Enhanced efficacy of recombinant Brucella abortus RB51 vaccines against B. melitensis infection in mice

Brucella abortus strain RB51 is an attenuated rough strain, currently being used as the official live vaccine for bovine brucellosis in the USA and several other countries. In strain RB51, the wboA gene, encoding a glycosyltransferase required for the O-side chain synthesis, is disrupted by an IS711 element. Recently, we have demonstrated that strain RB51WboA, RB51 complemented with a functional wboA gene, remains rough but expresses low quantities of O-side chain in the cytoplasm. Mice vaccinated with strain RB51WboA develop greatly enhanced resistance against challenge with B. abortus virulent strain 2308. We have also demonstrated that overexpression of Cu/Zn superoxide dismutase (SOD) in strain RB51 (RB51SOD) significantly increases its vaccine efficacy against strain 2308 challenge. In this study, we constructed a new recombinant strain, RB51SOD/WboA, that over expresses SOD with simultaneous expression of O-side chain in the cytoplasm. We tested the vaccine potential of strains RB51SOD, RB51WboA, RB51SOD/WboA against challenge with virulent Brucella melitensis 16M and B. abortus 2308 in mice. In comparison with strain RB51, strain RB51SOD induced better protection against strain 2308, but not strain 16M, challenge. Similar to strain RB51WboA, vaccination with strain RB51SOD/WboA resulted in complete protection of the mice from infection with strain 2308. When challenged with strain 16M, mice vaccinated with either strain RB51WboA or strain RB51SOD/WboA were significantly better protected than those vaccinated with strain RB51 or RB51SOD. These results suggest that strains RB51WboA and RB51SOD/WboA are good vaccine candidates for inducing enhanced protection against B. melitensis infection.     

Assessment of the effects of adjunctive gabapentin on postoperative pain after intervertebral disc surgery in dogs

ObjectiveTo assess the effect of adjunctive gabapentin (GBP) on pain after thoracolumbar intervertebral disc surgery in dogs.Study designProspective, randomized, controlled, clinical, ‘blinded’ trial.AnimalsSixty-three client owned dogs undergoing hemilaminectomyMethodsDogs were assigned to two treatment groups. The GBP group received gabapentin 10 mg kg^?1 orally every 12 hours starting before anaesthesia; the placebo (P) group received empty gelatin capsules. Background analgesia was initiated with intravenous levomethadone 0.6 mg kg^?1 (as the combination ‘L-Polamivet) at anaesthesia induction, followed by a fentanyl patch and levomethadone 0.2 mg kg^?1 subcutaneously every 8 hours for 24 hours. Pain was assessed by the short form of the Glasgow Composite Measure Pain Score (CMPS-SF) without the gait category, and by a Visual Analogue Scale (VAS). Serum GBP concentrations and cortisol concentrations were measured. Statistical analyses utilized chi square test, Kolmogorov-Smirnov test, two-way analysis of variances for repeated measurements, Wilcoxon test and Friedmann test as relevant. Correlations were tested by Spearman's and Pearson's correlation coefficient. p< 0.05 was considered significant.ResultsMedian CMPS-SF was lower in group GBP than in group P on days 0.5, 1, 4 and 5. However, CMPS-SF and VAS were not significantly different between groups. Both pain scores decreased significantly over time. Cortisol concentrations were not significantly different between groups. Minimum serum concentrations of GBP fell below the detection limit of 1 μg mL^?1 in 6 of 29 and 7 of 28 dogs at 24 and 72 hours, respectively.Conclusions and clinical relevance10 mg kg^?1 GBP orally twice a day did not result in a detectable reduction in pain behaviour compared to background opioid analgesia alone, although a trend to lower pain levels (p < 0.1) was present. Further studies are needed to determine if this is related to effective background analgesia or an ineffective dose of GBP.     

PULSED WAVE-DOPPLER ULTRASONOGRAPHIC EVALUATION OF THE COMMON CAROTID ARTERY IN THE RESTING HORSE: PHYSIOLOGIC DATA

A pulsed wave-Doppler ultrasonographic evaluation of common carotid arterial blood flow was carried out on 63 healthy Italian Saddlebred horses. Vessel diameter and tracing morphology were evaluated were evaluated and blood flow parameters (systolic, diastolic and mean velocity, acceleration and deceleration of the systolic wave, cartid pulse volume) were calculated and correlated with class variables (sex, age and body weight). On the basis of the presence of an incisure in the ascending branch of the systolic curve, subjects were divided in two groups: one with a bifid systolic curve and the other with a monophasic aspect. Correlations between: 1) diameter of the vessel and body weight and 2) carotid pulse volume and flow velocity (systolic, diastolic and mean) were found. A greater systolic pulse volume was found in male subjects, in subjects with greater body weight and in those which had a monophasic systolic wave.     

Procalcitonin gene expression after LPS stimulation in the porcine animal model

Procalcitonin (PCT), recognised as a marker of sepsis, was investigated in a porcine model of endotoxic shock. The results showed that continuous IV infusion (1-4 h) of LPS (40 μg/kg) in pigs was able to induce a generalised increase of PCT expression in lung, heart, kidney and liver. The increase in PCT was significant only in kidney and was accompanied by an increase in IL-6 gene expression. In vitro results demonstrated that peripheral blood mononuclear cells (PBMCs), as well as endothelial cells, were potentially capable of contributing to in vivo extrathyroidal PCT production. These findings support previous data from pigs concerning the occurrence of widespread activation of PCT extrathyroidal gene expression during endotoxic shock in pigs. Nevertheless, the levels of PCT detected were very low, suggesting the need for additional studies to validate the pig as a reliable animal model for investigating the role of PCT in sepsis.     

