Detection of Mycobacterium avium subsp. paratuberculosis in ovine faeces by direct quantitative PCR has similar or greater sensitivity compared to radiometric culture

The aims of this study were to develop a new real-time quantitative PCR (QPCR) assay based on IS900 for detection and quantification of Mycobacterium avium subsp. paratuberculosis (MAP) DNA in faeces, and to use this to detect infected sheep. Both the C and S strains of MAP were detected by the QPCR assay, and no cross reactions were detected with 51 other species of mycobacteria including 10 which contained IS900-like sequences. One copy of IS900 fragment cloned into plasmid pCR2.1 and 1 fg of MAP genomic DNA were consistently detected, while in spiked faecal samples the detection limit was 10 viable MAP per gram of ovine faeces. A total of 506 individual ovine faecal samples and 27 pooled ovine faecal samples with known culture results were tested. The QPCR assay detected 68 of 69 BACTEC culture positive individual faeces and there was a strong relation between time to detection in culture and DNA quantity measured by QPCR (r= -0.70). In pooled faecal samples, QPCR also agreed with culture (kappa=0.59). MAP DNA was detected from some culture negative faecal samples from sheep exposed to MAP, suggesting that the QPCR has very high analytical sensitivity for MAP in faecal samples and detects non-viable MAP in ovine faeces. None of the faecal samples from 176 sheep that were not exposed to MAP were positive in QPCR. This is the first report of a direct faecal QPCR assay that has similar sensitivity to a gold standard radiometric culture assay.     

Biological Degradation of Common Pharmaceuticals and Personal Care Products in Soils with High Water Content

Biological degradation rates of six pharmaceuticals and personal care products were examined in soil from a land application site and in adjacent soil with no prior history of effluent exposure. Microbial degradation rates were compared over 2 weeks under standing water or saturated conditions and draining conditions after having been saturated for 3 days. Biological degradation of 17??-estradiol exhibited rapid rates of biological degradation under both saturated and draining conditions. Half-lives for 17??-estradiol ranged from 1.5 to 4 days; 66?C97% was lost from the soils. Estriol showed a pattern of biological degradation in both saturated and draining conditions though the half-lives were longer (8.7?C25.9 days) than those observed for 17??-estradiol. Twenty-eight percent to 73% of estriol was lost over the 14 days treatment period. Estrone and 17??-ethinylestradiol exhibited slower rates of biological transformation under saturated and draining conditions. Half-lives for estrone ranged between 27.5 and 56.8 days with loss of at most 21%. 17??-ethinylestradiol exhibited half-lives of 22.6?C207 days. Half-life data for ibuprofen ranged from 30.4 to 1,706.4 days in this experiment. Losses of up to 17% were observed in draining soils. Triclosan loss was at most 10%, and half-lives were 70.9?C398.8 days. In all cases, soils that were draining from saturated conditions exhibited faster degradation rates than soils that remained saturated. Prior exposure of the soil to effluent did not always result in higher biological degradation rates.     

Relative Prevalence of Antibiotic Resistance among Heterotrophic Bacteria from Natural and Culture Environments of Freshwater Prawn,Macrobrachium Rosenbergii

As antibiotic residues and multidrug‐resistant (MDR) bacteria are of increasing concern to those involved in the seafood industry, there is an intense international focus to monitor MDR bacteria in seafood. There is also a trend to source seafood from organically raised farms in order to reduce antibiotic usage in culture operations. Hence this study was conducted to compare the antibiotic resistance among bacteria associated with various samples from the natural and culture environment of Macrobrachium rosenbergii. Samples from the natural environment included water, sediment, eggs, larvae, postlarvae (PL), feed items, and gut samples of M. rosenbergii and those from the culture environment were water, sediment, eggs, feed pellets, and gut samples of M. rosenbergii. Bacteria from water samples of natural and culture environments were frequently resistant to ampicillin, erythromycin, and penicillin, while more than 50% of the strains from sediment were resistant to erythromycin, nalidixic acid, and penicillin. Bacterial isolates from commercial feed had a relatively high degree of antibiotic resistance. Interestingly, occurrence of multiple drug resistance was higher among the bacteria associated with the samples from the natural environment of M. rosenbergii, pointing to more favorable selection pressure for drug resistant mutants in the natural environment.     

