Production of earthworm Allolobophora parva (Eisen) in cattle dung

Vermicomposting potential of Allolobophora parva is well proven in recent experiments but little is known about its growth and reproduction performance. Efforts were made in this study to assess the biological productivity of A. parva in cattle waste solids under laboratory conditions. The growth and reproduction performance of A. parva was monitored up to its termination state in experimental beddings. A. parva was weighed weekly and cocoons produced during the interval were also counted. The maximum individual biomass and maximum growth rate (wt. mg worm⁻¹ week⁻¹) was 190.9 ± 0.07 mg (after 13 weeks) and 2.66 (after 12th week), respectively. A. parva showed the maximum values of cocoon number (within a week) and reproduction rate: 26 ± 1.12 and 0.74 ± 0.05 cocoon worm⁻¹ week⁻¹, respectively, during the 8th week of vermiculture. Cocoon production in earthworms was terminated after the 17th week of culture. Data suggested that A. parva may acts as a potential candidate to convert negligible organic waste resources into worm biomass for sustainable environmental management.     

Nitrogen removal performance of a hybrid anammox reactor

The anaerobic ammonium oxidation (anammox) process has attracted considerable attention in recent years as an alternative to conventional nitrogen removal technologies. In this study, an innovative hybrid reactor combining fluidized and fixed beds for anammox treatment was developed. The fluidized bed was mechanically stirred and the gaseous product could be rapidly released from the anammox sludge to prevent washout of the sludge caused by floatation. The fixed bed comprising a non-woven biomass carrier could efficiently catch sludge to reduce washout. During the operation, nitrogen loading rates to the reactor were increased to 27.3 kg N/m³/d, with total nitrogen removal efficiencies of 75%. The biomass concentration in the fluidized bed reached 26-g VSS/L. Anammox granules were observed in the reactors, with settling velocities and sludge volumetric index of 27.3 ± 6.5 m/h and 23 mL/g, respectively. Quantification of extracellular polymeric substances revealed the anammox granules contained a significant amount of extracellular proteins.     

Chronic Opisthorchis felineus infection attenuates atherosclerosis – An autopsy study

Previously, we proposed a hypothesis that chronic helminthic infection may have beneficial effects on the development of atherosclerosis. The aim of this study was to investigate an association between Opisthorchis felineus chronic helminthic infections with aortic atherosclerosis and serum total cholesterol. A series of medico-legal autopsy specimens collected in Khanty-Mansiisk (the region in Russia endemic for O. felineus) were studied to assess O. felineus worm burden in cadaver livers. The areas of atherosclerotic lesions in the cadaver aortas were measured by visual planimetry. A family history of cardiovascular disease, smoking, hypertension or diabetes was elicited, and serum total cholesterol levels examined. Three hundred and nineteen cadavers (280 (87.8%) males and 39 (12.2%) females) aged 20–72 years were divided into five age groups: (i) 20–29, (ii) 30–39, (iii) 40–49, (iv) 50–59 and (v) >60 years old. The O. felineus mean worm burden was 257 ± 312 worms/liver. Infected subjects were categorised into three subgroups depending on the worm burden: mild (<100 worms), moderate (100–500 worms) and severe (>500 worms). Infected subjects had lower serum total cholesterol (mild worm burden, 186.4 ± 25.6 mg/dl; moderate worm burden, 183.4 ± 23.1 mg/dl, P = 0.002; severe worm burden, 170.6 ± 25.1 mg/dl, P < 0.001) than non-infected subjects (201.1 ± 21.2 mg/dl). The average percentage of aortic surface covered by fatty streaks, fibrotic plaques and complicated lesions was negatively related to worm burden in the infected subjects. Chronic helminthic infections was a negative predictor of aortic atherosclerosis; with an odds ratio of 1.72 (1.02–2.91), P = 0.041 for all subjects; and 3.19 (1.35–7.58), P = 0.008 for subjects aged >40 years old. Opisthorchis felineus chronic helminthic infectionswas found to be associated with lower serum total cholesterol levels and a significant attenuation of atherosclerosis.     

