Plasma endotoxin concentration in horses: a methods study

Background: Accurate determination of plasma endotoxin concentration is critical for ex vivo and in vitro cellular and molecular studies of endotoxemia in horses. However, reports are conflicting with respect to anticoagulant, handling, and sample preparation.

Objective:

Objective: The purpose of this study was to determine the effect of blood sample fraction and handling time on measurement of endotoxin concentration in horses.

Methods:

Methods: Whole blood, anticoagulated with 3.8% (0.12 M) sodium citrate (9:1), was collected from 5 healthy horses. Whole blood (WB), platelet-rich plasma (PRP), and platelet-poor plasma (PPP) were spiked with endotoxin (2 EU/mL). Endotoxin-spiked WB samples were centrifuged immediately to generate PRP for measurement. Endotoxin concentration was subsequently measured by Limulus amebocyte assay at 0, 15, 30, 45, and 60 minutes. Assays were performed in triplicate and results were analyzed using Student's t -test, with significance set at P < .05.

Results:

Results: Mean endotoxin concentrations in 2 EU/mL-spiked WB were significantly different from those in PPP at all time points tested. Recovery of endotoxin in PRP generated from WB was significantly diminished after just 15 minutes.

Conclusion:

Conclusion: PRP generated from WB is significantly more reliable than PPP in determining endotoxin concentration ex vivo. Measurement of endotoxin in PRP generated from WB was significantly diminished after 15 min, identifying a time frame within which to process blood samples for endotoxin analysis.     

Panniculitis in a horse with peripancreatitis and pancreatic fibrosis.

A 22-year-old pinto mixed breed mare was admitted for evaluation of severe colic signs and gastric reflux. Multiple nonpainful, variably sized hard masses were palpated in the subcutis over the thorax, abdomen, and hindquarters of the horse. The mare was diagnosed with sterile peritonitis and had systemically high gamma-glutamyltransferase, amylase, and lipase. Three days into treatment she became febrile with signs of persistent and mild abdominal discomfort; euthanasia was elected. Necropsy revealed peripancreatitis, pancreatic fibrosis, abdominal steatitis and panniculitis. Panniculitis associated with peripancreatitis has been described in humans and dogs but not in horses. Pathogenic models for this entity are discussed.     

Successful treatment of a squamous cell carcinoma in a white rhinoceros, Ceratotherium simum

A captive 33-year-old male white rhinoceros with seasonal dermatitis was diagnosed with a malignant squamous cell carcinoma on the right flank. Staphylococcus aureus was cultured from the skin lesions. No fungal or yeast was isolated. The dermatitis was treated with a combination of oral antibiotics (trimethoprim-sulphadiazine) and topically with weekly chlorhexidine washes and a mixture of a zinc oxide, balsam peru and bismuth oxide cream. Under azaperone and butorphanol anaesthesia, the skin tumour was surgically removed. The tumour was excised with wide margins and allowed to heal by secondary intention as primary wound closure was not possible. A post-mortem performed 2 years later for an unrelated condition revealed no metastases or recurrence of the skin tumour. It was presumed that chronic irritation or trauma may have contributed to the development of the skin tumour. This is the first detailed report of the successful treatment of a squamous cell carcinoma not associated with the horn in a rhinoceros.     

Comparison and phylogenetic analysis of the heat shock protein 70 gene of Babesia parasites from dogs

The heat shock protein 70 (hsp70) genes of Babesia gibsoni, B. canis canis, B. canis vogeli, and B. canis rossi isolated from infected dogs were cloned by polymerase chain reaction (PCR) and sequenced. In the nucleotide sequence and the predicted amino acid sequence of the gene, the parasites were very similar to each other. The nucleotide sequences of the hsp70 gene had more variety than those of 18S nuclear subunit ribosomal DNA (18S rDNA). A phylogenetic analysis of these sequences and comparisons with sequences from other Babesia and Theileria species revealed that all canine babesial isolates analyzed in the present study were closely related to each other and formed one cluster. Additionally, a phylogenetic analysis of Babesia and Theileria species showed that these parasites could be divided into three groups: group A including canine babesial isolates, B. divergens, B. odocoilei, B. bovis, B. caballi, and B. ovis; group B including Theileria annulata, T. orientalis, and T. cervi; and group C including B. microti and B. rodhaini. These results suggested that a phylogenetic analysis of the hsp70 gene sequence might be helpful in classifying Babesia and Theileria species, and that canine babesial isolates might be closely related to each other, indicating their evolution from the same ancestry.     

