Growth stage and ensiling: impact on chemical composition, conservation quality and in situ ruminal degradability of whole-crop oat.

BACKGROUND: The effects of growth stage (GS) and ensiling were assessed on whole-crop oat's (Avena sativa L. cv. Cantara) chemical composition, silage fermentation quality and in situ ruminal degradability. Oat was harvested and ensiled at six GS: boot, heading, water ripe, early milk, early dough and grain ripe (144, 151, 178, 234, 362 and 512 g kg-1 of dry matter (DM) of whole-crop forage, respectively). RESULTS: GS influenced chemical composition, silage fermentation quality and ruminal degradability of whole-crop oat. Lower DM and higher water-soluble carbohydrates (WSC) contents lead to adequate forage compaction and fermentation at early GS; however, effluent was produced until the dough stage (L and Q; P ≤ 0.003). Advancing in maturity increased (P < 0.001) crop yield (4.5 to 9.4 t DM ha-1 ), DM (144 to 512 g kg-1 ), neutral detergent fibre (NDF; 537 to 571 g kg-1 DM), lignin (44.6 to 71.3 g kg-1 DM) and starch contents (26.4 to 201 g kg-1 DM), and reduced (P < 0.001) crude protein (107 to 60 g kg-1 DM) and WSC (115 to 17.5 g kg-1 DM). DM and NDF ruminal degradability declined with maturity for fresh and ensiled forages (L and Q; P < 0.05). Density and buffering capacity decreased with GS (L and Q; P < 0.001), whereas pH and soluble protein increased (L and Q; P ≤ 0.004). CONCLUSION: The growth stage of oat influenced the nutritive value and ruminal degradation to a greater extent than ensiling, and thus it can play a paramount role in whole-crop oat silage quality. © 2021 Society of Chemical Industry.

Ruminal use of undegraded-feed soluble protein and accuracy of the estimations of the nutrient content in ruminal bacteria.

BACKGROUND: The possible escape from the rumen of undegraded-feed soluble proteins (SP) was investigated using nitrogen-15 (15 N) infusions (25 mg) in three rumen and duodenum cannulated wethers. Animals were fed three isoproteic diets differing in SP content and including protein concentrates either untreated (control) or treated with solutions of either malic acid (MHT) or orthophosphoric acid (OHT) and heat. RESULTS: Compared with control, MHT and OHT diets reduced ruminal concentrations of ammonia-nitrogen (NH3 -N) by 35.9% (P = 0.007), non-ammonia nitrogen (NAN) by 36.8% (P = 0.007), and SP-nitrogen (SP-N) by 45.2% (P = 0.072) over the post-feeding period. Both NAN and SP-N were lower (P ≤ 0.040) for OHT than for MHT diet. The 15 N enrichment of NAN and SP-N did not vary either among diets or with time, and both values were closely related (R2 = 0.965; P < 0.001). Estimations, either using solid-associated bacteria (SAB) or liquid-associated bacteria (LAB) as a reference, indicated that 0.983 and 0.894 of SP-N (values averaged across diets) was of microbial origin, respectively, which would indicate a practically negligible ruminal escape of feed SP-N. Values of 15 N-enrichment in SAB and LAB fitted well to previously published SAB-LAB relationships, indicating a 22% underevaluation of the N supply from SAB when only LAB is used as a reference. CONCLUSION: Both the negligible ruminal escape of feed SP and the underevaluation of the bacterial nutrient supply as a consequence of the use of LAB as the only bacterial reference should be considered to improve ruminant protein feeding systems. © 2019 Society of Chemical Industry.

Planning low-carbon electricity systems under uncertainty considering operational flexibility and smart grid technologies.

