First detection of Edwardsiella ictaluri (Proteobacteria: Enterobacteriaceae) in wild Australian catfish.

The bacterium Edwardsiella ictaluri is considered to be one of the most significant pathogens of farmed catfish in the United States of America and has also caused mortalities in farmed and wild fishes in many other parts of the world. E. ictaluri is not believed to be present in wild fish populations in Australia, although it has previously been detected in imported ornamental fishes held in quarantine facilities. In an attempt to confirm freedom from the bacterium in Australian native fishes, we undertook a risk-based survey of wild catfishes from 15 sites across northern Australia. E. ictaluri was detected by selective culturing, followed by DNA testing, in Wet Tropics tandan (Tandanus tropicanus) from the Tully River, at a prevalence of 0.40 (95% CI 0.21-0.61). The bacterium was not found in fishes sampled from any of the other 14 sites. This is the first report of E. ictaluri in wild fishes in Australia.

Isolation and screening of probiotic candidates from marron, Cherax cainii (Austin, 2002) gastrointestinal tract (GIT) and commercial probiotic products for the use in marron culture.

Six strains of bacteria including Bacillus mycoides (A10) and Shewanella species (A12) isolated from healthy marron intestine, Bacillus species (PM1), Bacillus subtilis (PM3), Bacillus sp. (PM4) and Bacillus sp. (AQ) from commercial probiotic products were investigated for probiotic potential in marron culture. Antibiotic susceptibility tests indicated PM3 and PM4 were susceptible to all nine antibiotics evaluated. A10, A12 and AQ were resistant to class penicillins, whereas PM1 was resistant to class penicillin and macrolides. All strains were non-pathogenic for marron. Strong inhibition against Vibrio mimicus and Vibrio cholerae non-01 was exhibited by PM4 and PM3. A10 inhibited V. mimicus at 72 h of growth, but not V. cholerae non-01, whereas A12 inhibited V. cholerae non-01 but not V. mimicus, and AQ showed no inhibition activity. A wide range of enzymes were produced by A10 and AQ using the API ZYM test. Protease enzymes were produced by PM3, PM4, AQ and PM1. In order of effectiveness, the following bacteria have probiotic potential: B. subtilis (PM3), Bacillus sp. (PM4) and B. mycoides (A10). Further study is required to determine the bacterium or any combination that gives a multibeneficial effect on marron.

Role of mast cells in colorectal cancer development, the jury is still out.

The link between inflammation and colorectal cancer development is becoming increasingly clear. It had long been recognized that patients with inflammatory bowel disease are at an increased risk of colon cancer. Evidence from experimental animals now also implicates the innate immune system in the development of sporadically occurring intestinal adenomas, the precursors to colorectal cancer. Here we discuss the interaction between the immune system and the adenoma to carcinoma sequence with a special emphasis on the role of mast cells which may play a key role in adenoma development. This article is part of a Special Issue entitled: Mast cells in inflammation.

Understanding the molecular epidemiology of the footrot pathogen Dichelobacter nodosus to support control and eradication programs.

The Gram-negative anaerobe Dichelobacter nodosus is the primary etiologic agent of ovine footrot. Few studies of the genetic diversity and epidemiology of D. nodosus have been done, despite the economic cost and welfare implications of the disease. This study examined a large collection of Australian isolates; 735 isolates from footrot-infected sheep from 247 farms in Western Australia (WA) were tested by pulsed-field gel electrophoresis (PFGE), and a subset of 616 isolates was tested by infrequent restriction site PCR (IRS-PCR). The genetic diversity of WA isolates was compared to that of 61 isolates from three other Australian states. WA isolates were genetically diverse, with 181 molecular types resolved by PFGE, resulting in a simple diversity ratio (SDR) of 1:4 and a Simpson's index of discrimination value (D) of 0.98. IRS-PCR resolved 77 molecular types (SDR = 1:8 and D = 0.95). The isolates were grouped into 67 clonal groups by PFGE (SDR = 1:11, D = 0.90) and 36 clonal groups by IRS-PCR (SDR = 1:17, D = 0.87). Despite the high genetic diversity, three common clonal groups predominated in WA and were found in other Australian states. On some farms, molecular type was stable over a number of years, whereas on other farms genetically diverse isolates occurred within a flock of sheep or within a hoof. This study provides a large database from which to appropriately interpret molecular types found in epidemiological investigations and to identify common and unknown types that may compromise footrot eradication or control programs.

