Factor-based assessment of continuous bio-H2 production from cheese whey.

Despite having been widely investigated, dark fermentative H2 production from organic residues is still limited by process-related issues which may hamper the perspectives of full-scale process implementation. Such constraints are mainly due to the process complexity, which is largely affected by multiple and often mutually interacting factors. In the present work, the results of continuous fermentative H2 production experiments using synthetic cheese whey as the input substrate were used to gain detailed knowledge of the process features and identify suitable and critical operating conditions. Specifically, innovative process interpretation involved a combination of analytical characterization of the fermentation broth, mass balance calculations and statistical methods (correlation and principal component analyses) to derive systematic considerations for process characterization and scale-up. The metabolic products mainly included acetate and butyrate, which however were likely to derive (in different proportions depending on the operating conditions) from both hydrogenogenic and competing pathways. For some tests, lactate and succinate were also found to have been formed. It was observed that the main features of the process (H2 yield and rate, stability condition) were correlated with the operational and analytical parameters. The first three principal components identified by the statistical analysis were able to account for: 1) the effect of retention time and total metabolites produced; 2) biogas (H2 and CO2) generation, butyrate production and stability condition; and 3) organic loading rate and propionate production. The results suggested that the main features of hydrogenogenic fermentation can be described by a reduced set of factors that may be usefully adopted for both process monitoring and prediction purposes.

Landfill gas generation after mechanical biological treatment of municipal solid waste. Estimation of gas generation rate constants.

Mechanical biological treatment (MBT) of residual municipal solid waste (RMSW) was investigated with respect to landfill gas generation. Mechanically treated RMSW was sampled at a full-scale plant and aerobically stabilized for 8 and 15 weeks. Anaerobic tests were performed on the aerobically treated waste (MBTW) in order to estimate the gas generation rate constants (k,y(-1)), the potential gas generation capacity (L(o), Nl/kg) and the amount of gasifiable organic carbon. Experimental results show how MBT allowed for a reduction of the non-methanogenic phase and of the landfill gas generation potential by, respectively, 67% and 83% (8 weeks treatment), 82% and 91% (15 weeks treatment), compared to the raw waste. The amount of gasified organic carbon after 8 weeks and 15 weeks of treatment was equal to 11.01+/-1.25kgC/t(MBTW) and 4.54+/-0.87kgC/t(MBTW), respectively, that is 81% and 93% less than the amount gasified from the raw waste. The values of gas generation rate constants obtained for MBTW anaerobic degradation (0.0347-0.0803y(-1)) resemble those usually reported for the slowly and moderately degradable fractions of raw MSW. Simulations performed using a prediction model support the hypothesis that due to the low production rate, gas production from MBTW landfills is well-suited to a passive management strategy.

Control of fermentation duration and pH to orient biochemicals and biofuels production from cheese whey.

Batch dark fermentation tests were performed on sheep cheese whey without inoculum addition at different operating pHs, relating the type and production yields of the observed gaseous and liquid by-products to the evolution of fermentation. Cheese whey fermentation evolved over time in two steps, involving an initial conversion of carbohydrates to lactic acid, followed by the degradation of this to soluble and gaseous products including short-chain fatty acids (mainly acetic, butyric and propionic acids) and hydrogen. The operating pH affected the production kinetics and yields, as well as the fermentation pathways. By varying the duration of the fermentation process, different cheese whey exploitation strategies may be applied and oriented to the main production of lactic acid, hydrogen or other organic acids.

Fermentative H2 production from food waste: Parametric analysis of factor effects.

Factorial fermentation experiments on food waste (FW) inoculated with activated sludge (AS) were conducted to investigate the effects of pH and the inoculum-to-substrate ratio (ISR [g VSAS/g TOCFW]) on biohydrogen production. The two parameters affected the H2 yield, the fermentation rate and the biochemical pathways. The minimum and maximum yields were 41 L H2/kg TOCFW (pH = 7.5, ISR = 1.74) and 156-160 L H2/kg TOCFW (pH = 5.5, ISR = 0.58 and 1.74). The range of carbohydrates conversion into H2 was 0.37-1.45 mol H2/mol hexose, corresponding to 9.4-36.2% of the theoretical threshold. A second-order predictive model for H2 production identified an optimum region at low pHs and high ISRs, with a theoretical maximum of 168 L H2/kg TOCFW at pH = 5.5 and ISR = 1.74. The Spearman's correlation method revealed several relationships between the variables, suggesting the potentially governing metabolic pathways, which turned out to involve both hydrogenogenic pathways and competing reactions.

