Molecular characterization of Sarcocystis spp. in seabirds from southern Brazil

Sarcocystis spp. are cyst forming apicomplexan parasites that infect many vertebrates including birds. Sarcocystis spp. infection was investigated in tissue samples (pectoral muscles, heart, and brain) of 47 dead seabirds collected from the coastline of Santa Catarina State SC – Brazil, between August 2019 and March 2020. A portion of each tissue was fixed in 10% buffered formalin for histopathologic analysis while DNA was extracted from another portion and screened using nested-PCR targeting ITS1. Based on molecular analysis, Sarcocystis spp. were identified in 15/47 (31.9%) seabirds of five species, kelp gull (Larus dominicanus), manx shearwater (Puffinus puffinus), neotropic cormorant (Phalacrocorax brasilianus), brown booby (Sula leucogaster) and great skua (Stercorarius skua). Microscopically visible sarcocysts were observed only in the pectoral muscle of four seabirds 8.5% (4/47), while in one brown booby, sarcocysts were seen in both pectoral and cardiac muscles. Two types of sarcocysts, thin walled (≤1 μm) and thick-walled (≥ 2 μm) were identified. Based on ITS1 sequence comparison, S. halieti, S. falcatula and three not yet described Sarcocystis spp. were detected. Phylogenetically, S. falcatula isolates were classified as two distinct clusters. This is the first confirmation of S. halieti in seabird's species in South America and S. falcatula in birds of the order Charadriiformes. Further molecular studies are needed to understand the epidemiology of the Sarcocystis spp. infection and its impact on the health of seabirds.     

Multidimensional niche differentiation might buffer invasion impacts: the case of oligohaline corophiids (Crustacea: Amphipoda) in the Baltic Sea

Biological Invasions - Multidimensional niche differentiation might increase the stability of coexistence by reducing overall niche overlap which might have implications on the dynamics of...     

Diatoms in northeast Pacific surface sediments as paleoceanographic proxies

Fossil diatom total abundances (# valves/g) in 54 surface-sediment samples from the northeast (NE) Pacific Ocean reflect the position of high primary production associated with coastal upwelling and that possible biases associated with dilution or dissolution are small. Diatom species assemblages, defined by Q-mode factor analysis in 30 samples with abundant diatoms, are related to modern oceanographic properties. Five statistical assemblages, given by five specific diatom species and/or groups, are related to upwelling (Chaetoceros spores), subtropical (Thalassionema nitzschioides), subarctic (Rhizosolenia hebetata), transitional (Neodenticula seminae) and freshwater (freshwater diatoms) ecological environments. These factors are significantly correlated with primary productivity, temperature, nutrient concentrations and salinity, although the strongest relationship is that between diatom assemblages and productivity. However, it is not possible to distinguish between coastal and open-ocean (curl-driven) upwelling based on Chaetoceros spores relative percentages by themselves or on the floral factors.     

Characterization of saxitoxin production and release and phylogeny of sxt genes in paralytic shellfish poisoning toxin-producing Aphanizomenon gracile

The freshwater cyanobacterium Aphanizomenon gracile is one of the most widely distributed producers of the potent neurotoxins saxitoxin (STX) and its derivatives (paralytic shellfish poisoning toxins, PSP toxins). However, the phylogeny of STX biosynthesis genes and the regulation of STX production and release remain poorly studied in the genus Aphanizomenon. In this study, two A. gracile strains from Spanish freshwaters were grown in semi-continuous cultures under three temperatures (15, 20 and 28 °C) and their STX production and release were determined by Enzyme-Linked ImmunoSorbent Assay (ELISA). STX production was stable along the temperature range, with 1.4–2.3-fold shifts in biomass-standardized STX contents, and maxima of 0.22 μg equivalent STX mg⁻¹ dry weight 15.3 fg equiv STX cell⁻¹ and 15.1 μg equiv STX mg⁻¹ Chl a. The extracellular fraction was remarkably high (13.6–35.3%), not clearly affected by temperature but with nitrate-depleted medium (BG11₀) inducing a 2-fold increase in extracellular content. STX production and release were not directly related to growth rates. The 16S rRNA phylogenetic analyses in sixteen A. gracile strains from Spanish and German freshwaters showed that PSP-producing A. gracile grouped within a monospecific and highly supported cluster, together with PSP-producing Aphanizomenon sp. NH-5 and clearly separated from a monospecific Aphanizomenon flos-aquae cluster. The sixteen A. gracile strains formed also monospecific and highly supported clusters for PSP-biosynthesis genes (sxtG, sxtI, sxtH and sxtX) together with Aphanizomenon sp. NH-5. This study evidences an elevated extracellular proportion of STX in A. gracile with importance for risk assessment, and supports the identification of Aphanizomenon sp. NH-5 as A. gracile.     

