Molecular characterization of Corynebacterium pseudotuberculosis isolates using ERIC-PCR

Caseous lymphadenitis is an infectious sheep and goats disease caused by Corynebacterium pseudotuberculosis and characterized by abscesses in superficial and visceral lymph nodes. C. pseudotuberculosis strains isolated from these hosts have been shown to be very difficult to type by the existing methods. The aim of this study is evaluating the potential of the Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) as a tool for molecular typing of C. pseudotuberculosis strains isolated in sheep. One hundred and twenty seven isolates of C. pseudotuberculosis were isolated from lesions suspected to have had caseous lymphadenitis collected from sheep at the slaughterhouse. Animals were from 24 flocks in 13 municipalities of the Minas Gerais State, Brazil. Species identification of the isolates was performed by routine biochemical tests and mPCR. Fingerprint was performed by RAPD using ERIC-1R, ERIC-2 and ERIC-1R+ERIC-2 primers. Seventeen different genotypes were generated by ERIC 1-PCR, 21 genotypes by ERIC 2-PCR and 21 genotypes by ERIC 1+2-PCR. Hunter-Gaston Discrimination Index (HGDI) found for ERIC 1, ERIC 2, ERIC 1+2 PCR were 0.69, 0.87, and 0.84, respectively. For most herds evaluated observed at most three different genotypes among isolates from animals of these property, in all ERIC-PCR assays. However a few flocks observed between four and nine genotypes per flock. The W Kendall value found for correlation among the three techniques of ERIC-PCR was 0.91 (P<5.0 x 10(-6)). The results show that ERIC-PCR has good discriminatory power and advantages over other DNA-based typing methods, making it a useful tool to discriminate C. pseudotuberculosis isolates.     

Farmed Gilthead Sea Bream (Sparus aurata) by-Products Valorization: Viscera Oil ω-3 Enrichment by Short-Path Distillation and In Vitro Bioactivity Evaluation

This study shows a pilot scale protocol aimed to obtain an omega 3-enriched oil after the processing of farmed gilthead sea bream viscera (SBV); this was oil was tested in vitro for bioactivity, attesting to the possibility to turn waste into profit The quality of the oil, in terms of requirements for animal and human consumption, was assessed by determining some chemical parameters, such as peroxide value (PV), thiobarbituric acid reactive substances (TBARS), ρ-anisidine (ρ-AV) content, total oxidation value (TOTOX), and phospholipids and free fatty acid (%), both in crude viscera oil (CVO) and refined viscera oil (RVO). Among the extraction conditions, the higher CVO yields were obtained at 60 °C for 10 min (57.89%) and at 80 °C for 10 min (67.5%), and the resulting oxidation levels were low when utilizing both extraction conditions. RVO, obtained from CVO extracted at 60 °C, showed the highest quality on the basis of the assessed parameters. The ethyl esters of the total fatty acid (TFA) contents extracted from RVO were enriched in the ω-3 polyunsaturated fatty acid fraction (PUFAE) up to almost 56% via short path distillation (SPD). Antioxidant activities and adipogenic properties were tested in vitro. PUFAE protected 3T3 L1 cells from oxidative stress and exerted an anti-adipogenic effect in Dicentrarchus labrax pre-adipocytes, attesting to the beneficial properties for both farmed fish and human health. These results could stimulate the adoption of solutions aimed to recover and utilize aquaculture by-products at a higher scale, turning “waste into profit” and indicating a strategy to reach more sustainable business models in aquaculture resource utilization according to the principles of the circular economy.     

Characterization of Fungal Biodiversity In Compost and Vermicompost

The ability of fungi to break down complex carbon sources makes them of vital importance in both the generation and application of compost. This paper illustrates the diversity and functions of the mycoflora of both a green compost (made solely thermophilically from plant debris) and a vermicompost (made mesophilically by the action of earthworms on plant and animal wastes). The soil dilution plate technique was applied on 3 media (PDA, CMC, PDA plus cycloheximide) and at 3 incubation temperatures (24°C, 37°C and 45°C) to isolate and identify fungal entities. Enzymatic activities (amylase, cellulase, chitinase, esterase, ligninase, pectinase, phosphatase, plastic degradation and xylanase) of most species from both composts were evaluated with a semiquantitative method on 15 substrates. There were substantial qualiquantitative differences in the species composition of the two composts. A total of 193 entities were isolated: 54 from green compost only, 77 from vermicompost only, 62 from both. This taxonomic diversity was reflected in the metabolic potential. Amilase, cellulase, protease and esterase activities were significantly higher in the green compost. Lignin and plastic polymer degradation were significantly higher in the vermicompost. Structural and functional characterization of this kind is of assistance in determining both the most appropriate application of composts and their hygienic quality.     

Basidiomycetes from Compost and their Dye Degradation and Enzyme Activities

The degree of decolorization of Poly R-478, a highly recalcitrant anthraquinone dye, by three basidiomycetes belonging to Polyporales isolated from compost was investigated together with the enzymes involved. Decolorization tests in two liquid cultures, one with a simple mycelium inoculum, the other with an inoculum of mycelium grown on straw, resulted in 70% and 87% decolorization respectively in 7 days. However, the efficiency did not increase significantly in the presence of the lignocellulose substrate. The three strains produced laccase and/or manganese peroxidase activity during the decolorization, whereas lignin peroxidase activity was not observed. Previous growth on straw enhanced the synthesis of ligninolytic enzymes, though there was no correlation between enzyme activity and decolorization. The three fungi can be proposed as promising candidates for the treatment of colored industrial effluents and probably for soils contaminated by complex polymers, such as polycyclic aromatic hydrocarbons.     

