Evaluation of a three-axial acceleromyography monitor in dogs compared with mechanomyography.

OBJECTIVE: To compare the ratio of the train-of-four (TOF) and double burst stimulation (DBS) obtained with three-axial acceleromyography (AMG) and mechanomyography (MMG) in dogs during recovery from a rocuronium-induced neuromuscular block. STUDY DESIGN: Prospective, randomized, experimental study. ANIMALS: A total of six intact healthy adult male Beagle dogs, weighing 9.1 ± 1.9 kg and aged 3-5 years. METHODS: Dogs were anesthetized with intravenous (IV) dexmedetomidine and propofol, and isoflurane in oxygen. Neuromuscular function was measured with AMG and MMG in the contralateral thoracic limbs. Rocuronium (0.5 mg kg-1) was administered IV, and the TOF and DBS ratios measured. During neuromuscular block offset, MMG values were recorded when AMG first reached ratios of 0.9 and 1.0. True recovery from neuromuscular block was determined as MMG ratio ≥ 0.9. The false-positive (AMG ≥ 0.9 or 1.0, and MMG ratio < 0.9) rate was determined. Paired values were compared, and bias and limits of agreement were calculated. Receiver operating characteristic (ROC) curves were created. RESULTS: When AMG first reached 0.9 and 1.0 during recovery, MMG values were lower (p < 0.040). When AMG reached 0.9, the false-positive rate was 29% with TOF and 27% with DBS. It decreased to 12% (TOF) and 11% (DBS) when a ratio of 1.0 was used. AMG values were higher than paired MMG values (p < 0.001). The AMG overestimated MMG by 24% and 22% for TOF and DBS, respectively. Areas under the ROC curves (95% confidence interval) were 0.91 (0.89, 0.94) and 0.86 (0.81, 0.94) for TOF and DBS, respectively. CONCLUSIONS: and clinical relevance The three-axial AMG monitor overestimated neuromuscular function and, in some cases, indicated adequate recovery despite the MMG ratio being < 0.9. A TOF or DBS ratio of at least 1.0 should be considered when monitoring recovery of neuromuscular block with this AMG device.

Postoperative oxygenation in healthy dogs following mechanical ventilation with fractions of inspired oxygen of 0.4 or >0.9.

OBJECTIVE: To evaluate arterial oxygenation during the first 4 postoperative hours in dogs administered different fractions of inspired oxygen (FiO2) during general anesthesia with mechanical ventilation. STUDY DESIGN: Prospective, randomized clinical trial. ANIMALS: A total of 20 healthy female dogs, weighing >15 kg and body condition scores 3-7/9, admitted for ovariohysterectomy. METHODS: Dogs were randomized to breathe an FiO2 >0.9 or 0.4 during isoflurane anesthesia with intermittent positive pressure ventilation. The intraoperative PaO2:FiO2 ratio was recorded during closure of the linea alba. Arterial blood was obtained 5, 60 and 240 minutes after extubation for measurement of PaO2 and PaCO2 (FiO2 = 0.21). Demographic characteristics, duration of anesthesia, PaO2:FiO2 ratio and anesthetic agents were compared between groups with Wilcoxon tests. The postoperative PaO2, PaCO2, rectal temperature, a visual sedation score and events of hypoxemia (PaO2 < 80 mmHg) were compared between groups with mixed-effects models or generalized linear mixed models. RESULTS: Groups were indistinguishable by demographic characteristics, duration of anesthesia, anesthetic agents administered and intraoperative PaO2:FiO2 ratio (all p > 0.08). Postoperative PaO2, PaCO2, rectal temperature or sedation score were not different between groups (all p > 0.07). During the first 4 postoperative hours, hypoxemia occurred in three and seven dogs that breathed FiO2 >0.9 or 0.4 during anesthesia, respectively (p = 0.04). CONCLUSIONS AND CLINICAL RELEVANCE: The results identified no advantage to decreasing FiO2 to 0.4 during anesthesia with mechanical ventilation with respect to postoperative oxygenation. Moreover, the incidence of hypoxemia in the first 4 hours after anesthesia was higher in these dogs than in dogs breathing FiO2 >0.9.

