Comparision of staining methods for two dimensional electrophoresis gel resolved with Puntius javanicus liver proteome

The aim of this study was to compare the various staining methods based on commassie briliant blue and silver nitrate stain for the two dimensional gel electrophoresis resolved with Puntius javanicus liver proteome. The staining methods were selected base on the previous reportabout their compatiblitywiththe mass spectrometry analysis. Sliver staining methodis known as the most sensitive method to visualize the maximum number of protein spots resolved in 2D gel but it is less sensitive(incompatible) toward mass spectrometry detection. Results of this study showed that a modifiedstaining method using colloidal coomassie blue G-250 (CCB) is roughly similarly sensitive but lower protein spot detected compared with silver staining (SS) as indicated at the number of 303±26 and 693±14of proteinresolved in both types of stained gels. The conventional methods of staining using commassie brilliant blue G-250 and R-250 only detected less number of protein spots(128±17and 78±11, respectively) compared to modified CCB staining method. As the commasie brilliant blue stain is known to be a very sensitive for mass spectrometry detection, the modified method of CCB was selected for further study on Puntius javanicus liver proteome

Comparision of staining methods for two dimensional electrophoresis gel resolved with Puntius javanicus liver proteome

The aim of this study was to compare the various staining methods based on commassie briliant blue and silver nitrate stain for the two dimensional gel electrophoresis resolved with Puntius javanicus liver proteome. The staining methods were selected base on the previous report about their compatibility with the mass spectrometry analysis. Silver staining method is known as the most sensitive method to visualize the maximum number of protein spots resolved in 2D gel but it is less sensitive(incompatible) toward mass spectrometry detection. Results of this study showed that a modified staining method using colloidal coomassie blue G-250 (CCB) is roughly similarly sensitive but lower protein spot detected compared with silver staining (SS) as indicated at the number of 303±26 and 693±14of protein resolved in both types of stained gels. The conventional methods of staining using commassie brilliant blue G-250 and R-250 only detected less number of protein spots(128±17and 78±11, respectively) compared to modified CCB staining method. As the commasie brilliant blue stain wasis known to be a very sensitive for mass spectrometry detection, the modified method of CCB was selected for further study on Puntius javanicus liver proteome

A proteomic based assessment on changes in myofirbrillar proteins of goat longissimus muscle as affected by heat treatments

The present study examined the effect of different heat treatments; (1) chilled, (2) boiled at 100°C for 30 min, and (3) autoclaved at 121°C at 15 psi for 20 min, on the expression of goat skeletal muscle proteins using two-dimensional gel electrophoresis. The molecular weight (MW) and isoelectric point (pI) of heat stable proteins were characterised followed by identification of the proteins by MALDI-TOF/TOF mass spectrometry. There were 153 protein spots obtained in the boiled samples, while only 46 protein spots were observed in the autoclaved samples. Thirteen spots that exhibited high intensity of protein were chosen from the autoclaved sample for MALDI-TOF/TOF mass spectrometry analysis. The putative heat stable proteins identified were myosin light chain (MLC), actin, tropomyosin (TPM), troponin T (TnT), myoglobin, and creatine kinase. The Proc-GLM analysis revealed that the heat treatments have resulted in significant differences in spot intensities of actin, troponin T (TnT), myoglobin, and creatine kinase with no significant changes noted in other proteins

Differences in thermostable actin profile of goat meat as observed in two-dimensional gel electrophoresis (2DE)

Different heat treatments, (1) chilled, 4°C (2) boiled at 100°C for 30 min and (3) autoclaved at 121°C at 15 psi for 20 min were employed on goat meat to mimic domestic and industrial cooking. The effects on intensity of actin proteins was observed using two-dimensional gel electrophoresis where significant differences (p>0.05) were observed in the spot intensity between chilled and boiled samples, similarly in chilled and autoclaved samples. However, no significant difference was observed between boiled and autoclaved samples. The slight changes observed in the cooking of meat confirmed that actin protein is susceptible to denaturation cause by heat. MALDI-TOF/TOF analysis revealed the peptide-mass fingerprint between positions 21 - 374 that not affected by heat treatment. Peptides from this position merit the candidature of actin as putative thermostable marker for detecting goat meat (chevon) in food product

Myofibrillar protein profile of Pectoralis major muscle in broiler chickens subjected to different freezing and thawing methods

The study examined the protein profile of Pectoralis major muscle in broiler chickens subjected to different freezing and thawing methods. Pectoralis major muscle was excised from the carcasses of twenty broiler chickens and split into left and right halves. The left half was subjected to slow freezing (-20°C) while the right half was rapidly frozen (-80°C). The samples were stored at their respective temperature for 2 weeks and assigned to either of tap water (27°C, 30 min), room temperature (26°C, 60 min), microwave (750W, 10 min) or chiller (4°C, 6 h) thawing. Changes in myofibrillar proteins following the thawing methods were monitored through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The electrophoretic profile indicated differences (p < 0.05) in intensities of the components of myofibrillar proteins among the thawing methods in both slow and rapidly frozen samples. Chiller thawing had significantly higher (p < 0.05) protein concentration than other methods in rapidly frozen samples. However, in slow freezing, there were no significant differences in protein concentration among the thawing methods. In rapidly frozen samples, the protein optical densities at molecular weight of 21, 27, 55 and 151kDa in tap water, chiller and room temperature thawing did not differ (p < 0.05). Similarly, in slowly frozen samples, protein optical densities at molecular weight of 21, 27, 85 and 151 kDa were not significantly different among chill, tap water and room temperature thawing. Microwave thawing consistently caused higher protein degradation resulting in significantly lower (p < 0.05) protein quality and quantity in both freezing methods

