Tuberculosis in wild seals and characterisation of the seal bacillus

SUMMARY Tuberculosis was diagnosed in 3 otariid seals found dead on beaches at 3 locations on the south coast of Western Australian between May 1990 and March 1991. This confirms that tuberculosis is present in the 2 native seals (Neophoca cinerea and Arctocephalus forsteri) in Western Australian waters. Mycobacterium sp isolated from the lungs of 2 of the seals were studied to determine the similarity of the strains to each other, to the strains isolated during 1986 from Australian sea lions and New Zealand fur seals kept in captivity at a marine park near Perth, Western Australia, and to a strain isolated in 1988 from a seal trainer who worked with the infected captive seals for 3 years. After restriction endonuclease analysis (REA) with the endonucleases Bst Ell, Bcl I and Pvu II, one of the wild seal strains appeared to have identical DNA fragment patterns to the strains from the captive seals and the seal trainer. The other wild seal isolate had identical REA profiles using Bst EII and Bcl I, but a minor difference was detected using Pvu II. Differences in these isolates were more clearly seen in restriction fragment length polymorphisms after hybridisation with two DNA probes. The secretory protein MPB70, present in M bovis, was not detected in wild seal isolates using sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blotting techniques. Analysis of protein and DNA fragment profiles indicated that seal tuberculosis isolates form a unique cluster within the M tuberculosis complex.     

Endometrial Oxytocin Receptor Concentration and Activity in Prepubertal Ewe Lambs

We have measured endometrial oxytocin receptor concentrations during prepuberty in ewe lambs, and have investigated the effect of progesterone on the activity of these receptors. In the first study, oxytocin receptor concentrations were undetectable in 2-week prenatal lambs but had increased immediately following birth and were then maintained throughout prepubertal life. Despite the presence of oxytocin receptors animals showed no prostaglandin F(2alpha) (PGF(2alpha)) release in response to exogenous oxytocin challenge at either 3 or 5 months of age. In a second study in 4-month-old ewe lambs treatment with exogenous progesterone resulted in the appearance of PGF(2alpha) release in response to oxytocin after 10 days of treatment. Thus, during the prepubertal life, ewe lambs possess the prerequisites of a luteolytic mechanism in that they have a dormant population of oxytocin receptors in which progesterone can induce oxytocin-stimulated PGF(2alpha) release.     

Effect of metoclopramide on gastro-esophageal reflux in anesthetized dogs

Administration of morphine before anesthesia leads to gastro-esophageal reflux (GER) in over 50% of dogs during the subsequent anesthetic. This GER is clinically silent but can lead to aspiration pneumonitis, esophagitis and esophageal stricture. In this prospective clinical study we aimed to determine the effect of metoclopramide on gastro-esophageal reflux (GER) in dogs undergoing elective orthopedic surgery. Dogs were admitted to the study if they were healthy, and had no history of vomiting or dysphagia. Dogs were fasted for an average of 18.2 ± 4.3 (mean ± SD) hours prior to induction of anesthesia. Anesthesia in all dogs included acepromazine, morphine, thiopental and isoflurane in oxygen. By random allocation, half the dogs received metoclopramide (M) as an IV bolus (0.4 mg kg^–1) and then infusion (0.3 mg kg^–1hour^–1), the others received equivalent volumes of saline (S). To measure esophageal pH a sensor-tipped catheter was placed with the tip 5–7 cm cranial to the lower esophageal sphincter, and connected to a computer for continual data collection. The pH of any fluid running from the mouth or nose was measured. Gastro-esophageal reflux was defined as a decrease in esophageal pH below 4 or an increase above 7.5. Fisher's Exact test was used to test significance of differences in incidence between groups. Separate multivariable logistic regression models were created for each outcome to assess the effects of risk factors on outcome. There were seven cases of GER in 16 dogs receiving M and 8/14 in those receiving S. There were no significant differences between M and S treated dogs in age, weight, duration of anesthesia and fasting, thiopental dose or incidence of vomiting. The administration of metoclopramide at this dose did not significantly reduce the incidence of GER in these anesthetized dogs.     

Quantification of saponins in aerial and subterranean tissues of Medicago truncatula

Triterpene saponins from aerial and subterranean organs of Medicago truncatula cv. Jemalong A-17 were qualitatively profiled and quantified using reverse-phase HPLC with on-line photodiode array detection and electrospray-ionization mass spectrometry (HPLC/PDA/ESI/MS). Absolute quantifications were performed for 3-O-beta-D-glucopyranosyl-medicagenic acid and soyasaponin 1 (3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-galactopyranosyl(1-->2)-beta-D-glucuronopyranoside] soyasapogenol B), whereas relative quantifications were determined for 29 other saponins in root, stem, leaf, seedpod, and seed. Roots contained the greatest total amount of saponins followed by leaf and seed, respectively. The quantitative data also reveal the differential accumulation of triterpene saponins in the various organs of M. truncatula. Specifically, relatively higher quantities of medicagenic acid conjugates accumulated in leaf and seed, whereas relatively higher levels of soysapogenol conjugates were observed in root. The differential accumulation of specific triterpene saponins is suggestive of spatially differentiated biosynthesis and/or biological function.     