Can we see fade? A survey of anesthesia providers and our ability to detect partial neuromuscular block in dogs.

OBJECTIVE: To assess the ability to visually detect fade during train-of-four (TOF) or double burst stimulation (DBS) in anesthetized dogs recovering from nondepolarizing neuromuscular block. STUDY DESIGN: Online anonymous survey. POPULATION: Data from 112 participants. METHODS: A web-based survey containing 12 videos of the response to ulnar nerve stimulation with TOF and 12 with DBS obtained at different levels of recovery from rocuronium-induced block was distributed to participants of the American College of Veterinary Anesthesia and Analgesia and the Academy of Veterinary Technicians in Anesthesia and Analgesia e-mail lists. Participants were asked to provide their highest training degree in anesthesiology, watch each video no more than twice, and determine whether fade was present. The probability to correctly recognize fade was calculated using binomial general linear models. General linear models and Tukey's tests were used to assess the effects of level of neuromuscular block, pattern of stimulation, and observers' training on the probability to detect fade. RESULTS: The survey was completed by 53 diplomates, 29 licensed veterinary technicians, 24 residents and six doctors of veterinary medicine (DVMs). The probability to detect fade decreased as partial neuromuscular block became more shallow (p < 0.0001). A TOF or DBS ratio of 0.7 had a 50% chance of being detected. DBS was superior to TOF for detecting fade when the ratio was 0.3-0.69. TOF was superior to DBS when the ratio was 0.7-0.9 (p < 0.0001). There were no differences among groups of observers when assessing fade with TOF or DBS. CONCLUSIONS AND CLINICAL RELEVANCE: Detection of fade from observations of the response to TOF in dogs is unreliable. Advance training in anesthesiology or the use of DBS confers little to no advantage for this subjective test.

Positive and negative staircase effect during single twitch and train-of-four stimulation: a laboratory study in dogs.

A positive staircase effect is well documented during neuromuscular monitoring. However, the increase in twitch amplitude may not remain stable over time. We compared the staircase phenomenon and twitch stability during single twitch (ST) or train-of-four (TOF) stimulation in anesthetized dogs. Force of contraction was measured in ten dogs. Each thoracic limb was stimulated with ST 0.1 Hz or TOF q 12 s for 25 min (random order). No neuromuscular blockers were administered. Every 5 min, ST and T1 amplitudes were compared within and between groups. Stability of twitch amplitude (<5 % change in 5 min) was also evaluated. ST and T1 amplitude increased over time without significant differences between groups. After 10 min of ST stimulation, the average ST amplitude had increased significantly to 107 %, and remained unchanged thereafter. T1 amplitude was significantly greater than baseline only at 5 (111 %) and 10 min (109 %); a decline towards baseline occurred thereafter. Stability was reached after 15 min for all dogs in the ST group, however, three dogs continued to have changes >5 % with TOF. An initial increase in ST amplitude remained stable over the observation period, but the increase in T1 amplitude during TOF was frequently followed by a decay. A stable twitch amplitude (variation <5 % in 5 min) was observed in all dogs with ST after 15 min of stimulation, which was not the case during TOF stimulation. Therefore, it appears at least in dogs, that ST might offer some advantages over T1 for measuring twitch amplitude.

Effects of neostigmine or edrophonium on force of contraction when administered at a train-of-four ratio of 0.9 in anesthetized dogs.

