Bacterium-bacterium inhibitory interactions among psychrotrophic bacteria isolated from Antarctic seawater (Terra Nova Bay, Ross Sea)

One hundred and forty bacteria isolated from Antarctic seawater samples were examined for their ability to inhibit the growth of indigenous isolates and their sensitivity to antibacterial activity expressed by one another. On the basis of 16S rRNA gene sequencing and analysis, bacterial isolates were assigned to five phylogenetically different taxa, Actinobacteria, alpha and gamma subclasses of Proteobacteria, Bacillaceae, and Bacteroidetes. Twenty-one isolates (15%), predominantly Actinobacteria, exhibited antagonistic properties against marine bacteria of Antarctic origin. Members of Bacteroidetes and Firmicutes did not show any inhibitory activity. Differences were observed among inhibition patterns of single isolates, suggesting that their activity was more likely strain-specific rather than dependent on phylogenetic affiliation. A novel analysis based on network theory confirmed these results, showing that the structure of this population is probably robust to perturbations, but also that it depends strongly on the most active strains. The determination of plasmid incidence in the bacterial strains investigated revealed that there was no correlation between their presence and the antagonistic activity. The data presented here provide evidence for the antagonistic interactions within bacterial strains inhabiting Antarctic seawater and suggest the potential exploitation of Antarctic bacteria as a novel source of antibiotics.     

Origin,diversity, and biogeography of Antarctic scale worms (Polychaeta: Polynoidae): a wide‐scale barcoding approach

The Antarctic marine environment hosts diversified and highly endemic benthos owing to its unique geologic and climatic history. Current warming trends have increased the urgency of understanding Antarctic species history to predict how environmental changes will impact ecosystem functioning. Antarctic benthic lineages have traditionally been examined under three hypotheses: (1) high endemism and local radiation, (2) emergence of deep‐sea taxa through thermohaline circulation, and (3) species migrations across the Polar Front. In this study, we investigated which hypotheses best describe benthic invertebrate origins by examining Antarctic scale worms (Polynoidae). We amassed 691 polynoid sequences from the Southern Ocean and neighboring areas: the Kerguelen and Tierra del Fuego (South America) archipelagos, the Indian Ocean, and waters around New Zealand. We performed phylogenetic reconstructions to identify lineages across geographic regions, aided by mitochondrial markers cytochrome c oxidase subunit I (Cox1) and 16S ribosomal RNA (16S). Additionally, we produced haplotype networks at the species scale to examine genetic diversity, biogeographic separations, and past demography. The Cox1 dataset provided the most illuminating insights into the evolution of polynoids, with a total of 36 lineages identified. Eunoe sp. was present at Tierra del Fuego and Kerguelen, in favor of the latter acting as a migration crossroads. Harmothoe fuligineum, widespread around the Antarctic continent, was also present but isolated at Kerguelen, possibly resulting from historical freeze–thaw cycles. The genus Polyeunoa appears to have diversified prior to colonizing the continent, leading to the co‐occurrence of at least three cryptic species around the Southern and Indian Oceans. Analyses identified that nearly all populations are presently expanding following a bottleneck event, possibly caused by habitat reduction from the last glacial episodes. Findings support multiple origins for contemporary Antarctic polynoids, and some species investigated here provide information on ancestral scenarios of (re)colonization. First, it is apparent that species collected from the Antarctic continent are endemic, as the absence of closely related species in the Kerguelen and Tierra del Fuego datasets for most lineages argues in favor of Hypothesis 1 of local origin. Next, Eunoe sp. and H. fuligineum, however, support the possibility of Kerguelen and other sub‐Antarctic islands acting as a crossroads for larvae of some species, in support of Hypothesis 3. Finally, the genus Polyeunoa, conversely, is found at depths greater than 150 m and may have a deep origin, in line with Hypothesis 2. These “non endemic” groups, nevertheless, have a distribution that is either north or south of the Antarctic Polar Front, indicating that there is still a barrier to dispersal, even in the deep sea.     

Retrotransposon‐based genetic diversity of Deschampsia antarctica Desv. from King George Island (Maritime Antarctic)

Deschampsia antarctica Desv. can be found in diverse Antarctic habitats which may vary considerably in terms of environmental conditions and soil properties. As a result, the species is characterized by wide ecotypic variation in terms of both morphological and anatomical traits. The species is a unique example of an organism that can successfully colonize inhospitable regions due to its phenomenal ability to adapt to both the local mosaic of microhabitats and to general climatic fluctuations. For this reason, D. antarctica has been widely investigated in studies analyzing morphophysiological and biochemical responses to various abiotic stresses (frost, drought, salinity, increased UV radiation). However, there is little evidence to indicate whether the observed polymorphism is accompanied by the corresponding genetic variation. In the present study, retrotransposon‐based iPBS markers were used to trace the genetic variation of D. antarctica collected in nine sites of the Arctowski oasis on King George Island (Western Antarctic). The genotyping of 165 individuals from nine populations with seven iPBS primers revealed 125 amplification products, 15 of which (12%) were polymorphic, with an average of 5.6% polymorphic fragments per population. Only one of the polymorphic fragments, observed in population 6, was represented as a private band. The analyzed specimens were characterized by low genetic diversity (uH_e = 0.021, I = 0.030) and high population differentiation (F_ST = 0.4874). An analysis of Fu's F_S statistics and mismatch distribution in most populations (excluding population 2, 6 and 9) revealed demographic/spatial expansion, whereas significant traces of reduction in effective population size were found in three populations (1, 3 and 5). The iPBS markers revealed genetic polymorphism of D. antarctica, which could be attributed to the mobilization of random transposable elements, unique features of reproductive biology, and/or geographic location of the examined populations.     

