Chitosan-Based Films with 2-Aminothiophene Derivative: Formulation,Characterization and Potential Antifungal Activity

In this study, films of chitosan and 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carbonitrile (6CN), a 2-aminothiophene derivative with great pharmacological potential, were prepared as a system for a topical formulation. 6CN-chitosan films were characterized by physicochemical analyses, such as Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), and scanning electronic microscopy (SEM). Additionally, the antifungal potential of the films was evaluated in vitro against three species of Candida (C. albicans, C. tropicalis, and C. parapsilosis). The results of the FTIR and thermal analysis showed the incorporation of 6CN in the polymer matrix. In the diffractogram, the 6CN-chitosan films exhibited diffraction halos that were characteristic of amorphous structures, while the micrographs showed that 6CN particles were dispersed in the chitosan matrix, exhibiting pores and cracks on the film surface. In addition, the results of antifungal investigation demonstrated that 6CN-chitosan films were effective against Candida species showing potential for application as a new antifungal drug.     

Effect of Fucoidan on the Mitochondrial Membrane Potential (ΔΨm) of Leukocytes from Patients with Active COVID-19 and Subjects That Recovered from SARS-CoV-2 Infection

Fucoidan is a polysaccharide obtained from marine brown algae, with anti-inflammatory, anti-viral, and immune-enhancing properties, thus, fucoidan may be used as an alternative treatment (complementary to prescribed medical therapy) for COVID-19 recovery. This work aimed to determine the ex-vivo effects of treatment with fucoidan (20 µg/mL) on mitochondrial membrane potential (ΔΨm, using a cationic cyanine dye, 3,3′-dihexyloxacarbocyanine iodide (DiOC₆(3)) on human peripheral blood mononuclear cells (HPBMC) isolated from healthy control (HC) subjects, COVID-19 patients (C-19), and subjects that recently recovered from COVID-19 (R1, 40 ± 13 days after infection). In addition, ex-vivo treatment with fucoidan (20 and 50 µg/mL) was evaluated on ΔΨm loss induced by carbonyl cyanide 3-chlorophenylhydrazone (CCCP, 150 µM) in HPBMC isolated from healthy subjects (H) and recovered subjects at 11 months post-COVID-19 (R2, 335 ± 20 days after infection). Data indicate that SARS-CoV-2 infection induces HPBMC loss of ΔΨm, even 11 months after infection, however, fucoidan promotes recovery of ΔΨm in PBMCs from COVID-19 recovered subjects. Therefore, fucoidan may be a potential treatment to diminish long-term sequelae from COVID-19, using mitochondria as a therapeutic target for the recovery of cellular homeostasis.     

Limnology of Macrobrachium amazonicum grow‐out ponds subject to high inflow of nutrient‐rich water and different stocking and harvest management

We evaluated the water characteristics and particle sedimentation in Macrobrachium amazonicum (Heller 1862) grow‐out ponds supplied with a high inflow of nutrient‐rich water. Prawns were subject to different stocking and harvesting strategies: upper‐graded juveniles, lower‐graded juveniles, non‐graded juveniles+selective harvesting and traditional farming (non‐grading juveniles and total harvest only). Dissolved oxygen, afternoon N‐ammonia and N‐nitrate and soluble orthophosphate were lower in the ponds in comparison with inflow water through the rearing cycle. Ponds stocked with the upper population fraction of graded prawns showed higher turbidity, total suspended solids and total Kjeldahl nitrogen than the remaining treatments. An increase in the chemical oxygen demand:biochemical oxygen demand ratio from inlet (4.9) to pond (7.1–8.0) waters indicated a non‐readily biodegradable fraction enhancement in ponds. The sedimentation mean rate ranged from 0.08 to 0.16 mm day^?1 and sediment contained >80% of organic matter. The major factors affecting pond ecosystem dynamic were the organic load (due to primary production and feed addition) and bioturbation caused by stocking larger animals. Data suggest that M. amazonicum grow‐out in ponds subjected to a high inflow of nutrient‐rich water produce changes in the water properties, huge accumulation of organic sediment at the pond bottom and non‐readily biodegradable material in the water column. However, the water quality remains suitable for aquaculture purposes. Therefore, nutrient‐rich waters, when available, may represent a source of unpaid nutrients, which may be incorporated into economically valued biomass if managed properly.     