Two-phase anaerobic digestion within a solid waste/wastewater integrated management system.

A two-phase, wet anaerobic digestion process was tested at laboratory scale using mechanically pre-treated municipal solid waste (MSW) as the substrate. The proposed process scheme differs from others due to the integration of the MSW and wastewater treatment cycles, which makes it possible to avoid the recirculation of process effluent. The results obtained show that the supplying of facultative biomass, drawn from the wastewater aeration tank, to the solid waste acidogenic reactor allows an improvement of the performance of the first phase of the process which is positively reflected on the second one. The proposed process performed successfully, adopting mesophilic conditions and a relatively short hydraulic retention time in the methanogenic reactor, as well as high values of organic loading rate. Significant VS removal efficiency and biogas production were achieved. Moreover, the methanogenic reactor quickly reached optimal conditions for a stable methanogenic phase. Studies conducted elsewhere also confirm the feasibility of integrating the treatment of the organic fraction of MSW with that of wastewater.

Dynamic transformations of nitrogen during mechanical-biological pre-treatment of municipal solid waste.

Mechanical-biological pre-treatment (MBP) of municipal solid waste (MSW) has gained evidence as a practice capable of accomplishing the requirements for environmental sustainable landfilling. In particular, MBP is effective in reducing the ammoniacal nitrogen content in the leachate. However, few data are available on the modifications of the nitrogen forms occurring during MBP and on the role played by processes such as nitrification and generation of refractory organic compounds. The dynamic transformations of nitrogen were investigated during the MBP. MSW was mechanically and biologically pre-treated; samples were collected at different stages of the process and analysed to investigate the evolution of nitrogen forms; batch and column leaching tests were performed as well. The results indicate that nitrification is negligible and volatilization can only partially explain the low ammoniacal nitrogen content in the leachate. Incorporation of ammoniacal nitrogen into a refractory organic form was assessed and is likely to play an important role. The maximum content of refractory organic nitrogen in the solid waste was achieved after about 60 days of aerobic pre-treatment; therefore, the minimal duration of the MBP should be about 8-9 weeks in order to optimize the ammoniacal nitrogen incorporation, unless the waste is characterized by a low C/N ratio.

Gamma-hydroxybutyric acid (GHB) and the mesoaccumbens reward circuit: evidence for GABA(B) receptor-mediated effects.

Gamma-hydroxybutyric acid (GHB) is a short-chain fatty acid naturally occurring in the mammalian brain, which recently emerged as a major recreational drug of abuse. GHB has multiple neuronal mechanisms including activation of both the GABA(B) receptor, and a distinct GHB-specific receptor. This complex GHB-GABA(B) receptor interaction is probably responsible for the multifaceted pharmacological, behavioral and toxicological profile of GHB. Drugs of abuse exert remarkably similar effects upon reward-related circuits, in particular the mesolimbic dopaminergic system and the nucleus accumbens (NAc). We used single unit recordings in vivo from urethane-anesthetized rats to characterize the effects of GHB on evoked firing in NAc "shell" neurons and on spontaneous activity of antidromically identified dopamine (DA) cells located in the ventral tegmental area. GHB was studied in comparison with the GABA(B) receptor agonist baclofen and antagonist (2S)(+)-5,5-dimethyl-2-morpholineacetic acid (SCH50911). Additionally, we utilized a GHB analog, gamma-(p-methoxybenzil)-gamma-hydroxybutyric acid (NCS-435), devoid of GABA(B) binding properties, but with high affinity for specific GHB binding sites. In common with other drugs of abuse, GHB depressed firing in NAc neurons evoked by the stimulation of the basolateral amygdala. On DA neurons, GHB exerted heterogeneous effects, which were correlated to the baseline firing rate of the cells but led to a moderate stimulation of the DA system. All GHB actions were mediated by GABA(B) receptors, since they were blocked by SCH50911 and were not mimicked by NCS-435. Our study indicates that the electrophysiological profile of GHB is close to typical drugs of abuse: both inhibition of NAc neurons and moderate to strong stimulation of DA transmission are distinctive features of diverse classes of abused drugs. Moreover, it is concluded that addictive and rewarding properties of GHB do not necessarily involve a putative high affinity GHB receptor.

