Specified Risk Material Research Articles

Development of Proteinaceous Plywood Adhesive and Optimization of Its Lap Shear Strength

AbstractThe primary goal of this research was to develop a waste protein based adhesive, and to optimize its lap shear strength under dry and wet conditions using Taguchi method. The feedstock for this work was specified risk material (SRM) that is considered the tissue potentially at risk of containing misfolded prions. SRM was hydrolyzed in accordance with government approved techniques and modified with a resin system to develop a renewable adhesive platform. Eight of the nine formulated adhesives met the minimum dry shear strength requirement of urea formaldehyde type adhesive, and three of the formulations also met the soak shear strength requirement. This work demonstrates a general valorization platform for the utilization of animal waste protein as a renewable adhesive feedstock.

Inactivation of infectious prions in the environment: a mini-review

Infectious prions (PrPTSE) contribute to horizontal transmission of transmissible spongiform encephalopathies. Specified risk material (SRM) wastes and wastewater generated from slaughterhouses and rendering plants represent a potential reservoir of PrPTSE. Approved disposal practices for SRM that ensure destruction of PrPTSE, including thermal or alkaline hydrolysis at 150°C–180°C under 4–12 atmospheres and incineration, gasification or combustion at 1000°C, are expensive and challenging to implement in commercial slaughter facilities. Consequently, a large portion of the SRM in Canada is rendered and buried in landfills, whereas they are incinerated at considerable cost in Europe. Alternatives for the disposal of these wastes have been proposed and include anaerobic digestion and composting. These biodegradative processes offer advantages in that they are less energy intensive, CH4 can be used in cogeneration and the end products used as fertiliser. In addition, advanced ozonation of the liquid fraction of SRM may allow its direct release into wastewater streams. For these practices to be adopted, assessment of the extent of PrPTSE inactivation during these processes is required. Moreover, multi-barrier use of these approaches (e.g., anaerobic digestion + composting or anaerobic digestion + ozone) may act synergistically to achieve an acceptable level of PrPTSE inactivation.

Biodegradation of prions in compost.

Composting may serve as a practical and economical means of disposing of specified risk materials (SRM) or animal mortalities potentially infected with prion diseases (transmissible spongiform encephalopathies, TSE). Our study investigated the degradation of prions associated with scrapie (PrP(263K)), chronic waste disease (PrP(CWD)), and bovine spongiform encephalopathy (PrP(BSE)) in lab-scale composters and PrP(263K) in field-scale compost piles. Western blotting (WB) indicated that PrP(263K), PrP(CWD), and PrP(BSE) were reduced by at least 2 log10, 1-2 log10, and 1 log10 after 28 days of lab-scale composting, respectively. Further analysis using protein misfolding cyclic amplification (PMCA) confirmed a reduction of 2 log10 in PrP(263K) and 3 log10 in PrP(CWD). Enrichment for proteolytic microorganisms through the addition of feather keratin to compost enhanced degradation of PrP(263K) and PrP(CWD). For field-scale composting, stainless steel beads coated with PrP(263K) were exposed to compost conditions and removed periodically for bioassays in Syrian hamsters. After 230 days of composting, only one in five hamsters succumbed to TSE disease, suggesting at least a 4.8 log10 reduction in PrP(263K) infectivity. Our findings show that composting reduces PrP(TSE), resulting in one 50% infectious dose (ID50) remaining in every 5600 kg of final compost for land application. With these considerations, composting may be a viable method for SRM disposal.

Subcritical hydrolysis and characterization of waste proteinaceous biomass for value added applications

AbstractBACKGROUNDAs a result of the bovine spongiform encephalopathy (BSE) emergence, certain tissues of cattle are categorized as specified risk material (SRM) and completely banned from their traditional applications as an ingredient in animal feed, pet food, or fertilizer applications. The goal of this study was to investigate the hydrolysis of such hazardous proteinacious biomass and extract a safe proteinacious fraction to produce industrial feedstock for value‐added applications.RESULTSThe SRM was hydrolyzed at subcritical temperatures of 180, 200, 220, 240, and 260 °C for 40 min according to government‐approved protocols. The recovery and cleavage of proteins and lipids, the generation and degradation of free and total amino acids, and the generation of organic acids were studied and found to be temperature‐dependent. The yield of the hydrolyzed and recovered proteinacious fractions varied between 71.6 and 87.6% by weight of the original protein content in the SRM. The molecular size was also significantly dependent on the hydrolysis temperature.CONCLUSIONThe valorization of an otherwise waste SRM by subcritical hydrolysis and recovery techniques resulted in protein hydrolyzates suitable for bio‐based chemicals and materials applications. The increase of temperature resulted in higher degrees of hydrolysis as shown by the smaller molecular size of hydrolyzed proteins. © 2014 Society of Chemical Industry

