Study of efficacy of miltefosine and allopurinol in dogs with leishmaniosis

Visceral leishmaniosis is a life-threatening disease of medical, social and economic importance in endemic areas. It is an opportunistic infection in immunocompromised patients, including human immunodeficiency virus-positive subjects. Dogs are the main reservoir of Leishmania infantum. The aim of this study was to evaluate the efficacy of miltefosine and allopurinol for the control of human leishmaniosis using the dog as a model. The study included 28 sick dogs treated with miltefosine (2 mg/kg/day PO) administered concurrently with allopurinol (10 mg/kg/day, PO) for 30 days, and then with allopurinol alone, at the same dosage, for 1 year. Eight dogs (four of which relapsed) received a second cycle of miltefosine within 6 months of the first cycle. Efficacy was measured by real-time polymerase chain reaction assay on whole blood samples and lymph node aspirates, collected at baseline and every 3 months for 12 months. Of the total number of animals (28), two showed renal insufficiency and died after the start of therapy with miltefosine. Two other dogs presented some side effects to treatment, such as nausea, vomiting and reduction in white and red blood cell counts, and these animals were excluded from the follow-up. The results showed that the first cycle of therapy with miltefosine and allopurinol induced a drastic and progressive reduction of L. infantum load in lymph node aspirates but the second cycle did not eliminate the parasite.     

Detection of genetically modified soybean using peptide nucleic acids (PNAs) and microarray technology

Peptide nucleic acid (PNA) microarrays for the detection of Roundup Ready soybeans in food have been prepared. PNA probes are known to be more efficient and selective in binding DNA sequences than the analogous oligonucleotides and are very suitable to be used for diagnostics in food. PNAs of different lengths were carefully designed and synthesized by solid-phase synthesis on an automatic synthesizer adopting the BOC strategy. PNAs were purified by HPLC and characterized by HPLC/MS. The probes were spotted on a functionalized surface to produce a microarray to be hybridized with PCR products. DNA extracted from reference material was amplified using Cy3- and Cy5-labeled primers, and the fluorescent PCR products obtained were hybridized on the microarray. Two protocols were adopted: the hybridization with dsDNA or with ssDNA obtained by digestion with the enzyme lambda exonuclease. The best results were obtained using a 15-mer PNA probe in combination with the ssPCR product derived from enzymatic digestion. The method was applied to the analysis of a sample of certified transgenic soybean flour.     

Biodegradation combined with ozone for the remediation of contaminated soils

The effectiveness of the SS-SBR (Soil Slurry – Sequencing Batch Reactor) process for the remediation of soils contaminated by several organic pollutants has been evaluated. Experimental tests have been performed on two different soils, a spiked one and an industrial aged soil. The spiked soil, artificially contaminated, has been prepared trying to simulate the pollution of an industrial aged soil in terms of number and kind of contaminants. PAHs (Polycyclic Aromatic Hydrocarbons) and phenols degradation has been particularly investigated because they are considered persistent and recalcitrant. Concerning the spiked soil, removal efficiencies higher than 95% in 6 to 9 weeks have been found for all the pollutants, except for five-rings PAHs; however, these compounds were partly removed in 11 to 13 weeks. Good results have been achieved also for the industrial aged soil with a maximum removal of about 80% in 7–8 weeks. To enhance the pollutants degradation, trying to obtain a faster remediation, the biological treatment has been combined with a chemical oxidation with ozone. The best degradation effectiveness of the combined process has been obtained applying the ozonation after few days of the biological treatment. Therefore, a combined biological and chemical treatment allowed to markedly improve the remediation of contaminated soils.     

Impact of Agro-industrial Byproducts on Bioconversion,Chemical Composition,in vitro Digestibility,and Microbiota of the Black Soldier Fly (Diptera: Stratiomyidae) Larvae