Effect of starch addition on the biological conversion and microbial community in a methanol-fed UASB reactor during long-term continuous operation

The effect of starch addition on the microbial composition and the biological conversion was investigated using two upflow anaerobic sludge bracket (UASB) reactors treating methanolic wastewater: one reactor was operated with starch addition, and another reactor was operated without starch addition. Approximately 300 days of operation were performed at 30 kg COD/m³/d, and then, the organic load of the reactors was gradually increased to 120 kg COD/m³/d. Successful operation was achieved at 30 kg COD/m³/d in both reactors; however, the methanol-fed reactor did not perform well at 120 kg COD/m³/d while the methanol-starch-fed reactor did. The granule analysis revealed the granule developed further only in the methanol-starch-fed reactor. The results of the microbial community analysis revealed more Methanosaeta cells were present in the methanol-starch-fed reactor, suggesting the degradation of starch produced acetate as an intermediate, which stimulated the growth of Methanosaeta cells responsible for the extension of granules.     

Sludge Reduction with a Novel Combined Worm-reactor

In China, as a result of economic, environmental and regulation factors, excess sludge treatment and disposal represents a rising challenge for small and mid-scale municipal wastewater treatment plants (WWTPs). Although the presence of worms in aerobic wastewater treatment may lead to substantial sludge reduction, the practical application is still uncontrollable because of unstable worm growth. To overcome unstable worm growth in activated sludge process, a combined worm-reactor, consisting of sections for both free-swimming and sessile worms, was developed to enhance worm growth. This plug flow type of worm-reactor has the sessile worm growing section filled with plastic carriers, and recirculation of sludge from the section for sludge settling was carried out in order to avoid worm washing out. Sludge reduction of treating the discharged excess sludge using this worm-reactor was investigated during 53 days. In the start-up phase the combined worm-reactor was inoculated with Tubificidae. Tubificidae mainly occurred in the carriers and on the bottom of the worm-reactor, however, neither Aeolosoma nor Nais was found in the worm-reactor. Results clearly showed that the average sludge reduction was 48%, and the COD removal was low, about 8%. High NH₄⁺ -N or NO₂⁻ -N concentrations or their combination may limit the growth of Tubificidae. Few PO ₄³⁻-P release into the effluent was observed in the worm-reactor. However, results obtained from this study cannot unequivocally be attributed to the presence of worms because it did not include a control system, and thus much more work will be needed in this concept in the further research.     

A new post-treatment system for anaerobic effluents containing a high Ca2+ content

A novel reactor concept has been developed that combines Ca²⁺ removal and BOD conversion under mesophilic (30 °C ) as well as under thermophilic (55 °C ) conditions. Soluble Ca²⁺, present in many industrial wastewaters, precipitates as a result of forced aeration in the aerobic bioreactor. The CaCO₃ precipitates, in turn, act as a carrier material for biofilm growth. This resulted in dense rapidly settling sludge granules covered with a biofilm on the surface. These sludge characteristics facilitate the application of hydraulic retention times of 1 h while maintaining a high biological activity in the reactor.     

Growth,reproductive biology and life cycle of the vermicomposting earthworm, Perionyx ceylanensis Mich. (Oligochaeta: Megascolecidae)

In the present study, an attempt has been made to study the growth, reproduction and life cycle of the earthworm, Perionyx ceylanensis Mich. in cowdung for the period of 340 days. Results showed that the overall mean growth rate was 1.79, 1.57 and 1.34 mg/worm/day respectively for the worms cultured singly, in batches of four and eight. Cocoon production rate was found between 0.85 and 0.94 cocoons/worm/day and the hatching success between 74.67% and 82.67%. The majority of the cocoons (95.16–96.77%) hatched only one hatchling. Worms raised singly also produced viable cocoons indicating that P. ceylanensis reproduce parthenogenetically. The life cycle of the worms cultured singly was ±57 days and it was ±50 days for the worms cultured in batches of four and eight. There is a vast scope to utilize P. ceylanensis for vermiculture practices due to short period of life cycle.     

