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Volatile fatty acids (VFAs) have become attractive and gained high research interest due to its significance for the chemical industry and economical advantage. These acids can be produced by utilizing organic waste such as food waste as substrate through anaerobic digestion. Anaerobic digestion is an environmental process that occurs naturally and produces biogas as the main product. VFAs are intermediate products formed during anaerobic digestion where acetic acid, a type of VFA, is the primary product. The main objective of this study was to utilize acetic acid as carbon and energy source for production of edible fungi, Rhizopus ologisporus, Mucor indicus and Volvariella volvacea. The first step was to evaluate if acetic acid could be used as carbon and energy source for edible fungi production. The results showed, that acetic acid is suitable as carbon and energy source for fungal biomass production. The second step was to optimize growth in liquid media. The cultivations were carried out by using five different conditions, where the liquid media contained different combinations of acetic acid, yeast extract and minerals as well as comparing orbital and linear oscillations. Fungal cultivation was possible regardless of the medium composition and type of water shaking baths. However, a linear water shaking bath with a combination of acetic acid yeast extract and/or minerals seems to be the best. Finally, as step three, acetic acid concentrations, 0.2 g/l and 2.0 g/l were used under similar conditions as in step two to see whether a higher concentration of acetic acid would be beneficial. Although the cultivation containing 2.0 g/l gave a higher value of dry weight, the value of yield is questionable. Further studies are needed to confirm if a higher concentration is beneficial or if it might act as an inhibitor for fungal cultivation. / Flyktiga fettsyror (VFAs) har ekonomiska fördelar och kan användas inom kemiska industrier i olika sammanhang, detta har lett till ett stort forskningsintresse för att kunna nyttja VFAs. Organiskt avfall, såsom matavfall, kan användas som substrat för att producera fettsyror genom anaerob rötning. Anaerob rötning är en miljövänlig process och VFAs bildas som intermediära produkter under den anaeroba nedbrytningen där annars bildas biogas som slutprodukt. Syftet med denna studie var att använda ättiksyra, (den vanligaste typen av VFAs), som kol- och energikälla vid odling av tre olika ätbara svampar, som Rhizopus oligosporus, Mucor indicus, och Volvariella volvacea. Först odlades dessa ätbara svampar i odlingsmedium innehållande ättiksyra. Resultatet visade att ättiksyra kan användas som kol- och energikälla vid produktion av svampbiomassa. Målet i de nästkommande stegen var att optimera tillväxtförhållande för svampodlingen. Fem olika odlingsmedier som innehöll olika kombinationer av ättiksyra, jästextrakt och mineraler användes. Det undersöktes dessutom hur två olika skakmetoder, orbitalt, eller linjärt, skakbad påverkar odlingen. Svamptillväxt var möjligt vid alla olika förhållanden oavsett sammansättningen av medium och typ av skakbad, däremot verkar odlingsmedium som innehåller ättiksyra, jästextrakt och/eller mineraler i kombination med linjär skakning vara de bästa förutsättningar för tillväxt av biomassa. I det sista steget kultiverades svamp med olika koncentrationer av ättiksyra, 0,2 g/l och 2,0 g/l, under liknande optimerade förhållanden som ovan, för att undersöka om en högre koncentration av ättiksyra skulle vara fördelaktig. Det producerades mer svampbiomassa (som torrvikt) vid koncentration av 2,0 g/l ättiksyra jämfört med när 0,2 g/l ättiksyra användes, dock var det svårt att säkerställa utbytet. Det behövs därför ytterligare fortsatta studier för att kunna bevisa om en högre koncentration av ättiksyra är fördelaktig för odlingen, eller om en högre koncentration skulle verka inhiberande för tillväxten.

