Analysis of gene expression patterns by microarray hybridization in blood mononuclear cells of SLA-DRB1 defined Canadian Yorkshire pigs

<p>Abstract</p> <p>Background</p> <p>The Swine Leukocyte Antigen (SLA) system encodes molecules for self-nonself discrimination and is associated with immune responses and disease resistance. Three lines of pigs defined by their SLA-DRB1 alleles were developed at the University of Guelph for xenotransplantation and immune response studies. The aim of this project was to explore the potential association between defined SLA-DRB1 alleles and gene transcriptional patterns of other immune-related genes in blood mononuclear cells.</p> <p>Findings</p> <p>Three SLA-DRB1 alleles were characterized using a RT-PCR-based sequencing method. The loci represented included a new allele, DRB1*04ns01. Next, microarray heterologous (bovine-porcine) hybridization together with qPCR were used to explore differential gene expression between SLA-DRB1-defined groups. Microarray analysis showed significant (p < 0.01) differential expression for 5 genes, mostly related to inflammation. Genes varied according to the comparison analyzed. Further testing with qPCR revealed the same trend of differential expression for 4 of the genes, although statistical significance was reached for only one.</p> <p>Conclusion</p> <p>A new SLA-DRB1 allele was characterized. A potential association was found between SLA-DRB1 alleles and inflammation-related genes. However, the influence of other genes cannot be ruled out. These preliminary findings agree with other studies linking MHC haplotypes and inflammation processes, including autoimmune disease. The study provides an initial view of the biological interactions between the SLA complex and other immune-related genes. Future studies will focus on characterization of SLA-haplotypes associated with these particular alleles and the dynamics of the immune response to antigenic challenges.</p

Qualitative investigation of meat species in meat products by real time polymerase chain reaction

Identification of meat species in processed meat products is a major problem in meat technology to produce safe and standard meat products. The aim of this study was identification of meat species in meat processed products including sausages and burgers. The study was carried out by targeting a conserved region of mitochondrial DNA and by using Real-Time polymerase chain reaction (RT-PCR) method. Sampling procedure consisted of different types of commercial meat products including susages, burgers, and salamis which were collected from different stores in Qazvin province-Iran. After DNA extraction, RT-PCR assay was performed to detect specific DNA sequences of animal species in meat product samples. Specific DNA sequences for chicken, horse, camel, beef, and turkey meat were detected successfully in collected samples. Consequently, we found in this study that RT-PCR method is a very powerful and easy method for monitoring commercial meat products based on conserved region of mitochondrial DNA. Practical applications Recently, adulteration in formulation of meat products such as burger and sausages is the principle problem in this industry. Meat species used in this category of products are varied according to type of adulteration and final product. Separately, safety of meat products is dependent on the formulation and the raw materials including meat species used for manufacturing of formulated meat products. In this study, identification of meat species in burger and sausage samples collected from different local stores in Qazvin Province, Iran was performed by Real-Time PCR. This molecular method is a very practical, rapid, precise, sensitive, and useful method for identification of meat species in these products. Chicken, horse, camel, beef, and turkey meat were detected successfully in the collected samples. Also, the quality and adulteration of the collected meat product samples have been determined for the future development of health and quality strategies and investigations

Comparison of real-time PCR and cultural method for detection of bacterial load in pasteurized milk

In this study, we used a TaqMan® probes through the real-time polymerase chain reaction (PCR) technique as a rapid and sensitive way to detect bacterial load of pasteurized milk. The reaction was optimized for enumeration of bacteria in pasteurized milk samples. In parallel, the same milk samples were assessed by conventional cultural-based method. The correlation between methods was evaluated by Bland Altman analysis. The minimum and maximum value for conventional culture-based method were 0 and 10,000 cfu/ml while qPCR gave us 55 and 7,071 (Bacteria/ml of pasteurized milk), respectively. Results indicated that Taq Man real-time PCR provides a useful tool for rapid and accurate quantification of bacterial load

Discrimination between some Mycoplasma spp. and Acholeplasma laidlawii in bovine milk using high resolution melting curve analysis

Abstract Objectives This study aimed to provide a rapid, accurate and cost-effective diagnostic real time polymerase chain reaction-high resolution melting curve assay (PCR-HRM) to identify and distinguish between four different mycoplasmas and Acholeplasma laidlawii isolated at cow-level from a single commercial dairy farm in South Australia. One set of genus-level universal primers was designed targeting the 16S ribosomal RNA gene. Results Real time PCR-HRM analysis was able to identify and distinguish between five different mollicutes, namely A. laidlawii, M. arginini, M. bovirhinis, M. bovis and uncultured Mycoplasma. Results were confirmed through sequencing. Our developed assay provides rapid and accurate screening for Mycoplasma mastitis detection

Influence of Two Various Durations of Resistance Exercise on Oxidative Stress in the Male Rat’s Hearts

