The REFLECT Statement: Methods and Processes of Creating Reporting Guidelines for Randomized Controlled Trials for Livestock and Food Safety

The conduct of randomized controlled trials in livestock with production, health, and food-safety outcomes presents unique challenges that might not be adequately reported in trial reports. The objective of this project was to modify the CONSORT (Consolidated Standards of Reporting Trials) statement to reflect the unique aspects of reporting these livestock trials. A 2-day consensus meeting was held on November 18–19, 2008 in Chicago, IL, to achieve the objective. Before the meeting, a Web-based survey was conducted to identify issues for discussion. The 24 attendees were biostatisticians, epidemiologists, food-safety researchers, livestock production specialists, journal editors, assistant editors, and associate editors. Before the meeting, the attendees completed a Web-based survey indicating which CONSORT statement items would need to be modified to address unique issues for livestock trials. The consensus meeting resulted in the production of the REFLECT (Reporting Guidelines for Randomized Control Trials) statement for livestock and food safety and 22-item checklist. Fourteen items were modified from the CONSORT checklist, and an additional subitem was proposed to address challenge trials. The REFLECT statement proposes new terminology, more consistent with common usage in livestock production, to describe study subjects. Evidence was not always available to support modification to or inclusion of an item. The use of the REFLECT statement, which addresses issues unique to livestock trials, should improve the quality of reporting and design for trials reporting production, health, and food-safety outcomes.     

Effect of anthelmintic treatment on sexual maturation in prepubertal beef heifers

Heifers treated with ivermectin at weaning have been reported to reach puberty at a younger age and lighter weight than untreated heifers. We tested the hypothesis that heifers administered ivermectin would respond with earlier follicular development and a greater LH response to a 1-mg estradiol-17beta challenge (E2C) than untreated heifers. Fall-born Angus heifers (n = 32) were randomly assigned on 284 +/- 9 d of age (215.5 +/- 20.8 kg) to receive ivermectin (IVR) or albendazole (ALB), IVR + ALB, or to remain as untreated controls (CONT). Each group (n = 8) was housed separately in adjacent pens throughout the trial and managed to gain .8 kg/heifer on a ration containing 13.2% CP, 58.8% TDN, and 49.9% DM. The CONT heifers received an additional 2.27 kg/heifer of corn silage and 1.59 kg/heifer of corn daily to maintain ADG at comparable levels. Individual body weight was recorded weekly, and nematode eggs per gram (EPG) of feces were measured every 21 d. Ultrasonography was performed on alternate days starting 2 wk prior to E2C to characterize follicular wave patterns. Follicles were separated into classes (C1 [3 to 5 mm], C2 [6 to 9 mm], and C3 [10 mm]) and sizes (largest [LF], second [SLF], third [TLF], and fourth largest follicles [FLF]). The sizes of the regressing dominant follicle 1 (DF1) and the progressing dominant follicle 2 (DF2) were also determined. Serum concentrations of LH were determined from hourly jugular blood samples collected 8 to 24 h after injection of E2C. The IVR + ALB treatment group had more C3 follicles than ALB and CONT (P < .07). The IVR-treated heifers had larger TLF than ALB and CONT (P < .04). The IVR- and IVR + ALB-treated heifers had larger FLF and DF2 than ALB and CONT (P < .1). Least squares means for DF2 were 9.5 +/- .5, 8.0 +/- .4, 9.5 +/- .3 and 8.3 +/- .3 mm, for IVR, ALB, IVR + ALB and CONT, respectively (P = .02 for treatment effect). The E2C-induced serum LH concentration did not differ with respect to treatment. We conclude that heifers administered IVR display increased follicular development, supporting our earlier investigations regarding reduced age at puberty in heifers treated with IVR near weaning.     

Rabies surveillance in the United States during 1989

In 1989, 4,808 cases of rabies in animals other than human beings were reported to the Centers for Disease Control, 1.8% more (4,724 to 4,808) than in 1988. Eighty-eight percent (4,224/4,808) of those affected were wild animals and 12% (584/4,808) were domestic animals. Cases were reported from 49 states, the District of Columbia, and Puerto Rico; Hawaii remained rabies-free. Skunks (1,657) continued to be the most commonly reported rabid wild animal. For the second consecutive year, more cats (212) were reported to be infected with rabies virus than any other domestic species. Compared with their 1988 reports, 5 states reported increases of greater than 100% (Alaska, 109%; New Jersey, 233%; Ohio, 133%; Oklahoma, 168%; and Washington, 125%), and 5 states reported decreases of greater than 50% (Connecticut, 63%; Mississippi, 56%; Montana, 67%; Nevada, 55%; and West Virginia, 53%) in 1989.     

