Real-time polymerase chain reaction testing for the detection of Mycobacterium genavense and Mycobacterium avium complex species in avian samples

Diagnosis of avian mycobacteriosis, caused by Mycobacterium genavense or species belonging to the Mycobacterium avium complex (MAC), is problematic. Polymerase chain reaction (PCR) offers rapid and sensitive detection of minute quantities of DNA, and conventional protocols have been used for evaluating avian specimens. The recent development of real-time PCR offers several advantages over conventional PCR. In attempts to improve diagnosing avian mycobacteriosis, a real-time TaqMan PCR assay was developed targeting the 65-kD heat shock protein gene of M. genavense and MAC spp. Nineteen reference isolates, 16 clinical isolates, and 32 avian tissue samples were used to evaluate the assay. When sufficient amplicons were produced, the species of mycobacteria was determined by standard sequencing of TaqMan PCR products and compared with results from commercial mycobacteriology laboratories and/or standard sequencing of conventional PCR products. The TaqMan PCR detected DNA from reference isolates of M. genavense, MAC spp., and Mycobacterium tuberculosis complex spp. Of the clinical isolates, the TaqMan PCR detected DNA from 10 of 12 Mycobacterium avium avium isolates and two of three Mycobacterium avium intracellulare isolates. For the tissue samples, the TaqMan PCR amplified DNA in six of nine samples that were identified by sequencing of conventional PCR products and/or by commercial mycobacteriology laboratories as being MAC spp. positive and three of four samples that were positive for M. genavense. There was some disagreement between speciation results from the TaqMan PCR and those from commercial mycobacteriology laboratories or conventional PCR or both. This disagreement was suspected to be because of relatively small numbers of base pairs in the TaqMan PCR products. The TaqMan PCR may provide a useful tool for evaluating clinical samples for DNA from mycobacteria species that most commonly infect birds; however, further refinement is needed in order to improve sensitivity and provide more accurate speciation.     

Use of the distomedial-proximolateral oblique radiographic view of the elbow joint for examination of the medial coronoid process in dogs

OBJECTIVE: To describe and evaluate a new radiographic view of the elbow joint in dogs that would potentially enhance observation of the medial coronoid process (MCP). SAMPLE POPULATION: 20 cadaver limbs from 10 dogs and clinical examination of 100 elbow joints of 53 dogs. PROCEDURE: Twenty elbow joints from 10 cadavers were imaged by use of mediolateral, flexed mediolateral, craniocaudal, craniolateral-caudomedial oblique (Cr15L-CdMO), and distomedial-proximolateral oblique (Di35M-PrLO) radiographic views before and after placement of 3 lead pellets placed on the cranial, medial, and craniodistal aspect of the MCP. Three examiners independently reviewed these radiographs. One hundred elbow joints of 53 dogs with forelimb lameness and signs of pain elicited on palpation of the elbow joint were examined. These joints were radiographed and treated by use of arthroscopy. Three examiners independently graded the radiographs. RESULTS: The MCP was identified on all Di35M-PrLO views made during the anatomic study. The Di35M-PrLO view had the largest area under the receiving operating characteristic (ROC) curve for detection of abnormalities of the MCP. Fractured and nonfractured MCP could only be significantly differentiated on Di35M-PrLO and mediolateral views. The Di35M-PrLO view had a higher agreement between examiners than other radiographic views for detection of fractures of the MCP. CONCLUSION AND CLINICAL RELEVANCE: The Di35M-PrLO view enhances the identification of anomalies and fragmentation of the MCP in dogs, compared with other radiographic views. The Di35M-PrLO view may be of benefit for early screening of dogs potentially affected with elbow dysplasia.     

Changes in the phytochemical composition and profile of raw, boiled, and roasted peanuts

Peanuts are consumed mostly as processed products. Although the effect of processing on isoflavone composition of legumes has been extensively studied, there has been no such study on peanuts. The objective of this study was to evaluate the effect of processing (boiling, oil- and dry-roasting) on the phytochemical composition of peanuts. Boiling had a significant effect on the phytochemical composition of peanuts compared to oil- and dry-roasting. Boiled peanuts had the highest total flavonoid and polyphenol content. The biochanin A and genistein content of boiled peanut extracts were two- and fourfold higher, respectively. trans-Resveratrol was detected only in the boiled peanuts, with the commercial product having a significantly (p < or = 0.05) higher concentration. Ultraviolet and mass spectrometry chromatograms for the boiled peanut extracts show the presence of four additional peaks that were not observed in the raw peanut extracts.     

