Availability and utilization of waste fish feed by mussels Mytilus edulis in a commercial integrated multi-trophic aquaculture (IMTA) system: A multi-indicator assessment approach

Fish feed waste enhancement of the particulate food supply and performance of mussels Mytilus edulis suspended near salmon cages at an integrated multi-trophic aquaculture (IMTA) site was assessed using a multi-indicator approach. Dietary indicators included bulk measurements of seston quantity and nutritional quality, proximate analysis (PA), fatty acid (FA) and stable isotope (SI) composition. Mussel tissue indicators consisted of PA and FA composition. Mussel performance was assessed from physiological integrations (scope for growth, SFG), growth efficiency (K₂) and condition index (CI). All measurements were made over 2 days at a commercial IMTA farm and a monoculture mussel farm in the Bay of Fundy (Canada). Significant differences detected in seston quantity and quality were within the range of natural spatial variability. The SFG of IMTA mussels was lower (28.71 J h⁻¹) than monoculture mussels (38.71 J h⁻¹) and reflected site differences in natural food availability and composition that affected absorption rate. PA of mussel organs didn't reflect a significant fish feed contribution to the mussel diet. However, dietary enhancement and assimilation of fish feed waste was demonstrated by significantly higher levels of feed FA biomarkers 20:1ω9, 18:2ω6, 18:1ω9 and low ω3/ω6 ratio in seston, mussel tissues and feces at the IMTA site than at the mussel farm. SI (δ¹³C and δ¹⁵N) in seston and mussel feces significantly differed among sites and IMTA mussels had significantly higher CI (21%) than monoculture individuals (16%). It was concluded that bulk indicators of the diet, short-term physiological integrations, and PA of mussel tissues have a limited capacity to detect dietary enhancement at IMTA sites. FA and SI tracers of fish feed waste were shown to be more sensitive for detecting the low-levels of diet enhancement within the large range of natural seston variation.     

Predation by the invasive American signal crayfish, Pacifastacus leniusculus Dana, on the invasive zebra mussel, Dreissena polymorpha Pallas: the potential for control and facilitation

Non-indigenous crayfish often have major ecological impacts on invaded water bodies, and have contributed to the decline of native crayfish species throughout Europe. The American signal crayfish, Pacifastacus leniusculus, is the most widespread invasive crayfish in Great Britain, where the zebra mussel, Dreissena polymorpha, is similarly an invasive pest species. The potential for the American signal crayfish to regulate zebra mussel populations was investigated through a series of laboratory experiments. Crayfish were found to be highly size selective, consuming significantly more of the smallest size class of zebra mussels offered (7–12 mm), over medium (16–21 mm) and large (25–30 mm). Crayfish feeding rate on zebra mussels was not altered when mussels were presented clumped together in natural druses compared with mussels in a disassembled druse. Crayfish spent significantly more time foraging when mussels were unattached, and a greater proportion of attacks were on medium and large than on small mussels (83% of attacks were on medium and large mussels when unattached as opposed to 47% when on druses). Individual crayfish feeding rate decreased significantly at densities of > ~5 crayfish m⁻². Signal crayfish are, therefore, unlikely to be able to significantly impact established populations of zebra mussels in the wild, although zebra mussels have the potential to provide crayfish with a substantial food source.     

Population genetic history of the dreissenid mussel invasions: expansion patterns across North America

This study tests population genetic patterns across the Eurasian dreissenid mussel invasions of North America—encompassing the zebra mussel Dreissena polymorpha (1986 detection) and the quagga mussel D. rostriformis bugensis (detected in 1990, which now has largely displaced the former in the Great Lakes). We evaluate their source-spread relationships and invasion genetics using 9–11 nuclear microsatellite loci for 583 zebra mussels (21 sites) and 269 quagga mussels (12 sites) from Eurasian and North American range locations, with the latter including the Great Lakes, Mississippi River basin, Atlantic coastal waterways, Colorado River system, and California reservoirs. Additionally, mtDNA cytochrome b gene sequences are used to verify species identity. Our results indicate that North American zebra mussels originate from multiple non-native northern European populations, whereas North American quagga mussels trace to native estuaries in the Southern Bug and Dnieper Rivers. Invasive populations of both species show considerable genetic diversity and structure (zebra F _ST = 0.006–0.263, quagga F _ST = 0.008–0.267), without founder effects. Most newer zebra mussel populations have appreciable genetic diversity, whereas quagga mussel populations from the Colorado River and California show some founder effects. The population genetic composition of both species changed over time at given sites; with some adding alleles from adjacent populations, some losing them, and all retaining closest similarity to their original composition. Zebra mussels from Kansas and California appear genetically similar and assign to a possible origin from the St. Lawrence River, whereas quagga mussels from Nevada and California assign to a possible origin from Lake Ontario. These assignments suggest that overland colonization pathways via recreational boats do not necessarily reflect the most proximate connections. In conclusion, our microsatellite results comprise a valuable baseline for resolving present and future dreissenid mussel invasion pathways.     

