Profiles of fatty acids and triacylglycerols and their influence on the anaerobic biodegradability of effluents from poultry slaughterhouse

The hydrolysis of effluent from a poultry slaughterhouse containing 800 mg oil and grease (O&G)/L was conducted with 1% (w/v) of an enzymatic pool obtained by solid-state fermentation with the fungus Penicillium restrictum. The chromatographic evaluation of the lipid profile during hydrolysis indicated a higher concentration of acids after 4 h of reaction (2954 mg/L), with a predominance of oleic, palmitic, and linoleic acids. Effluent aliquots were collected after 4, 8, and 24 h of hydrolysis and tested for anaerobic biodegradation in sequential batches. An adaptation of the biomass was observed, both in the control experiment (with non-hydrolyzed raw effluent) and in the experiments with enzymatically pre-treated effluent. The specific methane production in the control experiment was 0.248 L CH₄/g COD_consumed, and in the experiment with effluent pre-treated for 4 h, this production was 0.393 L CH₄/g COD_consumed, indicating a higher methane production after enzymatic hydrolysis.     

Long-chain fatty acyl-coenzyme A-induced inhibition of glucokinase in pancreatic islets from rats depleted in long-chain polyunsaturated omega3 fatty acids

The metabolism of D-glucose was recently reported to be impaired in pancreatic islets from second generation rats depleted in long-chain polyunsaturated omega3 fatty acids. Considering the increased clearance of circulating non-esterified fatty acids prevailing in these rats, a possible inhibition of glucokinase in insulin-producing cells by endogenous long-chain fatty acyl-CoA was considered. The present study was mainly aimed at assessing the validity of the latter proposal. The activity of glucokinase in islet homogenates, as judged from the increase in D-glucose phosphorylation rate in response to a rise in the concentration of the hexose represented, in the omega3-depleted rats, was only 81.8 +/- 4.8% (n = 11; p < 0.005) of the paired value recorded in control animals. This coincided with the fact that the inclusion of D-glucose 6-phosphate (3.0 mM) and D-fructose 1-phosphate (1.0 mM) in the assay medium resulted in a lesser fractional decrease of D-glucose phosphorylation in omega3-depleted rats than in control animals. Moreover, whereas palmitoyl-CoA (50 microM) decreased the activity of glucokinase by 38.0 +/- 6.0% (n = 4; p < 0.01) in islet homogenates from normal rats, the CoA ester failed to affect significantly the activity of glucokinase in islet homogenates from omega3-depleted rats. These findings afford direct support for the view that glucokinase is indeed inhibited by endogenous long-chain fatty acyl-CoA in islets from omega3-depleted rats, such an inhibition probably participating to the alteration of D-glucose catabolism prevailing in these islets.     

Protein degradation during anaerobic wastewater treatment: derivation of stoichiometry

The stoichiometry of reactions that describe protein degradation in anaerobic treatment systems were investigated. A methodology was developed to describe protein degradation to organic acids using a single reaction step. The reactions for individual amino acid fermentation and their mediating organisms were reviewed. The dominant fermentation pathways were selected based on a number of assumptions. Using the amino acid content of a model protein, it was then possible to determine stoichiometric coefficients for each major organic acid product in the overall degradation of the protein. The theoretical coefficients were then compared to those determined from two experimental runs on a continuously-fed, well-mixed, laboratory-scale anaerobic wastewater treatment system. In general, the coefficients compared well thus validating the use of a single reaction step for the overall catabolic reaction of protein degradation to organic acids. Furthermore, even when the protein concentration in feed or the feed flow rate was doubled, the amino acid fermentation pathways were found to occur predominantly by only one pathway. Although the choice of Stickland reactions over uncoupled degradation provided good comparisons, an electron balance showed that only about 40% of the amino acids could have proceeded coupled to other amino acid reactions. Uncoupled degradation of the remaining amino acids must have relied on the uptake of hydrogen produced from these reactions by hydrogen-consuming methane bacteria.     