Protection studies with a globin-enriched protein fraction of Ostertagia ostertagi

The protective capacity of an adult stage Ostertagia ostertagi globin antigen was tested in four vaccination experiments in cattle. In a preliminary experiment, calves were vaccinated three times intraperitoneally with 250 microg globin in Freund's adjuvant and challenged with a trickled infection of 25,000 infective larvae. In three subsequent field studies, calves were vaccinated twice or three times intramuscularly with 80-100 microg globin in Quil A and challenged with a natural gastrointestinal nematode infection on pasture. Higher globin-specific antibody levels were detected in the vaccinated calves than in the control animals in all vaccine trials. In the preliminary experiment, geometric mean cumulative egg counts in the globin group were reduced by 52% and total worm burdens were reduced by 28%, compared to the controls. In the first field trial cumulative faecal egg counts were reduced by 63% in the vaccinated calves. However, the reduction in faecal egg output in these two experiments was not statistically significant and no reduction in faecal egg counts was observed in the vaccinated animals in the two last field trials. In conclusion, vaccination of calves with O. ostertagi globin resulted in highly variable protection levels after challenge infection.     

Disseminated hemangiosarcoma in a cow

An antemortem diagnosis of disseminated hemangiosarcoma with extensive hepatic and splenic involvement was made in a 9-year-old cow evaluated for lethargy, weight loss, regenerative anemia, thrombocytopenia, and elevated liver enzymes. This is the first report of hemangiosarcoma in a cow with a suspected primary location of the liver and/or spleen.     

Bilateral Testicular Haemorrhage in a Bleu du Maine Ram: Clinical, Ultrasonographic and Histologic Features

Bilateral testicular haemorrhage was detected in a 5-year-old Bleu du Maine ram in an abattoir survey of male reproductive tracts in sheep; the ultrasonographic and histologic features of the lesions are described.     

Response of C2C12 mouse and turkey skeletal muscle cells to the beta-adrenergic agonist ractopamine

The effects of ractopamine (RAC) and ractopamine stereoisomers (RR, RS, SR, and SS) on cyclic AMP (cAMP) production, total protein, and DNA concentrations in mouse skeletal muscle cells (C2C12) were evaluated. The RAC (10 microM) caused an approximately 30% increase in cell number, protein, and DNA concentrations in myoblasts after 48 h; no differences were found in myotubes. The RAC-stimulated increase of these variables in myoblasts was blocked by the presence of equimolar concentrations of propranolol. At a later passage, myoblasts failed to exhibit an increase in cell number, protein, or DNA upon exposure to RAC. Both myoblasts and myotubes increased cAMP production in response to 10 microM RAC. The RAC isomers ranked RR > SR > RS approximately SS in ability to stimulate cAMP production, with essentially no response to SS. The SR produced about 50% of the RR response. Coincubation of propranolol (10 microM) and RAC (10 microM) prevented RAC-stimulated cAMP production in myotubes but not in myoblasts (approximately 35% of cAMP produced by RAC alone). Turkey satellite cells (derived from biceps femoris of 12-wk-old toms) produced essentially no increased cAMP when exposed to 10 microM RAC stereoisomers. Stability of RAC was evaluated under laboratory storage and culture conditions. The RAC was stable for more than 4 mo when stored in deuterated DMSO (>98% purity) at room temperature or in aqueous solutions at -80 degrees C, as determined from sequential nuclear magnetic resonance studies. Radiolabeled RAC was incubated for 72 h in the presence of serum-containing medium, with or without C2C12 cells. Ninety-eight percent of the parent compound found in the medium at time zero was present in the medium as parent at the end of 72 h. The cellular cAMP response to RAC through beta-adrenergic receptors seems to be stereospecific. If the state of myoblasts and myotubes in vitro reflects the in vivo state, then the ractopamine effect in vivo on cellular processes (including cell division and protein and DNA accumulation) may be independent of beta-adrenergic receptors in muscle.