Electricity grid operators and planners need to deal with both the rapidly increasing integration of renewables and an unprecedented level of uncertainty that originates from unknown generation outputs, changing commercial and regulatory frameworks aimed to foster low-carbon technologies, the evolving availability of market information on feasibility and costs of various technologies, etc. In this context, there is a significant risk of locking-in to inefficient investment planning solutions determined by current deterministic engineering practices that neither capture uncertainty nor represent the actual operation of the planned infrastructure under high penetration of renewables. We therefore present an alternative optimization framework to plan electricity grids that deals with uncertain scenarios and represents increased operational details. The presented framework is able to model the effects of an array of flexible, smart grid technologies that can efficiently displace the need for conventional solutions. We then argue, and demonstrate via the proposed framework and an illustrative example, that proper modelling of uncertainty and operational constraints in planning is key to valuing operationally flexible solutions leading to optimal investment in a smart grid context. Finally, we review the most used practices in power system planning under uncertainty, highlight the challenges of incorporating operational aspects and advocate the need for new and computationally effective optimization tools to properly value the benefits of flexible, smart grid solutions in planning. Such tools are essential to accelerate the development of a low-carbon energy system and investment in the most appropriate portfolio of renewable energy sources and complementary enabling smart technologies.This article is part of the themed issue 'Energy management: flexibility, risk and optimization'.

Ruminal degradation of cell wall associated nitrogenous compounds of several (15) N-labelled feeds.

BACKGROUND: Ruminal in situ effective degradability (ED) of dry matter (DM), neutral (NDF) and acid (ADF) detergent fibres, total-N and NDF (NDIN) and ADF (ADIN) bound-N in sunflower seed (SS), wheat grain (WG) and wheat straw (WS) were measured in three ruminally cannulated sheep, correcting microbial N-contamination using the (15) N dilution technique modified to consider the (15) N supply to adherent bacteria. RESULTS: The lack of correction for N-contamination under-evaluated ED estimates in 1.52% (total-N), 28.0% (NDIN) and 33.3% (ADIN) in SS and in 1.02% (total-N) and 4.43% (NDIN) in WG. In the remaining cases, this contamination prevented establishing apparent degradation kinetics and, therefore, errors were not measured. Microbial corrected ED estimates in SS were higher in total-N (0.917) than in NDIN (0.559) and ADIN (0.520), which showed similar values. This behaviour was also shown in WS (0.670, 0.386 and 0.426, respectively), whereas decreasing values were shown from total-N (0.917) to NDIN (0.830) and ADIN (0.482) in WG. CONCLUSION: Results confirm that NDF and ADF procedures failed to remove large fractions of particle adherent microorganisms, under-evaluating the ED of NDIN and ADIN. Degradation of NDIN represented a significant part of the degraded N, whereas ADIN contribution was only negligible in WG. © 2015 Society of Chemical Industry.

The role of pH on the resistance of resting- and active anammox bacteria to NO2- inhibition.

The anaerobic oxidation of ammonium (anammox) uses nitrite as terminal electron acceptor. The nitrite can cause inhibition to the bacteria that catalyze the anammox reaction. The literature shows a great divergence on the levels of NO2 (-) causing inhibition. Moreover, the conditions influencing the resistance of anammox bacteria to NO2 (-) inhibitory effect are not well understood. This work investigated the effect of the pH and the concentration of nitrite on the activity and metabolism of anammox granular sludge under different physiological conditions. Batch activity tests in a range of pH values were carried out in which either actively metabolizing cells or resting cells were exposed to nitrite in the presence or absence of the electron donating substrate ammonium, respectively. The response of the bacteria was evaluated by analyzing the specific anammox activity, the accumulation of nitric oxide, and the evolution of the ATP content in the biomass. Additionally, the effect of the pH on the tolerance of the biomass to single substrate feeding interruptions was evaluated in continuous anammox bioreactors. The results show that the influence of the pH on the NO2 (-) inhibition of anammox bacteria is greater under non-metabolizing conditions than during active metabolism. The exposure of resting cells to NO2 (-) (100 mg N L(-1) ) at pH values below 7.2 caused complete inhibition of the anammox activity. The inhibition was accompanied by accumulation of the intermediate, nitric oxide, in the gas phase. In contrast, just mild inhibition was observed for resting cells exposed to the same NO2 (-) concentration at pH values higher than 7.5 or any of the pH values tested in assays with actively metabolizing cells. ATP initially increased and subsequently decreased in time after resting cells were exposed to NO2 (-) suggesting an active response of the cells to nitrite stress. Furthermore, bioreactors operated at pH lower than 6.8 had greater sensitivity to NO2 (-) during an ammonium feed interruption than a bioreactor operated at pH 7.1. The results suggest that the ability of resting cells to tolerate NO2 (-) inhibition is seriously impeded at mildly acidic pH values; whereas actively metabolizing biomass is resistant to NO2 (-) toxicity over a wide range of pH values.