Rapid acclimation of methanogenic granular sludge into denitrifying sulfide removal granules

Rapid formation of denitrifying sulfide removal granules is of practical interest to start up an expanded granular sludge bed reactor for wastewater treatment. This study demonstrates that methanogenic granules can be easily acclimated into DSR granules in one day, removing all 1.30 kg m⁻³ d⁻¹ sulfide and converting >90% of 0.56 kg-N m⁻³d⁻¹ nitrate into di-nitrogen gas. Under high loadings, reactor performance, however, declined. Under high loading rates, sulfide first inhibited the heterotrophic denitrifier (Caldithrix sp.), thereby accumulating nitrite in the system; the autotrophic denitrifier (Pseudomonas sp. C23) was then inhibited by accumulated nitrite, leading to breakdown of the entire DSR process.     

Biological kinetics evaluation of anaerobic stabilization pond treatment of palm oil mill effluent

Biological kinetic (bio-kinetic) study of the anaerobic stabilization pond treatment of palm oil mill effluent (POME) was carried out in a laboratory anaerobic bench scale reactor (ABSR). The reactor was operated at different feed flow-rates of 0.63, 0.76, 0.95, 1.27, 1.9 and 3.8 l of raw POME for a day. Chemical oxygen demand (COD) as influent substrates was selected for bio-kinetic study. The investigation showed that the growth yield (Y_G), specific biomass decay (b), maximum specific biomass growth rate (μ_max), saturation constant (K_s) and critical retention time (Θ_c) were in the range of 0.990 g VSS/g COD_removed day, 0.024 day⁻¹, 0.524 day⁻¹, 203.433 g COD l⁻¹ and 1.908 day, respectively.     

Improvement strategy on enhanced biological phosphorus removal for municipal wastewater treatment plants: full-scale operating parameters, sludge activities, and microbial features

The poor quality of effluent discharged by municipal wastewater treatment plants (WWTPs) is threatening the safety of water ecology. This study, which integrated a field survey, batch tests, and microbial community identification, was designed to improve the effectiveness of the enhanced biological phosphorus removal (EBPR) process for WWTPs. Over two-thirds of the investigated WWTPs could not achieve total P in effluent lower than 0.5 mg/L, mainly due to the high ratio of chemical oxygen demand to P (28.6-196.2) in the influent. The rates of anaerobic P release and aerobic P uptake for the activated sludge varied from 0.22 to 7.9 mg/g VSS/h and 0.43 to 8.11 mg/g VSS/h, respectively. The fraction of Accumulibacter (PAOs: polyphosphate accumulating organisms) was 4.8 ± 2.0% of the total biomass, while Competibacter (GAOs: glycogen-accumulating organisms) accounted for 4.8 ± 6.4%. The anaerobic P-release rate was found to be an effective indicator of EBPR. Four classifications of the principal components were identified to improve the EBPR effluent quality and sludge activity.     

Effects of microwave and alkali induced pretreatment on sludge solubilization and subsequent aerobic digestion

Individual and combined effects of microwave (MW) and alkali pretreatments on sludge disintegration and subsequent aerobic digestion of waste activated sludge (WAS) were studied. Pretreatments with MW (600 W-85 °C-2 min), conventional heating (520 W-80 °C-12 min) and alkali (1.5 g NaOH/L - pH 12-30 min) achieved 8.5%, 7% and 18% COD solubilization, respectively. However, combined MW-alkali pretreatment synergistically enhanced sludge solubilization and achieved 46% COD solubilization, 20% greater than the additive value of MW alone and alkali alone (8.5 + 18% = 26.5%). Moreover, the results of the batch aerobic digestion study on MW-alkali pretreated sludge showed 93% and 63% reductions in SCOD and VSS concentrations, respectively, at 16 days of SRT. The VSS reduction was 20% higher than that of WAS without pretreatment.     