Growth,intake, diet digestibility,and nitrogen use in three hair sheep breeds fed alfalfa hay

Pen feeding and metabolism trials were conducted to determine intake, diet digestibility and nitrogen (N) use in three hair sheep breeds with differing growth potential offered an alfalfa hay diet. For pen feeding, 24 6-mo-old wether lambs, equally representing the Barbados Blackbelly, Katahdin, and St. Croix breeds, were paired by breed, placed in 12 enclosed cement-floor pens (2.5 m × 3.5 m), and offered chopped alfalfa (Medicago sativa L.; 17.6% CP, 50.4% NDF, and 36.4% ADF) ad libitum. Lambs were allowed a 14-d adaptation period to pens and diets, and remained on trial for 56 d. Starting body weight (BW) was different (P < 0.05) among breeds (Barbados Blackbelly: 23.8 kg; Katahdin: 36.5 kg; St. Croix: 29.1 kg). Daily gain was not different (P < 0.10) between Katahdin (131 g/d) and St. Croix (117 g/d), but both were higher (P < 0.05) than Barbados Blackbelly (87 g/d). Daily dry matter hay intake was similar among breeds (107–109 g/kg BW^0.75), and feed to gain ratio ranged from 8.7 in St. Croix and 9.1 in Katahdin to 10.5 in Barbados Blackbelly, but was not different (P > 0.10) among breeds. Six lambs per breed were used in the metabolism trial. Total DMI was greater (P < 0.01) for Katahdin (1196 g/d) than St. Croix (907 g/d) and Barbados Blackbelly (858 g/d), but was not different (P > 0.10) adjusted for body weight (mean: 71 g/kg BW^0.75). Coefficients of apparent digestibility for DM (P < 0.06), OM (P < 0.06), N (P < 0.05), NDF (P < 0.08), and ADF (P < 0.08) were higher for Katahdin compared to Barbados Blackbelly with St. Croix intermediate. Intake of N (P < 0.01), feces N (P < 0.08), urine N (P < 0.01), absorbed N (P < 0.01), and retained N (P < 0.05) also were greater for Katahdin compared to St. Croix and Barbados Blackbelly. Blood glucose, urea-N, and creatinine were not different (P > 0.10) among breeds. Breeds differed in their growth performance in line with breed mature size, and differences were observed in N digestibility and absorption.     

Variations in hemocyte counts in the mussel,Mytilus edulis: Similar reaction patterns occur in disappearance and return of molluscan hemocytes and vertebrate leukocytes