Assessment of changes in blood volume in response to resuscitative fluid administration in dogs

Objective: To determine the continuous changes in blood volume in response to fluid administration using an in‐line hematocrit monitor. Design: Prospective study. Setting: Research laboratory. Animals: Four healthy dogs. Interventions: Each dog received intravenous boluses of 80 mL/kg of 0.9% saline (S), 4 mL/kg of 7.5% saline (HS), 20 mL/kg of dextran 70 (D), 20 mL/kg of hetastarch (HES), or no fluids (control, C) on separate occasions. Fluids were administered at 150 mL/min in the S, D, and HES groups, and at 1 mL/kg/min in the HS group. Measurements and main results: Blood volume changes were measured every 20 seconds for 240 minutes using an in‐line hematocrit monitor. There was a rapid rise in blood volume during all infusions. Immediately after the administration of crystalloid fluids, the rapid rise in blood volume ceased. Subsequently, there was a steep decline in blood volume for 10 minutes, and a slower decline thereafter. In contrast, the rise in blood volume continued for at least 10 minutes after the infusion of the colloids was complete, and a plateau was observed for the remainder of the experiment. The blood volume effect, as measured by area under the curve, was significantly greater in the saline group than the other groups during the infusion time and for the 0–240 minutes time period. The areas under the curve for the two colloid solutions were not significantly different from each other during any time periods. The percent increase in blood volume immediately following the infusions was 76.4±10.0 in the S group, 17.1±3.2 in the HS group, 23.0±10.5 in the D group, and 27.2±6.4 in the HES group. At 30 minutes from the start of the infusion, the mean percent increases in blood volumes were 35.2±9.3 in the S group, 12.3±0.9 in the HS group, 35.9±7.3 in the D group, and 36.8±6.5 in the HES group. At 240 h post‐infusion, the mean percent increases in blood volume were 18.0±9.7 in the S group, 2.9±6.1 in the HS group, 25.6±16.1 in the D group, and 26.6±8.6 in the HES group. The C group had a mean percent change in blood volume of ?3.7±3.4 at the end of the experiment. Conclusions: This study indicates that the rapid administration of saline at clinically relevant doses leads to the largest immediate increase in blood volume, although this change is transient because of rapid redistribution of the fluid. Despite a brief increase in blood volume that was almost 3 times the volume administered, hypertonic saline led to the smallest increase in blood volume post‐infusion. The synthetic colloid solutions increased the blood volume by an amount greater than that infused and the effect was sustained for a longer period of time than seen following crystalloid administration, but the maximum increase in blood volume was significantly less than saline. The measurement of continuous changes in blood volume, using an in‐line hematocrit monitor, was a useful means of assessing the dynamic effects of fluid administration in dogs in a research setting.     

Occurrence of Piscirickettsiosis-like syndrome in tilapia in the continental United States.

From 2001 to 2003, tilapia (Oreochromis sp.) farms in Florida, California, and South Carolina experienced epizootics of a systemic disease causing mortality. The fish exhibited lethargy, occasional exophthalmia, and skin petechia. The gills were often necrotic, with a patchy white and red appearance. Grossly, the spleen and kidneys were granular with whitish irregular nodules throughout. Granulomatous infiltrates were observed in kidney, spleen, testes, and ovary tissues, but not in the liver. The granulomas contained pleomorphic coccoid bacteria, measuring 0.57 +/- 0.1 x 0.8 +/- 0.2 microm, that were Giemsa-positive, acid-fast-negative, and Gram-negative. The bacteria had a double cell wall, variable electron-dense and -lucent areas, and were present in the cytoplasm and within phagolysosomes. The syndrome was associated with cold stress and poor water conditions. These findings are consistent with an infectious process caused by a Piscirickettsia-like bacterium described previously in tilapia in Taiwan and Hawaii. This report involves the first identified cases of a piscirickettsiosis-like syndrome affecting tilapia in the continental United States.     