Effects of the comminution rate and microbial contamination of particles in the rumen on in situ estimates of protein and amino acid digestion of expeller palm kernel and rapeseed meal.

BACKGROUND: Microbial corrected effective in situ estimates of ruminal undegraded fraction (RU) and intestinal effective digestibility (IED) of dry matter (DM), crude protein (CP) and amino acids (AA) of expeller palm kernel (EPK) and rapeseed meal (RSM) were measured on three rumen- and duodenum-cannulated wethers using ¹5N labelling techniques and considering ruminal rates of comminution (k(c)) and outflow (k(p)) of particles. RESULTS: The lack of k(c) and microbial correction overestimated the RU of DM by 4.91% (EPK) and 9.88% (RSM). The lack of this correction also overestimated in both feeds the RU of CP, individual and total (TAA) AA as well as the IED of DM, CP, TAA and most AA. RU estimates were higher for CP than for TAA, but the opposite was observed for IED. The intestinal digested fraction was higher for CP than for TAA: 17.4% (EPK) and 13.8% (RSM). Digestion led to large changes in the essential AA profile in both feeds. CONCLUSION: The lack of k(c) and microbial correction as well as CP-based results leads to considerable overestimations in the protein use of both feeds. Digestion aggravates the lysine deficiency of EPK but has global positive effects in the absorbed profile of RSM.

Amino acid availability in ruminants of cereals and cereal co-products.

BACKGROUND: Microbial corrected in situ estimates of the ruminal undegraded fraction (RU) and intestinal effective digestibility (IED) of amino acids (AA), except tryptophan, of rye, wheat and corn grains, wheat bran, wheat and barley distilled dried grains and corn gluten feed were measured on three rumen- and duodenum-cannulated wethers using (15)N-labelling techniques and considering ruminal rates of particle comminution and outflow. RESULTS: The lack of microbial correction led to overestimations of the intestinal digested fraction that rose with the increase in ruminal degradability. Thus these overestimations varied widely among feeds (from 4.3 to 32.1% for total analysed AA) and among AA. Digestion led to large changes in the AA supply that were greater in the rumen than in the intestine. The impact of these changes on the protein value is conditioned by the magnitude of the undegraded protein fraction. CONCLUSION: Microbial contamination taking place in the rumen and changes in the AA supply with digestion should be considered to attain accurate estimates of AA digestion. Globally, digestion improved the AA supply in rye, wheat and wheat distilled dried grain and decreased it in corn and corn gluten feed by reducing the supply of valine and basic AA, especially lysine.

[Psychological study of the dysthymic disorder in the woman]. / Estudio psicológico del trastornodistímico en la mujer.

In this article, we study two dysthymic women who we are treating with psychotherapy in order to reveal the inner components that maintain depressive symptoms. The same findings have been confirmed in other dysthymic patients. The result of the study consisted in discovering a sentimental separation from their love object, while the woman still lives with her partner and while the depressive symptoms are appearing insidiously. This development leads them to the deterioration in the "ideal of love" they sought, that supported their lives and served as an "anchor of their personality. This point of view places classic notion about mourning into doubt.

Continuous H2/CO2 fermentation for acetic acid production under transient and continuous sulfide inhibition.