The focus of the current study was on the recovery of carbon from municipal bio-waste streams. Firstly, the relationship between methanogenic pathways and the properties of anaerobic granules was studied using two pilot-scale upflow anaerobic sludge blanket (UASB) reactors with different granule size distribution. UASB1 granules were larger (3-4 mm) with multi-layered internal structure and the archaeal community was dominated by acetoclastic methanogens, while the UASB2 granules were smaller (1-2 mm) without a layered internal structure and the archaeal community was predominated by hydrogenotrophic methanogens. The acetoclastic methanogenic activities of UASB1 (250-437 mL CH4/g VS·d) than UASB2 (150-260 mL CH4/g VS·d), confirming that acetoclastic methanogenesis was more dominant in UASB1. Temperature increase from 20oC to 28oC did not change the microbial community but resulted in increased and stable biogas production for both reactors. There was an increase in chemical oxygen demand (COD) with hydraulic retention time due to increased contact time. The second part of the study focused on anaerobic co-digestion to produce volatile fatty acid (VFA) due to its higher value. The impact of substrate ratio of primary sludge (PS) and external organic waste (OW) and the robustness of the VFA system in the long term were assessed. Lab-scale batch study with different proportions of PS and OW; 0%, 25%, 50%, 75%, 100% of OW in terms of COD and scaled up in a semi-continuous pilot reactor with substrate ratio of 50% OW based on the results of the lab-scale study were performed. There was an increase in VFA production with an increase in OW proportion due to the availability of biodegradable organics. Acetic acid was the most dominant VFA in the batch reactors while in the semi-continuous experiment, caproic acid was dominated (50%). As carbon sources, the VFA-rich liquid attained the highest specific denitrification rate when compared with acetate and methanol. / I kommunala avfallsströmmar finns det en stor potential för resursåtervinning på grund av det höga organiska innehållet vilket kan användas för biobaserade produkter. På grundval av detta så fokuserar denna studie på resursåtervinningen av kol från kommunala avfallsströmmar genom att undersöka nuvarande och nytillkomna tekniker för att få vattenreningssystem att också bli resursåtervinningssystem. Första delen av studien undersöker relationen mellan metanproduktion och karaktäriseringen av anaerobiska granuler med ändamålet att förbättra energiproduktionen genom direkt behandling av kommunalt avloppsvatten. För detta ändamål så användes två UASB (Upflow Anaerobic Sludge Blanket) pilotreaktorer. Reaktorerna kördes med olika stora granuler, olika temperaturinställningar och olika hydrauliska uppehållstider (HRTs). Storlek, mikrobiell struktur, intern mikrostruktur och specifik metanogenaktivitet av de anaerobiska granulerna analyserades. Granulerna i UASB1 var 3-4 mm stora med interna multilager av mikrostrukturer som bestod av acetoklastika-metanogen arkéer. Granulerna i UASB2 var 1- 2 mm stora utan interna multilager och bestod av hydrogenotrofa metanogener. Aktiviteten i granulerna i UASB1 var 250-437 mL CH4 /g VS·d och i UASB2 var det 150-260 mL CH4 /g VS·d, vilket bekräftar att de acetoklastisk-metanogesa var mer effektiva i UASB1 än de hydrogenotrofa i UASB2. Att öka temperaturen från 20 o C till 28 o C under försöket medförde ingen förändring av den mikrobiologiska strukturen, men gav en ökning i biogasproduktion i UASB1 och högre och stabilare biogasproduktionshastighet i UASB2. Ökningen av biogasproduktion berodde på en reduktion i metans löslighet i utflödet och stabilare metanogenes. Ökning i HRT resulterade i större reduktion av organiskt material mätt som kemisk syreförbrukning (COD) och större biogasproduktion på grund av längre kontakttid mellan substrat och mikroorganismer. Den andra delen av studien fokuserar på att utveckla ett tillvägagångssätt att få en anaerob rötning att producera flyktiga fettsyror (VFA) istället för biogas. Studien fokuserar på effekten som kvoten mellan substraten primärslam och externt matavfall (OW) har och hur robust VFA-systemet skulle vara i stor skala på lång sikt. Olika kvoter av primärslam och matavfall testades i labbskala i batchstudier med 0 %, 25 %, 50 %, 75 % och 100 % COD OW. Baserat på resultaten från labbskala så kördes 50% COD OW i ett semi-kontinuerligt pilotförsök. Batch-testerna i labbskala visade att högre % COD OW, gav högre VFA produktion på grund av högre koncentration av organiskt material. Ättiksyra var den mest förekommande VFAn i batch-testerna medan kapronsyra var högst förekommande (50%) i det semi-kontinuerliga försöket. Denitrifikationsförsök visade att VFA-rik vätska från pilotskalareaktorerna gav den högsta specifika denitrifikationshastigheten i jämförelse med acetat och metanol. Resultaten visar att värdefulla kolkällor kan återvinnas från kommunalt avfall genom anaerob behandling av kommunalt avloppsvatten och samfermentering av primärslam och matavfall. / <p>QC 2020-06-15</p>

