Characterization of an Insoluble and Soluble Form of Melanin Produced by Streptomyces cavourensis SV 21, a Sea Cucumber Associated Bacterium

Melanin is a widely distributed and striking dark-colored pigment produced by countless living organisms. Although a wide range of bioactivities have been recognized, there are still major constraints in using melanin for biotechnological applications such as its fragmentary known chemical structure and its insolubility in inorganic and organic solvents. In this study, a bacterial culture of Streptomyces cavourensis SV 21 produced two distinct forms of melanin: (1) a particulate, insoluble form as well as (2) a rarely observed water-soluble form. The here presented novel, acid-free purification protocol of purified particulate melanin (PPM) and purified dissolved melanin (PDM) represents the basis for an in-depth comparison of their physicochemical and biological properties, which were compared to the traditional acid-based precipitation of melanin (AM) and to a synthetic melanin standard (SM). Our data show that the differences in solubility between PDM and PPM in aqueous solutions may be a result of different adjoining cation species, since the soluble PDM polymer is largely composed of Mg²⁺ ions and the insoluble PPM is dominated by Ca²⁺ ions. Furthermore, AM shared most properties with SM, which is likely attributed to a similar, acid-based production protocol. The here presented gentler approach of purifying melanin facilitates a new perspective of an intact form of soluble and insoluble melanin that is less chemical altered and thus closer to its original biological form.     

P0 protein of cotton leafroll dwarf virus-atypical isolate is a weak RNA silencing suppressor and the avirulence determinant that breaks the cotton Cbd gene-based resistance

Cotton blue disease (CBD) is the most important disease present in cotton crops in South America and cotton leafroll dwarf virus (CLRDV) is the causal agent. The disease has been controlled by sowing cotton varieties resistant to CLRDV. However, in the 2009/10 growing season, an outbreak due to an atypical CLRDV isolate (CLRDV-at) occurred in northwest Argentina. Although CLRDV and CLRDV-at genomes are very closely related, the symptoms they produce in cotton plants are quite different. P0 is the most divergent protein between the isolates and in CLRDV is a silencing suppressor protein. This work characterized the silencing suppressor activity of the P0 protein encoded by CLRDV-at (P0^CL-at) and evaluated its role in Cbd-resistance break in cotton plants. It was demonstrated that P0^CL-at, despite having a mutation in the consensus of the F-box-like motif, was able to suppress local RNA silencing, but displayed lower activity than P0^CL. P0^CL and P0^CL-at showed no differences in the interaction with Gossypium hirsutum SKP1 orthologue (GSK1) and Nicotiana benthamiana SKP1 and both P0 proteins triggered destabilization of ARGONAUTE1. However, when the ability to enhance PVX symptoms was evaluated, P0^CL-at was shown to be a weaker pathogenicity factor than P0^CL in N. benthamiana. Interestingly, trans-expressed P0^CL-at enabled CLRDV to systemically infect CBD-resistant plants, and a chimeric CLRDV-P0^CL-at infectious clone succeeded in establishing infection in CBD-resistant cotton varieties with symptoms resembling those produced by CLRDV-at. These results strongly suggest that P0^CL-at is the avirulence (Avr) determinant involved in breaking cotton Cbd gene-based resistance.     

Distribution and genetic diversity among Aspergillus flavus isolates across three agro-ecologies essential for maize cultivation in Ghana

Fungal mycotoxins are important contaminants of agricultural commodities that pose serious concerns to producers, consumers and exporters. Aflatoxin is a carcinogenic metabolite produced by the fungi Aspergillus flavus and A. parasiticus. These fungi and other mould species living in the soil contaminate several crops including maize. This study was undertaken with the following objectives: (i) to assess the presence of A. flavus and other moulds in Ghanaian soil; (ii) to determine the distribution and density of A. flavus under three agro-ecologies noted for major maize production; and (iii) to assess the effect of percentage of carbon (C) and nitrogen (N) and soil pH on A. flavus population densities and diversity in the isolates found across the agro-ecologies. The data showed seven moulds that were common across the agro-ecologies. Significant differences (P < 0.05) were observed in the A. flavus density and distribution within and across these agro-ecologies. Fumesua soils recorded the highest levels of A. flavus (1.185 × 10³ cfu g⁻¹) while Akomadan recorded the least (9.76 × 10² cfu g⁻¹). Percentage available C, N and soil pH did not significantly influence A. flavus density. The A. flavus isolates identified in this study varied in genetic sequence within the aflatoxin gene cluster, but these differences were not distinguishable by origin.     