The interest in using byproducts from agro-food industries as a rearing substrate for insects is increasing rapidly. We investigated the influence of byproducts of vegetal origin (okara—a byproduct of soy milk production, maize distillers with solubles, brewer’s grains), used as rearing diet for black soldier fly larvae (BSFL), on the following parameters: biomass production, substrate reduction (SR), nutritional profile and in vitro digestibility, and larval gut microbiota. Hen diet was used as a control substrate. The highest larval biomass was collected on maize distillers, whereas the highest SR was observed on okara. The rearing substrate affected ash, ether extract, and chitin larval content. The BSFL reared on okara were characterized by a lower lauric acid content (17.6% of total fatty acids). Diets also influenced in vitro crude protein digestibility (%) for monogastrics, with the highest values for BSFL reared on maize distillers (87.8), intermediate for brewer’s grains and okara BSFL, and the lowest for hen BSFL (82.7). The nutritive value for ruminants showed a lower Net Energy for lactation for BSFL reared on hen diet than okara and dried maize distillers BSFL. The different byproducts showed an influence on the larval gut microbiota, with a major bacterial complexity observed on larvae fed with the hen diet. The neutral detergent fiber concentration of dietary substrate was negatively correlated with Firmicutes and Actinobacteria relative abundance. Insects valorized byproducts converting them into high-value larval biomass to be used for feed production. The results evidenced the effects of the tested byproducts on the measured parameters, underling the chemical composition importance on the final insect meal quality.     

A Metataxonomic Approach Reveals Diversified Bacterial Communities in Antarctic Sponges

Marine sponges commonly host a repertoire of bacterial-associated organisms, which significantly contribute to their health and survival by producing several anti-predatory molecules. Many of these compounds are produced by sponge-associated bacteria and represent an incredible source of novel bioactive metabolites with biotechnological relevance. Although most investigations are focused on tropical and temperate species, to date, few studies have described the composition of microbiota hosted by Antarctic sponges and the secondary metabolites that they produce. The investigation was conducted on four sponges collected from two different sites in the framework of the XXXIV Italian National Antarctic Research Program (PNRA) in November–December 2018. Collected species were characterized as Mycale (Oxymycale) acerata, Haliclona (Rhizoniera) dancoi, Hemigellius pilosus and Microxina sarai by morphological analysis of spicules and amplification of four molecular markers. Metataxonomic analysis of these four Antarctic sponges revealed a considerable abundance of Amplicon Sequence Variants (ASVs) belonging to the phyla Proteobacteria, Bacteroidetes, Actinobacteria and Verrucomicrobia. In particular, M. (Oxymycale) acerata, displayed several genera of great interest, such as Endozoicomonas, Rubritalea, Ulvibacter, Fulvivirga and Colwellia. On the other hand, the sponges H. pilosus and H. (Rhizoniera) dancoi hosted bacteria belonging to the genera Pseudhongella, Roseobacter and Bdellovibrio, whereas M. sarai was the sole species showing some strains affiliated to the genus Polaribacter. Considering that most of the bacteria identified in the present study are known to produce valuable secondary metabolites, the four Antarctic sponges could be proposed as potential tools for the discovery of novel pharmacologically active compounds.     

Spontaneous internal desynchronization of locomotor activity and body temperature rhythms from plasma melatonin rhythm in rats exposed to constant dim light

Background  

We have recently reported that spontaneous internal desynchronization between the locomotor activity rhythm and the melatonin rhythm may occur in rats (30% of tested animals) when they are maintained in constant dim red light (LL_dim) for 60 days. Previous work has also shown that melatonin plays an important role in the modulation of the circadian rhythms of running wheel activity (R_w) and body temperature (T_b). The aim of the present study was to investigate the effect that desynchronization of the melatonin rhythm may have on the coupling and expression of circadian rhythms in R_w and T_b.     

Effects of cultural cycles and nutrient solutions on plant growth,yield and fruit quality of alpine strawberry (Fragaria vesca L.) grown in hydroponics

Alpine strawberry (Fragaria vesca L.) was grown in hydroponics with the nutrient film technique, in order to evaluate the effects of four buffer concentrations (1.3, 1.6, 1.9, 2.2 mS cm⁻¹) and two cultural cycles (summer-spring versus autumn-spring) in terms of growth, yield and fruit quality (dry and optical residues, sugars, acids, antioxidants, mineral composition). The longer summer-spring cycle gave a correspondingly higher yield than the autumn-spring one. The 1.3 mS cm⁻¹ nutrient solution was the most effective in terms of overall and spring production. However, the autumn and winter yields were not affected by the buffer EC. Fruit quality did not change with the cultural cycle, but the berries harvested in the spring had higher vitamin C and sucrose content and lower nitrate content compared with berries picked up in the winter. Fruit quality was also improved when the nutrient solution concentration increased. From the productive point of view, the cultural cycle choice should be made considering that 71% of the yield of the more productive summer-spring cycle derived from the spring harvest. Moreover, as regards the nutrient solution strength, 1.3 mS cm⁻¹ EC should be preferred during the spring season, whereas the 2.2 mS cm⁻¹ EC proved to be best in the winter in terms of fruit quality.     