Alpha-amino acid phenolic ester derivatives: novel water-soluble general anesthetic agents which allosterically modulate GABA(A) receptors.

In the search for a novel water-soluble general anesthetic agent the activity of an alpha-amino acid phenolic ester lead, identified from patent literature, was markedly improved. In addition to improving in vivo activity in mice, good in vitro activity at GABA(A) receptors was also conferred. Within the series of compounds good enantioselectivity for both in vitro and in vivo activity was found, supporting a protein-mediated mechanism of action for anesthesia involving allosteric modulation of GABA(A) receptors. alpha-Amino acid phenolic ester 19, as the hydrobromide salt Org 25435, was selected for clinical evaluation since it retained the best overall anesthetic profile coupled with improved stability and water solubility. In the clinic it proved to be an effective intravenous anesthetic in man with rapid onset of and recovery from anesthesia at doses of 3 and 4 mg/kg.

Mesolimbic dopaminergic reduction outlasts ethanol withdrawal syndrome: evidence of protracted abstinence.

Rats chronically administered with ethanol every six hours for six consecutive days show, upon suspension of treatment, a marked somatic withdrawal syndrome characterized by classical neurological signs. The emergence of the behavioral syndrome coincides with a profound decline of dopaminergic mesolimbic neuronal activity which corresponds to a reduction of dopamine outflow in the nucleus accumbens [Diana et al. (1993) Proc. natn. Acad. Sci. U.S.A. 90, 7966-7969]. However, while the behavioral manifestation of the ethanol withdrawal syndrome recedes in about 48 h, electrophysiological indices of mesolimbic dopaminergic function are still reduced 72 h after ethanol discontinuation, thus outlasting the physical signs of ethanol withdrawal syndrome. Dopaminergic neuronal activity is reintegrated by anti-craving drugs such as ethanol itself and gamma-hydroxybutyric acid. It is postulated that the reduced spontaneous activity of mesolimbic dopaminergic neurons may form the neural basis of the dysphoric state which accompanies abrupt interruption of chronic ethanol administration. Pharmacological manipulations of dopaminergic activity targeted at restoring "normal" dopaminergic function after ethanol withdrawal may lead to way to the experimental basis of new therapeutic strategies of alcoholism.

Effects of acute, chronic ethanol and withdrawal on dorsal raphe neurons: electrophysiological studies.

The effect of a single intravenous administration of ethanol (0.25-1.0 g/kg) on the spontaneous activity of putative serotonin neurons of the dorsal raphe nucleus was studied in unanesthetized rats. Ethanol produced a slight but progressive decline in neuronal activity in 67% (six of nine) of all neurons tested. The remaining 33% (three of nine) were unresponsive. Upon withdrawal of chronic ethanol treatment (1-5 g/kg every 6 h for six consecutive days, 12 h from last ethanol administration), the mean firine rate of dorsal raphe neurons was found to be significantly reduced, by about 30% (n=71), as compared with the control group (n=83), whereas the cells/track index was unaltered. Under these conditions, ethanol administration further reduced firing rate in 67% (four of six) of all the neurons tested. In the remaining 33% (two of six), no response was observed. At 72 h after the last ethanol administration, the mean firing rate of dorsal raphe neurons was found to be within control values (n=90). Further, to evaluate the functional status of the autoreceptors under control conditions and after withdrawal from chronic ethanol, the selective serotonin-1A receptor agonist 8-hydroxy-(2-di-n-propylamino)tetralin was administered intravenously in cumulative doses (1-16 microg/kg) and dose-response curves were generated for both groups. Autoreceptor sensitivity of dorsal raphe neurons was found to be not statistically different in control and ethanol withdrawn rats (n=6 for both groups) as indexed by a similar potency displayed by 8-hydroxy-(2-di-n-propylamino)tetralin in reducing the spontaneous activity of dorsal raphe neurons. The results indicate that, in spite of the widespread use of serotonin transmission potentiating agents in the treatment of alcoholism, neither acute nor withdrawal from chronic ethanol administration produces drastic effects on dorsal raphe neurons. However, the inhibition of dorsal raphe neuronal activity after acute ethanol may be due to the reported ability of ethanol to increase serotonin release from terminal areas. This increased serotonin tone could, at the level of recurrent axon collaterals in the dorsal raphe nucleus, reduce the spontaneous activity of the cells. On the other hand, a similar reduction in spontaneous activity after withdrawal from ethanol correlates well with the reduction in serotonin levels observed under these conditions in microdialysis studies.