Waste gas fermentation powered by renewable H2 is reaching kiloton scale. The presence of sulfide, inherent to many waste gases, can cause inhibition, requiring additional gas treatment. In this work, acetogenesis and methanogenesis inhibition by sulfide were studied in a 10-L mixed-culture fermenter, supplied with CO2 and connected with a water electrolysis unit for electricity-powered H2 supply. Three cycles of inhibition (1.3 mM total dissolved sulfide (TDS)) and recovery were applied, then the fermenter was operated at 0.5 mM TDS for 35 days. During operation at 0.5 mM TDS the acetate production rate reached 7.1 ± 1.5 mmol C L-1 d-1. Furthermore, 43.7 ± 15.6% of the electrons, provided as H2, were distributed to acetate and 7.7 ± 4.1% to butyrate, the second most abundant fermentation product. Selectivity of sulfide as inhibitor was demonstrated by a 7 days lag-phase of methanogenesis recovery, compared to 48 h for acetogenesis and by the less than 1% electrons distribution to CH4, under 0.5 mM TDS. The microbial community was dominated by Eubacterium, Proteiniphilum and an unclassified member of the Eggerthellaceae family. The taxonomic diversity of the community decreased and conversely the phenotypic diversity increased, during operation. This work illustrated the scale-up potential of waste gas fermentations, by elucidating the effect of sulfide as a common gas impurity, and by demonstrating continuous, potentially renewable supply of electrons.

Lamb Fattening Under Intensive Pasture-Based Systems: A Review.

The benefits of pasture-based systems on the fatty acid composition of sheep meat appear to be achievable despite variability in the quality of the pastures. Lambs fed high levels of temperate pastures have an excess of N-ammonia derived from protein degradation. Furthermore, animal performance is highly variable depending on the quality of the pasture at the time of grazing, and high animal performance in these systems appears to be linked to the use of high-quality pastures with high availability, and is possibly added to by the inclusion of concentrates that allow increasing energy intake and a better use of the N in the pasture. The combination of high-quality pastures and total mixed ration offers a good alternative to the inclusion of concentrates in the diet, improving the use of N, and avoiding acidosis problems. However, information to determine the effect of a number of nutritional strategies on meat quality, and the minimum level of pasture intake necessary to achieve the benefits of pastoral systems is still lacking.

Mainstream partial nitritation/anammox with integrated fixed-film activated sludge: Combined aeration and floc retention time control strategies limit nitrate production.

Implementation of mainstream partial nitritation/anammox (PN/A) can lead to more sustainable and cost-effective sewage treatment. For mainstream PN/A reactor, an integrated fixed-film activated sludge (IFAS) was operated (26 °C). The effects of floccular aerobic sludge retention time (AerSRTfloc), a novel aeration strategy, and N-loading rate were tested to optimize the operational strategy. The best performance was observed with a low, but sufficient AerSRTfloc (~7d) and continuous aeration with two alternating dissolved oxygen setpoints: 10 min at 0.07-0.13 mg O2 L-1 and 5 min at 0.27-0.43 mg O2 L-1. Nitrogen removal rates were 122 ± 23 mg N L-1 d-1, and removal efficiencies 73 ± 13%. These conditions enabled flocs to act as nitrite sources while the carriers were nitrite sinks, with low abundance of nitrite oxidizing bacteria. The operational strategies in the source-sink framework can serve as a guideline for successful operation of mainstream PN/A reactors.

Microbial electrosynthesis from CO2: forever a promise?

Microbial electrosynthesis (MES) is an electrochemical process used to drive microbial metabolism for bio-production, such as the reduction of CO2 into industrially relevant organic products as an alternative to current fossil-fuel-derived commodities. After a decade of research on MES from CO2, figures of merit have increased significantly but are plateauing yet far from those expected to allow competitiveness for synthesis of commodity chemicals. Here we discuss the substantial technological shortcomings still associated with MES and evoke possible ways to mitigate them. It appears particularly challenging to obtain both relevant production rates (driven by high current densities) and energy conversion efficiency (i.e. low cell voltage) in microbial-compatible electrolytes. More competitive processes could arise by decoupling effective abiotic electroreductions (e.g. CO2 to CO or ethanol; H2 evolution) with subsequent fermentation processes.

Adaptation and characterization of thermophilic anammox in bioreactors.

