemic arterial pressure, plasma eicosanoid concentrations, colonic blood flow, vascular resistance, tissue pH, and morphologic features associated with large colon torsion and detorsion, and to evaluate the effects of sodium heparin (80 IU/kg of body weight, IV) treatment on these values. Values were determined in 20 anesthetized ponies that were randomly assigned into 4 equal groups: control; control/heparin; torsion; torsion/heparin. Torsions were created by a 720 degrees rotation of the cecum and colon around their long axes at the sternal and diaphragmatic flexures. After 1 hour of torsion, the torsion was corrected and the colon was allowed to reperfuse for 1 hour. Heparin was administered 30 minutes into the experiment. Parametric data were analyzed (P less than or equal to 0.05), using split-plot analysis of variance, with differences between means evaluated with a modified Bonferroni t test; histopathologic data were analyzed (P less than or equal to 0.05) with a Kruskal-Wallis one-way analysis of variance by ranks. Heparin prevented colonic detorsion-induced hypotension and increases in vascular resistance and thromboxane concentration, and it significantly increased colonic blood flow for 40 minutes during reperfusion. Heparin did not alter prostacyclin concentration or the histologic appearance of the large colon.

Enhancement of varicella-zoster virus plaquing efficiency with an agarose overlay medium.

The infectivity titers of varicella-zoster virus (VZV) are routinely estimated by plaque production in cell culture. In this report, we show that plaque counts for VZV (strain Oka/Merck), in MRC-5 cell cultures, are significantly enhanced (54% average enhancement) by the use of an agarose overlay medium, as compared to a fluid overlay medium. Evidence also is presented that less variability (P less than 0.05) in plaque counts occurs with the use of an agarose overlay medium.