Comparative performance of the highly invasive Mimosa invisa and its less invasive subspecies under different nutrient and light conditions

Invasive plants may be more plastic than non‐invasive plants and maintain high fitness under various environmental conditions. Previous studies mainly focused on the comparisons between invasive and native plants, and comparisons between highly invasive and less invasive exotic species are still relatively rare, especially for comparisons at the subspecies level. This study examined the effects of nutrient addition and shading on the performance of the highly invasive Mimosa invisa and its less invasive subspecies M. invisa var. inermis under either isolated or competitive conditions. Nutrient addition increased biomass and plant height and decreased root‐to‐shoot ratio (R/S). Shading decreased biomass and R/S and increased plant height. Under isolated conditions, the two invaders did not differ in R/S, plant height and plasticity of these traits in response to nutrient addition or shading, and the two invaders also did not differ in biomass production under each of the nutrient and light treatments. When the two invaders competed with each other, M. invisa outcompeted M. invisa var. inermis under high soil nutrient conditions, and the two invaders did not differ in performance under other growth conditions. Thus, only considering competition may we find out the difference between highly invasive species and their closely related, less invasive subspecies. Management of M. invisa should focus on habitats with high soil nutrient availability, in which M. invisa is more likely to dominate.     

Phytophagous insect fauna of Onopordum spp. thistles in the south of Syria

Thistles from the genus Onopordum (Asteraceae) are of Mediterranean and Eurasian origin. They are very common in the south of Syria, in particular in Damascus and Al Suwayda, and are found in fallow fields, pastures, roadsides and neglected areas. In 2015–2017, several sites with large populations of Onopordum spp. were identified. Weekly surveys of some of the sites, mainly in the regions of Damascus and Al Suwayda, revealed a significant diversity of phytophagous insect species, some of which had a very high population density. A total of 29 insect species were detected, belonging to four orders and 21 families. The most abundant species were from the order Coleoptera. These were observed to be feeding on different plant parts of Onopordum spp. Purely endophagous species comprised 38% of the species observed on Onopordum spp., ectophagous species comprised 45% with 17% of species mixed ectophagous and endophagous. Species that only fed on Onopordum spp. represented 34% of all the species observed.     

Effect of dietary protein reduction with lysine and methionnine supplementation on growth performance,body composition and total ammonia nitrogen excretion of juvenile grass carp,Ctenopharyngodon idella

A 63‐day growth trial was undertaken to estimate the effects of supplemented lysine and methionine with different dietary protein levels on growth performance and feed utilization in Grass Carp (Ctenopharyngodon idella). Six plant‐based practical diets were prepared, and 32CP, 30CP and 28CP diets were formulated to contain 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 crude protein without lysine and methionine supplementation. In the supplementary group, lysine and methionine were added to formulate 32AA, 30AA and 28AA diets with 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 dietary crude protein, respectively, according to the whole body amino acid composition of Grass Carp. In the groups without lysine and methionine supplementation, weight gain (WG, %) and specific growth rate (SGR, % day^?1) of the fish fed 32CP diet were significantly higher than that of fish fed 30CP and 28CP diets, but no significant differences were found between 30CP‐ and 28CP‐diet treatments. WG and SGR of the fish fed 32AA and 30AA diets were significantly higher than that of fish fed 28AA diets, and the performance of grass carp was also significantly improved when fed diets with lysine and methionine supplementation (P < 0.05), and the interaction between dietary protein level and amino acid supplementation was noted between WG and SGR (P < 0.05). Feed intake (FI) was significantly increased with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05), but feed conversion ratio (FCR) showed a significant decreasing trend (P < 0.05). Two days after total ammonia nitrogen (TAN) concentration test, the values of TAN discharged by the fish 8 h after feeding were 207.1, 187.5, 170.6, 157.3, 141.3 and 128.9 mg kg^?1 body weight for fish fed 32CP, 32AA, 30CP, 30AA, 28CP and 28AA diets, respectively. TAN excretion by grass carp was reduced in plant‐based practical diets with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05). The results indicated that lysine and methionine supplementation to the plant protein sources‐based practical diets can improve growth performance and feed utilization of grass carp, and the dietary crude protein can be reduced from 320 g kg^?1 to 300 g kg^?1 through balancing amino acids profile. The positive effect was not observed at 280 g kg^?1 crude protein level.     

Edwardsiellosis in farmed turbot,Scophthalmus maximus (L.), associated with an unusual variant of Edwardsiella tarda: a clinical,aetiological and histopathological study

During 2005 and 2010, a survey of edwardsiellosis on eight turbot, Scophthalmus maximus (L.), farms was conducted in China. This report presents the detailed results of the study on this disease. Diseased turbot displayed two distinct types of gross signs: black discoloration of the dorsal skin on the posterior portion of the body; and red cutaneous foci on the ventral side. Internally, the most pronounced clinical signs in all fish examined were enlarged kidneys. The causal agent of the disease was finally proved to be one species of bacterium that was identified as Edwardsiella tarda by physiological and biochemical tests, API 32E and 16S ribosomal RNA sequence analysis. It is noteworthy that unlike the commonly described E. tarda strains, the isolates in this study were non‐motile strains without flagella. A histopathological study revealed that E. tarda infection was systemic in turbot and that kidney showed the most significant pathological changes, including acute focal necrosis, an influx of macrophages and formation of granuloma. The most common histopathological characteristics of this disease are the proliferation of macrophage in various organs and formation of granuloma. In addition, this article also gave background information on the disease and presented the results of virulence tests with the E. tarda strain identified in this study.