COLLAGENASE CHEMONUCLEOLYSIS—A LONG TERM RADIOGRAPHIC STUDY IN NORMAL DOGS

Five clinically normal five year old dogs were used in this study. From a randomised table inter ver tebral discs were each injected with either collagenase or calcium chloride diluent. The surgically exposed cervical discs were injected with 50 units whereas thoracic and lumbar discs were injected under fluoroscopic guidance with 100 units of the enzyme. Postinjection radiographs revealed significant (p ≤ .05) disc space narrowing in enzyme injected discs. The cervical discs had the highest frequency of radiographic narrowing (87%) followed by the thoracic (70%) and lumbar (53%) discs. Spondylosis deformans developed at the sites of cervical enzyme injections. None of the dogs had neurologic abnormalities one year postinjection.     

USE OF ULTRASOUND-GUIDED FINE-NEEDLE ASPIRATION BIOPSY AND AUTOMATED CORE BIOPSY FOR THE DIAGNOSIS OF GASTROINTESTINAL DISEASES IN SMALL ANIMALS

The purpose of this study was to evaluate the usefulness of ultrasound-guided fine-needle aspiration biopsy and core biopsy in the diagnosis of infiltrative gastrointestinal diseases. Six dogs and seven cats with clinical signs of gastrointestinal disease underwent ultrasonography and intestinal lesions were identified. One or more ultrasound-guided fine-needle aspiration biopsy and/or core biopsy procedures were performed in each patient. Each patient also underwent one of the following additional procedures for comparison of results: 1) surgery (n=4), 2) endoscopy (n=2), 3) post mortem exam (n=3), and, 4) for lymphoma diagnosed with ultrasound-guided procedures, response to chemotherapy (n=4). Correct diagnoses were obtained in nine of the 13 patients, incorrect diagnoses in two of the 13 patients, and inconclusive diagnoses in two of the thirteen patients. Of the 16 total ultrasound-guided procedures performed, ten were confirmed as correct, three as incorrect, and three were non-diagnostic. Intestinal lesions with bowel wall thickness greater than 2.0 cm had a higher percentage of correct diagnoses than lesions of lesser wall thickness. Gastric lesions had a higher percentage of correct diagnoses than small and large intestinal lesions. Malignant lesions had a higher percentage of correct diagnoses than benign lesions. There were no complications. Ultrasound-guided gastrointestinal fine-needle aspiration biopsy and core biopsy appears to be a safe, accurate, and rapid procedure for use in the diagnosis of infiltrative gastrointestinal disease.     

Effects of tension of the girth strap on respiratory system mechanics in horses at rest and during hyperpnea induced by administration of lobeline hydrochloride

OBJECTIVE: To determine whether tension of the girth strap of a saddle would sufficiently affect rib motion and reduce lung volume to alter pulmonary resistance in horses. ANIMALS: 10 healthy adult horses. PROCEDURE: We used classical techniques to measure the effects of tightening a girth strap (15 kg of tension) on pulmonary dynamics during eupnea and hyperpnea in horses. Respiratory impedance was evaluated by use of oscillometry, and resistance and reactance data were partitioned into lung and chest wall components. Rib cage and abdominal contributions to tidal volume and minute ventilation were measured by use of respiratory inductance plethysmography. Effects of strap tension on functional residual capacity (FRC) were measured during eupnea by use of a helium-dilution technique. In a subgroup of 6 horses, we also measured transdiaphragmatic pressures during eupnea and hyperpnea induced by administration of lobeline hydrochloride (0.2 mg/kg, i.v.). RESULTS: Pulmonary resistance measured by use of oscillometry but not by use of classical methods was significantly increased by the tension of the girth strap. However, the increase in pulmonary resistance could not be explained by a decrease in FRC. Motion of the rib cage was significantly reduced during eupnea and hyperpnea. However, ventilatory variables (tidal volume, minute ventilation, and peak flows), FRC, and transdiaphragmatic pressures were unaltered by strap tension. CONCLUSIONS AND CLINICAL RELEVANCE: Although tension of the girth strap caused measurable changes in respiratory mechanics (loss of rib motion and increased pulmonary resistance), there was no evidence that ventilation was limited.     

