Detection of Microsatellite Instability by High-Resolution Melting Analysis in Colorectal Cancer

Background: Colorectal cancer is the third most common cancer worldwide. MSI is a molecular marker of a deficient MMR system and happens in almost 15% of CRCs. Because of a wide frequency of MSI⁺ CRC in Iran compared to other parts of the world, the importance of screening for this type of cancer is highlighted. Methods: The most common MSI detection technique is a fluorescent PCR-based method in which fragments are analyzed by CE. This technique is very time-consuming, difficult, and expensive. We sought to develop and evaluate a proper method with high accuracy, specificity, and sensitivity to screen the MSI⁺ CRC. A HRM analysis procedure is relying on the analysis of the melting curve attributes. Low cost, feasibility, high specificity, and sensitivity are outstanding attributes of HRM analysis. Results: Five mononucleotide microsatellite markers, including BAT-25, BAT-26, NR-21, NR-24, and NR-27, in 25 archival CRC tumor tissue samples were compared with normal tissue adjacent using HRM method. The specificity and sensitivity of BAT-25 with HRM method were 100% compared to CE, while other markers had lower sensitivity. However, when all the markers were considered together, the sensitivity and specificity became 100%. The number of MSI⁺ samples was 56%, which shows a higher ratio than previous Iranian studies. The highest MSI was related to BAT-26 (52%). Conclusion: The HRM method is much simpler and more cost-effective than current MSI techniques, and its sensitivity and accuracy are comparable. Therefore, it can serve as an alternative method in cases where CE is unavailable. Key Words: Biomarkers, Microsatellite instability, Colorectal cancer     

In vitro spasmolytic effect of aqueous extract of Calotropis procera on Guinea-pig trachea smooth muscle chain

The aqueous extract of Calotropis procera was evaluated for its spasmolytic effect using in vitro trachea smooth muscle chain of Guinea-pig. The extract (50, 100 and 200 microg/ml) showed a dose-dependent relaxant activity probably exhibited through the direct relaxant action on the smooth muscle.     

Antimicrobial resistance of Salmonella isolated from finishing swine and the environment of 60 Alberta swine farms

The study objective was to describe and evaluate antimicrobial resistance profiles in Salmonella isolated from Alberta swine finishing farms. Salmonella isolates (n = 322) were obtained from 192 fecal and 84 environmental samples of the 60 Salmonella-positive swine finishing farms. Isolates were classified susceptible, intermediate or resistant based on NCCLS guidelines. More than half of the isolates (53.4%) were susceptible to all of the 18 antimicrobials in the testing panel. No resistance was observed to amikacin, amoxicillin/clavulanic acid, cefoxitin, ceftiofur, ceftriaxone, cephalothin, ciprofloxacin, imipenem or nalidixic acid. Less than 1% of isolates were resistant to apramycin, gentamicin and trimethoprim/sulfamethoxazole. Higher frequencies of resistance were observed for chloramphenicol (4.7%), ampicillin (7.8%), kanamycin (11.8%), sulfamethoxazole (21.1%), streptomycin (25.5%) and tetracycline (38.8%). Eleven Salmonella serovars had isolates with resistance to > or =3 antimicrobials. The most frequently resistant serovar was Salmonella Derby, with 27 (38.0%) isolates resistant to > or =3 antimicrobials, including resistance to five and six antimicrobials. An absence of resistance to cephalosporins and fluoroquniolones and a low proportion of isolates resistant to amikacin, amoxicillin/clavulanic acid, apramycin, gentamicin and trimethoprim/sulfamethoxazole are encouraging findings from public health and animal health perspectives. Frequent resistance observed for ampicillin, kanamycin, sulfamethoxazole, streptomycin and tetracycline, antimicrobials commonly used in veterinary medicine for decades, indicates an urgent need to utilize these antimicrobials more prudently if their benefits are to be preserved.     

Normal cross-sectional anatomy of the bovine digit: comparison of computed tomography and limb anatomy

The purpose of this study was to define the structures of the digits and hoof in Holstein dairy cattle by using computed tomography scan (CT scan). Transverse, sagittal and dorsoplantar CT images of two isolated cattle cadaver digits were obtained using a Siemens ARTX2 Somatom. The CT images were compared to corresponding frozen cross-sections. Relevant anatomical structures were identified and labelled at each level. The CT images provided anatomical detail of the digits and hoof in Holstein dairy cattle. Transversal images provided excellent depiction of anatomical structures when compared to corresponding frozen cross-sections. The information presented in this paper would serve as an initial reference to the evaluation of CT images of the digits and hoof in Holstein dairy cattle.     

Prevalence of antimicrobial resistance in fecal generic Escherichia coil isolated in western Canadian cow-calf herds. Part I--beef calves.

