Molecular epidemiology of Salmonella Heidelberg in an equine hospital

From 1992 to 1997, multi-drug resistant (MDR) Salmonella Heidelberg isolates were cultured from a number of horses hospitalised in a veterinary hospital in Victoria, Australia. To examine the relationships between the cases, 28 isolates from the hospital were compared by pulsed field gel electrophoresis (PFGE), IS200 element profiles, antimicrobial resistance patterns, plasmid profiles and phage typing. The PFGE patterns following digestion with XbaI and BlnI restriction endonucleases showed that the isolates from the veterinary hospital originated from a common source. These isolates also had indistinguishable IS200 profiles. However, PFGE was more discriminatory than IS200 profiles. All the veterinary hospital isolates and one independent isolate had the same antimicrobial resistance pattern and had at least one plasmid in common. Localisation of antimicrobial resistance genes indicated that the veterinary hospital isolates had more than one plasmid carrying resistance genes and that the genes encoding sulphathiazole and trimethoprim resistance were not on these plasmids. Phage typing was ineffective as 22 of the 28 isolates were untypeable. In conclusion, the combination of different methods used for epidemiological studies suggested that a single strain of MDR S. Heidelberg was isolated from horses admitted to the hospital for 6 years and caused salmonellosis in susceptible horses within that period with no apparent correlation between the antimicrobials used and retention of its MDR phenotype.     

Thyrotropin-releasing hormone stimulation test to assess thyroid function in severely sick cats

Basal serum thyroxine (T4) concentration and the thyrotropin-releasing hormone (TRH) stimulation test were used to assess thyroid function in 36 critically ill cats examined between July 1996 and October 1998. Of the 36 cats. hyperthyroidism (as underlying or complicating disease) was suspected in 22 based on clinical signs, palpable thyroid nodules, and abnormal thyroid gland histology (study group). Hyperthyroidism was not suspected in the remaining 14 cats, which served as the control group. Based on serum T4 concentrations, suppression of thyroid function was documented in 14 (64%) cats of the study group and in 10 (71%) cats of the control group. The TRH stimulation test revealed an increase in serum T4 of less than 50% of the baseline concentration in 18 (82%) cats of the study group, and in 6 (43%) cats of the control group. In conclusion, based on the results of serum T4 determinations and the TRH stimulation tests, it was not possible to differentiate between cats with clinical and histologic evidence of thyroid dysfunction (hyperthyroidism) and cats with severe nonthyroidal illnesses.     

Evaluation of urinary carnitine and taurine excretion in 5 cystinuric dogs with carnitine and taurine deficiency

Five client owned dogs with cystinuria were diagnosed with carnitine and taurine deficiency while participating in a clinical trial that used dietary management of their urolithiasis. Stored 24-hour urine samples collected from the cystinuric dogs before enrollment in the clinical diet trial were quantitatively evaluated for carnitine and taurine. These results were compared to those obtained from 18 healthy Beagles. Both groups of dogs were fed the same maintenance diet for a minimum of 2 weeks before 24-hour urine collection. The protocol used for 24-hour urine collections was the same for cystinuric dogs and healthy Beagles except that cystinuric dogs were catheterized at baseline, 8 hours, 12 hours, and at the end of the collection, whereas Beagles were catheterized at baseline, 8 hours, and at the end of the collection. Three of 5 dogs with cystinuria had increased renal excretion of carnitine. None of the cystinuric dogs had increased renal excretion of taurine, but cystinuric dogs excreted significantly less (P < .05) taurine in their urine than the healthy Beagles. Carnitinuria has not been recognized previously in either humans or dogs with cystinuria, and it may be 1 risk factor for developing carnitine deficiency. Cystinuric dogs in this study were not taurinuric; however, cystine is a precursor amino acid for taurine synthesis. Therefore, cystinuria may be 1 risk factor for developing taurine deficiency in dogs. We suggest that dogs with cystinuria be monitored for carnitine and taurine deficiency or supplemented with carnitine and taurine.     

Detection of Mycobacterium avium subsp. paratuberculosis in tissue samples by single, fluorescent and nested PCR based on the IS900 gene