It was asked whether variations in hemocyte counts in a mussel can be explained by mechanisms known to govern the leukocyte number in vertebrates. Hemolymph of 25 freshly collected Mytilus edulis contained (4.2 ± 1.75) × 10⁶ cells/mL including basophilic and eosinophilic granulocytes and 6.6 ± 5.5% hyalinocytes (15 animals). After 12 or 30 days under optimal laboratory conditions, hemocytes in circulation decreased to less than 1 × 10⁶/mL, the lowest number observed being 5 × 10⁵ cells/mL. Within 2 min of a stressful stimulus, cell numbers doubled or increased by a factor of 3 or 4. After stressing mussels by keeping them out of water for 1 h, cell counts were as high as 1.2 × 10⁷ cells/mL. The quick rate of increase in cell counts is not due to hemocyte proliferation. In mussels, returned to optimal water conditions, cell numbers dropped following an exponential decay curve (y = 5.6865 · (0.9936^X). Not all hemocyte types decreased in number to the same extent. After a strong decrease in the total cell count induced by injection of LPS, the remaining hemocyte population contained a larger percentage of basophils. This indicated the disappearance of eosinophilic cells from the circulation. Stress situations caused their return. Hemocytopenia or stress-induced hemocytosis in M. edulis, both in conjunction with changes in the percentage of granulocytes present, resembles margination/demargination processes in mammals where the concentration of circulating leukocyte subsets depends on the expression of adhesive receptor–ligand molecules on the surface of specific leukocyte types and vascular endothelial cells. In Mytilus edulis, variations in the concentration of distinct cell groups excluded heart activity to explain cell fluctuations. Furthermore, in this mussel, where hemocyte proliferation is not the reason for rapid hemocytosis, cell divisions were nevertheless demonstrated; they seem to be important in maintaining hemocyte homeostasis as 10–20% of cells in circulation possess the capacity to proliferate. They belong to the group of basophilic granulocytes.     

Toxic strains of the Alexandrium ostenfeldii complex in southern South America (Beagle Channel,Argentina)

During phytoplankton monitoring in the Beagle Channel (≈54°52′ S, 67°32′ W) a previously undetected Alexandrium species was observed in coincidence with mouse bioassay toxicity. Detailed thecal plates analysis using epifluorescence and scanning electron microscopy revealed the presence of the Alexandrium ostenfeldii species complex, showing a mixture of the diagnostic features usually used to discriminate between the morphospecies A. ostenfeldii and A. peruvianum. Cells of the A. ostenfeldii complex were commonly observed during spring after the main annual diatom bloom, when temperatures and salinities were respectively around 7.5–10 °C and 30–30.5 psu, and nutrients showed a seasonal decrease. Toxin analysis by liquid chromatography–mass spectrometry revealed the production of 13-desmethyl spirolide C and 20-methyl spirolide G in cell cultures. The cellular contain of spirolides during exponential phase growth was 0.5906 ± 0.0032 and 0.1577 ± 0.0023 pg cell⁻¹ for 13-desMe-C and 20-Me-G, respectively. A third unknown compound, with a structure resembling that of spirolides was also detected in culture. Moreover, an additional compound with a similar m/z (692) than that of 13-desMe-C but presenting a higher retention time (Rt = 40.5 min) was found in high proportions in mussel samples. PSP toxins were present at low concentration in mussels but were not detected in cultures. These results extend the world-wide distribution of toxic strains of the A. ostenfeldii complex to the Beagle Channel (southern South America), where toxic events have been traditionally linked to the presence of Alexandrium catenella. This is the first confirmed occurrence of spirolides in mussels and plankton from Argentina, which highlights the importance of monitoring these toxins and their producing organisms to protect public health and improve the management of shellfish resources.     

Invasive quagga mussels can be attenuated by redear sunfish (Lepomis microlophus) in the Southwestern United States

The degree to which invasive quagga mussels have spread across the Southwestern United States is unprecedented, and until now, none of the controls currently available is risk-free to the environment or human health. Here we demonstrate that stocking high-density (0.42 fish/m³ or 1.90 fish/m²) redear sunfish (Lepomis microlophus) in an infested lake enclosure removed adult quagga mussels efficiently and appeared to suppress their growth and recruitment. However, mussels were not eradicated during the short term experimental course. Long-term study needs to be conducted to determine whether mussels can be completely eliminated by redear sunfish and what optimum density would be required for balancing high efficacy mussel control with cost-effectiveness. Redear sunfish is a species native to the Southeastern United States that has become a popular, stocked sportfish in the Southwest. Targeted stocking of redear sunfish may be an efficient and environmentally friendly approach to mitigate the colonization of invasive quagga mussels in suitable open water ecosystems, especially those with new dreissenid infestations or those threatened by invasion.     