Infection with Photobacterium damselae subspecies damselae and Vibrio harveyi in snapper, Pagrus auratus with bloat.

OBJECTIVE: To diagnose the cause of chronic, low mortality associated with bloat in tanks of snapper at an aquaculture facility. DESIGN: A clinical, pathological and microbiological investigation into the cause of a low number of ongoing mortalities associated with bloat in snapper at an aquaculture facility is outlined. Necropsy, histology, microbiology and a comparison of haematology and water analysis from affected and unaffected fish and holding tanks, respectively were conducted. RESULTS: Affected moribund fish were found in lateral or dorsal recumbency floating on the water surface within 24 hours of death. Photobacterium damselae subspecies damselae was isolated from intestinal contents and Vibrio harveyi from the blood of affected fish and both were isolated from culture water. Both V harveyi and P damselae subspecies damselae isolates were sensitive to tetracycline, ciprofloxacin and sulphamethoxazole plus trimethoprim. Environmental parameters such as pH and dissolved oxygen were similar in tanks of affected and unaffected fish. Affected fish had gas distended swimbladders, anaemia, and the intestines were diffusely distended with a clear, pale yellowish fluid. Livers were mottled tan and green in a zonal pattern. Histologically the intestines of fish from tanks suffering mortality had a moderate granulocytic enteritis with oedema and infiltrations with eosinophilic granule cells that were also present as an infiltrate in the gills. There were elevated numbers of melanomacrophage centres and haemosiderin deposits in the spleen, kidney and liver of affected fish. CONCLUSION: Vibrio harveyi and Photobacterium damselae subspecies damselae infection should be recognised as potential pathogens of snapper held in water of less than optimal quality.

Percutaneous transluminal coronary angioplasty in chronic coronary artery occlusion.

OBJECTIVES: This study was conducted to determine the procedural success rate, complication rate and long-term outcome of percutaneous transluminal coronary angioplasty in chronically occluded coronary arteries. BACKGROUND: Coronary angioplasty of chronically occluded vessels has a lower success rate than has angioplasty of nonoccluded vessels, but it is frequently considered safe because the target vessel is already occluded. The purpose of this study was to determine the reliability of these assumptions at our institution, with the objectives stated above. METHODS: We identified from the angioplasty data base at our institution 100 consecutive coronary angioplasty procedures performed between 1987 and 1991 for chronic total occlusion, defined as complete occlusion (Thrombolysis in Myocardial Infarction [TIMI] grades 0 and 1 flow) for > or = 3 months. The records of the 95 patients who underwent these procedures were reviewed to determine procedural outcome and medium-term results. RESULTS: Procedural success was obtained in 47 occluded vessels (47%). Significantly fewer successes were obtained in the right coronary artery (26.8%) than in either the left anterior descending (57.1%) or the left circumflex (45%) coronary artery (p < 0.05). A procedural failure without serious adverse consequences occurred in 45 procedures (45%), but in eight patients (right coronary artery in five, left anterior descending artery in three) attempted recanalization was complicated by extensive coronary dissection with acute myocardial ischemia, and one of these patients died. There were no emergency operations, but elective coronary artery bypass surgery was undertaken in 26 patients (in 3 after extensive dissection, in 7 after an apparently good result and in 16 in whom the procedure failed). At 12 months after the procedure, 64.1% of those with a procedural success were event free compared with 32.6% of those whose procedure was both unsuccessful and uncomplicated (p < 0.025) and 25% of those in whom it was unsuccessful and complicated by coronary dissection (p < 0.025). CONCLUSIONS: In this series of recanalization of chronically occluded coronary arteries, there was a low procedural success rate, particularly for the right coronary artery. However, when procedural success was obtained, the long-term outlook was good. The overall risk of coronary dissection was comparable to the risk in nonoccluded vessels but was particularly high in the right coronary artery (13%).

Causes of symptoms in chronic congestive heart failure and implications for treatment.