The production of volatile fatty acids (VFAs) from waste stream is gaining high attention because of their high market value and wide range of applications. In this study, the production of VFA from co-fermentation of primary sludge from wastewater treatment plant and food waste without pH control was evaluated using a pilot-scale reactor in a semi-continuous mode of operation. In addition, the influence of substrate and inoculum on VFA production and composition was assessed using a batch fermentation experiment. The pilot-scale reactor was operated at a retention time of 7 days and 10 days in phase 1 (126 days) and phase 2 (25 days) respectively. A maximum VFA production of (687 mg COD/g VS) was obtained when the pilot-scale reactor was operated at a retention time of 7 days on day 107. The change in retention time from 7 to 10 days led to a higher hydrolysis rate; however, no improvement in VFA production was observed. The most abundant VFA produced after the reactor stabilized was caproic acid (50 %), followed by acetic acid (23%) and butyric acid (20%). Higher amount ammonium nitrogen (1.3 to 14.32 mg/g VS) compared to soluble phosphorus (0.69 to 7 mg/g VS) was released during the co-fermentation process. Furthermore, the loss of the VFA due to the production of methane was highly reduced because the pH of the reactor adjusted by itself in the range of (5 – 5.7). The batch fermentation experiment revealed that VFA production without pH control is highly influenced by the type of substrate and inoculum used. While the distribution of VFAs, is highly influenced by the inoculum type compared to the substrate used. Finding from this study indicates that there is a potential to produce VFA from co-fermentation of primary sludge and food waste without pH control. / Lättflyktiga fettsyror (VFA; eng. Volatile Fatty Acids) är viktiga byggstenar för produktionen av en mängd kommersiellt viktiga kemikalier. VFA produceras för närvarande från icke-förnybara petrokemiska källor, som kan orsaka miljöproblem på grund av utsläpp av växthusgaser. VFA kan också produceras som en mellanprodukt vid den anaeroba nedbrytningsprocessen. Produktionen av VFA från avfallsströmmar har i nuläget fått stort intresse på grund av dess höga marknadsvärde och breda applikationsområde jämfört med biogas. Det mesta av forskningen som hittills genomförts har dock baserats på justeringen av pH genom tillsatts av HCl eller NaOH. Denna metod har dock mindre storskalig praktisk användning på grund av hög konsumtion av kemikalier. I denna studie undersöktes produktionen av VFA från samjäsning av primärslam och matavfall utan pH-kontroll med hjälp av en pilotskalereaktor i ett semikontinuerligt driftläge. Dessutom mättes den påverkan substrat och inokulum hade på produktion och av VFA genom batchjäsningsexperiment. Pilotskalereaktorn drevs med uppehållstid på 7 dagar respektive 10 dagar i fas 1 (126 dagar) och fas 2 (25 dagar). Maximal VFA produktion (på 687 mg COD/g VS) uppnåddes på dag 107, när pilotreaktorn hade en uppehållstid på 7 dagar. Ändringen i uppehållstid från 7 till 10 dagar ledde till en högre hydrolystakt; dock observerades ingen förbättring i VFA-produktion. Det vanligast förekommande VFA som produceras efter att reaktorn stabiliserades var caproinsyra (50%), följt av ättiksyra (23%) och smörsyra (20%). Högre mängd ammoniumkväve (1,3 till 14,32 mg/g VS) jämfört med löslig fosfor (0,69 till 7 mg/g VS) erhölls under samjäsningsprocessen. Förlusten av VFA på grund av produktion av metan minskade markant eftersom pH i reaktorn justerade sig själv i intervallet 5 – 5,7. Batchjäsningsexperiment genomfördes i 15 dagar med användning av endast primärslam, endast matavfall och en blandning av primärslam och matavfall. Två inokulum med ursprung från en anaerob rötkammare och dels från pilotskalereaktorn användes i experimentet. Resultaten visade att VFA-produktion utan pH kontroll påverkas betydligt av vilket slags substrat och inokulum som används. Sammansättning av VFA påverkas mer av vilket slags inokulum som användes jämfört med vilket substrat som används. Sammanfattningsvis visar resultatet av denna studie att det finns potential för att producera VFA genom samjäsning av primarslam och matavfall utan pH kontroll.