Treatment of fresh leachate with high-strength organics and calcium from municipal solid waste incineration plant using UASB reactor

Treatment of a fresh leachate with high-strength organics and calcium from municipal solid waste (MSW) incineration plant by an up-flow anaerobic sludge blanket (UASB) reactor was investigated under mesophilic conditions, emphasizing the influence of organic loading rate (OLR). When the reactor was fed with the raw leachate (COD as high as 70,390-75,480 mg/L) at an OLR of 12.5 kg COD/(m³ d), up to ∼82.4% of COD was removed suggesting the feasibility of UASB process for treating fresh leachates from incineration plants. The ratio of volatile solids/total solids (VS/TS) of the anaerobic sludge in the UASB decreased significantly after a long-term operation due to the precipitation of calcium carbonate in the granules. Scanning electron microscopy (SEM) observation shows that Methanosaeta-like species were in abundance, accompanied by a variety of other species. The result was further confirmed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and sequencing.     

Cultivation of shear stress sensitive microorganisms in disposable bag reactor systems

Technical scale (≥5 l) cultivations of shear stress sensitive microorganisms are often difficult to perform, as common bioreactors are usually designed to maximize the oxygen input into the culture medium. This is achieved by mechanical stirrers, causing high shear stress. Examples for shear stress sensitive microorganisms, for which no specific cultivation systems exist, are many anaerobic bacteria and fungi, such as basidiomycetes. In this work a disposable bag bioreactor developed for cultivation of mammalian cells was investigated to evaluate its potential to cultivate shear stress sensitive anaerobic Eubacterium ramulus and shear stress sensitive basidiomycetes Flammulina velutipes and Pleurotus sapidus. All cultivations were compared with conventional stainless steel stirred tank reactors (STR) cultivations. Good growth of all investigated microorganisms cultivated in the bag reactor was found. E. ramulus showed growth rates of μ = 0.56 h⁻¹ (bag) and μ = 0.53 h⁻¹ (STR). Differences concerning morphology, enzymatic activities and growth in fungal cultivations were observed. In the bag reactor growth in form of small, independent pellets was observed while STR cultivations showed intense aggregation. F. velutipes reached higher biomass concentrations (21.2 g l⁻¹ DCW vs. 16.8 g l⁻¹ DCW) and up to 2-fold higher peptidolytic activities in comparison to cell cultivation in stirred tank reactors.     

Performance of enhanced biological SBR process for aniline treatment by mycelial pellet as biomass carrier

Mycelial pellet of Aspergillus niger Y3 was used as a biomass carrier to immobilize the aniline-degrading bacterium, Acinetobacter calcoaceticus JH-9 and the mix culture of the COD rapid degradation bacteria. In order to investigate its removal effect on aniline and COD, the combined mycelial pellets were applied in the SBR. Comparison of the performances was conducted between another SBR inoculated with sole strain JH-9 and the above SBR. The results showed that the stable degradations of aniline and COD were observed in both reactors. In the SBR with combined mycelial pellet, the biological removal efficiency was about 0.9 mg aniline/(L·d). It was much higher than that in the activated sludge reactor. Meanwhile, the performances of the sedimentation velocity, liquid-solid phase separation and the effluent quality were better in the SBR. According to SEM images and PCR-DGGE analysis, the species immobilized on the biomass carrier were more predominant in this system.     