Treatment of slaughterhouse wastewater in a UASB reactor and an anaerobic filter

A study was performed to assess the feasibility of anaerobic treatment of slaughterhouse wastewaters in a UASB (Upflow Anaerobic Sludge Blanket) reactor and in an AF (Anaerobic Filter). Among the different streams generated, the slaughter line showed the highest organic content with an average COD of 8000 mg/l, of which 70% was proteins. The suspended solids content represented between 15 and 30% of the COD. Both reactors had a working volume of 21. They were operated at 37°C. The UASB reactor was run at OLR (Organic Loading Rates) of 1–6.5 kg COD/m³/day. The COD removal was 90% for OLR up to 5 kg COD/m³/day and 60% for an OLR of 6.5 kg COD/m³/day. For similar organic loading rates, the AF showed lower removal efficiencies and lower percentages of methanization. At higher OLR sludge, flotation occurred and consequently the active biomass was washed out from the filter. The results indicated that anaerobic treatment systems are applicable to slaughterhouse wastewaters and that the UASB reactor shows a better performance, giving higher COD removal efficiencies than the AF.     

Gene transfer of the Caenorhabditis elegans n-3 fatty acid desaturase inhibits neuronal apoptosis

Previous studies have shown that n-3 polyunsaturated fatty acids (PUFAs) can exert an antiapoptotic effect on neurons. The present study was designed to investigate whether the Caenorhabditis elegans fat-1 gene encoding an n-3 fatty acid desaturase (an enzyme that converts n-6 PUFAs to corresponding n-3 PUFAs) can be expressed functionally in rat cortical neurons and whether its expression can change the ratio of n-6 : n-3 fatty acids in the cell membrane and exert an effect on neuronal apoptosis. Infection of primary rat cortical cultures with Ad-fat-1 resulted in high expression of the fat-1 gene. Lipid analysis indicated a decrease in the ratio of n-6 : n-3 PUFAs from 5.9 : 1 in control cells, to 1.45 : 1 in cells expressing the n-3 fatty acid desaturase. Accordingly, the levels of prostaglandin E2, an eicosanoid derived from n-6 PUFA, were significantly lower in cells infected with Ad-fat-1 when compared with control cells. Finally, there was a significant inhibition of growth factor withdrawal-induced apoptotic cell death in neurons expressing the fat-1 gene. These results demonstrate that expression of the fat-1 gene can inhibit apoptotic cell death in neurons and suggest that the change in the n-6 : n-3 fatty acid ratio may play a key role in this protective effect.     

Long-chain fatty acid transport in bacteria andyeast. Paradigms for defining the mechanism underlying this protein-mediated process

Protein-mediated transport of exogenous long-chain fatty acids across the membrane has been defined in a number of different systems. Central to understanding the mechanism underlying this process is the development of the appropriate experimental systems which can be manipulated using the tools of molecular genetics. Escherichia coli and Saccharomyces cerevisiae are ideally suited as model systems to study this process in that both [1] exhibit saturable long-chain fatty acid transport at low ligand concentration; [2] have specific membrane-bound and membrane-associated proteins that are components of the transport apparatus; and [3] can be easily manipulated using the tools of molecular genetics. In E. coli, this process requires the outer membrane-bound fatty acid transport protein FadL and the inner membrane associated fatty acyl CoA synthetase (FACS). FadL appears to represent a substrate specific channel for long-chain fatty acids while FACS activates these compounds to CoA thioesters thereby rendering this process unidirectional. This process requires both ATP generated from either substrate-level or oxidative phosphorylation and the proton electrochemical gradient across the inner membrane. In S. cerevisiae, the process of long-chain fatty acid transport requires at least the membrane-bound protein Fat1p. Exogenously supplied fatty acids are activated by the fatty acyl CoA synthetases Faa1p and Faa4p but unlike the case in E. coli, there is not a tight linkage between transport and activation. Studies evaluating the growth parameters in the presence of long-chain fatty acids and long-chain fatty acid transport profiles of a fat1 strain support the hypothesis that Fat1p is required for optimal levels of long-chain fatty acid transport.     