Diagnostic procedures and surgical treatment of craniodorsal coxofemoral luxation in calves

OBJECTIVE: To report use of ultrasonographic examination of the coxofemoral joint, a surgical technique for repair of craniodorsal coxofemoral dislocation, and outcome. STUDY DESIGN: Clinical case reports. ANIMALS: Calves (n=4) with coxofemoral luxation. METHODS: Craniodorsal coxofemoral luxation was diagnosed by physical examination, radiographic, and ultrasonographic findings. Open surgical reduction of the femoral head was performed using a modified caudal approach. RESULTS: Craniodorsal luxation of the femoral head and the presence of an intact femoral neck were confirmed by ultrasonography. All luxations were successfully reduced and reluxation did not occur. At follow-up, 1 heifer had calved and 1 was 5 months pregnant. One calf died of bronchopneumonia 6 days after surgery. One calf had severe coxofemoral degenerative joint disease diagnosed (ultrasonography, radiography, and arthrocentesis) 3 months after surgery and confirmed by necropsy. CONCLUSION: Ultrasonography proved to be a simple and effective non-invasive technique for diagnosis of coxofemoral luxation. Immediate surgical intervention in hip dislocation in calves is necessary to avoid unnecessary trauma to subchondral structures. In calves, open instead of closed surgical reduction appears preferable because it allows access to the acetabular cavity for removal of debris. CLINICAL RELEVANCE: Ultrasonography should be considered a supplementary but not an alternative to radiographic examination for diagnosis of coxofemoral luxation and for follow-up examinations after reduction.     

Expert opinion based modelling of the risk of human infection with H5N1 through the consumption of poultry meat in Germany

According to current scientific opinion the risk of human infection with H5N1 via preparation and consumption of poultry meat is negligible.This opinion has not yet been challenged by a formal risk assessment, due to the lack of empirical data. We have developed a scenario pathway model as a conceptual framework for a formal assessment of the H5N1 risk to humans through consumption of poultry meat and parameterise the model using information derived from expert opinions. The aim of this study was to investigate whether the notion of an overall negligible risk via the oral infection route is consistent with ad hoc data and expert opinions on the relevant parameters of the model.The model is mainly based on expert opinion. A stochastic Monte-Carlo simulation was conducted which took into consideration (amongst others) the exposure and infection of chicken (broiler and layer), turkeys, ducks and geese, the probabilities of detection prior to slaughter, virus survival and contamination during slaughter, as well as during the cutting and preparation of meat in commercial plants and in private households, respectively. The empirical consumption pattern for poultry meat in Germany was taken into account in the simulation.The results show that the risk for the individual consumer is practically zero whereas up to 23 cases per year in Germany might occur if the upper (more pessimistic) ranges of the expert opinions apply.The finding of a low but non-negligible risk to the population is discussed in relation to the epidemiological information available from recent outbreaks in South East Asia.     

Ochratoxin a content of urine samples of healthy humans in Hungary

The ochratoxin A (OTA) content of urine samples from 88 healthy humans living at five settlements in three counties of Hungary was determined by immunoaffinity column cleanup and high-performance liquid chromatography (HPLC). OTA was detected in 61% of the samples in an average concentration of 0.013 ng/ml (range: 0.006-0.065 ng/ml). OTA concentrations measured in urine samples from men and women were not significantly different. The OTA concentration of samples from Heves county was significantly (t-test; p < 0.003) higher than that of samples from Hajdú-Bihar and Somogy counties. The regional differences in OTA concentration of urine samples indicate regional differences in the OTA exposure of the human population. Further studies are necessary to determine the cause of the regional differences in the OTA intake. The studies allow us to conclude that the OTA intake of the majority of the Hungarian population is low (< 1 ng/kg of body weight per day) but a certain part of the rural population may take up higher levels of OTA.     

In vitro antiviral activity of dehydroepiandrosterone and its synthetic derivatives against vesicular stomatitis virus

In this work the antiviral activity of 20 dehydroepiandrosterone (DHEA) analogs with different substituents at positions C-3, C-15, C-16 and C-17 were evaluated against vesicular stomatitis virus (VSV) in Vero cell cultures. The selectivity indexes (SI) obtained with DHEA and epiandrosterone (EA) were 50 and 72.6, respectively. The work showed that the compounds 21-norpregna-5,17(20)-dien-3β,16α-diyl-diacetate, 17,17-ethylendioxyandrostan-5,15-dien-3β-ol and 3β-hydroxypregn-17(20)-en-16-one had higher SI values than ribavirin, which was used as a reference drug. The antiviral mode of action of DHEA was also investigated against VSV replication in Vero cells, and time of addition experiments showed that DHEA mainly affected a late event in the virus growth cycle. Analysis of RNA and protein synthesis indicated that DHEA adversely affected positive strand RNA synthesis and viral mature particle formation.