The aim of this study was to determine if fluorescent PCR could be used instead of nested PCR, for the detection of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) in clinical specimens, to improve the sensitivity without increasing the risk for cross-contamination. We investigated and compared the sensitivity of single PCR, fluorescent PCR and nested PCR for the detection of IS900, an insertion sequence specific for M. paratuberculosis. A previously described extraction method for clinical specimens, based on xylene, was evaluated regarding its suitability for routine diagnostic work. The sensitivity of each PCR system was assessed by analysing a serial dilution of M. paratuberculosis DNA. To improve the reliability of the PCR and to facilitate the interpretation of the PCR results, a positive internal control molecule ("mimic") was developed and used for single and fluorescent PCR. In nested PCR, an existing mimic was used. The efficiency of recovering DNA of M. paratuberculosis from clinical specimens by the extraction method and detection of the organism by PCR was studied by analysing spiked ileum mucosa specimens. The final evaluation was performed on seventeen ileum mucosa specimens, previously found positive for M. paratuberculosis by bacterial culture. Twelve of the samples were positive by fluorescent PCR and nested PCR, and 10 samples were positive by single PCR. The use of mimics showed inhibition in specimens harbouring few M. paratuberculosis organisms, illustrating the effect of inhibitory substances in combination with small amounts of M. paratuberculosis DNA. We conclude that the extraction method was not adequate to recover small amounts of M. paratuberculosis and that inhibitory substances were still present in the processed specimens, but that the method is useful for identifying positive samples. Fluorescent PCR was a suitable alternative to both single PCR and nested PCR for the detection of M. paratuberculosis.     

Effects of chromium nicotinate on performance, carcase characteristics and blood chemistry of growing turkeys

1. The objective of this trial was to study the effects of chromium nicotinate (Cr) supplementation on the performance, carcase characteristics and blood constituents of 9 to 22 week-old male turkeys. 2. Fifty-four 8-week-old male turkeys weighing 2.9 +/- 0.28 kg were allocated at random into 9 groups with 3 dietary treatments and 3 replicates. The dietary treatment was different dietary concentrations of chromium in the basal diet, to provide 0 (control), 1 or 3 mg/kg. The feeding trial lasted for 14 weeks. 3. Results showed that 1 mg/kg chromium supplementation significantly improved weight gain and food intake at 9 to 18 weeks of age but did not significantly influence performance at 19 to 22 week of age. The breast and thigh muscle were significantly increased in birds receiving 1 mg/kg chromium supplementation but were decreased by 3 mg/kg of supplementation. 4. Dietary chromium supplementation did not significantly influence other serum constituents, including insulin, HDL, VLDL+LDL, HDL-C, VLDL-C+LDL-C, total protein, albumin, and gamma-globulin at 18 and 22 weeks of age. 5. The serum triacylglycerol (TG) and uric acid contents were significantly increased, while glycerol and alpha-globulin were significantly reduced by 3 mg/kg chromium supplementation. However, 1 mg/kg chromium supplementation significantly reduced serum cholesterol and glycerol at 18-weeks old. At 22 weeks of age, 1 mg/kg chromium supplementation significantly increased serum glucose and decreased the uric acid concentration whereas 3 mg/kg chromium supplementation significantly increased the creatinine concentration and decreased beta-globulin concentration.     

Changes in ascorbic acid content in primary cultured rat hepatocytes exposed to 2,2'-azobis (2-amidinopropane) dihydrochloride, a radical initiator

Changes in ascorbate content in primary cultured rat hepatocytes exposed to oxidative stress derived from water soluble radical initiator 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) were examined. Cells were exposed to 0.05 and 5 mg/ml of AAPH as 'mild' and severe' oxidative stresses, respectively. Lipid peroxidation in hepatocytes was induced by 'severe' oxidative stress, but not by 'mild' oxidative stress. Ascorbate decreased at 6 hr after administration of both mild' and severe' oxidative stresses, and recovered to the control level after a further 6 hr. In cells treated with 'severe oxidative stress, however, total ascorbate (reduced form plus oxidized form) had increased 24 hr after administration. These results indicated that consumption alone did not account for the increase of ascorbate in hepatocytes under oxidative stress.     

Experimental reproduction of severe disease in CD/CD pigs concurrently infected with type 2 porcine circovirus and porcine reproductive and respiratory syndrome virus