Bacterial Exopolysaccharides from Extreme Marine Habitats: Production,Characterization and Biological Activities

Many marine bacteria produce exopolysaccharides (EPS) as a strategy for growth, adhering to solid surfaces, and to survive adverse conditions. There is growing interest in isolating new EPS producing bacteria from marine environments, particularly from extreme marine environments such as deep-sea hydrothermal vents characterized by high pressure and temperature and heavy metal presence. Marine EPS-producing microorganisms have been also isolated from several extreme niches such as the cold marine environments typically of Arctic and Antarctic sea ice, characterized by low temperature and low nutrient concentration, and the hypersaline marine environment found in a wide variety of aquatic and terrestrial ecosystems such as salt lakes and salterns. Most of their EPSs are heteropolysaccharides containing three or four different monosaccharides arranged in groups of 10 or less to form the repeating units. These polymers are often linear with an average molecular weight ranging from 1 × 10⁵ to 3 × 10⁵ Da. Some EPS are neutral macromolecules, but the majority of them are polyanionic for the presence of uronic acids or ketal-linked pyruvate or inorganic residues such as phosphate or sulfate. EPSs, forming a layer surrounding the cell, provide an effective protection against high or low temperature and salinity, or against possible predators. By examining their structure and chemical-physical characteristics it is possible to gain insight into their commercial application, and they are employed in several industries. Indeed EPSs produced by microorganisms from extreme habitats show biotechnological promise ranging from pharmaceutical industries, for their immunomodulatory and antiviral effects, bone regeneration and cicatrizing capacity, to food-processing industries for their peculiar gelling and thickening properties. Moreover, some EPSs are employed as biosurfactants and in detoxification mechanisms of petrochemical oil-polluted areas. The aim of this paper is to give an overview of current knowledge on EPSs produced by marine bacteria including symbiotic marine EPS-producing bacteria isolated from some marine annelid worms that live in extreme niches.     

Cloning and expression analysis of kenaf (Hibiscus cannabinus L.) major lignin and cellulose biosynthesis gene sequences and polymer quantification during plant development

In order to apply state-of-the-art molecular breeding techniques in fibre crop it is necessary to have a good knowledge of major polymer biosynthesis gene sequences and their expression pattern. Polymerase chain reaction was employed to isolate sequences of the major genes for lignin and cellulose biosynthesis in a kenaf cultivar. CeSA, 4cl, c4h, cad, and ccr gene primers were designed according to their conservative regions; partial sequences of lignin and cellulose biosynthesis genes were obtained. One actin II gene sequence was also isolated from the kenaf genome as a housekeeping gene to be employed in qPCR analysis. Expression levels of genes c4h, cad and CeSA in bark and core from plants harvested at three different growth stages were evaluated. Using qPCR analyses it was found that the expression levels of the two biosynthesis lignin genes in bark tissues increased during plant growth, while a negative trend was recorded in core tissues. In both bark and core, the quantity of lignin was positively correlated to plant growth while cellulose content decreased.     

Multiple buds are an alternative target to genetic manipulation of kenaf (Hibiscus cannabinus L.)

Kenaf (Hibiscus cannabinus L.) is an annual fiber crop cultivated as an important source of cellulose and lignin. In the recent years, fibers have increased their economic importance as organic component in bio-composite materials. However, industry requires rheological improvements of these fibers. Genetic manipulation has been considered a feasible approach for fiber improvement; however, kenaf is considered a crop species difficult to manipulate. Here we propose meristematic cells from mature embryos as target for genetic modification driven by Agrobacterium tumefaciens. In vitro meristems proliferation and multiple shoots regeneration were evaluated by sowing kenaf mature seeds on medium containing three different concentration of thidiazuron, a substituted urea compound. The highest number of regenerated shoots was obtained from mature seeds germinated in presence of 10 μM thidiazuron after 14 days of culture. Interactions between two A. tumefaciens strains and two kenaf varieties were assessed. Transgene stable integration and its inheritance in T₁ generation were also verified, demonstrating that kenaf meristematic cells are an useful target for genetic manipulation by agroinfection.