OBJECTIVE: Anticholinesterase drugs may produce paradoxical neuromuscular block when administered at shallow levels of neuromuscular block. The objective of this study was to evaluate the effects of neostigmine and edrophonium when administered at near-complete reversal from nondepolarizing block in anesthetized dogs. STUDY DESIGN: Incomplete crossover, randomized, blinded experimental study. ANIMALS: A total of 12 Beagle dogs. METHODS: Each dog was anesthetized twice with propofol and maintained with isoflurane and dexmedetomidine. Intravenous (IV) vecuronium (0.1 mg kg-1) was administered. When the mechanographic train-of-four (TOF) ratio had spontaneously recovered to ≥0.9, either neostigmine (0.04 mg kg-1) or edrophonium (0.5 mg kg-1) was administered IV, preceeded by atropine. Changes in twitch height or TOF ratio were measured for the next 10 minutes. Recurarization was considered to be present if values decreased by ≥10%. RESULTS: Data from four dogs in each treatment were excluded from analysis, resulting in data from five dogs administered both treatments, three dogs administered neostigmine and three dogs administered edrophonium. There was no difference between groups for age, weight, T1 and T4 twitch heights or TOF ratio values, before or after anticholinesterase administration. The TOF ratio decreased by 17% and 18% in two of the eight dogs administered neostigmine, resulting from a larger increase in T1 relative to T4. No reductions in individual twitch amplitudes were recorded in those dogs. When edrophonium was used, no cases of recurarization were observed. CONCLUSIONS AND CLINICAL RELEVANCE: The results support use of edrophonium for reversal of shallow neuromuscular block. The decreases in TOF ratio recorded after neostigmine does not necessarily indicate muscular weakness. Although the clinical implications are uncertain, the results suggest that, at these doses, edrophonium may be preferable to neostigmine for reversal of shallow neuromuscular block in dogs.

Recovery from rocuronium-induced neuromuscular block was longer in the larynx than in the pelvic limb of anesthetized dogs.

OBJECTIVE: To determine if neuromuscular monitoring at the pelvic limb accurately reflects neuromuscular function in the larynx after administration of rocuronium in anesthetized dogs. STUDY DESIGN: Prospective experimental study. ANIMALS: Six healthy Beagle dogs. METHODS: Anesthesia was maintained in dogs with isoflurane and a continuous infusion of dexmedetomidine. Rocuronium (0.6 mg kg-1) was administered intravenously to induce neuromuscular block. Train-of-four (TOF) impulses were applied to the left recurrent laryngeal nerve (RLn) and the peroneal nerve (Pn). The evoked TOF ratio (TOFR; T4:T1) was measured with electromyography (EMG) simultaneously at the larynx and at the pelvic limb. Spontaneous recoveries of T1 to 25% (T125%) and 75% (T175%) of twitch height, and to TOFR of 0.70 and 0.90 (TOFR0.90) at each EMG site were compared. RESULTS: Data from five dogs were analyzed. Times to T125% were similar at the pelvic limb and larynx when measured by EMG; time to T175% was slower at the larynx by 6±4 minutes (p=0.012). The larynx had a slower recovery to TOFR0.70 (41±13 minutes) and TOFR0.90 (45±13 minutes) than did the pelvic limb [29±8 minutes (p=0.011) and 33±9 minutes (p=0.003), respectively]. When the pelvic limb EMG returned to TOFR0.70 and TOFR0.90, the larynx EMG TOFR0.70 and TOFR0.90 values were 0.32±0.12 (p=0.001) and 0.38±0.13 (p=0.001), respectively. CONCLUSIONS AND CLINICAL RELEVANCE: After administration of rocuronium, neuromuscular function assessed by EMG recovered approximately 36% slower at the larynx than at the pelvic limb. The results in these dogs suggest that quantitative neuromuscular monitoring instrumented at a pelvic limb may be unable to exclude residual block at the larynx in anesthetized dogs.

Partial neuromuscular block impairs arytenoid abduction during hypercarbic challenge in anesthetized dogs.

OBJECTIVE: To evaluate the effect of two levels of partial neuromuscular block (NMB) on arytenoid abduction, tidal volume (VT) and peak inspiratory flow (PIF) in response to a hypercarbic challenge in anesthetized dogs. STUDY DESIGN: Prospective laboratory study. ANIMALS: Eleven healthy male Beagle dogs aged 3-5 years. METHODS: Dogs were anesthetized with propofol and dexmedetomidine infusions. The rima glottidis was observed via an endoscope placed through a laryngeal mask airway. Atracurium infusion was titrated to obtain two levels of partial NMB. The normalized glottal gap area (NGGA; glottal gap area normalized to height squared of rima glottidis) at peak inspiration during a hypercarbic challenge (10% CO2 inspired for 1 minute) was measured at baseline, during mild [train-of-four (TOF) ratio 0.4-0.6] and shallow (TOF ratio 0.7-0.9) NMB, and 30 minutes after spontaneous recovery from NMB. The VT and PIF were measured at the same time points and compared using anova for repeated measures and Tukey's post hoc tests. RESULTS: The NGGA and VT were significantly lower than baseline during both levels of partial NMB with no difference between mild and shallow NMB (p < 0.05). They returned to baseline values after spontaneous recovery from NMB. PIF was not altered significantly during partial NMB. CONCLUSIONS AND CLINICAL RELEVANCE: The NGGA and VT at peak inspiration in response to a hypercarbic challenge were reduced during partial NMB block, with decreased abduction of the arytenoid cartilages. This dysfunction was present even at shallow levels of NMB.