Genomic data reveal international lineages of critical priority Escherichia coli harbouring wide resistome in Andean condors (Vultur gryphus Linnaeus, 1758)

Critical priority pathogens have globally disseminated beyond clinical settings, thereby threatening wildlife. Andean Condors (Vultur gryphus) are essential for ecosystem health and functioning, but their populations are globally near threatened and declining due to anthropogenic activities. During a microbiological and genomic surveillance study of critical priority antibiotic‐resistant pathogens, we identified pandemic lineages of multidrug‐resistant extended‐spectrum β‐lactamase (ESBL)‐producing Escherichia coli colonizing Andean Condors admitted at two wildlife rehabilitation centres in South America. Genomic analysis revealed the presence of genes encoding resistance to hospital and healthcare agents among international E. coli clones belonging to sequence types (STs) ST162, ST602, ST1196 and ST1485. In this regard, the resistome included genes conferring resistance to clinically important cephalosporins (i.e., CTX‐M‐14, CTX‐M‐55 and CTX‐M‐65 ESBL genes), heavy metals (arsenic, mercury, lead, cadmium, copper, silver), pesticides (glyphosate) and domestic/hospital disinfectants, suggesting a link with anthropogenic environmental pollution. On the other hand, the presence of virulence factors, including the astA gene associated with outbreak of childhood diarrhoea and extra‐intestinal disease in animals, was identified, whereas virulent behaviour was confirmed using the Galleria mellonella infection model. E. coli ST162, ST602, ST1196 and ST1485 have been previously identified in humans and food‐producing animals worldwide, indicating that a wide resistome could contribute to rapid adaptation and dissemination of these clones at the human–animal–environment interface. Therefore, these results highlight that Andean Condors have been colonized by critical priority pathogens, becoming potential environmental reservoirs and/or vectors for dissemination of virulent and antimicrobial‐resistant bacteria and/or their genes, in associated ecosystems and wildlife.     

Health risk perspectives of metal(loid) exposure via consumption of striped venus clam (Chamelea gallina Linnaeus, 1758)

The metal(loid)s are mainly transported to the Black Sea via rivers and affect the coastal area where fisheries are intensive. Chamelea gallina (striped venus clam) is the most abundant mollusk species in the Black Sea and used as a bioindicator for monitoring of metal(loid) pollution. In this respect, the concentrations of As, Cd, Cr, Cu, Pb, and Zn in C. gallina collected from 16 different locations along the Black Sea coast (Turkey, Bulgaria, and Russia) were determined and evaluated for the potential human health risk. The results obtained were found to be below the maximum permissible limits indicated in the food safety guidelines. Similarly, the analyzed metal(loid)s did not pose a potential hazard to humans for consumption of the C. gallina, regarding provisional tolerable weekly intake described by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Moreover, target hazard quotient and hazard index were found to be lower than 1 for not only average consumer but also the consumer who consumed a portion (160 g) once a week. In contrast, consumption of C. gallina could cause health risks for Cd and As in case of increasing portion sizes.     

Triadic interactions in captive Barbary macaques (Macaca sylvanus,Linnaeus, 1758): “Agonistic buffering”?

This study presents data on the expression of male-immature triadic interactions, previously termed agonistic buffering, in a captive Macaca sylvanus group. Agonistic buffering has been hypothesized as inhibiting or modifying the expression of aggression. This was tested by examining (1) the dominance ranks of the animals involved in the triadic interactions, (2) the events preceding and following the triadic interactions, and (3) the presence of an infant in nonagonistic encounters between juvenile, subadult, and adult males. The results obtained do not support the hypothesis of agonistic buffering as the single explanation for triadic interactions, but emphasize the contextual variability in the expression of these triadic interactions.     