The most common symptoms of patients with heart failure are shortness of breath and fatigue. The causes of these symptoms may be different in various entities encompassed by the general term heart failure, such as acute pulmonary edema, circulatory collapse and chronic heart failure. In patients with acute heart failure, shortness of breath is closely related to left atrial pressure. In patients with chronic heart failure, optimally treated with diuretics, the body fluid compartments are usually of normal size. Recent work strongly suggests that, in such patients, central hemodynamic abnormalities are not the sole determinants of symptoms. Impaired vasodilation and altered metabolism in skeletal muscle, circulating metabolites and pulmonary ventilation-perfusion mismatch with consequent increased physiologic dead space may all contribute to the genesis of symptoms. Consequently, it may be possible to alleviate symptoms by treatments that are not aimed directly at improving central hemodynamics. Whether such an approach could also modify prognosis is unknown.

Regional blood flow, muscle strength and skeletal muscle histology in severe congestive heart failure.

Regional blood flow to exercising skeletal muscle is reduced in patients who have undergone treatment for severe congestive heart failure, and is a key factor determining the limitation of exercise capacity. Recent studies have shown that the histology, contractile function and biochemistry of skeletal muscle are also abnormal. The mechanisms for both the reduced blood flow and the intrinsic abnormality of skeletal muscle are unknown. The interpretation of experimental data is complicated by different etiologies of heart failure, drug treatment, exercise protocols, the limitations of methods for the measurement of blood flow and metabolism in intact humans, and by the selection of particular groups of muscles for study that may not reflect changes in other muscles in the body.

Effectiveness and safety of a single intravenous bolus injection of tissue-type plasminogen activator in acute myocardial infarction. Bolus Dose-Escalation Study of Tissue-Type Plasminogen Activator (BEST) Investigators.

The efficacy of multiple intravenous bolus injections of tissue-type plasminogen activator (t-PA) in inducing rapid coronary recanalization in patients with acute myocardial infarction was previously demonstrated. In this Bolus Dose-Escalation Study of Tissue-Type Plasminogen Activator (BEST), the efficacy of 3 different doses of a single rapid intravenous bolus injection of t-PA (dute-plase, Wellcome Foundation, London) in inducing coronary patency (Thrombolysis In Myocardial Infarction perfusion grade 2 or 3) in 64 patients with acute myocardial infarction presenting less than 6 hours after onset of symptoms was investigated. At 60 minutes after administration of t-PA, the infarct-related coronary artery was patent in 9 of 17 patients (53%; 95% confidence interval [CI] 28 to 77%) after 0.3 MU/kg, in 14 of 23 (61%; 95% CI 39 to 80%) after 0.45 MU/kg and in 10 of 14 (71%; 95% CI 42 to 92%) after 0.6 MU/kg. At 90 minutes after t-PA, coronary patency was present in 9 of 17 cases (53%; 95% CI 28 to 77%) after 0.3 MU/kg, in 12 of 24 (50%; 95% CI 29 to 71%) after 0.45 MU/kg and in 10 of 13 (77%; 95% CI 46 to 95%) after 0.6 MU/kg. One patient in each dose group had a silent reoccluded infarct-related artery by 24 hours, and there were 2 clinical reinfarctions before discharge. No major bleeding events were observed. There were 5 hospital deaths, all unrelated to t-PA. A single intravenous bolus injection of 0.6 MU/kg of t-PA appears to be effective in inducing rapid coronary patency and to be safe in patients with acute myocardial infarction.

Blood flow and skeletal muscle in patients with heart failure.

The factors that contribute to the symptoms of breathlessness and fatigue, and that limit exercise capacity in patients with chronic heart failure are poorly understood. Recent evidence suggests that the major mechanism is not related to central hemodynamics but to a reduction of skeletal muscle mass and diminished blood flow to skeletal muscle on exercise.

In vitro validation of the luminal measurement of a novel catheter based moulding technique.

OBJECTIVE: To investigate a modified angioplasty balloon catheter, which uses a novel balloon polymer to produce luminal moulds. DESIGN: The catheter was tested in polyurethane phantoms of diameter 1.5 to 4.0 mm. Inflations were to 1.4 atmospheres for 20 seconds at 37 degrees C. The moulds were viewed by reinflating the balloon to 0. 34 atmospheres and quantified using macrophotography and caliper measurement. RESULTS: Evidence of systematic error was found with lumen diameters

Genetic differences between nitrate-negative and nitrate-positive C. pseudotuberculosis strains using restriction fragment length polymorphisms.