Kinetics modeling of the acidolysis with immobilized Rhizomucor miehei lipases for production of structured lipids from sunflower oil

Sunflower oil modification for production of semisolid fats was carried out via acidolysis using palmitic and stearic acids (P + St), hexane and a developed biocatalyst from Rhizomucor miehei lipases. Its kinetic behavior was studied by employing three mathematical models proposed in the literature. Furthermore, a new model was proposed to describe not only the variation of triacylglycerols (TAG), diacylglycerols (DAG), and free fatty acids groups but also the acyl migration reaction occurrence. The effect of the reaction temperature on the kinetic and equilibrium parameters, as well as TAG and reaction intermediates profiles was analyzed. Increasing reaction temperature generated major changes in the overall composition of acylglycerols and gave rise to the highest composition of P + St in the obtained structured lipids (58%, 70 h, 60 °C). P + St incorporation was successfully adjusted by an empirical model (Model I) and a lumped parameter model (Model II) for all the studied reaction times, while the model based on a Ping Pong Bi Bi mechanism (Model III) was only able to describe the kinetics behavior (through the variation of reactant saturated fatty acids concentration) until 24 h. Experimental data were fit satisfactorily by the proposed model (Model IV), showing that the increment in the disaturated TAG formation achieved by the increment in temperature was principally related to the favored DAG formation from triunsaturated TAG.     

Field-to-laboratory transport protocol impacts subsequent physiological biomarker response in the marine mussel,Perna canaliculus

The transfer of mussels from field to laboratory, or transplantation between clean and contaminated field settings, is a common protocol in ecotoxicology. However, collection and transport of mussels could lead to stress that may impact biomarker responses, and thus confound interpretation of results. Physiological responses (clearance rate, absorption efficiency, excretion rate, respiration rate and scope-for-growth) of green-lipped mussels (Perna canaliculus) exposed to four different transportation protocols were investigated. These protocols included immersion in site seawater (SSW), immersion in artificial seawater (ASW), and emersion (aerial transport; EMS) at two temperatures (15 °C and 5 °C). Physiological measurements were conducted after a simulated 24 h “transport” phase and a 48 h “recovery” phase. Clearance rates were significantly inhibited by the EMS 5 °C and ASW protocols relative to SSW treatment, although the clearance rate of the latter recovered after 48 h. A similar pattern was observed for excretion and respiration rates for ASW. Decreased excretion rates for EMS 15 °C and respiration rates for EMS 5 °C were also recorded relative to values for SSW following “recovery”. Negative scope-for-growth was observed for all treatments except EMS 15 °C. These data suggest transport emersed at ambient air temperatures is the best method to maintain physiological health of green-lipped mussels.     

The CCKB antagonist CI988 reduces food intake in fasted rats via a dopamine mediated pathway

Studies have shown a reduction of food intake following peripheral and brain injection of CCK. However, it remains to be established whether endogenous central CCK is involved in the regulation of food intake. We investigated the role of central CCK in the regulation of food intake by pharmacological manipulation of the CCK_B (CCK₂) receptor system. Intracerebroventricularly (ICV) cannulated male Sprague Dawley rats were fasted for 24 h and received an ICV injection of the CCK_B receptor antagonist CI988 at a dose of 10 nmol or 49 nmol or vehicle. Another group received two consecutive ICV injections consisting of the corticotropin-releasing factor (CRF) receptor-1 (CRF₁) antagonist, CP376395 (3 nmol) or the CRF₂ receptor antagonist, K41498 (2 nmol) alone, or followed by CI988 (49 nmol). Lastly, another group of rats received an intraperitoneal (IP) injection of the dopamine antagonist, flupentixol (∼197 and ∼493 nmol/kg) alone, or followed by CI988 (49 nmol, ICV). Cumulative food intake was assessed for 11 h. Vehicle injected rats showed a robust feeding response. CI988 at 49 nmol reduced food intake by 30% starting at 2 h post injection. CP376395 and K41498 had no effect on food intake. Flupentixol injected IP at a dose of 197 and 493 nmol/kg alone did not modulate food intake whereas the higher dose blocked the CI988-induced reduction of feeding. During the dark phase, CI988 had no effect on food intake in unfasted rats. In summary, CCK_B signaling is involved in the regulation of food intake after a fast likely by downstream dopamine signaling.     