Thermosetting Proteinaceous Plastics from Hydrolyzed Specified Risk Material

AbstractIn response to the bovine spongiform encephalopathy (BSE) outbreak in the last decade, the US, Canada and several other countries implemented an enhanced feed ban that eliminates specified risk material (SRM) from all animal feed, pet foods and fertilizer applications. The environmental risks and severe loss of profitability by the rendering industry associated with landfilling of several million tons of SRM demand immediate and economically viable solutions of converting such materials into value added applications. In this research, SRM was hydrolyzed, the protein hydrolyzates were extracted and further modified through chemical crosslinking to develop a novel protein‐based plastics. The plastics developed in this research exhibited promising thermal and solvent resistance, and tensile strength.

Recovery and characterization of proteinacious material recovered from thermal and alkaline hydrolyzed specified risk materials

The purpose of this study was to characterize proteinacious material recovered from cattle tissues known as specified risk materials (SRM) hydrolyzed according to two protocols approved by the Canadian Food Inspection Agency and Food and Drug Administration and to evaluate its potential for non-food/feed value-added applications. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), size exclusion high performance liquid chromatography (SEC-HPLC), and free amino acid determination methods were adopted to measure the extent of protein cleavage and the average molecular weight of hydrolyzates, and to identify the reactive functionalities available. Molecular weight analysis revealed that the alkaline hydrolysis protocol cleaved the proteins more severely than thermal hydrolysis. Both protocols narrowed the molecular weight distribution of the proteins and increased their solubility in water. Several active functional groups amenable to further chemical processing were identified in the protein hydrolyzates. In conclusion, this work demonstrated that both hydrolytic protocols can be adopted as pre-processing steps to produce safe and renewable proteinacious feedstock for industrial applications.

Microbial communities and greenhouse gas emissions associated with the biodegradation of specified risk material in compost

Provided that infectious prions (PrPSc) are inactivated, composting of specified risk material (SRM) may be a viable alternative to rendering and landfilling. In this study, bacterial and fungal communities as well as greenhouse gas emissions associated with the degradation of SRM were examined in laboratory composters over two 14day composting cycles. Chicken feathers were mixed into compost to enrich for microbial communities involved in the degradation of keratin and other recalcitrant proteins such as prions. Feathers altered the composition of bacterial and fungal communities primarily during the first cycle. The bacterial genera Saccharomonospora, Thermobifida, Thermoactinomycetaceae, Thiohalospira, Pseudomonas, Actinomadura, and Enterobacter, and the fungal genera Dothideomycetes, Cladosporium, Chaetomium, and Trichaptum were identified as candidates involved in SRM degradation. Feathers increased (P<0.05) headspace concentrations of CH4 primarily during the early stages of the first cycle and N2O during the second. Although inclusion of feathers in compost increases greenhouse gas emissions, it may promote the establishment of microbial communities that are more adept at degrading SRM and recalcitrant proteins such as keratin and PrPSc.

Biodegradation of specified risk material and fate of scrapie prions in compost

Composting may be a viable alternative to rendering and land filling for the disposal of specified risk material (SRM) provided that infectious prion proteins (PrPTSE) are inactivated. This study investigated the degradation of SRM and the fate of scrapie prions (PrPSc) over 28 days in laboratory-scale composters, with and without feathers in the compost matrices. Compost was mixed at day 14 to generate a second heating cycle, with temperatures exceeding 65°C in the first cycle and 50°C in the second cycle. Approximately 63% and 77% of SRM was degraded after the first and second cycles, respectively. Inclusion of feathers in the compost matrices did not alter compost properties during composting other than increasing (P < 0.05) total nitrogen and reducing (P < 0.05) the C/N ratio. However, addition of feathers enhanced (P < 0.05) SRM degradation by 10% upon completion of experiment. Scrapie brain homogenates were spiked into manure at the start of composting and extracted using sodium dodecyl sulphate followed by detection using Western blotting (WB). Prior to composting, PrPSc was detectable in manure with 1–2 log10 sensitivity, but was not observable after 14 or 28 days of composting. This may have been due to either biological degradation of PrPSc or the formation of complexes with compost components that precluded its detection.