Cyanide toxicosis in Asian small-clawed otters (Amblonyx cinereus) secondary to ingestion of loquat (Eriobotrya japonica).

Two Asian small-clawed otters (Amblonyx cinereus) died acutely in their exhibit within a 4-day period. Neither animal had significant gross lesions at necropsy. Histologic findings were consistent with acute vascular shock and disseminated intravascular coagulation, and a toxic etiology was suspected. The animals' exhibit contained opened, uneaten loquat (Eriobotrya japonica) fruit with the seeds removed and large quantities of undigested loquat seed material in the feces of the remaining animals. Cyanide was detected in the stomach contents collected at necropsy and in fresh loquat seeds from the exhibit. Loquat is related to other cyanide-containing fruit trees, including cherries, peaches, and almonds.     

A molecular epidemiologic investigation of Salmonella from a meat source to the feces of captive cheetah (Acinonyx jubatus).

Low cheetah (Acinonyx jubatus) birth rates were observed for a long time in a captive breeding facility in which Salmonella, which was possibly present in contaminated beef, was isolated from still-born lion (Panthera leo) cubs. Salmonella, including 14 isolates of Salmonella serovar typhimurium and 19 isolates of Salmonella serovar muenchen, was subsequently isolated 47 times from 378 meat samples at the facility during a 13-mo period. Salmonella, including 26 isolates of S. serovar typhimurium, 10 of S. serovar muenchen, and 11 other serovars, also was isolated 54 times from 119 fecal samples. Only three plasmid profiles were identified in 59 S. typhimurium isolates from both meat and fecal samples. Although random-amplified polymorphic DNA fingerprinting using different primers in the polymerase chain reaction was able to distinguish between S. typhimurium and S. muenchen and to demonstrate similar chromosomal DNA fingerprints in some of the isolates from meat and feces, the results were not consistent enough to prove that the Salmonella in the feces originated from contaminated meat. However, the predominance of only two serovars in the meat fed to carnivores and in the feces of these animals suggests that the meat was the source of the Salmonella organisms in the feces.     

Blind inlets: conservation practices to reduce herbicide losses from closed depressional areas

Purpose

In a 6-year study, we investigated the effectiveness of blind inlets as a conservation practice in reducing pesticide losses compared to tile risers from two closed farmed depressional areas (potholes) in the US Midwest under a 4-year cropping rotation.

Materials and methods

In two adjacent potholes within the same farm and having similar soils, a conventional tile riser and blind inlet were installed. Each draining practice could be operated independent of each other in order to drain and monitor each depression with either practice. Sampling events (runoff events) were collected from the potholes from 2008 to 2013 using autosamplers. The samples were analyzed for atrazine, metolachlor, 2,4-D, glyphosate, and deethylatrazine.

Results and discussion

The results of this study demonstrated that the blind inlet reduced analyzed pesticide losses; however, the level of reduction was compound dependent: atrazine (57 %), 2,4-D (58 %), metolachlor (53 %), and glyphosate (11 %).

Conclusions

Results from this study corroborate previous research findings that blind inlets are an effective conservation practice to reduce discharge and pollutants, including pesticides from farmed pothole surface runoff in the US Midwest.

     

Seismic anisotropy beneath Ruapehu volcano: a possible eruption forecasting tool

The orientation of crustal seismic anisotropy changed at least twice by up to 80 degrees because of volcanic eruptions at Ruapehu Volcano, New Zealand. These changes provide the basis for a new monitoring technique and possibly for future midterm eruption forecasting at volcanoes. The fast anisotropic direction was measured during three seismometer deployments in 1994, 1998, and 2002, providing an in situ measurement of the stress in the crust under the volcano. The stress direction changed because of an eruption in 1995-1996. Our 2002 measurements revealed a partial return to the pre-eruption stress state. These changes were probably caused by repeated filling and depressurizing of a magmatic dike system.