Clonidine fails to modify dopaminergic neuronal activity during morphine withdrawal.

RATIONALE: Cellular substrates of opiate withdrawal syndrome involve several brain areas, in particular the mesolimbic dopaminergic and noradrenergic systems, but the interactions between the two pathways remain unclear. OBJECTIVES: The aim of the present work was to investigate the effects of the alpha2-agonist clonidine on ventral tegmental area dopamine neurons during morphine withdrawal syndrome by recording their neuronal activity before and after the administration of low and relatively high doses of clonidine (from 5 to 100 microg/kg). METHODS: The spontaneous neuronal activity of meso-accumbens dopaminergic neurons, identified by antidromical stimulation from the nucleus accumbens, was recorded by use of in vivo extracellular single-unit recordings in control and morphine-withdrawn rats after chronic administration (15 days). RESULTS: Control rats showed a mean spontaneous firing frequency of 2.47+/-0.48 Hz, percentage of burst firing of 22+/-12 and an increase in firing after the administration of cumulative doses of clonidine (5, 10, 20, 40, 100 microg/kg). Conversely, both spontaneous firing rate (1.55+/-0.25 Hz) and the percentage of burst firing (5+/-2) were found to be significantly reduced in rats abstinent for 24 h, and increasing doses of clonidine did not re-establish electrophysiological activity observed in the controls. CONCLUSION: The results indicate that: 1) clonidine did not restore the decreased firing activity of DA neurons in morphine-withdrawn rats, and 2) high doses of clonidine increased firing in control rats but not in morphine-withdrawn rats.

Ethanol withdrawal does not induce a reduction in the number of spontaneously active dopaminergic neurons in the mesolimbic system.

The effect of ethanol withdrawal, after chronic administration, on the electrophysiological properties of antidromically identified mesoaccumbens dopaminergic neurons was studied in two groups of rats with relative controls (withdrawal from chronic saline). The first group was anesthetized with chloral hydrate whereas the second was immobilized with D-tubocurarine. In chloral hydrate anesthetized rats, a significant reduction in the number of spontaneously active dopamine neurons was observed as compared with chronic saline withdrawn controls. In contrast, in ethanol-withdrawn D-tubocurarine treated rats, the number of spontaneously active dopamine neurons, as measured by the cells/track index, was found not different than chronic saline withdrawn controls. Further, intravenous administration of apomorphine, did not reverse the reduced cells/track index in chloral-hydrate anesthetized rats but consistently inhibited dopaminergic firing. Apomorphine-induced inhibition of firing was significantly more pronounced in ethanol withdrawn chloral-hydrate anesthetized rats. Firing rate and firing pattern were found decreased during ethanol withdrawal irrespective of experimental conditions. The results do not support the possibility that dopaminergic neurons of the mesoaccumbens pathway might be affected by depolarization inactivation during ethanol withdrawal. Rather, they confirm a reduction of neuronal activity already reported by previous studies. The reduced cells/track index observed in chloral hydrate anesthetized rats during ethanol withdrawal awaits an alternative explanation to the depolarization inactivation mechanism.

Repeated naltrexone administration accelerates resolution of morphine somatic withdrawal signs in morphine-dependent rats.