Field evaluation of moxidectin/praziquantel oral gel in horses

The safety and efficacy of 2% moxidectin/12.5% praziquantel oral gel administered at a rate of 0.4 mg moxidectin and 2.5 mg praziquantel/kg was studied in client-owned horses under field use conditions. Four hundred horses (300 treated with moxidectin/praziquantel oral gel and 100 treated with vehicle) were enrolled, feces were collected, and eggs were counted. Investigators as well as horse owners were masked to treatment assignment. No adverse reactions to treatment were observed in any horses. Moxidectin/praziquantel gel reduced Anoplocephala spp by more than 99% and provided a significant (P <.05) reduction (> 98%) in the strongyle egg count of treated horses.     

Evaluation of urine sulfated and nonsulfated bile acids as a diagnostic test for liver disease in dogs

OBJECTIVE: To evaluate 3 methods for measuring urine bile acids (UBA) and compare their diagnostic performance with that of the serum bile acids (SBA) test and other routine screening tests in dogs with hepatic disorders. DESIGN: Prospective study. ANIMALS: 15 healthy dogs, 102 dogs with hepatic disorders, and 9 dogs with clinical signs of hepatic disorders that were found to have nonhepatic disorders. PROCEDURES: Blood and urine samples were collected from sick dogs and healthy dogs for serum biochemical analyses, and determination of concentrations of SBA and UBA. Urine samples were obtained from 15 healthy dogs to establish an upper cutoff value for UBA concentrations. The UBA were measured by use of a quantitative-linked enzymatic colorimetric method. Three analytical modifications were evaluated; 1 quantified only urine sulfated bile acids (USBA), 1 only urine nonsulfated bile acids (UNSBA), and 1 quantified both (USBA plus UNSBA). The UBA values were standardized with the urine creatinine concentration. RESULTS: The UNSBA-to-creatinine ratio and USBA plus UNSBA-to-creatinine ratio tests had the best diagnostic performance of the UBA tests; each had a substantially higher specificity, slightly higher positive predictive value, slightly lower negative predictive value, and lower sensitivity than the SBA test. These UBA-to-creatinine values were positively correlated with SBA values. The USBA-to-creatinine ratio had poor sensitivity, indicating a low rate of bile acid sulfation in dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The UBA can be measured in dogs with sufficient repeatability and accuracy for clinical application. The UNSBA-to-creatinine ratio and USBA plus UNSBA-to-creatinine ratio identified dogs with hepatic disorders nearly as well as the SBA test.     

Usefulness, completeness, and accuracy of Web sites providing information on osteoarthritis in dogs

OBJECTIVE: To evaluate the quality of information regarding osteoarthritis (OA) in dogs currently available on the World Wide Web. DESIGN: Survey study. PROCEDURE: 5 search engines were searched with the keywords "dog," "degenerative joint disease," "canine," and "osteoarthritis," and the first 50 sites listed by each search engine were analyzed. Unique Web site addresses were distributed to 3 diplomates of the American College of Veterinary Surgeons, who provided a standardized evaluation of each site. RESULTS: 30 unique Web sites were evaluated. Twenty (66%) provided information consistent with conventional knowledge as outlined in textbooks and peer-reviewed literature, 8 (27%) provided experimental or anecdotal information in addition to conventional knowledge, and 2 (7%) provided misleading information. Mean scores for overall usefulness of the information provided in regard to clinical features of and treatment for OA were 1.3 and 1.5, respectively (1 = information of minimal use; 5 = very useful information). Twenty-three (77%) sites encouraged pet owners to seek the advice of a veterinarian. Twenty-three (77%) sites were given overall quality scores < 2, and 7 (23%) were given scores between 2 and 3 (1 = site was counterproductive; 5 = site was very valuable). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the quality of information currently available on the Web that addresses OA in dogs is questionable. Although most of the sites conveyed some conventional information with reasonable accuracy, the information was incomplete, of minimal use, and often considered counterproductive.