Anammox, the oxidation of ammonium with nitrite, is a key microbial process in the nitrogen cycle. Under mesophilic conditions (below 40 °C), it is widely implemented to remove nitrogen from wastewaters lacking organic carbon. Despite evidence of the presence of anammox bacteria in high-temperature environments, reports on the cultivation of thermophilic anammox bacteria are limited to a short-term experiment of 2 weeks. This study showcases the adaptation of a mesophilic inoculum to thermophilic conditions, and its characterization. First, an attached growth technology was chosen to obtain the process. In an anoxic fixed-bed biofilm bioreactor (FBBR), a slow linear temperature increase from 38 to over 48 °C (0.05-0.07 °C d-1) was imposed to the community over 220 days, after which the reactor was operated at 48 °C for over 200 days. Maximum total nitrogen removal rates reached up to 0.62 g N L-1 d-1. Given this promising performance, a suspended growth system was tested. The obtained enrichment culture served as inoculum for membrane bioreactors (MBR) operated at 50 °C, reaching a maximum total nitrogen removal rate of 1.7 g N L-1 d-1 after 35 days. The biomass in the MBR had a maximum specific anammox activity of 1.1 ± 0.1 g NH4+-N g-1 VSS d-1, and the growth rate was estimated at 0.075-0.19 d-1. The thermophilic cultures displayed nitrogen stoichiometry ratios typical for mesophilic anammox: 0.93-1.42 g NO2--Nremoved g-1 NH4+-Nremoved and 0.16-0.35 g NO3--Nproduced g-1 NH4+-Nremoved. Amplicon and Sanger sequencing of the 16S rRNA genes revealed a disappearance of the original "Ca. Brocadia" and "Ca. Jettenia" taxa, yielding Planctomycetes members with only 94-95% similarity to "Ca. Brocadia anammoxidans" and "Ca. B. caroliniensis", accounting for 45% of the bacterial FBBR community. The long-term operation of thermophilic anammox reactors and snapshot views on the nitrogen stoichiometry, kinetics and microbial community open up the development path of thermophilic partial nitritation/anammox. A first economic assessment highlighted that treatment of sludge reject water from thermophilic anaerobic digestion of sewage sludge may become attractive.

Real-life management of patients with breakthrough cancer pain caused by bone metastases in Spain.

Purpose: We aimed to explore the characteristics, and real-life therapeutic management of patients with breakthrough cancer pain (BTcP) caused by bone metastases in Spain, and to evaluate physicians' opinion of and satisfaction with prescribed BTcP therapy. Participants and methods: For the purposes of this study, an ad-hoc questionnaire was developed consisting of two domains: a) organizational aspects and care standards; b) clinical and treatment variables of bone metastatic BTcP patients. In addition, physicians' satisfaction with their prescribed BTcP therapy was assessed. Specialists collected data from up to five patients receiving treatment for BTcP caused by bone metastasis, all patients gave their consent to participate prior to inclusion. Results: A total of 103 cancer pain specialists (radiation oncologists [38.8%], pain specialists [33.0%], and palliative care (PC) specialists [21.4%]) were polled, and data on 386 BTcP patients with bone metastatic disease were collected. Only 33% of the specialists had implemented specific protocols for BTcP management, and 19.4% had established referral protocols for this group of patients. Half of all participants (50.5%) address quality of life and quality of care in their patients; however, only 27.0% did so from the patient's perspective, as they should do. Most patients had multiple metastases and were prescribed rapid-onset fentanyl preparations (71.2%), followed by immediate-release morphine (9.3%) for the treatment of BTcP. Rapid-onset fentanyl was prescribed more often in PC units (79.0%) than in pain units (75.9%) and radiation oncology units (61.1%) (p<0.01). Furthermore, most physicians (71.8%) were satisfied with the BTcP therapy prescribed. Conclusions: Our results demonstrate the need for routine assessment of quality of life in patients with bone BTcP. These findings also underscore the necessity for a multidisciplinary therapeutic strategy for breakthrough pain in clinical practice in Spain.

Association of residual feed intake with abundance of ruminal bacteria and biopolymer hydrolyzing enzyme activities during the peripartal period and early lactation in Holstein dairy cows.