Body size and food thresholds for zero growth in Dreissena polymorpha: a mechanism underlying intraspecific competition

1. Dreissena polymorpha is an extraordinarily successful invasive species that shows high recruitment of small juvenile mussels on established mussel banks. Such juvenile settlement on, and overgrowth of, large adult mussels; however, leads to competition with adults, and often at high densities and low‐food concentrations. 2. The concept of food thresholds for zero growth has been a powerful approach to explaining size‐related exploitative competition in different zooplankton species. We applied it to investigate whether food threshold concentrations for zero growth (C₀) differ between juvenile and adult zebra mussels. 3. By determining body mass growth at various concentrations of a diet mixture (Nannochloropsis limnetica and Isochrysis aff. galbana) we demonstrate that the threshold food concentration for growth of juvenile mussels (C₀ = 0.08 mg C L⁻¹) is substantially lower than that for adults (C₀ = 0.36 mg C L⁻¹). 4. This indicates that, at low food availability, juvenile zebra mussels are competitively superior to their larger conspecifics. Within zebra mussel banks plankton food is substantially depleted and so the observed mechanism might ensure juvenile success and therefore the regeneration of mussel banks in nature.     

Fouling of European freshwater bivalves (Unionidae) by the invasive zebra mussel (Dreissena polymorpha)

1. The zebra mussel (Dreissena polymorpha) is well known for its invasive success and its ecological and economic impacts. Of particular concern has been the regional extinction of North American freshwater mussels (Order Unionoida) on whose exposed shells the zebra mussels settle. Surprisingly, relatively little attention has been given to the fouling of European unionoids. 2. We investigated interspecific patterns in fouling at six United Kingdom localities between 1998 and 2008. To quantify the effect on two pan‐European unionoids (Anodonta anatina and Unio pictorum), we used two measures of physiological status: tissue mass : shell mass and tissue glycogen content. 3. The proportion of fouled mussels increased between 1998 and 2008, reflecting the recent, rapid increase in zebra mussels in the U.K. Anodonta anatina was consistently more heavily fouled than U. pictorum and had a greater surface area of shell exposed in the water column. 4. Fouled mussels had a lower physiological condition than unfouled mussels. Unlike tissue mass : shell mass ratio, tissue glycogen content was independent of mussel size, making it a particularly useful measure of condition. Unio pictorum showed a stronger decline in glycogen with increasing zebra mussel load, but had a broadly higher condition than A. anatina at the time of study (July). 5. Given the high conservation status and important ecological roles of unionoids, the increased spatial distribution and fouling rates by D. polymorpha in Europe should receive more attention.     

Phosphorus recycling by zebra mussels in relation to density and food resource availability

Using flow-through microcosms, we examined phosphorus (P) recycling by zebra mussels under conditions of nearly constant food resource supply and varying zebra mussel population densities (600–5200 ind./m²). At all density levels, zebra mussels filtered substantial algae, measured as chlorophyll biomass. Because chlorophyll biomass inputs were low throughout the study, zebra mussel biomass-specific rates of chlorophyll filtration declined with increasing density, suggesting food resource limitation at the higher densities. We observed net total P export and high zebra mussel biomass-specific rates of P recycling over time in microcosms at high zebra mussel densities. In systems with a low zebra mussel density, net total P export did not occur over time. Our results suggest the occurrence of P remineralization by zebra mussels and net loss associated with emaciation during periods of temporary starvation. These findings have implications for P dynamics since zebra mussels can be subjected to periods of starvation over seasonal and annual time scales.     

The metabolic cost of bicarbonate use in the submerged plant Elodea nuttallii

The aquatic plant Elodea nuttallii (Planch.) St. John has been shown to express plasticity in the source of inorganic carbon it uses for photosynthesis. An investigation was undertaken to determine what effect the switch from CO₂ to HCO₃⁻ use had on the growth of E. nuttallii. Plants were grown under reduced CO₂ availability that favoured the switch, together with control plants (CO₂ at equilibrium with air) that continued to use CO₂ only. The extent to which both sets of plants could utilise HCO₃⁻ was determined (as the ratio of oxygen evolution at pH 9 and 6.5), and several measures of growth were made. Although reduced CO₂ availability produced an increase in HCO₃⁻ utilisation, no differences were found in the measured growth of the plants. Therefore, it was possible to estimate, from the difference between the estimated rate of photosynthesis of the plants utilising HCO₃⁻ and those using CO₂ only, the approximate cost of constructing, maintaining and running the bicarbonate utilisation mechanism in this species as 69 μmol photons m⁻² s⁻¹. This value can be used to estimate an irradiance of circa 80 μmol m⁻² s⁻¹ below which HCO₃⁻ use would not be expected in this species, an irradiance commonly experienced by submerged macrophytes in the field.     