Aurantoside J: a new tetramic acid glycoside from Theonella swinhoei. Insights into the antifungal potential of aurantosides

The chemical investigation of an Indonesian specimen of Theonella swinhoei afforded four aurantosides, one of which, aurantoside J (5), is a new compound. The structure of this metabolite, exhibiting the unprecedented N-α-glycosidic linkage between the pentose and the tetramate units, has been determined through detailed spectroscopic analysis. The four obtained aurantosides have been tested against five fungal strains (four Candida and one Fusarium) responsible of invasive infections in immuno-compromised patients. The non-cytotoxic aurantoside I (4) was the single compound to show an excellent potency against all the tested strains, thus providing valuable insights about the antifungal potential of this class of compounds and the structure-activity relationships.     

New cholinesterase inhibiting bisbenzylisoquinoline alkaloids from Abuta grandifolia

The phytochemical study of the stem bark and wood of Abuta grandifolia (Mart.) Sandwith led to the identification of four bisbenzylisoquinoline alkaloids (BBIQs), namely (R,S)-2 N-norberbamunine (1), (R,R)-isochondodendrine (2), (S-S)-O4″-methyl, Nb-nor-O6'-demethyl-(+)-curine (3), and (S-S)-O4″-methyl, O6'-demethyl-(+)-curine (4), together with the aporphine alkaloid R-nornuciferine (5), all obtained by countercurrent distribution separation (CCD) and identified on the basis of their spectroscopic data. Alkaloids 3 and 4 were new. All the isolated compounds were tested for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities. 1 was the most active against AChE, whereas 3 and 4 were the most potent against BChE. Interestingly, all tested alkaloids are more potent against BChE than against AChE. This selectivity of cholinesterase (ChE) inhibition could be important in order to speculate on their potential therapeutic relevance.     

Identification of virulence factors by multiplex PCR in Escherichia coli isolated from calves in Minas Gerais, Brazil

In this study, multiplex PCR was employed to investigate the virulence factors of Escherichia coli strains isolated from 60-day-old calves. Faecal samples were collected from 54 calves at 12 dairy farms in the state of Minas Gerais, Brazil. A total of 156 isolates were obtained after culture and microbiological isolation and were tested by multiplex PCR for the presence of genes encoding toxins (Stx1, Stx2 and STa) and adherence factors (intimin, F41 and F5). Seventy of 156 isolates were positive for at least one virulence factor: ten (14.3?%) from diarrhoeic animals and 60 (85.7?%) from healthy calves. The virulence markers identified were: Stx1 (82.8?%), eae (24.3?%), F41 (11.4?%), F5 (10?%), STa (4.28?%) and Stx2 (4?%). In diarrhoeic animals, Stx1 (70?%) and F41 (30?%) were identified, while Stx1 (83.3?%), eae (28.3?%), F41 (8.3?%), F5 (11.6?%), STa (5?%) and Stx2 (1.6?%) were detected in isolates from healthy calves. Mixed infections with pathotypes Shiga toxin-producing E. coli (STEC)/enteropathogenic E. coli, STEC/enterohaemorrhagic E. coli and STEC/other (eae/F5, Stx1/STa) were detected in five healthy calves. Pathogenic E. coli were identified in 59.26?% of all calves and on 75?% of the dairy farms studied, not only in diarrhoeic (five of six) but also in healthy calves (27 of 48), which demonstrates the importance of this agent in the aetiology of diarrhoea in calves in the state of Minas Gerais.     

Effect of a climate change scenario on <Emphasis Type="Italic">Fusarium equiseti</Emphasis> leaf spot on wild rocket and radish under phytotron simulation

This study was undertaken by simulating the effects of increasing the temperature and CO₂ values on the incidence and severity of F. equiseti on wild rocket (Diplotaxis tenuifolia) and radish (Raphanus sativus), under phytotron conditions. Two sets of 3 trials were carried out in which eight different temperature and CO₂ combinations were tested:1) 400–450 ppm CO₂, 18–22 °C; 2) 800–850 ppm CO₂, 18–22 °C; 3) 400–450 ppm CO₂, 22–26 °C, 4) 800–850 ppm CO₂, 22–26 °C, 5) 400–450 ppm CO₂, 26–30 °C; 6) 800–850 ppm CO₂, 26–30 °C; 7) 400–450 ppm CO₂, 14–18 °C; 8) 800–850 ppm CO₂, 14–18 °C. The temperature and CO₂ levels were significant factors of influence on disease incidence (DI) and severity (DS) in all the trials, and their combination significantly influenced the DI and DS of F. equiseti leaf spot on both hosts. Disease incidence and severity increased on wild rocket at 850 ppm of CO_2, in comparison to 450 ppm, in each tested temperature range. The highest CO₂ value on radish, for all the tested temperature regimes, caused an increase in DI and DS, which resulted statistically significant at the highest tested temperature range. The results obtained in this study add more concern to the possible negative effects of the spread of F. equiseti on vegetables in Italy as well as in other areas suffering from increased temperatures as a consequence of climate changes.