BACKGROUND: Residual feed intake (RFI) in dairy cattle typically calculated at peak lactation is a measure of feed efficiency independent of milk production level. The objective of this study was to evaluate differences in ruminal bacteria, biopolymer hydrolyzing enzyme activities, and overall performance between the most- and the least-efficient dairy cows during the peripartal period. Twenty multiparous Holstein dairy cows with daily ad libitum access to a total mixed ration from d - 10 to d 60 relative to the calving date were used. Cows were classified into most-efficient (i.e. with low RFI, n = 10) and least-efficient (i.e. with high RFI, n = 10) based on a linear regression model involving dry matter intake (DMI), fat-corrected milk (FCM), changes in body weight (BW), and metabolic BW. RESULTS: The most-efficient cows had ~ 2.6 kg/d lower DMI at wk 4, 6, 7, and 8 compared with the least-efficient cows. In addition, the most-efficient cows had greater relative abundance of total ruminal bacterial community during the peripartal period. Compared with the least-efficient cows, the most-efficient cows had 4-fold greater relative abundance of Succinivibrio dextrinosolvens at d - 10 and d 10 around parturition and tended to have greater abundance of Fibrobacter succinogenes and Megaspheara elsdenii. In contrast, the relative abundance of Butyrivibrio proteoclasticus and Streptococcus bovis was lower and Succinimonas amylolytica and Prevotella bryantii tended to be lower in the most-efficient cows around calving. During the peripartal period, the most-efficient cows had lower enzymatic activities of cellulase, amylase, and protease compared with the least-efficient cows. CONCLUSIONS: The results suggest that shifts in ruminal bacteria and digestive enzyme activities during the peripartal period could, at least in part, be part of the mechanism associated with better feed efficiency in dairy cows.

Nitrogen cycle microorganisms can be reactivated after Space exposure.

Long-term human Space missions depend on regenerative life support systems (RLSS) to produce food, water and oxygen from waste and metabolic products. Microbial biotechnology is efficient for nitrogen conversion, with nitrate or nitrogen gas as desirable products. A prerequisite to bioreactor operation in Space is the feasibility to reactivate cells exposed to microgravity and radiation. In this study, microorganisms capable of essential nitrogen cycle conversions were sent on a 44-days FOTON-M4 flight to Low Earth Orbit (LEO) and exposed to 10-3-10-4 g (gravitational constant) and 687 ± 170 µGy (Gray) d-1 (20 ± 4 °C), about the double of the radiation prevailing in the International Space Station (ISS). After return to Earth, axenic cultures, defined and reactor communities of ureolytic bacteria, ammonia oxidizing archaea and bacteria, nitrite oxidizing bacteria, denitrifiers and anammox bacteria could all be reactivated. Space exposure generally yielded similar or even higher nitrogen conversion rates as terrestrial preservation at a similar temperature, while terrestrial storage at 4 °C mostly resulted in the highest rates. Refrigerated Space exposure is proposed as a strategy to maximize the reactivation potential. For the first time, the combined potential of ureolysis, nitritation, nitratation, denitrification (nitrate reducing activity) and anammox is demonstrated as key enabler for resource recovery in human Space exploration.

Smart operation of nitritation/denitritation virtually abolishes nitrous oxide emission during treatment of co-digested pig slurry centrate.

The implementation of nitritation/denitritation (Nit/DNit) as alternative to nitrification/denitrification (N/DN) is driven by operational cost savings, e.g. 1.0-1.8 EUR/ton slurry treated. However, as for any biological nitrogen removal process, Nit/DNit can emit the potent greenhouse gas nitrous oxide (N2O). Challenges remain in understanding formation mechanisms and in mitigating the emissions, particularly at a low ratio of organic carbon consumption to nitrogen removal (CODrem/Nrem). In this study, the centrate (centrifuge supernatant) from anaerobic co-digestion of pig slurry was treated in a sequencing batch reactor. The process removed approximately 100% of ammonium a satisfactory nitrogen loading rate (0.4 g N/L/d), with minimum nitrite and nitrate in the effluent. Substantial N2O emission (around 17% of the ammonium nitrogen loading) was observed at the baseline operational condition (dissolved oxygen, DO, levels averaged at 0.85 mg O2/L; CODrem/Nrem of 2.8) with ∼68% of the total emission contributed by nitritation. Emissions increased with higher nitrite accumulation and lower organic carbon to nitrogen ratio. Yet, higher DO levels (∼2.2 mg O2/L) lowered the aerobic N2O emission and weakened the dependency on nitrite concentration, suggesting a shift in N2O production pathway. The most effective N2O mitigation strategy combined intermittent patterns of aeration, anoxic feeding and anoxic carbon dosage, decreasing emission by over 99% (down to ∼0.12% of the ammonium nitrogen loading). Without anaerobic digestion, mitigated Nit/DNit decreases the operational carbon footprint with about 80% compared to N/DN. With anaerobic digestion included, about 4 times more carbon is sequestered. In conclusion, the low CODrem/Nrem feature of Nit/DNit no longer offsets its environmental sustainability provided the process is smartly operated.