Testing a typology system of running waters for conservation planning in Hungary

The zebra mussel (Dreissena polymorpha) and its congener the quagga mussel (Dreissena rostriformis bugensis) are both invaders in freshwater, but have very different invasion histories, with zebra mussels attaining substantially faster rates of spread at virtually all spatial scales. However, in waterbodies where they co-occur, D. r. bugensis can displace D. polymorpha. To determine if the mechanisms for this displacement are associated with different survival and growth, we kept mussels in flow-through tanks for 289 days with two temperature regimes that mimicked the natural surface water (littoral zone) and hypolimnion conditions of Lake Erie. For the littoral zone regime, we used water directly from the surface of Lake Erie (range 4–25°C, average 11.9 ± 0.6°C). For the profundal zone treatment, Lake Erie surface water was chilled to about 6°C (range 5–8°C, average 6.2 ± 0.6°C) for the full duration of the experiment. For each of these temperature regimes, we used three replicate tanks with only zebra mussels present and three replicate tanks with only quagga mussels (150 ind./tank each), and three replicate tanks with both species (75 ind./tank of each species). Quagga mussels had higher survivorship and grew more than zebra mussels in all treatments. For both species, the size of the mussel entering the winter was critical for survivorship. Larger mussels had a higher survival over the winter in all treatments. For both species, there was a survivorship and growth tradeoff. In the warmer littoral zone treatment both species had higher growth, but lower survival than in the colder profundal zone treatment. Surprisingly, although quagga mussels outperformed zebra mussels, zebra mussel survivorship was better when they were faced with competition by quagga mussels than with just intraspecific competition. In addition, quagga mussels suffered size-specific mortality during the growing season only when facing interspecific competition with zebra mussels. Further experiments are needed to determine the possible mechanisms for these interspecific effects.     

Digestive capacities allow the Mexican long-nosed bat (Leptonycteris nivalis) to live in cold environments

Digestive capabilities of nectar-feeding vertebrates to assimilate sugars affect their ability to acquire and store energy and could determine the minimal temperatures at which these animals can survive. Here, we described the sugar digestive capability of Leptonycteris nivalis and related it with its capacity to live in cold environments. We measured the enzymatic activity, food intake rate and changes in body mass of bats feeding at four different sucrose concentrations (from 5 to 35% wt./vol.). Additionally, we used a mathematical model to predict food intake and compared it with the food intake of bats. L. nivalis was able to obtain ~ 111.3 kJ of energy regardless of the sugar concentration of their food. Also, bats gained ~ 2.57 g of mass during the experimental trials and this gain was independent of sugar concentration. The affinity (1 / K_m) of sucrase (EC 3.2.1.48) was one order of magnitude higher relative to that reported for its sister species Leptonycteris yerbabuenae (0.250 and 0.0189 mmol^? 1 L, respectively), allowing this species to have a higher energy intake rate. We propose that the high ability to acquire energy conferred L. nivalis the faculty to invade cold environments, avoiding in this way the ecological competition with its sympatric species L. yerbabuenae.     

Mechanisms and impact of differential fouling of the zebra mussel Dreissena polymorpha on different unionid bivalves

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The influence of two differently sized dams on mussel assemblages and growth

Dams have been shown to impact freshwater mussels. We examined how mussels respond to differently sized dams (18 vs. 4 m) on the St. Croix River and its tributary, the Sunrise River. We hypothesized that: mussel density and growth rate would be greater downstream of the smaller dam due to the relatively greater food subsidies and temperature effects of the reservoir above it; and the effects of the small dam would moderate downstream as the localized impacts of the dam were reduced. We quantitatively sampled mussels upstream and downstream of the dams. For a common species, Actinonaias ligamentina, we ascertained growth rates by measuring successive growth rings. The highest mussel richness and diversity were upstream and downstream of the large dam. Higher mussel density was found immediately below the small dam but declined downstream. A. ligamentina downstream of the small dam grew faster and were larger than individuals in other reaches. Food availability and temperature appeared to influence mussel density and growth rate for A. ligamentina downstream of the small dam. This study provides information that may help managers decide whether to remove small dams or to maintain them because of the unique mussel habitats below these structures.     