Effect of selection for intramuscular fat on the fatty acid composition of rabbit meat

Intramuscular fat (IMF) content and composition are relevant for the meat industry due to their effect on human health and meat organoleptic properties. A divergent selection experiment for IMF of Longissimus dorsi (LD) muscle was performed in rabbits during eight generations. The aim of this study is to estimate the correlated responses to selection for IMF on the fatty acid composition of LD. Response to selection for IMF was 0.34 g/100 g of LD, representing 2.4 phenotypic SD of the trait. High-IMF line showed 9.20% more monounsaturated fatty acids (MUFA) and 0.39%, 9.97% and 10.3% less n-3, n-6 and polyunsaturated fatty acids (PUFA), respectively, than low-IMF line. The main MUFA and PUFA individual fatty acids followed a similar pattern, except for C18:3n-3 that was greater in the high-IMF line. We did not observe differences between lines for the percentage of total saturated fatty acids, although high-IMF line showed greater C14:0 and C16:0 and lower C18:0 percentages than low-IMF line. Heritability estimates were generally high for all fatty acids percentages, ranging from 0.43 to 0.59 with a SD around 0.08, showing an important genetic component on these traits. Genetic correlations between IMF and LD fatty acid percentages were strong and positive for C14:0, C16:1, C18:1n-9, and MUFA, ranging from 0.88 to 0.97, and strong and negative for C18:0, C18:2n-6, C20:4n-6, n-6 and PUFA, ranging from −0.83 to −0.91. These correlations were accurately estimated, with SD ranging from 0.02 to 0.06. The genetic correlations between IMF and other fatty acids were estimated with lower accuracy. In general, phenotypic and genetic correlations were of the same order. Our experiment shows that selection for IMF strongly affects the fatty acid composition of meat, due the high heritabilities of fatty acids and their high genetic correlations with IMF.     

Volatile fatty acid production during anaerobic mesophilic digestion of tea and vegetable market wastes

Extraction of the organic content from vegetable market waste and tea waste was carried out in a packed digester for 24 and 300 h respectively. The sequence of appearance of volatile fatty acids during digestion of both the substrates was found to be different. The sequence was (Acetic, Propionic) > (Isobutyric, Butyric) > Valeric for digestion of vegetable market waste while it was Isovaleric > (Isobutyric, Acetic) > Propionic during digestion of tea waste. During the course of digestion, the early appearance of an acid did not relate to its high concentration. The rate of production of acetic acid and propionic acid was found to be higher than other volatile acids during digestion of both the substrates, although it was approximately ten times higher for vegetable market waste compared to tea waste. The acids can be arranged in four groups according to their rate of production as Acetic > Propionic > Butyric > (Valeric, Isobutyric) for vegetable market waste and Acetic > Isobutyric > Isovaleric > Propionic for tea waste.     

Differentiation-related changes in lipid classes with long-chain and very long-chain polyenoic fatty acids in rat spermatogenic cells

In rat seminiferous tubules (ST), cells that contain polar and neutral lipids with long-chain polyenoic fatty acids (PUFA) and sphingomyelins (SM) and ceramides (Cer) with very long chain (VLC) PUFA of the n-6 series coexist. In this study, pachytene spermatocytes and round spermatids were isolated to determine how these lipids change during spermatogenesis. As the amount per cell of PUFA-rich glycerophospholipids (GPL) decreased with cell size, the 22:5/20:4 ratio increased with cell differentiation. The elovl2 and elovl5 genes, required for 22:5 formation, were expressed (mRNA) in both cell types. Residual bodies- particles with compacted organelles and materials discarded from late spermatids-concentrated cholesterol, 22:5-rich triacylglycerols, and GPL, including plasmalogens and phosphatidylserine. Species of SM and Cer with nonhydroxylated (n-) VLCPUFA (28:4, 30:5, and 32:5) predominated in pachytene spermatocytes, whereas species with the corresponding 2-hydroxy (2-OH) VLCPUFA prevailed in round spermatids. Thus, a dramatic increase in the 2-OH/n-VLCPUFA ratio in SM and Cer was a hallmark of differentiation. A substantial decrease of 2-OH SM occurred between spermatids and mature spermatozoa and 2-OH SM species were collected in residual bodies “en route” to Sertoli cells. Notably, spermatids and spermatozoa gained a significant amount of ceramides devoid of n-VLCPUFA but having 2-OH VLCPUFA as their main fatty acids.     

