Animals Used in these Published Studies on CES Clark, Nancy, Mills, Daniel, Marchant, Jeremy. Evaluation of the potential efficacy of the Alpha-Stim SCS in the Horse. DeMontfort University Equestrian Centre and Field Station, Caythorpe, Lincolnshire, United Kingdom. 2002. (Publication in process) After completing a successful pilot study of the stress reduction effects of the Alpha- Stim SCS in a 6 horse pilot study, a double-blind study was completed with 8 thoroughbred horses (2 fillies, 3 mares, 3 geldings) at the De Montfort University Equestiran Centre and Field Station at Caythorpe, Lincolnshire, United Kingdom. Alpha-Stim AS-Trode brand self adhesive electrodes were attached to shaved areas on each side of the neck beneath the collar. Alpha-Stim SCS devices were set to provide 0.5 Hz biphasic stimulation at 200 microamperes, or sham stimulation. Heart rate measurements (HR) were provided by an attached Polar Horse Trainer transmitter belt and a Polar Vantage receiver. Observers blind to the treatment condition recorded duration of behaviors of body locomotion, head motion, ear position, oral behavior and the state of the lower lip at 15 second intervals for 15 seconds duration. Four trials were carried out each day for a total of 8 days (4 testing days per week). The frequency of each behavior was calculated and divided by the total number of observations for statistical analysis. The time standing alert decreased significantly from trial 1 to trial 4 (p<0.05). The time spent standing alert also continued to decrease after the stimulation had stopped, suggesting an ongoing effect of CES following the period of actual stimulation. The horses spent significantly more time standing dozing between trials 1 and 4 (p<0.05), less time with lower lip tense between trials 1 and 2 (p<0.05), had less ear flicking from trial 1 to 3 (p<0.05), increased head wobbling between phases 1 and 4 (p<0.05), less time vocalizing between phase 1 and 4 (p<0.05), and less time shaking the head between phases 1 and 3 (p<0.05). All of these changes were highly intercorrelated and strongly indicated a reduction in the horses’ state of arousal following treatment but not sham treatment with Alpha-Stim CES. Finally, although not significant, there was a trend in which the variation in mean HR values decreased from the first to the latter phases of the study. HR fell more in the three horses with the highest mean HR going into the study. The authors concluded that taken together these results are consistent with potentially beneficial effects using the Alpha-Stim SCS for horses. Effects were seen on the behaviors of greatest relevance to assessing anxious arousal in the given circumstances, namely time CES Studies of Animals 1 spent alert and dozing, and a number of other parameters consistent with relaxation. Specifically, there was no significant increase in any parameter associated with excitement nor is there any evidence that Alpha-Stim has any detrimental effects on the horse’s wellbeing. A number of parameters, which may also be indication of relaxation, were not significantly effected by the Alpha-Stim SCS but this could be explained by their rarity. The results further suggest that CES effects extend beyond the period of immediate stimulation. Other CES Device Reports Culp, L.P., Skarda, R.T. and Muir, W.W. Comparisons of the effects of acupuncture, electroacupuncture, and transcutaneous cranial electrical stimulation on the minimum alveolar concentration of isoflurane in dogs. Am J Vet Res. 66(8):1364-70, 2005. This study at the College of Veterinary Medicine, Ohio State University, Columbus, compared the effects of acupuncture (AP), electroacupuncture (EA), and CES with high- frequency intermittent currents on the minimum alveolar concentration (MAC) of isoflurane and associated cardiovascular variables in dogs. 8 healthy adult female Beagles were anesthetized with isoflurane on 4 occasions, allowing a minimum of 10 days between experiments. Isoflurane MAC values were determined for each dog without treatment (controls) and after treatment with AP and EA (AP points included LI 4, LU 7, GV 20, GV 14, San Tai and Baihui) and CES. Isoflurane MAC values were determined by use of noxious electrical buccal stimulation. Heart rate, mean arterial blood pressure (MAP), arterial blood oxygen saturation (SpO2) measured by use of pulse oximetry, esophageal body temperature, inspired and expired end-tidal isoflurane concentrations, end-tidal carbon dioxide concentration, and bispectral index (BIS) were monitored. Blood samples were collected for determination of plasma cortisol concentration. Mean +/- SD baseline MAC of isoflurane was 1.19 +/- 0.1%. Acupuncture did not significantly change MAC of isoflurane. Treatments with EA and CES significantly lowered the MAC of isoflurane by 10.1% and 13.4%, respectively. The SpO2, heart rate, MAP, BIS, esophageal body temperature, and plasma cortisol concentration were not significantly different after AP, EA, CES, and control treatments at any time interval. The authors concluded that the use of EA and CES decreased MAC of isoflurane in dogs without inducing adverse hemodynamic effects. However, the reduction in isoflurane MAC by EA and CES treatments was not considered clinically relevant. Robert, Claude, Limoge, Aymé and Stinus, Luis. Transcranial electrical stimulation (Limoge's currents) potentiates the inhibition of righting reflex induced by droperidol in rats. Brain Research. 