Morphine-dependent rats were divided into two subgroups. The first was allowed to develop a spontaneous withdrawal syndrome which was still present 33 h after the last morphine administration. The second subgroup received five injections of naltrexone. While the first two injections of naltrexone precipitated a marked somatic withdrawal syndrome, subsequent naltrexone failed to worsen the somatic signs. 24 h after the last morphine administration, naltrexone was completely ineffective. It is concluded that naltrexone shortens the duration of the morphine somatic withdrawal signs in dependent rats.

Heterogeneous responses of substantia nigra pars reticulata neurons to gamma-hydroxybutyric acid administration.

The effect of intravenous administration of gamma-hydroxybutyric acid (GHB) (50-400 mg/kg) on the firing rate of substantia nigra pars reticulata (SN-PR) neurons was studied by making single cell extracellular recordings in unanesthetized rats. For comparison, the effect of intravenous muscimol (0.5-2 mg/kg) and ethanol (0.5-2 g/kg) was also studied. GHB produced variable effects: dose-related inhibition in 7 out of 18 (38.8%) neurons and no significant change in 11 out of 18 (61.2%) neurons tested. In contrast, muscimol and ethanol produced a dose-related inhibition of the SN-PR firing rate. The results indicate that GHB, unlike muscimol and ethanol, has no profound effect on the activity of SN-PR neurons, and thus disinhibition of dopaminergic units, through inhibition of SN-PR neurons, is probably not the mechanism by which GHB stimulates the firing rate of dopaminergic neurons.

Marked decrease of A10 dopamine neuronal firing during ethanol withdrawal syndrome in rats.

The electrophysiological activity of mesoaccumbens dopaminergic neurons was monitored during the ethanol-withdrawal syndrome in ethanol-dependent and in control rats. Spontaneous firing was reduced by about half in ethanol-dependent rats as compared to controls. Likewise, the number of spikes/burst was also reduced in ethanol-dependent rats. These results are consistent with the reduction in dopamine release observed during ethanol-withdrawal syndrome and may provide the basis for the aversive effects of the ethanol-withdrawal syndrome.

Cannabinoids activate mesolimbic dopamine neurons by an action on cannabinoid CB1 receptors.

The present study was designed to determine if cannabinoids share with other drugs of abuse the ability to stimulate mesolimbic dopaminergic neurons and if this effect is mediated by cannabinoid receptors. To this end, the effects of the prototypical cannabinoid, delta9 tetrahydrocannabinol ¿(-)-trans-(6aR,10aR)-6a,7,8,10a-tetrahydro-6,6,9-trimethyl- 3-pentyl-6H-dibenzo[b,d]pyran-1-ol¿, and the two highly potent synthetic cannabinoids, ¿(R)-(+)-[2,3-dihydro-5-methyl-3-[(4-morpholinyl)-methyl]pyrrolo[1,2,3-d e]-1,4-benzoxazin-6-yl, +(1-naphtalenyl)methanone¿ WIN 55,212-2 and ¿(-)-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-4-(3-hydroxypropyl )-cicloexan-1-ol¿ CP 55,940, on the spontaneous discharge rate of meso-accumbens dopamine (A10 dopamine) neurons were studied in rats. The intravenous administration of delta9-tetrahydrocannabinol, WIN 55,212-2 and CP 55,940 (0.0625-1.0 mg/kg) produced a dose-dependent increase in the spontaneous firing of A10 dopamine neurons both in non-anesthetized and anesthetized rats, with a maximal percent increase of 120, 187 and 155 in non-anesthetized and 33, 102 and 52, respectively, in anesthetized rats. The stimulant response to cannabinoids was suppressed by the specific cannabinoid receptor antagonist ¿N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-me thyl-1H-pyrazole-3-carboxamide¿ SR 141716A, indicating a cannabinoid receptor-mediated effect. These findings support the contention that cannabinoids regulate mesolimbic dopamine transmission and may help to explain the addictive properties of marijuana.

Chronic administration of l-sulpiride at low doses reduces A10 but not A9 somatodentritic dopamine autoreceptor sensitivity.