Metabolic engineering of plant cells for biotransformation of hesperedin into neohesperidin, a substrate for production of the low-calorie sweetener and flavor enhancer NHDC

Neohesperidin dihydrochalcone (NHDC) is a seminatural, safe, low-calorie sweetener, bitterness blocker, and flavor enhancer with unique properties and applications for the food, beverage, pharmaceutical, and animal feed industries. Current production is limited by the availability of the substrate neohesperidin, a flavonoid that accumulates to significant levels only in the inedible bitter citrus species. We propose a process to convert hesperidin, a tasteless flavonoid extracted from orange peels that are abundant byproducts of the vast orange juice industry, into neohesperidin using metabolic engineering and biotransformation via three steps: (i) extraction of hesperidin from orange peels, (ii) hydrolysis of sugar moieties, and (iii) biotransformation of hesperidin hydrolysis products into neohesperidin. We overcame the current technological bottleneck in biotransformation of hesperidin hydrolysis products into neohesperidin using metabolically engineered plant cell cultures expressing a recombinant flavanone-7-O-glucoside-2-O-rhamnosyltransferase. A small-scale production experiment established the feasibility of the proposed process.     

Efficient and sensitive method for quantitative analysis of alkaloids in hardinggrass (Phalaris aquatica L.)

An efficient high-performance thin-layer chromatography (HPTLC) method for the analysis of alkaloids in hardinggrass (Phalaris aquatica L.) was developed. The method employed HPTLC glass plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of ethyl acetate/chloroform/7 N NH4OH in methanol (8:2:1, v/v/v). Using unidimensional double-development, bands were well separated for 10 alkaloid standards as well as alkaloids observed in hardinggrass plant extracts. Identities of compounds observed using HPTLC were validated by high-performance liquid chromatography-mass spectrometry (HPLC-MS). Software was used to quantify individual alkaloids in plant samples based on HPTLC retention factors and intensities relative to standards of known concentration. Correlation coefficients of 0.99 were obtained between estimated and actual concentrations for four standards (methyltyramine, hordenine, gramine, and 5-methoxydimethyltryptamine), with linearity in the range of 120-3840 ng/spot. The HPTLC method is repeatable and specific for beta-carboline, tryptamine, gramine, and tyramine type alkaloids in mixed standard and plant extracts. Initial results indicate substantial variation in alkaloid composition among and within hardinggrass populations.     

Retrospective analysis of detomidine infusion for standing chemical restraint in 51 horses

Objective To assess the effectiveness of a detomidine infusion technique to provide standing chemical restraint in the horse. Design Retrospective study. Animals Fifty‐one adult horses aged 9.5 ± 6.9 years (range 1–23 years) and weighing 575 ± 290.3 kg. Methods Records of horses presented to our clinic over a 3‐year period in which a detomidine infusion was used to provide standing chemical restraint were reviewed. Information relating to the types of procedure performed, duration of infusion, drug dosages and adjunct drugs administered was retrieved. Results Detomidine was administered as an initial bolus loading dose (mean ± SD) of 7.5 ± 1.87 µg kg^?1. The initial infusion rate was 0.6 µg kg^?1 minute^?1, and this was halved every 15 minutes. The duration of the infusion ranged from 20 to 135 minutes. Twenty horses received additional detomidine or butorphanol during the procedure. All horses undergoing surgery received local anesthesia or epidural analgesia in addition to the detomidine infusion. A wide variety of procedures were performed in these horses. Conclusions Detomidine administered by infusion provides prolonged periods of chemical restraint in standing horses. Supplemental sedatives or analgesics may be needed in horses undergoing surgery. Clinical relevance An effective method that provides prolonged periods of chemical restraint in standing horses is described. The infusion alone did not provide sufficient analgesia for surgery and a significant proportion of animals required supplemental sedatives and analgesics.     