Three-week-old cesarean-derived colostrum-deprived (CD/CD) pigs were inoculated with porcine circovirus type 2 (PCV2, n = 19), porcine reproductive and respiratory syndrome virus (PRRSV, n = 13), concurrent PCV2 and PRRSV (PCV2/PRRSV, n = 17), or a sham inoculum (n = 12) to compare the independent and combined effects of these agents. Necropsies were performed at 7, 10, 14, 21, 35, and 49 days postinoculation (dpi) or when pigs became moribund. By 10 dpi, PCV2/PRRSV-inoculated pigs had severe dyspnea, lethargy, and occasional icterus; after 10 dpi, mortality in this group was 10/11 (91%), and all PCV2/ PRRSV-inoculated pigs were dead by 20 dpi. PCV2-inoculated pigs developed lethargy and sporadic icterus, and 8/19 (42%) developed exudative epidermitis; mortality was 5/19 (26%). PRRSV-inoculated pigs developed dyspnea and mild lethargy that resolved by 28 dpi. Microscopic lesions consistent with postweaning multisystemic wasting syndrome (PMWS) were present in both PCV2- and PCV2/PRRSV-inoculated pigs and included lymphoid depletion, necrotizing hepatitis, mild necrotizing bronchiolitis, and infiltrates of macrophages that occasionally contained basophilic intracytoplasmic inclusion bodies in lymphoid and other tissues. PCV2/ PRRSV-inoculated pigs also had severe proliferative interstitial pneumonia and more consistent hepatic lesions. The most severe lesions contained the greatest number of PCV2 antigen-containing cells. PRRSV-inoculated pigs had moderate proliferative interstitial pneumonia but did not develop bronchiolar or hepatic lesions or lymphoid depletion. All groups remained seronegative to porcine parvovirus. The results indicate that 1) PCV2 coinfection increases the severity of PRRSV-induced interstitial pneumonia in CD/CD pigs and 2) PCV2 but not PRRSV induces the lymphoid depletion, granulomatous inflammation, and necrotizing hepatitis characteristic of PMWS.     

Incidence and prognostic value of low plasma ionized calcium concentration in cats with acute pancreatitis: 46 cases (1996-1998).

OBJECTIVE: To determine the incidence and prognostic significance of low plasma ionized calcium concentration in cats with clinical signs of acute pancreatitis (AP). DESIGN: Retrospective study. ANIMALS: 46 cats with AP and 92 control cats with nonpancreatic diseases. PROCEDURE: Medical records were reviewed, and results of clinicopathologic testing, including plasma ionized and total calcium concentrations, acid-base values, and electrolyte concentrations, were recorded. Cats with AP were grouped on the basis of outcome (survived vs died or were euthanatized), and plasma ionized calcium concentrations, acid-base values, and electrolyte concentrations were compared between groups. RESULTS: Serum total calcium concentration was low in 19 (41%) cats with AP, and plasma ionized calcium concentration was low in 28 (61%). Cats with AP had a significantly lower median plasma ionized calcium concentration (1.07 mmol/L) than did control cats (1.12 mmol/L). Nineteen (41%) cats with AP died or were euthanatized; these cats had a significantly lower median plasma ionized calcium concentration (1.00 mmol/L) than did cats that survived (1.12 mmol/L). Ten of the 13 cats with AP that had plasma ionized calcium concentrations < or = 1.00 mmol/L died or were euthanatized. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that low plasma ionized calcium concentration is common in cats with AP and is associated with a poorer outcome. A grave prognosis and aggressive medical treatment are warranted for cats with AP that have a plasma ionized calcium concentration < or = 1.00 mmol/L.     

Contamination of pigs by nose-to-nose contact or airborne transmission of Salmonella Typhimurium.

The aim of this study is to assess the risk of contamination by Salmonella Typhimurium of pigs by nose-to-nose contact or the airborne route. Thirty twelve-week-old SPF pigs were divided into 4 groups housed in 4 different rooms: the first room contained Salmonella-free control pigs (n = 4), the second room had 10(3) CFU S. Typhimurium inoculated pigs (n = 5) and non-inoculated "contact" pigs (n = 4), the third room had pigs (n = 8) receiving potentially contaminated air from the following room through a hole (4 pigs housed in the pen situated near the hole and 4 pigs in the pen at the opposite side of the room), and the fourth room had pigs (n = 5) inoculated with 10(6) CFU Salmonella Typhimurium and also non inoculated "contact" pigs (n = 4). The "contact" and the inoculated pigs were housed in adjacent pens allowing nose-to-nose contact. The 5 pigs orally inoculated with 10(6) CFU S. Typhimurium were bacteriologically and serologically positive 1 week later and their environment was contaminated as early as 1 day pi. The faecal samples of 4 nose-to-nose contact pigs were bacteriologically positive and one of them was seropositive 5 weeks pi before the pigs were commingled. The 8 pigs housed in the third room received S. Typhimurium by an active airflow coming from the contaminated room (1000 m3/hour). Their faecal samples remained negative until 8 weeks pi but the environmental swabs taken in the room close to the airinlet were contaminated 2 days pi and positive swabs were found elsewhere in the room 5 weeks pi. Two seropositive pigs were encountered 8 weeks pi in the pen situated near the hole. Only one among the 5 pigs inoculated with 10(3) CFU had bacteriologically positive faeces 1-week pi and the 4 pigs kept in nose-to-nose contact with them remained negative. A dose of 10(3) CFU was too small to induce persistent excretion and to stimulate a humoral immune response. However, the dose of 10(6) CFU induced contamination of nose-to-nose contact pigs and contamination of the environment by airflow.