Biocomposites from hydrolyzed waste proteinaceous biomass: mechanical, thermal and moisture absorption performances

The main goal of this research is maximizing the utilization of renewable materials in both the matrix and reinforcement, more importantly exploitation of waste material for biocomposite development and evaluating the mechanical, thermal and water resistance performances. Woven roving and chopped strand mat fiber glass, and hemp fiber mats are incorporated to an epoxy resin based matrix cured with novel hydrolyzed specified risk material (SRM) extracts. Aminophenyl sulphone (APS) is used as a control crosslinking agent for the epoxy resin. Results show that the biocomposites developed in this research exhibit promising flexural strength, tensile strength and tensile modulus; despite relatively poor moisture resistance. The use of waste protein hydrolyzate extracts, hydrolyzed proteins, as crosslinking agent of epoxy resins in making biocomposites is novel and promising and results can be extended to other proteinaceous biomasses as curing agent of epoxy resins.

A Quantitative Assessment of the Amount of Prion Diverted to Category 1 Materials and Wastewater During Processing

In this article the development and parameterization of a quantitative assessment is described that estimates the amount of TSE infectivity that is present in a whole animal carcass (bovine spongiform encephalopathy [BSE] for cattle and classical/atypical scrapie for sheep and lambs) and the amounts that subsequently fall to the floor during processing at facilities that handle specified risk material (SRM). BSE in cattle was found to contain the most oral doses, with a mean of 9864 BO ID50 s (310, 38840) in a whole carcass compared to a mean of 1851 OO ID50 s (600, 4070) and 614 OO ID50 s (155, 1509) for a sheep infected with classical and atypical scrapie, respectively. Lambs contained the least infectivity with a mean of 251 OO ID50 s (83, 548) for classical scrapie and 1 OO ID50 s (0.2, 2) for atypical scrapie. The highest amounts of infectivity falling to the floor and entering the drains from slaughtering a whole carcass at SRM facilities were found to be from cattle infected with BSE at rendering and large incineration facilities with 7.4 BO ID50 s (0.1, 29), intermediate plants and small incinerators with a mean of 4.5 BO ID50 s (0.1, 18), and collection centers, 3.6 BO ID50 s (0.1, 14). The lowest amounts entering drains are from lambs infected with classical and atypical scrapie at intermediate plants and atypical scrapie at collection centers with a mean of 3 × 10(-7) OO ID50 s (2 × 10(-8) , 1 × 10(-6) ) per carcass. The results of this model provide key inputs for the model in the companion paper published here.

Specified risk material removal practices: Can we reduce the BSE hazard to human health?

Following the bovine spongiform encephalopathy (BSE) epidemics across Europe in the early 1990s, the removal of designated BSE specified risk material (SRM) became mandatory to minimize the risk of consumer's exposure to the infectious agent. Despite this preventive measure, cross-contamination of edible meat with SRM can occur during conventional slaughter, primarily by carcass splitting, captive bolt stunning, and head removal. BSE-affected subjects accumulate the pathological feature in the central nervous system (CNS). Currently, there are no markers that can identify the presence of SRM in meat as a whole. Nevertheless, some assays are able to detect traces of CNS, hence this parameter is officially taken as indicative of SRM contamination.In this two-stage study, we carried out a survey to estimate the prevalence of carcass contamination at two slaughterhouses, one large and the other medium-sized; we then compared three different methods (conventional vs. suction vs. water-jet) for spinal cord removal employed at the large slaughterhouse to assess their performance in preventing the spread of central nervous tissue (CNT) over the carcass.The prevalence of carcass contamination was 68.4%. Compared to the other two spinal cord removal techniques (conventional and suction), the water-jet system was associated with less CNS contamination (62 vs. 60 vs. 36%, respectively; P = 0.0047).