Ultrasound-guided posterior extraconal block in the dog: anatomical study in cadavers

Objective

To describe a novel ultrasound-guided posterior extraconal block in the dog.

The effects of femoral stem and neck length on cement strains in a canine total hip replacement model

OBJECTIVE: To determine the effects of femoral prosthesis stem length and head size on cement strains in a canine hip replacement system. STUDY DESIGN: An in vitro experimental model. SAMPLE POPULATION: (1) Three standard and 3 1-cm shortened femoral implants with +3 femoral heads. (2) Two standard implants with +0, +3, and +6 femoral heads. METHODS: Femoral stems were embedded in polymethylmethacrylate cement. A uniaxial proximodistal-oriented strain gauge was applied to the cement on the medial and lateral aspects of the construct 1.5 cm, 6.0 cm and 7.0 cm distal to the collar. Each construct with a +3 femoral head was mounted in a materials testing system. An axial compressive load (0-200 N) was applied to the femoral head and cement strains were recorded. Additionally, 2 standard length constructs were also tested with +0 and +6 femoral heads. The effects of stem length and neck length on cement strains were assessed with analysis of variance. RESULTS: Strains increased at all locations with increasing loads for all constructs. Shorter implants had higher strains by 152% and 171%, lateral (P =.003) and medial (P =.0025) to the stem tip. No significant strain differences were noted, at any strain gauge location, between different neck lengths (P values ranged from.20 to.67). CONCLUSIONS: Although a shorter implant stem has a potential to improve implant fit, it led to significantly higher cement strains that may increase the risk for aseptic loosening. Changes in femoral neck length did not significantly affect cement strains under the conditions tested. CLINICAL RELEVANCE: Shortening of the femoral stem currently cannot be recommended in canine total hip replacement. The existing use of variable neck lengths likely does not increase the risk of failure of the femoral stem.     

Flash electroretinography in standing horses using the DTL microfiber electrode

Purpose The goal of our study was the evaluation of a practical method for the recording of flash electroretinograms (ERGs) in sedated, standing horses with the DTL? microfiber electrode. Methods The horses were sedated intravenously with detomidine hydrochloride (0.015 mg/kg). The pupil was dilated and the auriculopalpebral nerve was blocked. The ERGs were recorded with the active electrode on the cornea (DTL?), the reference electrode near the lateral canthus, and the ground electrode over the occipital bone. The light intensities of the white strobe light were 0.03 cd·s/m² (scotopic) and 3 cd·s/m² (scotopic and photopic). Photopic and scotopic single flash and flicker responses to Ganzfeld stimulation were recorded. During the 20‐min dark adaptation period the retina was stimulated every 5 min with the 0.03 cd·s/m² single flash. Results The median b‐wave amplitudes and implicit times were 38 µV and 33 ms (photopic cone‐dominated response), 43 µV and 63 ms (5‐min dark adaptation), 72 µV and 89 ms (10 min), 147 µV and 103 ms (15 min), 188 µV and 109 ms (20 min, 0.03 cd·s/m², rod response), and 186 µV and 77 ms (20 min, 3 cd·s/m², maximal combined rod‐cone response). A steady increase in amplitude and implicit time was noted during dark adaptation. No oscillatory potentials could be isolated. Conclusions The use of detomidine hydrochloride sedation and the DTL? microfiber electrode allowed the recording of good quality ERGs. This protocol should permit the detection of functional problems in the retina without the risk involved with general anesthesia.