Corynebacterium pseudotuberculosis has been classified into two biotypes according to ability to breakdown nitrate (Biberstein et al., 1971). Restriction enzyme analysis (REA) has shown to reflect this differentiation, but numerous bands generated by this technique make interpretation difficult (Songer et al., 1988). Restriction fragment length polymorphism's (RFLP's) has become an accepted genetic tool and was used in this study to determine if differences in nitrate reduction and other phenotypic characteristics could be identified genetically. Thirteen C. pseudotuberculosis isolates from four species of domestic animals from different parts of the world were investigated for phenotypic and genetic differences. Three closely related bacteria, Corynebacterium ulcerans, Actinomyces pyogenes (previously C. pyogenes),and Rhodococcus equi (previously C. equi) were included in the study to determine if the RFLP bands were unique to C. pseudotuberculosis. All C. pseudotuberculosis isolates were positive for urease production. Some differences in maltose and sucrose fermentation ability and nitrate reduction were recorded. Genetic differences were identified between the nitrate-positive group and the nitrate-negative group using non-radioactive ribosomal RNA (rRNA) probes Southern blotted to restriction digests of ApaI, PstI, and SstI. A small number of bands were seen, with distinct differences between the nitrate-positive and the nitrate-negative strains. No genetic variations were seen between strains which reflected differences in carbohydrate fermentation. Strains isolated from different animal species and from different parts of the world could not be differentiated genetically using these three restriction enzymes.

A spirochete isolated from a case of severe virulent ovine foot disease is closely related to a Treponeme isolated from human periodontitis and bovine digital dermatitis.

The isolation of spirochetes from severe ovine foot disease has been reported recently by our research group. In this study we describe the preliminary classification of this spirochete based on nucleotide sequence analysis of the PCR-amplified 16S rRNA gene. Phylogenetic analysis of this sequence in comparison with other previously reported 16S rRNA gene sequences showed that the spirochete belonged to the treponemal phylotype Treponema vincentii which has been associated with bovine digital dermatitis and human periodontal disease. Further work is required to define the common virulence determinants of these closely related treponemes in the aetiology of these tissue destructive diseases.

Recurrent laryngeal nerve paralysis during transvenous insertion of a permanent endocardial pacemaker.

We present 2 patients with sudden onset of dysphonia due to acute unilateral vocal cord paralysis that occurred during transvenous insertion of a permanent endocardial pacemaker. Acute paralysis of the vocal cord due to this kind of medical intervention is a very unusual complication that has been rarely reported. We believe that traumatic insertion of a pacemaker through the subclavian vein triggered a dynamic process that led to vocal cord paralysis. Three hypothetic explanations are presented: 1) a direct vagus nerve lesion, 2) a direct inferior laryngeal nerve lesion, and 3) a central nervous system lesion caused by a cerebral microembolus. Physicians should be aware of the possibility that associated acute paralysis of the vocal cord may occur after transvenous insertion of an endocardial pacemaker.

Conservative surgical management of invasive differentiated thyroid cancer.

The majority of well differentiated thyroid carcinoma are tumours of low grade malignancy. Laryngotracheal invasion by well differentiated thyroid carcinoma is an uncommon occurrence. The surgical management of patients with thyroid cancer invading the upper airway has primarily been by total laryngectomy. Other surgeons recommend in selected cases partial laryngeal and/or tracheal resection. A total of 122 patients with thyroid carcinoma were treated in our department between 1967 and 1982. Only seven patients with well differentiated tumours had airway invasion. In these seven patients we used a partial laryngeal and/or tracheal resection. In three of the patients with tracheal invasion a myoperichondrial flap was used for closing the tracheal defect. A partial resection of the larynx and trachea, and end to end anastomosis between the trachea and the remaining part of the larynx was performed in another four patients. The techniques used and a long-term follow-up are presented (Table I).

Delayed brainstem auditory evoked responses in experimental diabetes mellitus.