Evaluation of an aerobic composting process for the management of Specified Risk Materials (SRM)

In Nova Scotia (NS), approximately 2700 tonnes of Specified Risk Materials (SRM) are produced annually. SRM disposal is a serious concern for abattoirs and the beef industry. Composting offers a low risk and simple means to transform raw SRM into a more stable and easily managed material. In this project, wheat straw and sawdust were used to compost with SRM on a pilot scale. The study evaluated changes over time in total carbon, total nitrogen, pH, temperature, moisture content and electrical conductivity. Compost temperatures in all treatments met the Canadian Council of Ministers of the Environment (CCME) guidelines for pathogen kill. The compost maturity tests showed that the evolution of CO2–C in all the final compost products was less than 1mgg−1 organic matter day−1. Wheat straw performed well as a composting feedstock for raw SRM as sawdust. While the wheat straw has advantages including greater availability, lower cost and easily decomposable carbon compounds more management is required to maintain adequate compost temperatures. The influences of seasonal variations due to temperate climatic conditions on SRM composting were also studied with wheat straw. The results suggest no significant differences in composting effectiveness between the two seasons.

Inactivation of Template-Directed Misfolding of Infectious Prion Protein by Ozone

Misfolded prions (PrP(Sc)) are well known for their resistance to conventional decontamination processes. The potential risk of contamination of the water environment, as a result of disposal of specified risk materials (SRM), has raised public concerns. Ozone is commonly utilized in the water industry for inactivation of microbial contaminants and was tested in this study for its ability to inactivate prions (263K hamster scrapie = PrP(Sc)). Treatment variables included initial ozone dose (7.6 to 25.7 mg/liter), contact time (5 s and 5 min), temperature (4°C and 20°C), and pH (pH 4.4, 6.0, and 8.0). Exposure of dilute suspensions of the infected 263K hamster brain homogenates (IBH) (0.01%) to ozone resulted in the in vitro destruction of the templating properties of PrP(Sc), as measured by the protein misfolding cyclic amplification (PMCA) assay. The highest levels of prion inactivation (≥4 log(10)) were observed with ozone doses of 13.0 mg/liter, at pH 4.4 and 20°C, resulting in a CT (the product of residual ozone concentration and contact time) value as low as 0.59 mg · liter(-1) min. A comparison of ozone CT requirements among various pathogens suggests that prions are more susceptible to ozone degradation than some model bacteria and protozoa and that ozone treatment may be an effective solution for inactivating prions in water and wastewater.

대중매체 영향에 따른 일부 대학생의 식품안전 지식 조사

This study was conducted to assess food safety knowledge of college students according to mass-media reports. To accomplish this, a survey was conducted to investigate the reliability, of primary mass-media sources on Bovine Spongiform Encephalopathy(BSE) as well as the preferences of college students. The most common sources of media consumed by male and female students were terrestrial television(TV)(60%), the internet(21%), and newspapers(11%) and terrestrial TV(60%), newspapers (18%), and the internet(17%), respectively. The preferences for media of major and non-major students were terrestrial TV(64%), newspapers(17%), and the internet(14%) and terrestrial TV(51%), the internet(22%), and newspapers(19%), respectively. The reliability of media as viewed by male students were as follows: terrestrial TV(53%) and the internet(40%), whereas for female students, these values were terrestrial TV(55%) and the internet(37%). The reliability of media as viewed by major and non-major students was as follows: terrestrial TV(67%) and the internet(28%), and terrestrial TV(44%) and the internet(44%), respectively. In the case of male students, the primary sources for information regarding BSE were terrestrial TV(54%) and the internet(38%), whereas for female students, the primary sources were terrestrial TV(57%) and internet (37%). In the case of major students, the primary sources for information regarding BSE were terrestrial TV(52%) and the Internet(40%), and for non-major students, the internet(47%) and terrestrial TV(39%). Both male and female students were found to have less knowledge regarding prions, SRM(Specified Risk Materials), MM(methionine-methionine) type genes, and cow above 30 months associated with BSE compared to the other factors associated with this disease. The BSE-related knowledge held by major and non-major students was revealed unknown more than the majority of responses. Based on these results, greater effort should be made to provide meaningful information to improve the food-safety knowledge of college students according to mass-media.

Biodegradation of specified risk material and characterization of actinobacterial communities in laboratory-scale composters