Effect of ciprofibrate on the activation and oxidation of very long chain fatty acids

The effect of ciprofibrate, a hypolipidemic drug, was examined in the metabolism of palmitic (C_16:0) and lignoceric (C_24:0) acids in rat liver. Ciprofibrate is a peroxisomal proliferating drug which increases the number of peroxisomes. The palmitoyl-CoA ligase activity in peroxisomes, mitochondria and microsomes from ciprofibrate treated liver was 3.2, 1.9 and 1.5-fold higher respectively and the activity for oxidation of palmitic acid in peroxisomes and mitochondria was 8.5 and 2.3-fold higher respectively. Similarly, ciprofibrate had a higher effect on the metabolism of lignoceric acid. Treatment with ciprofibrate increased lignoceroyl-CoA ligase activity in peroxisomes, mitochondria and microsomes by 5.3, 3.3 and 2.3-fold respectively and that of oxidation of lignoceric acid was increased in peroxisomes and mitochondria by 13.4 and 2.3-fold respectively. The peroxisomal rates of oxidation of palmitic acid (8.5-fold) and lignoceric acid (13.4-fold) were increased to a different degree by ciprofibrate treatment. This differential effect of ciprofibrate suggests that different enzymes may be responsible for the oxidation of fatty acids of different chain length, at least at one or more step(s) of the peroxisomal fatty acid -oxidation pathway.     

Influence of surfactants on anaerobic digestion of waste activated sludge: acid and methane production and pollution removal

The objective of this study is to summarize the effects of surfactants on anaerobic digestion (AD) of waste activated sludge (WAS). The increasing amount of WAS has caused serious environmental problems. Anaerobic digestion, as the main treatment for WAS containing three stages (i.e. hydrolysis, acidogenesis, and methanogenesis), has been widely investigated. Surfactant addition has been demonstrated to improve the efficiency of AD. Surfactant, as an amphipathic substance, can enhance the efficiency of hydrolysis by separating large sludge and releasing the encapsulated hydrolase, providing more substance for subsequent acidogenesis. Afterwards, the short chain fatty acids (SCFAs), as the major product, have been produced. Previous investigations revealed that surfactant could affect the transformation of SCFA. They changed the types of acidification products by promoting changes in microbial activity and in the ratio of carbon to nitrogen (C/N), especially the ratio of acetic and propionic acid, which were applied for either the removal of nutrient or the production of polyhydroxyalkanoate (PHA). In addition, the activity of microorganisms can be affected by surfactant, which mainly leads to the activity changes of methanogens. Besides, the solubilization of surfactant will promote the solubility of contaminants in sludge, such as organic contaminants and heavy metals, by increasing the bioavailability or desorbing of the sludge.     

CPT1alpha over-expression increases long-chain fatty acid oxidation and reduces cell viability with incremental palmitic acid concentration in 293T cells

To test the cellular response to an increased fatty acid oxidation, we generated a vector for an inducible expression of the rate-limiting enzyme carnitine palmitoyl-transferase 1alpha (CPT1alpha). Human embryonic 293T kidney cells were transiently transfected and expression of the CPT1alpha transgene in the tet-on vector was activated with doxycycline. Fatty acid oxidation was measured by determining the conversion of supplemented, synthetic cis-10-heptadecenoic acid (C17:1n-7) to C15:ln-7. CPT1alpha over-expression increased mitochondrial long-chain fatty acid oxidation about 6-fold. Addition of palmitic acid (PA) decreased viability of CPT1alpha over-expressing cells in a concentration-dependent manner. Both, PA and CPT1alpha over-expression increased cell death. Interestingly, PA reduced total cell number only in cells over-expressing CPT1alpha, suggesting an effect on cell proliferation that requires PA translocation across the mitochondrial inner membrane. This inducible expression system should be well suited to study the roles of CPT1 and fatty acid oxidation in lipotoxicity and metabolism in vivo.     