Skeletal muscle proteome and meat quality of broiler chickens subjected to gas stunning prior slaughter or slaughtered without stunning

The study examined the effects of pre-slaughter gas stunning and slaughter without stunning on meat quality and skeletal muscle proteome of broiler chickens. Fifty Cobb broiler chickens were randomly assigned to either a neck cut without pre-slaughter stunning (Halal slaughter) or pre-slaughter gas stunning followed by a neck cut. Samples of Pectoralis major muscle at 7 min, 4 h and 24 h postmortem were analyzed for pH, shear force, color, drip and cooking losses. Proteome profile of the 7 min samples was examined by two-dimensional polyacrylamide gel electrophoresis. Birds subjected to Halal slaughter had higher (P < 0.05) redness than those gas stunned at 4 and 24 h postmortem. Gas-stunned birds had lower (P < 0.05) muscle pH and shear force and higher (P < 0.05) drip and cooking losses compared with those subjected to Halal slaughter throughout postmortem storage. Gas stunning up-regulated (P < 0.05) the expression of beta-enolase, pyruvate kinase and creatine kinase compared with Halal slaughter. Results indicate that pre-slaughter gas stunning hastened postmortem energy metabolism and had detrimental effects on the water holding capacity and redness of broiler breast muscles

Transforming agriculture research into commercialisation: experience of Universiti Putra Malaysia

One of the major goals of any high impact research and development is an overall improvement in the well-being and sustainable quality of life through innovations. As universities continuously disseminate innovations from R&D activities, many prototypes and lab-scale products, whether tangible or intangible, can be made available for public use. The success of bringing these innovations to the marketplace depends on the quality and capability of the technology transfer office to lead different types of activities, engagements, negotiation and inclusiveness towards fulfilling the needs of commercialisation partners and the market. This paper presented a general overview of transforming research output into commercialisation in the context of Universiti Putra Malaysia (UPM). Throughout this paper, different commercialization channels, the roles of technology transfer offices and multiple agencies are further discussed with a special focus on agricultural innovations and technologies. This review contributes to both academic and agricultural industry research, development and commercialization activities by illustrating current innovation produced by UPM and industry-university collaboration, conducted at a leading agriculture university

LC–QTOF-MS identification of porcine-specific peptide in heat treated pork identifies candidate markers for meat species determination

The purpose of this study was to identify porcine-specific peptide markers from thermally processed meat that could differentiate pork from beef, chevon and chicken meat. In the initial stage, markers from tryptic digested protein of chilled, boiled and autoclaved pork were identified using LC–QTOF-MS. An MRM method was then established for verification. A thorough investigation of LC–QTOF-MS data showed that only seven porcine-specific peptides were consistently detected. Among these peptides, two were derived from lactate dehydrogenase, one from creatine kinase, and four from serum albumin protein. However, MRM could only detect four peptides (EVTEFAK, LVVITAGAR, FVIER and TVLGNFAAFVQK) that were consistently present in pork samples. In conclusion, meat species determination through a tandem mass spectrometry platform shows high potential in providing scientifically valid and reliable results even at peptide level. Besides, the specificity and selectivity offered by the proteomics approach also provide a robust platform for Halal authentication

The relationship between dietary calcium intakes contain in goats feed with calcium serum concentration in pregnant boer goats

The aim of this research is to identify the relationship present between calcium concentrations in goats’ feed with calcium serum level for pregnant meat goat. The calcium was tested from both samples. There were between feed and blood serum. The biochemical parameters were studied using 40 pregnant does (10 animals in each group). The present study indicates those hematological calcium deficiencies are likely to happen due to low level of calcium concentration content in feed

Development of microbial spoilage and lipid and protein oxidation in rabbit meat

This experiment aimed to determine microbial spoilage and lipid and protein oxidation during aerobic refrigerated (4 °C) storage of rabbit meat. Forty male New Zealand white rabbits were slaughtered according to the Halal slaughter procedure. The hind limbs were used for microbial analysis while the Longissimus lumborum m. was used for determination of lipid and protein oxidation. Bacterial counts generally increased with aging time and the limit for fresh meat (108 cfu/g) was reached at d 7 postmortem. Significant differences in malondialdehyde content were observed after 3 d of storage. The thiol concentration significantly decreased with increase in aging time. The band intensities of myosin heavy chain and troponin T significantly reduced with increased refrigerated storage while actin remained relatively stable. This study thus proposes protein oxidation as a potential deteriorative change in refrigerated rabbit meat along with microbial spoilage and lipid oxidation