As a result of bovine spongiform encephalopathy in Canada, specific tissues at risk of harbouring prions are not allowed to enter the food chain. Composting may be a viable alternative to rendering and land filling for the disposal of specified risk material (SRM). Two types of laboratory-scale composters, actively-heated and ambient systems were constructed to assess the biodegradation of SRM over 30days. A second heating cycle was generated by mixing the compost after 15days. Compared to ambient composters, temperature profiles in actively-heated composters were above 50°C for 5 and 4days longer in the first and second composting cycles, respectively. Degradation of SRM was similar between two composter types during two composting cycles, averaging 52.2% in the first cycle and 43.9% in second cycle. Denaturing gradient gel electrophoresis (DGGE) revealed that changes in the actinobacteria populations in the first composting cycle were of a temporal nature, whereas alterations in populations in the second composting cycle were more related to active heating of compost. Sequencing of the dominant DGGE bands showed the predominance of Corynebacterium, Promicromonospora, Pseudonocardia, and Thermobifida in the first composting cycle and Corynebacterium, Mycobacterium, Nocardia, Saccharomonospora, and Streptomyces in the second composting cycle. Active heating can alter the nature of actinobacteria populations in compost, but does not appear to have a major impact on the extent of degradation of SRM.

BSE infectivity in jejunum, ileum and ileocaecal junction of incubating cattle

To establish bovine spongiform encephalopathy (BSE) public health protection measures it is important to precisely define the cattle tissues considered as specified risk materials (SRM). To date, in pre-clinical BSE infected cattle, no evidence of the BSE agent had been found in the gut outside of the ileal Peyer's Patches. This study was undertaken to determine when and where the pathological prion protein (PrPSc) and/or BSE infectivity can be found in the small intestine of cattle 4 to 6 months of age, orally challenged with BSE. Samples of the jejunum, the ileum and the ileocaecal junction from 46 BSE infected cattle, culled from 1 up to 44 months post infection (mpi) were examined by immunohistochemistry. Samples from cattle 8 mpi to 20 mpi were additionally studied by PTA Western blot, rapid tests, and by mouse (TgbovXV) bioassay. In doing so nearly all of the cattle, from 4 up to 44 mpi, had detectable amounts of PrPSc and/or infectivity in the distal ileum. In the distal ileum clear time-dependent variations were visible concerning the amount of PrPSc, the tissue structures affected, and the cells involved. BSE infectivity was found not only in the ileum and ileocaecal junction but also in the jejunum. The systematic approach of this study provides new data for qualitative and quantitative risk assessments and allows defining bovine SRM more precisely.

Scientific Opinion on BSE/TSE infectivity in small ruminant tissues

The objectives addressed were i) to provide an update on TSE (Transmissible Spongiform Encephalopathy) infectivity distribution in small ruminant tissues; and ii) to indicate based on the current epidemiological situation as regards to BSE (Bovine Spongiform Encephalopathy) in the small ruminant population in the EU (European Union), whether a review of the existing SRM (Specified Risk Materials) list for small ruminants should be envisaged with regard to the potential exposure to the BSE agent. The appraisal was addressed by reviewing for Classical scrapie, BSE and Atypical scrapie in small ruminants aspects related to: i) tissue infectivity distribution according to the age and the genotype of sheep and goats; and ii) the infectious load in the different tissues. In order to perform the assessment all the currently available scientific results were reviewed, and data on TSE monitoring in small ruminants in the EU and on small ruminants slaughtered by species and age category in each EU Member State were considered. The reduction of the infectivity associated to the carcass of an infected individual achieved by the current SRM policy in small ruminants for Classical scrapie and BSE was estimated. The total number of Classical scrapie infected sheep and goats that could enter yearly into the food chain was provided. Moreover, considerations about Atypical scrapie were given. A set of simulations allowing estimating the impact of different policy options on the BSE infectious load potentially present in an infected sheep was provided.

GC/MS detection of central nervous tissues as specified BSE risk material in meat products: validation by an externally controlled blind trial