Drosophila chem mutations disrupt epithelial polarity in Drosophila embryos.

Drosophila embryogenesis has proven to be an extremely powerful system for developmental gene discovery and characterization. We isolated five new EMS-induced alleles that do not complement the l(3R)5G83 lethal line isolated in the Nüsslein-Volhard and Wieschaus screens. We have named this locus chem. Lethality of the new alleles as homozygous zygotic mutants is not completely penetrant, and they have an extended phenocritical period. Like the original allele, a fraction of mutant embryos die with cuticular defects, notably head involution and dorsal closure defects. Embryonic defects are much more extreme in germline clones, where the majority of mutant embryos die during embryogenesis and do not form cuticle, implying a strong chem maternal contribution. chem mutations genetically interact with mutations in cytoskeletal genes (arm) and with mutations in the epithelial polarity genes coracle, crumbs, and yurt. chem mutants dorsal open defects are similar to those present in yurt mutants, and, likewise, they have epithelial polarity defects. chem1 and chem3 mutations suppress yurt3 , and chem3 mutants suppress crumbs1 mutations. In contrast, chem1 and coracle2 mutations enhance each other. Compared to controls, in chem mutants in embryonic lateral epithelia Crumbs expression is mislocalized and reduced, Coracle is increased and mislocalized basally at embryonic stages 13-14, then reduced at stage 16. Arm expression has a similar pattern but levels are reduced.

Starved anammox cells are less resistant to NO2⁻ inhibition.

Anaerobic ammonium oxidizing (anammox) bacteria are be inhibited by their terminal electron acceptor, nitrite. Serious nitrite inhibition of the anammox bacteria occurs if the exposure coincides with the absence of the electron donating substrate, ammonium and pH < 7.2. Starvation of biomass occurs during underloading of bioreactors or biomass storage. This work investigated the effect of starvation on the sensitivity of anammox bacteria to nitrite exposure. Batch activity tests were carried out evaluating the response of anammox biomass subjected to different levels of starvation upon exposure to nitrite in the presence and absence of ammonium (active- and resting-cells, respectively). The response of the bacteria was evaluated by measuring the specific anammox activity and the evolution of the ATP content in the biomass over time. The 50% inhibitory concentrations of nitrite in starved- and fresh-resting-cells was 7 mg N L(-1) and 52 mg N L(-1), respectively. By contrast, only moderate nitrite inhibition occurred to starved anammox biomass when exposed to nitrite and ammonium simultaneously. Maximum ATP levels were observed in fresh cells. The ATP content in starved resting cells peaked 2-3 h after addition of NO2(-)(-). The response was hindered in cells starved for long periods. These findings agreed with a bioreactor study in which underloading of anammox biomass (0.10 g N L(-1) d(-1)) decreased its tolerance to a nitrite (only) exposure (101 mg NO2(-)-N L(-1)) and completely disrupted the N removal capacity of the biomass. A similar accumulation of 108 mg NO2(-)-N L(-1) after operation at 0.95 g N L(-1) d(-1) did not cause observable inhibition of the bacteria. The results taken as a whole demonstrate that starved anammox biomass is highly sensitive to nitrite toxicity. An explanation is proposed based on energy requirements to translocate nitrite in the cell.

The intracellular proton gradient enables anaerobic ammonia oxidizing (anammox) bacteria to tolerate NO2 - inhibition.

Anammox bacteria are inhibited by nitrite, which is one of their substrates. By utilizing 2,4 dinitrophenol and carbonyl cyanide m-chlorophenyl hydrazone, two uncouplers of respiration, we demonstrate that nitrite tolerance of anammox cells is strongly dependent on their ability to maintain a proton gradient, which may be the driving force for active nitrite transport system.