Host genetic influence on papillomavirus-induced tumors in the horse.

The common equine skin tumors known as sarcoids have been causally associated with infection by bovine papillomavirus (BPV). Additionally, there is evidence for host genetic susceptibility to sarcoids. We investigated the genetic basis of susceptibility to sarcoid tumors on a cohort of 82 affected horses and 270 controls genotyped on a genome-wide platform and two custom panels. A Genome Wide Association Study (GWAS) identified candidate regions on six chromosomes. Bayesian probability analysis of the same dataset verified only the regions on equine chromosomes (ECA) 20 and 22. Fine mapping using custom-produced SNP arrays for ECA20 and ECA22 regions identified two marker loci with high levels of significance: SNP BIEC2-530826 (map position 32,787,619) on ECA20 in an intron of the DQA1 gene in the Major Histocompatibility Complex (MHC) class II region (p = 4.6e-06), and SNP BIEC2-589604 (map position 25,951,536) on ECA22 in a 200 kb region containing four candidate genes: PROCR, EDEM2, EIF6 and MMP24 (p = 2.14e-06). The marker loci yielded odds ratios of 5.05 and 4.02 for ECA20 and ECA22, respectively. Associations between genetic MHC class II variants and papillomavirus-induced tumors have been reported for human papillomavirus and cottontail rabbit papillomavirus infections. This suggests a common mechanism for susceptibility to tumor progression that may involve subversion of the host immune response. This study also identified a genomic region other than MHC that influenced papillomavirus-induced tumor development in the studied population.

Bone marrow transcriptome and epigenome profiles of equine common variable immunodeficiency patients unveil block of B lymphocyte differentiation.

Common variable immunodeficiency (CVID) is a late-onset humoral deficiency characterized by B lymphocyte dysfunction or loss, decreased immunoglobulin production, and recurrent bacterial infections. CVID is the most frequent human primary immunodeficiency but still presents challenges in the understanding of its etiology and treatment. CVID in equine patients manifests with a natural impairment of B lymphocyte differentiation, and is a unique model to identify genetic and epigenetic mechanisms of disease. Bone marrow transcriptome analyses revealed decreased expression of genes indicative of the pro-B cell differentiation stage, importantly PAX5 (p≤0.023). We hypothesized that aberrant epigenetic regulation caused PAX5 gene silencing, resulting in the late-onset and non-familial manifestation of CVID. A significant increase in PAX5 enhancer region methylation was identified in equine CVID patients by genome-wide reduced-representation bisulfite sequencing and bisulfite PCR sequencing (p=0.000). Thus, we demonstrate that integrating transcriptomics and epigenetics in CVID enlightens potential mechanisms of dysfunctional B lymphopoiesis or function.

The same ELA class II risk factors confer equine insect bite hypersensitivity in two distinct populations.