The addition of central nervous tissues (CNT), such as brain and spinal cord, in the manufacturing of meat products is either forbidden--if the material falls under the legal definition of specified risk material (SRM)--or must be labelled on the packed product. To foster official food control, several CNT detection methods were developed, but only fatty acid patterns as detected by gas chromatography/mass spectrometry (GC/MS) allow the further characterization of the detected CNT as to both the animal species and--surprisingly--the age of the animal from which the CNT was derived in accordance with the legal definition. Complementing a previous report in this journal by Lücker et al. 2010 (doi: 10.1007/s00216-010-3956-5) on CNT quantification by GC/MS, we now report results of the validation of this new analytical approach by an externally controlled blind trial elucidating its potential to identify the species and age of the CNT detected. The 72 samples (24 standards of emulsion-type sausage, each heated in three different batches: 75°C, 30 min; 115°C, 25 min; 133°C, 40 min) containing porcine, ovine or bovine muscle tissue and differing amounts of CNT (bovine or ovine brain: 0, 0.1, 0.5, 1.0, 3.0% m/m) were produced externally and provided blind for analyses to our laboratory. In accordance with the previous study, heating had no detectable effect on the GC/MS analysis. Judged by the present sensitivity of this method (cut-off 0.2% CNT), all of the samples containing 0.5% or more CNT (n = 57, 100%) were identified correctly as CNT-positive. The CNT species was identified correctly in 54 samples (94.7%), with three samples of one specific standard (0.5% ovine CNT) falsely classified as bovine CNT. However, the CNT age of these samples was correctly classified (more than 12 months). Overall, 57 samples (100%) were correctly classified as SRM-positive and 15 samples (100%) as SRM-negative. To the best of our knowledge, this is the first time that a legal demand for the (1) detection of traces of a specific tissue in a food matrix, (2) the identification of its taxonomic origin and (3) the classification of its age has been shown to be analytically possible in totally blind samples. The very positive validation results of this externally controlled blind trial recommend the present GC/MS approach for the detection of CNT in meat products as a reference method. However, our results also demonstrate the need for further studies, in particular to increase sensitivity and to conduct ring trials including more than one laboratory.

Assessment of Microbial Communities In Decomposition of Specified Risk Material Using a Passively Aerated Laboratory-Scale Composter

The occurrence of bovine spongiform encephalopathy (BSE) in Canada has resulted in the implementation of regulations to remove specified risk material (SRM) from the food chain. SRM includes the distal ileum of all cattle, and the skull, brain, trigeminal ganglia, eyes, palatine tonsils, and spinal cord and dorsal root ganglia of cattle ≥30 months of age. Composting may be a viable alternative to rendering for SRM disposal. In our study, two bulking agents, barley straw and wood shavings, were composted with beef manure along with SRM in passively aerated, laboratory-scale composters. Both composts heated rapidly, exceeding 55°C after 3 days with oxygen declining in the early composting stage with wood-shaving compost, but returning to near-original levels after 5 days. During composting the two matrices differed (P <0.05) only in water content, TC and bulk density. In the final compost, water content, TC and C/N ratio were higher (P < 0.05), while EC was lower (P < 0.05) in the wood shavings as compared to the straw compost. Approximately 50% of SRM was decomposed after 15 days of composting, with 30% of SRM being decomposed within the first 5 days. Phospholipid fatty acid (PLFA) profiles were used to characterize the microbial communities and showed that Gram positive bacteria were predominant in compost at day 5, a point that coincided with a rapid increase in temperature. Gram negative bacteria and anaerobes declined at day 5 but populations recovered by day 15. Fungi appeared to be suppressed as temperatures exceeded 55°C and did not appear to recover over the remainder of the composting period, with the exception of the straw compost at day 15. On day 5, Actinomycetes increased in the straw compost, but declined in the wood shavings compost, with this group increasing in both types of compost at day 15. Although temporal changes were evident, compost matrices or depth within the composter did not obviously influence microbial communities. Decomposition of SRM also did not differ between compost matrices or with depth in the composters. These results suggest that SRM decompose rapidly during composting and that both mesophilic and thermophilic microbial communities play a role in its decomposition.

Anaerobic digestion of specified risk materials with cattle manure for biogas production

Biogas production from anaerobic digestion (AD) of specified risk materials (SRM) co-digested with cattle manure was assessed in a 3×2 factorial design. SRM replaced manure at 0 (control), 10% or 25% (w/w) as the substrate fed to six 2-L biodigesters maintained at 37°C or 55°C. Digesters were fed substrate (30gL−1 total volatile solids) at 6-d intervals for 90d, with a retention time of 30d. Keratin (<20mg) was added to each digester to model the degradation of β-sheet rich proteins. Methane production was measured daily, and effluent was collected at feeding to monitor SRM degradation using real-time PCR analysis of bovine-specific DNA fragments. Compared with control, methane production increased by 83% or 161% (P<0.05) with 10% or 25% SRM at 37°C, and by 45% and 87%, respectively, at 55°C (P<0.05). Bovine DNA degradation over 6d was higher (P<0.05) at 37°C as compared to 55°C. Dry matter degradation of keratin at 37°C decreased with increasing SRM concentration (P<0.05), whereas at 55°C no difference between treatments was observed (P>0.05). Inclusion of SRM increases the production of methane during the anaerobic digestion of manure and may offer a means of deriving economic value from the disposal of SRM.