Introduction: The previous studies have suggested that alteration in oxidative stress and antioxidant defense depends on various factors, such as mode, intensity, frequency and duration of exercise. In this study, we compared the effects of two various durations of resistance exercise (1 month and 4 month) on oxidative stress and antioxidant status in cardiac tissue. Methods: Thirty Wistar male rats divided into 3 groups: control (sedentary), exercise-1 (regular exercise for 1 month) and exercise-2 group (regular exercise for 4 months). After the final to the experiment, the rats were anesthetized, and then blood and heart samples were obtained and used to determine glutathione peroxidase (GPX), superoxide dismutase (SOD), malondialdehyde (MDA) and biochemical estimation. Results: MDA levels between control and exercise-2 groups showed no significant difference, hence, MDA level in exercise-1 group was higher compared to control group (P<.01). The heart GPX activity increased significantly in exercise-2 group regarding other groups (P<.01). The SOD activities of groups were similar. Creatine kinase (CK) and lactate dehydrogenase (LDH) concentrations increased in the exercise-1 compared to the other groups (P<.01). Conclusion: Our results indicate that in heart, the adaptation and alteration in oxidative stress and cell injury level depend on duration of exercise

Analysis of gene expression patterns by microarray hybridization in blood mononuclear cells of SLA-DRB1 defined Canadian Yorkshire pigs-0

numbers) and *04ns01 (new allele). The black arrows mark (a) the point mutation and (b) the corresponding amino acid residue substitution.<p><b>Copyright information:</b></p><p>Taken from "Analysis of gene expression patterns by microarray hybridization in blood mononuclear cells of SLA-DRB1 defined Canadian Yorkshire pigs"</p><p>http://www.biomedcentral.com/1756-0500/1/31</p><p>BMC Research Notes 2008;1():31-31.</p><p>Published online 23 Jun 2008</p><p>PMCID:PMC2529311.</p><p></p

Evaluation of three cryoprotectants used with bovine milk affected with Mycoplasma bovis in different freezing conditions

Abstract Objectives Currently, there is no consensus protocols regarding the combination of glycerol (GLY), gelatin or foetal bovine serum (FBS) with dimethyl sulphoxide (DMSO) as cryoprotectants for Mycoplasma bovis in bovine milk samples. This study aimed to compare different cryopreservation compounds and storage temperatures for M. bovis. Results There were significant differences in the survival of M. bovis on different media. Differences were also observed between different storage conditions. All additives improved the survival of M. bovis in comparison to control (CON). The combination of GLY and DMSO was shown to be significantly different to CON with 57.1% (95% CI = 21.43–133.34) and 19.1% (95% CI = 11.73–60.27), respectively at week 16, and its use should be encouraged as a cryoprotectant for M. bovis at − 20 and − 80 °C. GEL/DMSO showed the highest survival rate for M. bovis with 57.14% (95% CI = 21.43–133.34) at 4 °C in comparison with CON 14.29% (95% CI = 9.60–50.39). FBS/DMSO showed the highest survival rate for the short-term preservation similarly to other additives. The evaluated cryopreservative compounds would improve survivability of M. bovis in milk for both transport and long-term storage. Hence, it is recommended to use the mentioned methods for routine transportation or storage purposes for suspicious M. bovis milk samples

Analysis of gene expression patterns by microarray hybridization in blood mononuclear cells of SLA-DRB1 defined Canadian Yorkshire pigs-1

Up) and qPCR (*0502, n = 9; *04ns01, n = 14) results for the comparison between SLA-DRB1*0502 and *04ns01. b) Microarray (n = 2 per group) and qPCR (*0502, n = 9; *0701, n = 6) results for the comparison between SLA-DRB1*0502 and *0701 alleles. c) Microarray (n = 2 per group) and qPCR (*0701, n = 6; *04ns01, n = 14) results for the comparison between SLA-DRB1*0701 and *04ns01.<p><b>Copyright information:</b></p><p>Taken from "Analysis of gene expression patterns by microarray hybridization in blood mononuclear cells of SLA-DRB1 defined Canadian Yorkshire pigs"</p><p>http://www.biomedcentral.com/1756-0500/1/31</p><p>BMC Research Notes 2008;1():31-31.</p><p>Published online 23 Jun 2008</p><p>PMCID:PMC2529311.</p><p></p

Diagnostic Challenges in Canine Parvovirus 2c in Vaccine Failure Cases

In this study, three different diagnostic tests for parvovirus were compared with vaccination status and parvovirus genotype in suspected canine parvovirus cases. Faecal samples from vaccinated (N17) and unvaccinated or unknown vaccination status (N41) dogs that had clinical signs of parvovirus infection were tested using three different assays of antigen tests, conventional and quantitative PCR tests. The genotype of each sample was determined by sequencing. In addition to the suspected parvovirus samples, 21 faecal samples from apparently healthy dogs were tested in three diagnostic tests to evaluate the sensitivity and specificity of the tests. The antigen test was positive in 41.2% of vaccinated dogs and 73.2% of unvaccinated diseased dogs. Conventional PCR and qPCR were positive for canine parvovirus (CPV) in 82.4% of vaccinated dogs and 92.7% of unvaccinated dogs. CPV type-2c (CPV-2c) was detected in 82.75% of dogs (12 vaccinated and 36 unvaccinated dogs), CPV-2b was detected in 5.17% dogs (one vaccinated and two unvaccinated) and CPV-2a in 1.72% vaccinated dog. Mean Ct values in qPCR for vaccinated dogs were higher than the unvaccinated dogs (p = 0.049), suggesting that vaccinated dogs shed less virus, even in clinical forms of CPV. CPV-2c was the dominant subtype infecting dogs in both vaccinated and unvaccinated cases. Faecal antigen testing failed to identify a substantial proportion of CPV-2c infected dogs, likely due to low sensitivity. The faecal samples from apparently healthy dogs (n = 21) showed negative results in all three tests. Negative CPV faecal antigen results should be viewed with caution until they are confirmed by molecular methods