The use of lingual venous blood to determine the acid‐base and blood‐gas status of dogs under anesthesia

ObjectiveTo assess the suitability of lingual venous blood (LBG) as an alternative to arterial blood (ABG) samples in determining acid–base balance and blood–gas status in dogs anesthetized for elective procedures and with medetomidine and isoflurane administration under experimental conditions.Study designProspective, randomized clinical and experimental study.AnimalsClinical population of 18 ASA I/II dogs for elective surgery and five healthy Beagles (3 females and 2 males) for the experimental study.MethodsBlood sampling was simultaneously performed at dorsal pedal arterial and lingual venous sites, generating paired data. Two paired samples were collected from each dog in the clinical part and four from each dog in the experimental part (two during isoflurane anesthesia and two during isoflurane plus medetomidine). A modified Bland and Altman method was used to examine data from the clinical part and the experimental data were subjected to a paired sign's test following transformation where appropriate.ResultsThe pH of LBG overestimated ABG, with limits of agreement of (?0.01, 0.02). The partial pressure of carbon dioxide (PCO₂) of LBG overestimated ABG by 0.6 mmHg [0.1 kPa], with limits of agreement of (?3.5, 4.6) mmHg [?0.5, 0.6 kPa]. The partial pressure of oxygen (PO₂) of LBG underestimated ABG by 86.3 mmHg [?11.5 kPa], with limits of agreement of (?199.8, 27.3) mmHg [?26.6, 3.6 kPa]. During medetomidine administration values for PO₂ (p = 0.03) and lactate (p = 0.03) were lower for LBG when compared with ABG. The LBG value of PO₂ was lower (p = 0.03) during medetomidine and isoflurane administration versus isoflurane alone.Conclusions and clinical relevanceThe pH and PCO₂ of LBG samples provide clinically acceptable substitutes of ABG samples in the dog population studied. The wider limits of agreement for PO₂ render it less reliable as a substitute for ABG. The difference in PO₂ identified between LBG and ABG during medetomidine administration may not preclude the use of LBG as substitutes for ABG samples.     

Influence of Changes in Body Weight on Peak Vertical Force in Osteoarthritic Dogs: A Possible Bias in Study Outcome

Objective— Force platform gait analysis is a recognized clinical evaluation tool that captures and documents the in vivo pathomechanics of osteoarthritis (OA). In a clinical trial designed to evaluate the impact of 2 specific diets, an increase in body weight (BW) was observed in lame client-owned dogs. Covariance analysis was used to evaluate the interference of BW changes toward the evolution of peak vertical force (PVF) values. These secondary findings are reported in this study.
Study Design— Prospective study.
Animals— Lame dogs (n=26).
Methods— Dogs with radiographic evidence of OA and low PVF values were fed with 2 specific diets for 30 and 60 days. PVF and BW were recorded at baseline, day 30 (D30), and D90.
Results— Mean (±SD) PVF values (%BW) did not differ significantly over time (D0: 63.9±17.2; D30: 65.5±17.4; and D90: 66.5±20.1). In contrast, BW (kg) was significantly higher at D90 (41.3±7.9) when compared with D30 (39.9±8.4) and D0 (40.0±8.7). Upon covariance analyses, BW changes interfere significantly with PVF values already normalized in %BW ( P =.013). Values of PVF adjusted using BW as a covariate were then 63.4±17.1 (D0), 65.0±17.3 (D30), and 67.6±20.5 (D90), whereas D90 was significantly higher than D0.
Conclusion— These findings highlighted the interference of changes in BW toward locomotor function of OA dogs when using PVF values normalized in %BW. Exacerbation of lameness when a gain in BW occurred was also sustained, raising a possible bias in clinical study outcomes.
Clinical Relevance— A BW increase in dogs with OA could exacerbate a preexisting lameness and induce a bias in clinical trials.     

Effect of pre-anesthetic meperidine on gastro-esophageal reflux in anesthetized dogs

Meperidine has been shown to decrease lower esophageal sphincter tone in monkeys and people, to have little effect in cats, and to physically increase it in dogs. We hypothesized that administration of meperidine to dogs before anesthesia would decrease the probability of GER during the subsequent anesthetic. In this randomized, prospective clinical trial we aimed to determine the incidence of GER in dogs undergoing elective orthopedic surgery and receiving either meperidine or morphine prior to anesthesia. Dogs were admitted to the study, if they were healthy, with no history of vomiting or dysphagia. Dogs were fasted overnight. Dogs were received either morphine (0.66 mg kg^–1 IM) or meperidine (8.8 mg kg^–1 IM) with acepromazine. Anesthesia in all dogs included thiopental and isoflurane in oxygen. To measure esophageal pH a sensor-tipped catheter was placed with the tip 5–7 cm cranial to the lower esophageal sphincter, and connected to a computer for continual data collection. Dogs were observed for vomiting after pre-medication, and the pH of any fluid running from the mouth or nose during anesthesia was measured. Gastro-esophageal reflux was defined as a decrease in esophageal pH below 4 or an increase above 7.5 for greater than 15 seconds. One-way anova was used to test significance of differences between groups in parametric variables. Fisher's Exact test was used to test significance of differences in incidence between groups. In dogs receiving meperidine the incidence of vomiting was 0, and of GER was 31% (4/13), compared to 60% (6/10) and 60% (6/10), respectively in dogs receiving morphine. In this preliminary study, the administration of pre-anesthetic meperidine was associated with a 29% reduction in the absolute risk of GER compared to morphine.