Influence of light intensity on biofilm formation and the performance of pink shrimp Farfantepenaeus paulensis juveniles reared in cages

A reduction in light intensity may increase shrimp activity, although it may also negatively affect the development of photoautotrophic organisms present in biofilm, an important natural food source for shrimp. This experiment evaluated the influence of light intensity on biofilm development and on the growth of cultured Farfantepenaeus paulensis juveniles. Six cages were deployed in a shrimp culture pond and stocked with 60 juveniles (0.72 ± 0.03 g). Three cages were randomly chosen and covered with five layers of polyamide net to reduce light incidence (shaded treatment), while the other three cages were not covered (control treatment). Biofilm chlorophyll a and microorganism abundance, as well as shrimp growth, were monitored during the experiment. Chlorophyll a concentration and the abundance of bacteria and flagellates were significantly lower in the shaded treatment (P<0.05). After 75 rearing days, shrimp stocked in control treatment achieved significantly higher (5.98 g) (P<0.05) mean weight than shrimp reared in shaded treatment cages (5.13 g). Similarly, the biomass produced was also significantly higher (P<0.05) in the control treatment (322.92 g) than in the shaded treatment (287.31 g). The results of this study demonstrate that light intensity has a huge influence on shrimp performance mainly due to the increase in natural food availability.     

Associations between morphometric variables and weight and yields carcass in Pirapitinga Piaractus brachypomus

The objective of this work was to evaluate the association between morphometric variables and carcass characteristics in Pirapitinga. We used a thousand specimens of Pirapitinga with an average weight of 1,200 g, which were stunned, slaughtered, weighed, measured, and processed for morphometric and processing yield analysis, to obtain weights, carcass and fillet yields. Initially, the linearity of the variables was verified. Pearson's simple and partial correlation tests were performed between all metrics. Track analysis was performed considering the weights and yields of carcass and filet as dependent variables and the others as independent variables. Ridge regression models were used to eliminate the effects of multicollinearity among the independent variables. Observations showed that the simple correlations between body weights and yields were superior to the partial ones in terms of magnitude. The length and circumference of the specimens were the measures most linearly associated with weight, carcass weight and fillet with rib weight. As for carcass yield and fillet yield, linear correlation coefficients were low and not significant when associated with body weights. With the path analysis, we could observe significant positive correlations between the morphometric measurements for weight, carcass weight and fillet with rib weight. The standard length, body circumference and body circumference/body height ratio are the most correlated. The latter are the most important variables in the evaluation of body weights in Pirapitinga fish. They can serve as criteria for indirect selection in searching for fish with better carcass characteristics. As for fillet and carcass yields, the morphometric variables have not shown to be efficient for indirect selection.     

Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Natural Stable Isotopes for Determination of Gastrointestinal Transit Time in Fish

This study evaluated the application of stable isotopes of carbon as an alternative and more accurate method to determine gastrointestinal transit time (GTT) in fish by comparing it to the inert marker method. The stable isotope method detects alterations of the normal carbon flow in a biological system by analyzing naturally occurring isotopes of carbon, contrary to studies based on conventional techniques that apply external markers to the diet to determine GTT through visual observation of the color change in feces. Therefore, 320 pacu, Piaractus mesopotamicus juveniles were reared in 32 tanks under two different temperatures (25 and 29 C). The pacu juveniles received two different diets, one based on ingredients derived from C₃ photosynthetic cycle plants and the other based on C₄ plant ingredients, both containing titanium oxide (TiO₂) as a marker. After 40 d, the isotopic signature of the diets was changed, and the marker was replaced by chromic oxide (Cr₂O₃). In the isotopic technique, the feces were analyzed to determine the exchange in the isotopic ratio of carbon δ¹³C. Both methods found that GTT was faster (nearly 6 h) in fish at 29 C when using the C₄/C₃ feeding strategy and slower in fish at 25 C using the C₃/C₄ strategy (15 h by inert marker and 18 h by the isotopic method). In conclusion, GTT determination in pacu juveniles using the stable isotope technique exhibits the same accuracy obtained with the inert marker method at temperatures suitable (nearly 29 C) for the metabolism of these animals.     