Ruminant livestock play a vital role in fulfilling the nutrient requirements of humans by providing protein, energy, and essential microminerals. With the increasing demand for meat and dairy products, the ruminant industry must continue to improve the productivity and efficiency of ruminant animals with limited resources while minimizing the environmental impact. Rumen fermentation is the focal point of the productivity and efficiency of the animal and numerous chemical, physical and biochemical interactions make the rumen a complex ecosystem. Therefore, improving the understanding of fermentation dynamics in a holistic manner and characterizing how fermentation varies in response to different nutrient supplies can greatly expand our knowledge on rumen fermentation to develop better engineered rumen manipulation strategies. The central aim of these investigations was to employ alternative analytical strategies for holistic exploration of complex relationships among rumen, animal, and dietary variables and to estimate rumen volatile fatty acid (VFA) dynamics under different nutrient supplies. The objective of the first study was to explore the strengths and limitations of mixed-model meta-analysis, recursive feature elimination (RFE), and additive Bayesian networking (ABN) in identifying relationships among diet, rumen, and milk performance variables. Both mixed-models and ABN agreed upon most of the variables and relationships identified while RFE failed to capture interactions. Given the capacity of mixed models for quantitative inquiry and the potential of ABN to illustrate complex associations in a more intuitive way, future investigations combining both approaches hold potential to explore intercorrelated data in a holistic manner. Followed by the successful use of ABN in the first study, the goal of a follow up study was to investigate the potential of two different network approaches to explore rumen level interactions using data generated in continuous culture experiments. Two network analysis approaches, EBIC-LASSO network (ELN) and Bayesian learning network (BLN) were leveraged to explore the relationships among rumen fermentation parameters in continuous culture experiments. Unidirectional ELN illustrated prominent variables while BLN, which produces a directed acyclic graph, identified directional relationships implying causality. Overall, both networking approaches demonstrate strengths in capturing connectedness and directionality of rumen fermentation variables. In a complementary line of work, the next experiment focused on developing an alternative method for iso-tope based assessments to produce less expensive, and more efficient screening of fermentation conditions driven by diet. Cannulated wethers were used in this study and 4 dietary treatments combining lowly and highly degradable fiber (timothy hay and beet pulp, respectively) and protein (heat-treated soybean meal and soybean meal, respectively) were tested. Results indicated that fluid volume of the rumen and the rate of passage were influenced by protein, but not fiber, source. Higher rumen volumes and lower passage rates were associated with heat-treated soybean meals. The effect of dietary treatments on VFA absorption dynamics was prominent compared to the minimal changes in production dynamics. Overall, heat-treated soybean meal appears to influence VFA disappearance resulting in low concentrations within the rumen, but greater flux of VFA disappearance. In conclusion, this method demonstrated the capacity to estimate VFA dynamics beyond concentrations and molar proportions while being cost effective and more physiologically relevant. In a fourth study, we sought to investigate the growth performance and rumen VFA profile in response to different planes of nutrients and naturally occurring coccidiosis. Coccidiosis infection altered rumen isobutyrate concentrations and tended to alter major VFA concentrations suggesting the need of future work to explore coccidiosis effects on rumen fermentation. The first two investigations highlighted the potential and strength of leveraging alternative analytical tools to complement statistical approaches generally used in ruminant nutrition while concurrently improving ability to explain complex associations in the rumen. The third and fourth projects characterized the rumen VFA dynamics and profile in response to the different nutrient degradability and health status, respectively. Collectively, these investigations contribute to better understanding of rumen dynamics through novel analytical and experimental approaches. / Doctor of Philosophy / With increasing global population, income, urbanization, and changes in dietary habits, the demand for meat and milk continues to grow. The ruminant animal industries (beef cattle, dairy cattle, sheep, goat, and buffalo) carry the burden of increasing production utilizing limited resources while minimizing the negative environmental impact caused by ruminant operations. To achieve this goal the productivity of the animal must be increased, and in order to increase the efficiency of production a better understanding of factors driving