Tissue fatty acid composition of pigs fed different fat sources

Dietary fat influences the physico-chemical properties of meat, and fatty acid (FA) composition may have implications on human health. The objectives of the experiment were to study tissue FA partitioning and the effect of dietary fat source on tissue FA composition. Seventy crossbred gilts (61.8 ± 5.2 kg BW average) were fed one of seven treatments: a diet containing a very low level of fat (no fat (NF)) and six fat-supplemented diets (10%: tallow (T), high-oleic sunflower oil (HOSF), sunflower oil (SFO), linseed oil (LO), fat blend (FB: 55% tallow, 35% SFO, 10% LO) and fish oil blend (FO: 40% fish oil, 60% LO). Differential tissue FA depositions were observed, with flare fat being the most saturated, followed by intermuscular, and subcutaneous being the least saturated. Monounsaturated fatty acid (MUFA) deposition showed an opposite tissue pattern. Subcutaneous fat showed the highest MUFAs, intermuscular fat showed intermediate values and flare fat showed the lowest MUFAs. Intramuscular polyunsaturated fatty acid (PUFA) content was less susceptible to dietary treatment modifications compared with other depots. Significant tissue FA modifications were observed due to dietary treatments, mainly in diets rich in PUFA. The saturated fatty acids (SFA) were high in NF-fed and low in HOSF-fed animals, MUFA were high in HOSF-fed and low in SFO-, LO- and FO-fed animals, while PUFA were high in SFO- and LO-fed and low in HOSF-, T- and NF-fed animals. Pigs fed LO and FB showed detectable levels of EPA, which depended on the linolenic content of the diet. The only effective way to increase tissue DHA contents was to add DHA in the diet through FO feeding. Araquidonic acid was high in SFO diets and low in LO and FB diets, and also high in intramuscular fat compared with other tissues. EPA and DHA were also high in intramuscular fat compared with other fat depots. The deposition of oleic and linoleic acids depended on the composition of dietary fat, as their deposition varied between diets, even at similar levels of intake of each FA. The NF diet resulted in the greatest proportion of SFAs (palmitic and stearic) of all treatments tested. SFAs were less susceptible to modification than MUFA in response to the different PUFA levels supplemented in the diet. T resulted in less fat deposition in some of the fat depots and more in others, suggesting that T could partition fat differently among fat depots.     

Lipid composition of lactational diets influences the fatty acid profile of the progeny before and after suckling

The purpose of the present study was to compare the influence of adding no or 8% fat of varying sources (coconut oil, fish oil, rapeseed oil and sunflower oil) to diets for sows 1 week prior to farrowing and during lactation on the composition of fatty acids in plasma and tissues of the progeny while sucking and 3 weeks after weaning from the sow. A control diet without supplemental fat and four diets supplemented with 8% of coconut oil, rapeseed oil, fish oil or sunflower oil were provided to lactating sows (n = 15), and during the post-weaning period the same weaner diet was provided to all piglets (n = 15 litters), which were housed litterwise. The dietary ratio of n-6:n-3 fatty acids of the maternal diets largely influenced the progeny, as the ratio varying from 1.2 (fish oil) to 12.2 (sunflower oil) in the sow milk was reflected in plasma and adipose tissues of the sucking progeny. The liver showed similar variations according to dietary treatments, but a lower n-6:n-3 fatty acids ratio. From day 4 to later on during the suckling period, the concentration of C14:0, C16:0 and C18:1 in the liver of the piglets decreased, irrespective of the dietary treatments of sows. In plasma and liver, the total concentration of saturated fatty acids (SAFA), monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) did not differ markedly in piglets sucking sows fed different dietary fatty acids, whereas the adipose tissue of piglets sucking sows fed sunflower oil and coconut oil showed the highest proportion of PUFA and SAFA, respectively. Weaning lowered the concentration of lipid-soluble extracts in plasma and the concentration of fatty acids in the liver of the piglets. Within the post-weaning period, dietary treatments of sows, rather than age of piglets, influenced the fatty acid composition of plasma and adipose tissue of the piglets, whereas the hepatic fatty acid profile was more affected by the age of the piglets during the post-weaning period. This study shows that the fatty acid profile of plasma and tissues of the progeny is highly dependent on the maternal dietary composition, and that the dietary impact persists for up to 3 weeks after the suckling period.     