Green and economical production of propionic acid by Propionibacterium freudenreichii CCTCC M207015 in plant fibrous-bed bioreactor

Propionic acid production by Propionibacterium freudenreichii from molasses and waste propionibacterium cells was studied in plant fibrous-bed bioreactor (PFB). With non-treated molasses as carbon source, 12.69 ± 0.40 g l^-1 of propionic acid was attained at 120 h in free-cell fermentation, whereas the PFB fermentation yielded 41.22 ± 2.06 g l^-1 at 120 h and faster cells growth was observed. In order to optimize the fermentation outcomes, fed-batch fermentation was performed with hydrolyzed molasses in PFB, giving 91.89 ± 4.59 g l^-1 of propionic acid at 254 h. Further studies were carried out using hydrolyzed waste propionibacterium cells as substitute nitrogen source, resulting in a propionic acid concentration of 79.81 ± 3.99 g l^-1 at 302 h. The present study suggests that the low-cost molasses and waste propionibacterium cells can be utilized for the green and economical production of propionic acid by P. freudenreichii.     

Fasciolagigantica: Parasitological and scanning electron microscopy study of the in vitro effects of ivermectin and/or artemether

Objective

To evaluate the in vitro effects of different concentrations of ivermectin and/or artemether on Fasciolagigantica worms and to study the parasitological changes and tegumental alterations using scanning electron microscopy (SEM).

Methods

Fasciola gigantica worms were incubated in vitro for 24 and 48 h with three concentrations of either ivermectin or artemether (10, 20 and 50 μg/ml) or both in half concentration of either (5, 10 and 25 μg/ml).

Results

Exposure of Fasciola worms to 25 + 25 μg/ml of combined drug regimens or to 50 μg/ml of either ivermectin or artemether for 48 h led to 100%, 41.7% and 75% worm killing which were accompanied by a significant reduction in egg laying capacity and significant increase in dead eggs maximally recorded in combined drug regimens. SEM of the flukes incubated for 48 h with combined drug regimens showed maximal tegumental disruption with swelling of the worm body, roughness, blebbing, sloughing and complete loss of spines. Disruption to the tegument of the flukes induced by artemether was more than that of ivermectin.

Conclusions

Artemether alone or combined with ivermectin in half doses had potent fasciocidal activities. Besides, half doses of combined drug regimens had higher ovicidal effects than each drug alone. In vivo studies are recommended to explore the efficacy of combined regimens against Fasciola infection.     

Biogeochemical processes in intensive zero-effluent marine fish culture with recirculating aerobic and anaerobic biofilters

The biogeochemical processes that drive nutrient transformations and recycling in organic marine sediment-water environments were studied for 17 months in a zero-effluent intensive recirculating culture system. The system consisted of a 10 m³ gilthead seabream (Sparus aurata) tank coupled to aerobic and anaerobic water treatment elements. Nutrients and alkalinity were measured in the system to quantify the main biogeochemical processes. Fractions of the carbon fed in feed were found in fish (18.3%) and in sludge (11%); the missing carbon was respired by fish (45%) and by aerobic (8.4%) and anaerobic (7.7%) microorganisms. Fractions of the nitrogen fed in feed were found in fish (15.4%) and in sludge (14.3%); the missing nitrogen was eliminated by nitrification-denitrification. Most of the phosphorus and ash fed in feed and not found in fish accumulated within the sludge in the system. The rates of nitrification, denitrification and sulphate reduction increased with time, reaching 0.3 g N m^− 2 d^− 1, 53 g N m^− 2 d^− 1 and 145 g S m^− 2 d^− 1, respectively. Nitrification developed more rapidly than denitrification, leading at first to nitrate accumulation (to 20 mmol NO₃ l^− 1 by day 200) and a decrease in alkalinity. Once denitrification surpassed nitrification, nitrate concentrations decreased, eventually being reduced to < 0.3 mmol NO₃ l^− 1 by day 510, and alkalinity stabilized. Toxic hydrogen sulphide, generated within the anaerobic sludge, was oxidized by oxygen and nitrate as it diffused through the anaerobic-aerobic sediment-water interface. When nitrate levels in the water above the sludge dropped below 2 mmol l^− 1, sulphide was also oxidized in the fluidized bed reactor. Denitrification reduced nitrate in the water, respired (jointly with sulphate reduction) carbon in the sludge, oxidized the hydrogen sulphide, and contributed to stabilization of alkalinity and accumulation of polyphosphate in bacteria as a major sink of labile P.     