the production is critical. Ruminant animals have the unique ability to convert plant fiber into human edible milk and meat through a process that predominantly occurs in the special gut compartment called the rumen. In this process several compounds are produced, and among those volatile fatty acid (VFA) is of utmost importance because it fulfills energy demands for growth, production and reproduction. The rumen is a complex ecosystem consisting of numerous variables and associations. Understanding those relationships is crucial to manipulate rumen mechanisms. The overall objective of this work was to evaluate the potential of alternative statistical approaches, which demonstrated success in other disciplines, for better depiction of complex associations and characterization of production and absorption mechanisms of rumen VFA in response to different nutrient supplies. The objective of first investigation was to evaluate a feature selection method (recursive feature elimination; RFE) and a network approach (Additive Bayesian network; ABN) concurrently with a standard variable selection method (mixed model meta-analysis) commonly used to develop animal nutrition models. We attempted to find out the most important dietary, rumen, and animal variables for milk yield, milk fat and protein content as an example. Results indicate that the network approach was well aligned with the standard tool and can be used as a complementary approach. In our second investigation, we leveraged two networking analyses, a frequentist network which was unidirectional and a Bayesian network which was directional to explore rumen level interactions. The unidirectional network approach highlighted the most important variables in the rumen and numerous relationships among these variables. The directional network was more useful in understanding of causal associations within the system. In the third experiment we estimated the production and absorption of VFA in response to the different protein (heat-treated and regular soybean meal) and fiber (timothy hay and beet pulp) sources. The results revealed that the production of VFA was minimally affected by the diet, but the absorption was higher with heat-treated soybean meal. Our last project investigated the effect of parasitic infection, i.e., coccidiosis, and high and low levels of nutrition on growth and rumen VFA of growing lambs. Infection of coccidiosis altered a minor VFA (isobutyrate) and tended to alter total and major VFA (acetate and propionate). All these findings help to improve our understanding of rumen fermentation and subsequently develop strategies to manipulate rumen fermentation to enhance efficiency and productivity.

Mathematical modeling in animal agriculture can be applied at various levels including at the tissue, organ, animal, farm, regional and global levels. The purposes of this research were i) to evaluate models used to estimate volatile fatty acid (VFA) and methane (CH4) production and assess their impact on regional enteric CH4 inventory, and ii) to develop a process-based, whole-farm model to estimate net farm GHG emissions. In the first study, four VFA stoichiometric models were evaluated for their prediction accuracy of rumen VFA and enteric CH4 production. Comparison of measured and model predicted values demonstrated that predictive capacity of the VFA models varied with respect to the type of VFA in rumen fluid which impacted estimated enteric CH4 production. Moving to a larger scale assessment, we examined the enteric CH4 inventory from Manitoba beef cattle (from 1990 to 2008) using two mechanistic rumen models that incorporate VFA stoichiometric models: COWPOLL and MOLLY, and two empirical models: Intergovernmental Panel on Climate Change (IPCC) Tier 2 and a nonlinear equation (Ellis). The estimated absolute enteric CH4 production varied among models (7 to 63%) indicating that estimates of GHG inventory depend on model selection. This is an important consideration if the values are to be used for management and/or policy-related decisions. Development of models at the individual farm component level (animal, soil, crop) does not accurately reflect net GHG emissions generated from the whole production system. We developed a process-based, whole-farm model (Integrated Components Model, ICM), using the existing farm component models COWPOLL, manure-DNDC and some aspects of IPCC to integrate farm components and their associated GHG emissions. Estimates of total farm GHG emissions and their relative contribution using the ICM were comparable to estimates using two other whole-farm models (Integrated Farm System Model and Holos model). Variation was observed among models both in estimating whole-farm GHG emissions and the relative contribution of the different sources in the production system. Overall, whole-farm models are required to explore management options that will mitigate GHG emissions and promote best management practices. However, for full assessment of the production system, other benefits of the system (e.g., carbon sequestration, ecosystem services), which are not part of current whole-farm models, must be considered.