Wastewater effects on Phaeodactylum tricornutum (Bohlin): Setting up a classification system

Phytoplankton is highly productive in marine coastal ecosystems that generally present high levels of anthropogenic pressures. Microalgae presents very high surface-to-volume ratio and may respond promptly to contaminants showing either an increase in growth or inhibition effects. Thus phytoplankton organisms provide information on the potential impacts of contaminants on the supported marine-coastal food webs. The diatom Phaeodactylum tricornutum Bohlin is commonly used in toxicity testing according to the ISO 10253:2006 standardised protocol, but a comprehensive inventory of tested substances has not been compiled yet. The aim of this study is to establish a wastewater effect score based on P. tricornutum exposed to domestic, municipal and industrial wastewaters from activated sludge sequencing batch reactor and ultra-filtration membrane biological reactors. Wastewater samples produced stimulation and inhibition effects identified by biostimulation unit (BU₅₀) and toxicity unit (TU₅₀) both at 50% effect, respectively. Within the stimulation scenario, toxicity was low if 0 < BU₅₀ ≤ 0.31, medium if 0.31 < BU₅₀ ≤ 1.05, high if 1.05 < BU₅₀ ≤ 1.64, and very high if BU₅₀ > 1.64. Within the inhibition scenario, toxicity was low if 0 < TU₅₀ ≤ 0.07, medium if 0.07 < TU₅₀ ≤ 2.67, high if 2.67 < TU₅₀ ≤ 5.86, and very high if TU₅₀ > 5.86. Results evidenced that nitrogen and phosphorus concentrations were not correlated to ecotoxicological values, probably due to the presence of undetected micronutrients, confirming the importance of toxicity-based hazard assessment.     

Dihydropyrimidones: As novel class of β-glucuronidase inhibitors

Dihydropyrimidones 1–37 were synthesized via a ‘one-pot’ three component reaction according to well-known Biginelli reaction by utilizing Cu(NO₃)₂·3H₂O as catalyst, and screened for their in vitro β-glucuronidase inhibitory activity. It is worth mentioning that amongst the active molecules, compounds 8 (IC₅₀ = 28.16 ± .056 μM), 9 (IC₅₀ = 18.16 ± 0.41 μM), 10 (IC₅₀ = 22.14 ± 0.43 μM), 13 (IC₅₀ = 34.16 ± 0.65 μM), 14 (IC₅₀ = 17.60 ± 0.35 μM), 15 (IC₅₀ = 15.19 ± 0.30 μM), 16 (IC₅₀ = 27.16 ± 0.48 μM), 17 (IC₅₀ = 48.16 ± 1.06 μM), 22 (IC₅₀ = 40.16 ± 0.85 μM), 23 (IC₅₀ = 44.16 ± 0.86 μM), 24 (IC₅₀ = 47.16 ± 0.92 μM), 25 (IC₅₀ = 18.19 ± 0.34 μM), 26 (IC₅₀ = 33.14 ± 0.68 μM), 27 (IC₅₀ = 44.16 ± 0.94 μM), 28 (IC₅₀ = 24.16 ± 0.50 μM), 29 (IC₅₀ = 34.24 ± 0.47 μM), 31 (IC₅₀ = 14.11 ± 0.21 μM) and 32 (IC₅₀ = 9.38 ± 0.15 μM) found to be more potent than the standard d-saccharic acid 1,4-lactone (IC₅₀ = 48.4 ± 1.25 μM). Molecular docking study was conducted to establish the structure–activity relationship (SAR) which demonstrated that a number of structural features of dihydropyrimidone derivatives were involved to exhibit the inhibitory potential. All compounds were characterized by spectroscopic techniques such as ¹H, ¹³C NMR, EIMS and HREI-MS.