Fatty acid oxidation in anoxic marine sediments: the importance of hydrogen sensitive reactions

In anoxic marine sediments fatty acids may be oxidized directly by sulfate reducing bacteria, or may be oxidized by pathways which result in hydrogen production. Some of these latter reactions are quite sensitive to hydrogen concentrations ... in other words if hydrogen concentrations become elevated, fatty acid oxidation will cease. Thus sulfate reducers may actually play two important roles in the metabolism of fatty acids in marine sediments. The sulfate reducers both can utilize fatty acids directly, and also can oxidize hydrogen and thus control hydrogen partial pressures in the sediments. Therefore sulfate reducers may act indirectly to facilitate fatty acid oxidation by hydrogen-producing pathways. We carried out a series of incubations of slurried salt marsh sediment under high and low hydrogen partial pressures and in the presence and absence of molybdate to investigate the relative importance of sulfate reducers and other bacteria mediating hydrogen-sensitive reactions. Our results suggest that both classes of bacteria contribute significantly to fatty acid turnover in marine sediments. Studies of low molecular weight fatty acid turnover in sediment must explicitly recognize that manipulation of sediment (including addition of molydbate to inhibit sulfate reducers) may have a large impact on hydrogen partial pressures in sediment, and may thus significantly alter the pathways and/or rates of fatty acid turnover.     

Effect of volatile fatty acids on Shigella dysenteriae during anaerobic digestion of human night soil

Thein vitro toxic effect of different volatile fatty acids (VFA) on Shigella dysenteriae was studied in pure culture. Volatile fatty acids viz., acetate, propionate, butyrate, valerate, caproate and heptanoate, exerted pH dependent toxic effect on the pathogen, with minimum inhibitory concentration in the range of 10–3000 mg l⁻¹. The effect of high levels of VFA on S. dysenteriae was studied during anaerobic digestion of human night soil in an experimental digester with VFA level ≅ 9000 mg l⁻¹ and pH ≅ 6.5. Another digester, with VFA level ≅ 700 mg l⁻¹ and pH 7.4, served as the control. In the experimental digester, S. dysenteriae was completely eliminated within 18 days. In the control digester, a four-log reduction in pathogen count was achieved however the pathogen was not completely eliminated. T ₉₀ values for the experimental and control digesters were 2.2 and 3.7 days respectively. This revised version was published online in July 2006 with corrections to the Cover Date.     

Myocardial fatty acid oxidation during ischemia and reperfusion

Inhibition of fatty acid oxidation is an early event in myocardial ischemia that most likely contributes to tissue injury by the accumulation of potentially toxic intermediates such as acylCoA and acylcarnitine. After reperfusion both myocardial oxygen consumption and fatty acid oxidation may rapidly recover to preischemic levels, even when contractile function remains depressed. The mechanisms underlying the apparent dissociation between contractile function and oxidative metabolism early during reperfusion are still controversial. In isolated rat hearts subjected to 60 min of no-flow ischemia myocardial oxygen consumption and oxidation of palmitate were lowered during reperfusion by 3 mM of NiCl₂ and by 6 µM of ruthenium red. The results provide indirect evidence for the hypothesis that intracellular calcium transport may be involved in the mechanisms responsible for the high oxidative metabolic rate early after reperfusion     

Inclusion/exclusion of fatty acids in amylose complexes as a function of the fatty acid chain length

Structural models are proposed for amylose-fatty acid complexes depending on the respective chain lengths of their constituents. The three studied fatty acids induce the Vh amylose crystalline type. However, in contrast to lauric and palmitic acids, caprylic acid is not present in crystals. On the basis of the relative amounts of amylose and fatty acid determined in complexes and previous results of molecular modelling, inclusion of lauric and palmitic acids inside the amylose helices is proposed; the acyl chains are included in crystalline areas and the car☐ylic groups in amorphous areas. The absence of caprylic acid in crystals could be due to the solubility of this compound in the crystallization medium.