The effect of chronic treatment (twice daily for 21 days) with low doses of l-sulpiride (2 mg/kg i.p.) on the apomorphine-induced inhibition of A10 and A9 dopaminergic neurons was compared with the effect of chronic administration of the classic antidepressant desipramine (20 mg/kg i.p. daily for 21 days). Intravenous administration of apomorphine (0.01-0.04 mg/kg), to rats treated chronically with l-sulpiride, produced a reduction of the spontaneous firing rate of A9 dopaminergic neurons not significantly different from that observed in control (saline-treated) rats. In contrast, apomorphine at the same doses was more potent in inhibiting A10 firing in control rats than in l-sulpiride-treated subjects. On the other hand, desipramine-treated rats were found normosensitive (as compared to saline-treated rats) to the inhibitory properties of apomorphine in both A9 and A10 dopaminergic neurons. It is suggested that chronic l-sulpiride-induced reduction of autoreceptor sensitivity in the A10 region may contribute to its clinical antidepressant effect.

Failure of chronic haloperidol to induce depolarization inactivation of dopamine neurons in unanesthetized rats.

Chronic treatment with neuroleptics has been reported to induce a status of depolarization inactivation of the majority of midbrain dopamine neurons. The present study was aimed at determining whether general anesthesia might be a contributory cause of depolarization inactivation of substantia nigra dopamine neurons. In agreement with previous studies, where neuronal sampling was carried out in animals under chloral hydrate anesthesia, chronic treatment with haloperidol (0.5 mg/kg daily for 21-28 days) produced a marked reduction (about 80%) in the number of spontaneously active dopamine neurons. However, when neuronal sampling was performed in unanesthetized rats, chronic administration of haloperidol (daily for 21-28 days) failed to reduce the incidence of active dopaminergic neurons. The results suggest that depolarization inactivation of dopamine neurons is not present in the intact animal but is probably produced during the neuronal sampling procedure as a consequence of neuroleptic-induced hyperexcitability of dopamine neurons combined with their stimulation by general anesthetics.

Engineering and environmental properties of thermally treated mixtures containing MSWI fly ash and low-cost additives.

An experimental work was carried out to investigate the feasibility of application of a sintering process to mixtures composed of Municipal Solid Waste Incinerator (MSWI) fly ash and low-cost additives (waste from feldspar production and cullet). The proportions of the three constituents were varied to adjust the mixture compositions to within the optimal range for sintering. The material was compacted in cylindrical specimens and treated at 1100 and 1150 degrees C for 30 and 60 min. Engineering and environmental characteristics including weight loss, dimensional changes, density, open porosity, mechanical strength, chemical stability and leaching behavior were determined for the treated material, allowing the relationship between the degree of sintering and both mixture composition and treatment conditions to be singled out. Mineralogical analyses detected the presence of neo-formation minerals from the pyroxene group. Estimation of the extent of metal loss from the samples indicated that the potential for volatilization of species of Pb, Cd and Zn is still a matter of major concern when dealing with thermal treatment of incinerator ash.

An experimental study on fermentative H2 production from food waste as affected by pH.

Batch dark fermentation experiments were performed on food waste and mixtures of food waste and wastewater activated sludge to evaluate the influence of pH on biological H2 production and compare the process performance with and without inoculum addition. The effect of a preliminary thermal shock treatment of the inoculum was also investigated as a means to harvest the hydrogenogenic biomass. The best performance in terms of both H2 generation potential and process kinetics was observed at pH=6.5 under all experimental conditions (no inoculum, and untreated or thermally treated inoculum added). H2 production from food waste was found to be feasible even without inoculum addition, although thermal pre-treatment of the inoculum notably increased the maximum production and reduced the lag phase duration. The analysis of the fermentation products indicated that the biological hydrogen production could be mainly ascribed to a mixed acetate/butyrate-type fermentation. However, the presence of additional metabolites in the digestate, including propionate and ethanol, also indicated that other metabolic pathways were active during the process, reducing substrate conversion into hydrogen. The plateau in H2 generation was found to mirror the condition at which soluble carbohydrates were depleted. Beyond this condition, homoacetogenesis probably started to play a role in the degradation process.