The Comparative Analysis of Carboxylic Acid Composition of Four Iris Species from Ukraine

The present article reports results of analysis of carboxylic acids in leaves of Iris species from Ukraine using a gas chromatography (GC) method with mass spectrometric (MS) detection (GC/MS). Carboxylic acids play significant roles in contemporary society as evidenced by multiple applications in fields of medicine, agriculture, pharmacy, food, and other industries. Study of natural plant products as a source of organic acids is of particular interest. Carboxylic acid composition of leaves of Iris hungarica Waldst. & Kit., Iris germanica L., Iris pallida Lam., and Iris variegate L. was studied for the first time applying GC/MS method. The mass spectrums of compounds were matched with NIST and WILEY Libraries. The GC/MS analysis revealed the presence of 26 common acids in the plant raw materials studied. The short-chain carboxylic acids, such as citric (1337.5–12364.4 mg/kg), malic (50.8–4558.0 mg/kg) and oxalic (1199.0–3435.2 mg/kg) acids were contained in significantly high quantity in all samples. Ferulic, p-coumaric and vanillic acids were the most abundant among phenolic acids. α-Linolenic acid was dominant in the leaves of I. germanica (869.5 mg/kg), I. pallida (753.3 mg/kg), and I. variegate (250.3 mg/kg) among polyunsaturated fatty acids, however, linoleic acid prevailed in the plant raw material of I. hungarica (1150.7 mg/kg). Since the leaves of Iris species studied contain carboxylic acids with diverse pharmacological activity, extracts of these raw materials are perspective for development food supplements and medicines.     

Expansion of bloom-forming Dolichospermum lemmermannii (Nostocales,Cyanobacteria) to the deep lakes south of the Alps: Colonization patterns,driving forces and implications for water use

Since the beginning of the 1990s, the largest lakes south of the Alps (Garda, Iseo, Como and Maggiore) showed a progressive colonization of Dolichospermum lemmermannii (Cyanobacteria). The appearance of surface blooms of this species raised serious concerns because of the impacts on the tourist economy and the potential toxigenic effects. Nevertheless, no detailed investigations were done to clarify the taxonomic position, ecology and toxicity of this species. In this work, phylogenetic analyses based on the 16S rRNA and rpoB genes demonstrated how the strains isolated from the Italian lakes were clustered together with other D. lemmermannii strains isolated in Northern Europe. The expansion of this species in the southern subalpine lakes contrasted with the prevailing south to north dispersion paths typical of other Nostocales (e.g. Cylindrospermopsis and Aphanizomenon). Conversely, the spread was consistent with the geographical areal of this cyanobacterium, which appears circumscribed between the 40th parallel and the Arctic Circle. This aspect highlights the ecological heterogeneity that characterizes the order Nostocales. In Lake Garda, D. lemmermannii always developed in the warmest months (>15 °C) with low abundances (generally <200 cell mL⁻¹). Nevertheless, owing to its ability to form surface water blooms, this species is considered as one of the most nuisance algae in the subalpine lake district. From the other side, different strains isolated from these large lakes tested negative for the biosynthesis of microcystins, anatoxin-a, nodularins and cylindrospermopsins, and for the presence of mcyE and anaC genes of the microcystins and anatoxin-a gene clusters.     

Effects of a great cormorant colony on wood-inhabiting fungal communities in a coastal Scots pine forest

We studied the changes in the wood-inhabiting fungal communities on dead wood of Scots pine (Pinus sylvestris) under varying duration and intensity of cormorant nesting. The activities of the birds created large amounts of dead wood, changed wood chemistry and altered stand characteristics. Analyses based on records of fruiting structures of fungi showed that fungal communities in the cormorant colony differed from communities in unaffected forest. Ornithogenic factors impacted least on generalist species and promoted the occurrence of fungi uncharacteristic of coniferous wood or wood in general. The rate of ornithogenic change was rapid: a shift in fungal assemblages was evident by the third year of expansion of the bird colony into unaffected forest. It featured the disappearance of tremelloid fungi, a decline of corticioid fungi and an increase of hyphomycetoid fungi. The proportion of coniferous wood specialists decreased and that of generalists increased. It is expected that the changes in fungal communities will persist until the forest is fully regenerated.     

High-Yield Expression in Escherichia coli,Purification and Application of Budding Yeast K2 Killer Protein

Saccharomyces cerevisiae K2 toxin is a highly active extracellular protein, important as a biocontrol agent for biotechnological applications in the wine industry. This protein is produced at negligible levels in yeast, making difficult to isolate it in amounts sufficient for investigation and generation of analysis tools. In this work, we demonstrate the use of a bacterial system for expression of the recombinant K2 protein, suitable for generation of antibodies specific for toxin of the yeast origin. Synthesis of the full-length S. cerevisiae K2 preprotoxin in Escherichia coli was found to be toxic to the host cell, resulting in diminished growth. Such effect was abolished by the introduction of the C-terminal truncation into K2 protein, directing it into non-toxic inclusion body fraction. The obtained protein is of limited solubility thus, facilitating the purification by simple and efficient chromatography-free procedure. The protein aggregates were successfully refolded into a soluble form yielding sufficient amounts of a tag-less truncated K2 protein suitable for polyclonal antibody production. Antibodies were raised in rabbit and found to be specific for detection of both antigen and native S. cerevisiae K2 toxin.