Mathematical modeling in animal agriculture can be applied at various levels including at the tissue, organ, animal, farm, regional and global levels. The purposes of this research were i) to evaluate models used to estimate volatile fatty acid (VFA) and methane (CH4) production and assess their impact on regional enteric CH4 inventory, and ii) to develop a process-based, whole-farm model to estimate net farm GHG emissions. In the first study, four VFA stoichiometric models were evaluated for their prediction accuracy of rumen VFA and enteric CH4 production. Comparison of measured and model predicted values demonstrated that predictive capacity of the VFA models varied with respect to the type of VFA in rumen fluid which impacted estimated enteric CH4 production. Moving to a larger scale assessment, we examined the enteric CH4 inventory from Manitoba beef cattle (from 1990 to 2008) using two mechanistic rumen models that incorporate VFA stoichiometric models: COWPOLL and MOLLY, and two empirical models: Intergovernmental Panel on Climate Change (IPCC) Tier 2 and a nonlinear equation (Ellis). The estimated absolute enteric CH4 production varied among models (7 to 63%) indicating that estimates of GHG inventory depend on model selection. This is an important consideration if the values are to be used for management and/or policy-related decisions. Development of models at the individual farm component level (animal, soil, crop) does not accurately reflect net GHG emissions generated from the whole production system. We developed a process-based, whole-farm model (Integrated Components Model, ICM), using the existing farm component models COWPOLL, manure-DNDC and some aspects of IPCC to integrate farm components and their associated GHG emissions. Estimates of total farm GHG emissions and their relative contribution using the ICM were comparable to estimates using two other whole-farm models (Integrated Farm System Model and Holos model). Variation was observed among models both in estimating whole-farm GHG emissions and the relative contribution of the different sources in the production system. Overall, whole-farm models are required to explore management options that will mitigate GHG emissions and promote best management practices. However, for full assessment of the production system, other benefits of the system (e.g., carbon sequestration, ecosystem services), which are not part of current whole-farm models, must be considered.

The objective of this study is to illustrate that phase separation improves the efficiency of an anaerobic system which digests semi-solid organic wastes. Organic fraction of municipal solid waste (OFMSW) was the semi-solid organic waste investigated. In the first part of the study, the optimum operational conditions for acidified reactor were determined by considering the volatile solid (VS) reductions and average acidification percentages at the end of two experimental sets conducted. Organic loading rate (OLR) of 15 g VS/L.day, pH value of 5.5 and hydraulic retention time (HRT) of 2 days were determined to be the optimum operational conditions for the acidification step. Maximum total volatile fatty acid and average acidification percentage were determined as 12405 mg as HAc/L and 28%, respectively in the reactor operated at optimum conditions. In the second part, an acidification reactor was operated at the optimum conditions determined in the first part. The effluents taken from this reactor as well as the waste stock used to feed this reactor were used as substrate in the biochemical methane potential (BMP) test. The results of BMP test revealed that the reactors fed by acidified samples indicated higher total chemical oxygen demand (tCOD) removals (39%), VS reductions (67%) and cumulative gas productions (265 mL). The result of this study indicated that the separation of the reactors could lead efficiency enhancement in the systems providing that effective control was achieved on acidified reactors.