Does reverse growth dominance develop in old plantations of Eucalyptus saligna?

Patterns of stand growth and dominance among individual trees change through stand development. We tested a prediction that large, old plantations of Eucalyptus saligna (Sm.) in Hawaii and Brazil would show a pattern of “reverse” growth dominance, with the largest trees contributing more to stand biomass than to current stand increment. This pattern might develop if the largest trees accounted for a large portion of the total resource use of the stand, but were less efficient at utilizing resources to produce stemwood. The 70-year-old plantation in Hawaii had a stem mass of 610 Mg/ha, compared with 325 Mg/ha in the 66-year-old Brazil plantation. Cumulative frequency plots for both sites showed that dominant trees comprised a very large proportion of total stand mass; the largest 20% of the trees accounted for 50% and 30% of the total stand mass in Hawaii and Brazil, respectively. Contrary to expectations, the largest trees continued to produce a disproportionately large amount of total stand growth, with the largest 20% of trees accounting for 60% (Hawaii) and 40% (Brazil) of stand growth. The frequency distributions resulted in positive growth dominance coefficients (similar to Gini coefficients) of 0.14 for Hawaii and 0.09 for Brazil, rather than negative values that would indicate that reverse growth dominance. The failure of these monoculture Eucalyptus stands to develop reverse growth dominance contrasts starkly with a variety of studies with pine species, which tend to show neutral-to-reverse growth dominance. Indeed, continued dominance of large trees suggests the trees might respond well to silvicultural treatments such as thinning or fertilization. Future work that contrasts patterns of growth dominance among species will be important as a foundation for developing generalizations about trends in growth dominance with stand development.     

Characterization of symptoms of senescence and chilling injury on inflorescences of Heliconia bihai (L.) cv. Lobster Claw and cv. Halloween

Inadequate temperatures during the shipping and commercialization of cut tropical flowers may accelerate the senescence process and cause chilling injury, leading to symptoms that have not yet been described for Heliconia bihai. The aim of the present study was to evaluate physiological responses in cut inflorescences of H. bihai cv. Lobster Claw (LC) and cv. Halloween (HW) as well as symptoms of senescence and chilling injury. For such, changes in fresh weight, bract color (L*, a* and b*), percentage of absolute integrity (PAI) of cell membranes and leakage of potassium ions (LPI) were determined. The flowering stems were evaluated at five different intervals after harvest (0, 2, 4, 6 and 8 d). A refrigerated treatment (RT) with a temperature of 6.5 °C and 85% relative humidity was compared to a control treatment (CT) at room temperature of 24 °C and 66% relative humidity. Both cultivars stored at 6.5 °C exhibited dryness of bract tissue (symptom of senescence) and dark stains that became brownish and evolved to necrosis (symptom of chilling injury). The visual quality of inflorescences decreased with time in both cultivars maintained without refrigeration. The severity of chilling injury increased with the length of storage time in both cultivars. There was a significant reduction in the fresh weight of inflorescences in both treatments (RT and CT) and both cultivars (LC and HW). Bract color changed in both cultivars at 6.5 °C. There was no change in PAI throughout the evaluation period in the inflorescences stored at room temperature, whereas those stored at 6.5 °C for 6 and 8 d had lower PAI values. The inflorescences in the control treatment underwent no change in LPI values, whereas those stored under refrigeration had increased LPI values after the sixth day of storage. The physiological responses of cut Heliconia flowers were influenced by storage period and temperature, as demonstrated by visual symptoms of chilling injury and senescence.     

Synthesis and toxicological properties of some alkyl and aryl 3,4-methylenedioxyphenyl N,N′-thio-bis-N-methylcarbamates

A series of asymmetrical thio-bis-N-methylcarbamate derivatives were synthesised by combining 3,4-methylenedioxyphenyl N-methylcarbamate with different N-methylcarbamates. Some of these derivatives combined good insecticidal activity against the housefly with reduced acute oral toxicity to mice. The thio-bis-N-methylcarbamates obtained exhibited a lower anti-cholinesterase activity than the more toxic of the two carbamate moieties contained in each combination. The weak synergism of piperonyl butoxide on the insecticidal activity of the derivatives suggests the possibility of a self synergism phenomenon caused by the 3,4-methylenedioxyphenyl group.