822(10-2):132-141, 1999. (France) The effects of CES on droperidol-treated rats were evaluated using the righting reflex latency (RRL) test. CES was shown to potentiate the inhibition of righting reflex induced by droperidol. This potentiation was found to depend on the dose of the drug, the characteristics of the current delivered and the duration of stimulation. The author’s also observed that CES- induced potentiation of inhibition of righting reflex produced by droperidol injection was not reversed after naltrexone administration, or when measures were performed on p- chlorophenylalanine (pCPA)-treated animals. These results suggest that, under the experimental conditions: (i) CES does not interact with endogenous opioids to potentiate droperidol effects, (ii) the effect of CES on dopaminergic system prevails against CES action CES Studies of Animals 2 on serotonergic system. Though these findings enlarge the comprehension of CES effects on the central nervous system, further investigations are necessary to elucidate CES mechanisms. Little, Bert and Patterson, Margaret A. Embryofetal effects of neuroelectric therapy (NET). Electro and Magnetobiology. 15(1):1-8, 1996. After obtaining IRB approval by the University of Texas Southwestern Medical Center, virgin Spraque-Dawley female rats were mated with Spraque-Dawley male rats. All rats were fitted with 2 ear identification tags, including controls who did not receive any CES, to provide equal nociception and stress. All rats were permitted to move freely within their cages. The treated rats were divided into 3 groups and given CES 1 hour daily throughout their pregnancy at either 10, 100, or 1,000 Hz, 1 volt, 0.125 mA, at a 0.22 mS pulse width via alligator clips attached to the ear tags. On day 18 of pregnancy, the dams were killed by guillotine or with ether, and cesarean section was performed immediately. Fetuses were counted and the uteri were examined for evidence of embryo resorption. The fetuses were immediately placed in 10% fomalin. Xeroradiographic surveys of posteroanterior and lateral views of each fetus were taken and evaluated for cranial, rib, vertebral, or long bone abnormalities. After thorough external examination, autopsies were performed under a dissecting microscope to evaluate the palate, heart and major vessels, lungs, liver, kidneys, ureters, and bladder. Statistical analysis included the use of x2, analysis of variance, and Newman-Keuls multiple group comparisons test. 844 fetal rats were evaluated. The detailed external examinations under light microscopy revealed no obvious neural tube defects, limb reduction deformities, or anterior abdominal wall abnormalities in the controls or any of the treatment groups. Skeletal surveys of the fetal rats revealed no vertebral column, rib, or long bone deformities. Comparison between groups revealed more pregnancy resorptions and fewer offspring in all treatment groups as compared to the control group, with the difference reaching significance in the 1,000 Hz treatment group. Average fetal weights were inversely proportional to frequency and were significantly different among groups. Fetal brain weight followed a similar pattern of reduction, except that weights were not significantly different between the medium and highest frequency treatment groups. In their discussion, the authors stated that while the incidence of congenital anomalies was zero, the reason pregnancy resorptions were increased may be due to the CES treated rats being more complacent. Their behavior resembled the effects of CES in humans, even in this aggressive population well known for their violence. The treated rats were not as active as the controls, accordingly it is possible that food intake was lowered in the treatment group, a reasonable implication given the reduction in fetal weights. The authors concluded that CES may be embryolethal in the very early stages of pregnancy in the rat and might cause some miscarriages, but there is no evidence of fetotoxic effects. The relevance of these findings to humans is unknown. Mantz, J.,