Simultaneous study of particle reworking, irrigation transport and reaction rates in sediment bioturbated by the polychaetes Heteromastus and Marenzelleria

The present study employed simultaneously methods to investigate particle and solute transport and reaction rates in sandy sediments inhabited by two worms (2500 m^− 2) with different feeding modes. Heteromastus filiformis is a head-down deposit-feeder and the main activities exerted by this worm are transport of particles as faecal pellets from subsurface to surface sediments and burrow ventilation. Marenzelleria viridis is a surface deposit-feeder that actively searches for food by burrowing near the sediment surface, producing a network of ventilated galleries in this zone. M. viridis exhibited 1.5 to 2.2 times higher particle mixing rates (D_b = 3.3 to 4 × 10^− 3 cm^− 2 d^− 1) compared to H. filiformis. In M. viridis treatments, continuous advection (eddy diffusion) was the major factor influencing solute transport resulting in apparent diffusion rates (D_a = 2.2 cm^− 2 d^− 1), which were 3 times higher than molecular diffusion within the sediment. In H. filiformis inhabited sediments, the transport of solutes was discontinuous and driven by a surprisingly high nonlocal exchange (α = 1.1-1.3 d^− 1), emphasizing its strong irrigation effects. Accordingly, the enhancement of solute fluxes was more pronounced for H. filiformis compared to M. viridis. Depth integrated TCO₂ production derived from diagenetic modelling, which takes into account three reaction zones, is in good agreement with rates obtained from measured fluxes, indicating the applicability of both approaches to get reliable rates. However, the reaction rates showed that the presence of animals had a modest effect on microbial carbon oxidation. The results proved that transport conditions are deeply related to feeding modes. Exchange of solutes was the most important transport process by H. filiformis, while M. viridis affected both mixing and solute transport.     

Nutrients release and phosphorus distribution during oligochaetes predation on activated sludge

Release of nitrogen and phosphorus into effluent of activated sludge process was reported in the recent researches of sludge reduction induced by oligochaetes. In this study, batch test and radioisotope ³²P tracer test were therefore carried out to further investigate nutrients release and phosphorus distribution among supernatant, sludge and worm during predation of oligochaetes on sludge. Results showed that more nutrients release into supernatant occurred in the tests of worms with sterilized sludge than that of worms with activated sludge, and release of nitrogen and phosphorus was few in the tests of worms with activated sludge. Statistical analysis showed that no factor was significantly correlated with phosphorus concentration in the supernatant, but time and worm were only two factors significantly affecting total nitrogen (TN) concentration in the supernatant in the test of worms with activated sludge. Through test of radioisotope ³²P tracer, ³²P mainly distributed in activated sludge and supernatant, and more release of ³²P into the supernatant occurred in the test of sludge with worms. After 24 h, ³²P concentration of supernatant in the test of sludge with worms was 9% higher than that in the test of sludge without worms, and ³²P concentration of worm increased by 2.7%. Additionally, the release rate of phosphorus into supernatant caused by worm's predation on activated sludge was 0.1211 mgTP/gWorm (wet weight) h.     

Aerobic granular sludge formation for high strength agro-based wastewater treatment

The present study investigates the formation of aerobic granular sludge in sequencing batch reactor (SBR) fed with palm oil mill effluent (POME). Stable granules were observed in the reactor with diameters between 2.0 and 4.0 mm at a chemical oxygen demand (COD) loading rate of 2.5 kg COD m⁻³ d⁻¹. The biomass concentration was 7600 mg L⁻¹ while the sludge volume index (SVI) was 31.3 mL g SS⁻¹ indicating good biomass accumulation in the reactor and good settling properties of granular sludge, respectively. COD and ammonia removals were achieved at a maximum of 91.1% and 97.6%, respectively while color removal averaged at only 38%. This study provides insights on the development and the capabilities of aerobic granular sludge in POME treatment.