The brainstem auditory evoked responses (BAER) were utilized for the evaluation of central neural transmission in alloxan-induced diabetes in rats. The mean latencies of waves I, III, V and the interpeak latencies III-V and I-V were prolonged in diabetic rats as compared to the same rats before alloxan administration. The incidence of abnormal BAER was more frequent in the group of rats with severe diabetes (82 per cent) than in mildly diabetic animals (42 per cent). Our results may suggest the presence of a central neuropathy in experimental diabetes, which can be detected by the method of BAER.

Delayed brainstem auditory evoked responses in diabetic patients.

The incidence of subclinical central diabetic neuropathy is unclear due to difficulty in detecting latent alterations of central neural transmission process. The aim of this study was to evaluate a central neuroconductive mechanism in diabetics by brainstem auditory evoked responses (BAER). We found increased latencies of peaks I, III, V in diabetics as compared to control subjects. These BAER abnormalities were demonstrated in 62 per cent of insulin-controlled diabetics and in 33 per cent o patients treated by diet, or peroral drugs. No alterations in brainstem responses were observed in patients with latent diabetes. We did not find any correlation between the BAER abnormalities and the duration of the disease, the blood glucose level or the level of control of the diabetes.

Antimicrobial susceptibility testing of Serpulina hyodysenteriae.

The macrobroth dilution technique was used to test the in-vitro effectiveness of 4 commonly used antimicrobial agents against 23 Australian isolates and 7 overseas strains of Serpulina hyodysenteriae. Minimum inhibitory concentrations and minimum bactericidal concentrations were determined. The growth of 90% of isolates was inhibited by dimetridazole at a concentration of 4 micrograms/mL, and by tiamulin at 8 micrograms/mL. Australian isolates resistant to both antimicrobial agents were identified. Lincomycin was less effective than these antimicrobial agents, with 90% of isolates requiring a concentration of 128 micrograms/mL for inhibition of growth, and 54% being susceptible at 64 micrograms/mL. Tylosin did not prevent the growth of the majority of S hyodysenteriae isolates tested, and 90% were resistant to concentrations of > or = 128 micrograms/mL. Resistant isolates came from different geographical areas. Resistance was not related to overall genetic background of the spirochaetes, and was not correlated with the presence of plasmids or the serogroup of the isolates.

Effect of the interval between shearing and dipping on the spread of Corynebacterium pseudotuberculosis infection in sheep.

OBJECTIVE: To determine whether the spread of Corynebacterium pseudotuberculosis infection to sheep in dips could be controlled by increasing the time between shearing and dipping. DESIGN: A controlled treatment trial where only the time between shearing and dipping was varied. ANIMALS AND PROCEDURE: One hundred and ninety-five sheep were found to be negative for C. pseudotuberculosis exposure by assay of CLA toxin antibody, were divided into four treatment groups. Each was shorn at either 0, 2, 4 or 8 weeks before dipping in a solution containing C. pseudotuberculosis. Blood samples were taken 6 weeks after dipping and sheep were slaughtered 12 weeks after dipping. A fifth smaller group of 14 sheep shorn 26 weeks before dipping, was also exposed to C. pseudotuberculosis and was slaughtered with the other sheep. RESULTS: The occurrence of caseous lymphadenitis abscesses did not differ between groups or with sheep shorn 26 weeks before dipping. The proportion of sheep that seroconverted to the C. pseudotuberculosis toxin and cell wall ELISA was larger in sheep dipped immediately after shearing than in sheep in the other groups. CONCLUSIONS: Delaying dipping until 8 weeks after shearing did not decrease the C. pseudotuberculosis infection rate due to dipping. Sheep dipped immediately after shearing developed higher concentrations of antibody to C. pseudotuberculosis than sheep when dipping occurred between 2 and 8 weeks and later after shearing.

Rage signalling promotes intestinal tumourigenesis.

Development of colon cancer is a multistep process that is regulated by intrinsic and extrinsic cellular signals. Extrinsic factors include molecular patterns that are derived from either pathogens (PAMPs) or cellular damage (DAMPs). These molecules can promote tumourigenesis by activation of the innate immune system, but the individual contribution of ligands and their receptors remains elusive. The receptor for advanced glycation end products (Rage) is a pattern recognition receptor that binds multiple ligands derived from a damaged cell environment such as Hmgb1 and S100 protein. Here we show that Rage signalling has a critical role in sporadic development of intestinal adenomas, as Apc(Min/+) Rage(-/-) mice are protected against tumourigenesis.