The objective of this study was to describe antimicrobial resistance patterns in fecal generic Escherichia coli obtained from calves in western Canadian cow-calf herds. Susceptibility testing was completed on 1677 isolates obtained from 480 beef calves in 91 herds in the spring, and from 1187 isolates obtained from 394 calves in 45 herds in the fall of 2002. Resistance was rare to antimicrobials classified as being of very high importance to human health. Isolates were most commonly resistant to tetracycline, sulphamethoxazole, and streptomycin. Resistance to at least one antimicrobial was identified in 48.8% of the isolates collected in the spring, and 7.0% of those collected in the fall. Spring or fall calf resistance status was not associated with calf gender, breed, age of the dam, or if the calf was treated before sample collection. Calves < or = 3 days of age were less likely to be resistance positive than calves > 10 days of age.     

Effect of selected water temperatures used in Mycoplasma gallisepticum vaccine reconstitution on titer at selected time intervals

Numerous methods are currently used throughout the poultry industry for the administration of vaccines. Each utilizes water for vaccine reconstitution and/or administration, including two of the three commercially available live Mycoplasma gallisepticum (MG) vaccines. Selected water temperatures were used to reconstitute and/or dilute the three commercially available live MG vaccines. Water temperatures included 4 C, 22 C (room temperature), and 32 C, and titer (color change units) was recorded at four time intervals, at point of reconstitution (time 0), 15, 30, and 60 min postreconstitution of the vaccines (time periods 15, 30, and 60, respectively). Results for F strain MG (FMG) vaccine showed significant decreases in titer from time 0 to time 15 for the 22 C and 32 C water temperatures but no significant decrease for any time period for FMG reconstituted with 4 C water. For 6/85 strain MG no significant difference in titer was noted for any of four time periods within any of the three water temperatures. For ts-11 strain MG a significant decrease was observed in titer at each of the four postdilution time periods when diluted with 32 C water. There was no significant decrease in titer at any time period for ts-11 MG vaccine when diluted with either 4 C or 22 C water.     

Magnetic resonance imaging of the normal bovine digit

The purpose of this study was defining the normal structures of the digits and hoof in Holstein dairy cattle using Magnetic Resonance Image (MRI). Transverse, Sagital and Dorsoplantar MRI images of three isolated cattle cadaver digits were obtained using Gyroscan T5-NT a magnet of 0.5 Tesla and T1 Weighted sequence. The MRI images were compared to corresponding frozen cross-sections and dissect specimens of the cadaver digits. Relevant anatomical structures were identified and labeled at each level. The MRI images provided anatomical detail of the digits and hoof in Holstein dairy cattle. Transversal images provided excellent depiction of anatomical structures when compared to corresponding frozen cross-sections. The information presented in this paper would serve as an initial reference to the evaluation of MRI images of the digits and hoof in Holstein dairy cattle, that can be used by radiologist, clinicians, surgeon or for research propose in bovine lameness.     

Erratum to: Antimicrobial resistance and virulence profile of enterococci isolated from poultry and cattle sources in Nigeria

Tropical Animal Health and Production -     

Comparison of bacterial culture and real-time PCR for the detection of Salmonella in grow-finish pigs in Western Canada using a Bayesian approach

The study objective was to evaluate the accuracy of a real-time polymerase chain reaction (RT-PCR) and a culture protocol used to detect Salmonella in the faeces of grow-finish pigs using a Bayesian approach. The RT-PCR was invA-gene-based assay, while the culture protocol included pre-enrichment in buffered peptone water, selective enrichment in tetrathionate and Rappaport-Vassiliadis broths, and isolation on semi-solid (modified semi-solid RV) or solid (XLT4, Rambach) agar plates. Bayesian analysis was performed using a two-test, two-population model with dependence between culture and RT-PCR and compared to a second model with conditional independence between these two tests. Two hundred and ninety three individual faecal and 294 pooled pen samples from grow-finish pig collected from 10 farms were tested and results were divided into two groups according to herd size (five herds <250 sows, five herds with >400 sows). In the dependence model, RT-PCR sensitivity (Se) and specificity (Sp) were estimated to be 90% (95% probability interval 74, 97) and 99% (98, 99), respectively. Culture Se was 92% (75, 99), while culture Sp was considered 100% as all culture-positive samples were confirmed by serotyping. In the conditional independence model, RT-PCR Se and Sp, and culture Se, were 96% (93, 98), 99% (98, 100) and 97% (94, 100), respectively. The dependence model resulted in posterior estimates of Se that were lower and with broader probability intervals than the independence model, indicating that when RT-PCR and culture are evaluated relative to each other, the correlation between these tests is an important source of bias and should be adjusted for during analysis. The RT-PCR evaluated in this study performed almost comparably to culture; given the cost savings associated with using this test and more timely results, the RT-PCR may be a useful alternative to culture for screening large numbers of samples, particularly when Salmonella prevalence is low.