Insect bite hypersensitivity (IBH) is a chronic allergic dermatitis common in horses. Affected horses mainly react against antigens present in the saliva from the biting midges, Culicoides ssp, and occasionally black flies, Simulium ssp. Because of this insect dependency, the disease is clearly seasonal and prevalence varies between geographical locations. For two distinct horse breeds, we genotyped four microsatellite markers positioned within the MHC class II region and sequenced the highly polymorphic exons two from DRA and DRB3, respectively. Initially, 94 IBH-affected and 93 unaffected Swedish born Icelandic horses were tested for genetic association. These horses had previously been genotyped on the Illumina Equine SNP50 BeadChip, which made it possible to ensure that our study did not suffer from the effects of stratification. The second population consisted of 106 unaffected and 80 IBH-affected Exmoor ponies. We show that variants in the MHC class II region are associated with disease susceptibility (p (raw) = 2.34 × 10(-5)), with the same allele (COR112:274) associated in two separate populations. In addition, we combined microsatellite and sequencing data in order to investigate the pattern of homozygosity and show that homozygosity across the entire MHC class II region is associated with a higher risk of developing IBH (p = 0.0013). To our knowledge this is the first time in any atopic dermatitis suffering species, including man, where the same risk allele has been identified in two distinct populations.

Effects of two fractions of inspired oxygen during anesthesia on early postanesthesia oxygenation in healthy dogs.

OBJECTIVE To evaluate the effects of 2 fractions of inspired oxygen (Fio2s) during anesthesia on postanesthesia Pao2 and other measures of oxygen exchange. ANIMALS 22 healthy adult sexually intact female dogs undergoing ovariohysterectomy by ventral midline celiotomy. PROCEDURES Dogs were randomly assigned to receive either oxygen (Fio2 > 0.9 [100% oxygen]; n = 11; control group) or a mixture of nitrogen and oxygen (Fio2 = 0.4; 11; 40% oxygen group) as the carrier gas for isoflurane while anesthetized. All dogs were allowed to breathe spontaneously while anesthetized. For each dog, the Pao2, Paco2, other indices of oxygenation, and extent of sedation were monitored at predetermined times during and for 1 hour after anesthesia. Measured variables were compared between the 2 treatment groups and over time within each treatment group. RESULTS None of the measured variables differed significantly between the control and 40% oxygen groups at any time during the postanesthesia period. Within each treatment group, the Paco2 and extent of sedation decreased over time during the postanesthesia period. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that indices of oxygenation did not differ significantly between healthy dogs in which the Fio2 was maintained at > 0.9 and those in which the Fio2 was maintained at 0.4 while anesthetized for ovariohysterectomy. Thus, the addition of nitrogen to the carrier gas for an inhalant anesthetic conferred neither an advantage nor disadvantage in regard to oxygenation during the first hour of anesthesia recovery.

Diversity of immunoglobulin lambda light chain gene usage over developmental stages in the horse.

To further studies of neonatal immune responses to pathogens and vaccination, we investigated the dynamics of B lymphocyte development and immunoglobulin (Ig) gene diversity. Previously we demonstrated that equine fetal Ig VDJ sequences exhibit combinatorial and junctional diversity levels comparable to those of adult Ig VDJ sequences. Herein, RACE clones from fetal, neonatal, foal, and adult lymphoid tissue were assessed for Ig lambda light chain combinatorial, junctional, and sequence diversity. Remarkably, more lambda variable genes (IGLV) were used during fetal life than later stages and IGLV gene usage differed significantly with time, in contrast to the Ig heavy chain. Junctional diversity measured by CDR3L length was constant over time. Comparison of Ig lambda transcripts to germline revealed significant increases in nucleotide diversity over time, even during fetal life. These results suggest that the Ig lambda light chain provides an additional dimension of diversity to the equine Ig repertoire.

Developmental progression of equine immunoglobulin heavy chain variable region diversity.

Humoral immunity is a critical component of the immune system that is established during fetal life and expands upon exposure to pathogens. The extensive humoral immune response repertoire is generated in large part via immunoglobulin (Ig) heavy chain variable region diversity. The horse is a useful model to study the development of humoral diversity because the placenta does not transfer maternal antibodies; therefore, Igs detected in the fetus and pre-suckle neonate were generated in utero. The goal of this study was to compare the equine fetal Ig VDJ repertoire to that of neonatal, foal, and adult horse stages of life. We found similar profiles of IGHV, IGHD, and IGHJ gene usage throughout life, including predominant usage of IGHV2S3, IGHD18S1, and IGHJ1S5. CDR3H lengths were also comparable throughout life. Unexpectedly, Ig sequence diversity significantly increased between the fetal and neonatal age, and, as expected, between the foal and adult age.