Preparation and anti-osteoporotic activities in vivo of phosphorylated peptides from Antarctic krill (Euphausia superba)

Antarctic krill (Euphausia superba) protein serves as a novel sustainable protein source for human. Krill protein isolate was phosphorylated by the dry-heating method with sodium pyrophosphate. Phosphorylated peptides from Antarctic krill (PP-AKP) were obtained from phosphorylated protein through tryptic hydrolysis. Two types of phosphate bonds were introduced by phosphorylation, i.e. PO and PO bonds. The anti-osteoporotic activities of PP-AKP at two doses (400 and 800 mg/kg body weight) were investigated with an osteoporotic rat model, which was established with bilateral ovariectomy surgery. Different doses of PP-AKP were given intraperitoneal injections to rats once a day with alendronate as a positive control. Phosphorylated peptides from Antarctic krill dose-dependently preserved bone mineral density in osteoporotic rats by increasing the degree of bone mineralization. Both trabecular and cortical bone strength in osteoporotic rats was significantly improved with PP-AKP treatment. The mechanism by which PP-AKP augmented bone mineral density and bone strength was relation to the reduction in osteoclast-mediated bone remodeling, as was supported by the decrease in bone resorption markers. Phosphorylated peptides from Antarctic krill could be developed as functional food or nutritional supplements.     

A dipteran from south of the Antarctic Circle: Belgica antarctica (Chironomidae) with a description of its larva

Belgica antarctica Jacobs is recorded for the first time from two localities south of the Antarctic circle: Orford Cliff (66°55'S) on the Continental mainland and the Refuge Islands (68°21'S). The larva is described, and measurements of head capsule lengths indicate four instars. The larvae south of the Antarctic circle appear to be slightly smaller than those from within the main distributional range. Belgica antarctica is certainly the earth's southernmost free-living holometabolous insect, but the flea Glaciopsyllus antarcticus has been recorded 14 minutes of latitude farther south than the southernmost records of B. antarctica.     

Macromolecular interactions with cadmium and the effects of zinc,copper, lead,and mercury ions

Interactions of cadmium (Cd) ions with bovine serum albumin (BSA), bovine hepatic metallothionein (MT), calf thymus histone and deoxyribonucleic acid (DNA), and bovine hepatic chromatins were studied in the presence and absence of divalent zinc (Zn), copper (Cu), mercury (Hg), or lead (Pb) ions, using equilibrium dialysis at pH 7 and at 37°C. The BSA had 3.5 Cd-binding sites with an apparent affinity constant of 1×10⁵. The other metal ions inhibited the binding by reducing the affinity constant and the number of Cd-binding sites in BSA. There were 6 high affinity and 13 low affinity Cd-binding sites in the MT. Zinc ions had poor efficacy in reducing the binding of Cd to the MT. However, the Cu²⁺ and Hg²⁺ ions inhibited the Cd binding to a considerable extent, the former ions being more potent in this respect. Histone did not bind Cd. There were two kinds of Cd-binding sites in DNA: One mole of Cd per four moles DNA-phosphorus at low affinity sites, and one mole of Cd per 6.7 moles DNA-phosphorus at high affinity sites. Their apparent association constants were 8.3×10⁵ and 4.4×10⁶ M, respectively. The other metal ions had inhibitory effects on the binding of Cd to DNA. Histone reduced the Cd-DNA interactions to only a minor extent. The other metal ions reduced the binding of Cd to DNA-histone complex to a small extent. Cadmium binds to the euchromatin (Euch), heterochromatin (Het), and Euch-Het mixture almost equally. The other metal ions reduced the binding maximally in Euch-Het followed next in order by Het and Euch. Cupric ions were the most potent inhibitors of the interactions of Cd with the nuclear materials.     

Heterolobosean amoebae from Arctic and Antarctic extremes: 18 novel strains of Allovahlkampfia,Vahlkampfia and Naegleria

The diversity of heterolobosean amoebae, important members of soil, marine and freshwater microeukaryote communities in the temperate zones, is greatly under-explored in high latitudes. To address this imbalance, we studied the diversity of this group of free-living amoebae in the Arctic and the Antarctic using culture dependent methods. Eighteen strain representatives of three heterolobosean genera, Allovahlkampfia Walochnik et Mulec, 2009 (1 strain), Vahlkampfia Chatton et Lalung-Bonnaier, 1912 (2) and Naegleria Alexeieff, 1912 (15) were isolated from 179 samples of wet soil and fresh water with sediments collected in 6 localities. The Allovahkampfia strain is the first representative of the genus from the Antarctic; 14 strains (7 from the Arctic, 7 from the Antarctic) of the highly represented genus Naegleria complete the ‘polar’ cluster of five Naegleria species previously known from the Arctic and Sub-Antarctic regions, whereas one strain enriches the ‘dobsoni’ cluster of Naegleria strains of diverse origin. Present isolations of Naegleria polaris De Jonckheere, 2006 from Svalbard, in the Arctic and Vega Island, in the Antarctic and N. neopolaris De Jonckheere, 2006 from Svalbard and Greenland in the Arctic, and James Ross Island, the Antarctic demonstrate their bipolar distribution, which in free-living amoebae has so far only been known for Vermistella Morand et Anderson, 2007.     

New hyperbenthic species of Misophriopsis and Misophriella, first record of misophrioid copepods (Crustacea) from Antarctic waters

  Misophrioid copepods are reported for the first time from Antarctic waters. A species of each of the genera Misophriopsis and Misophriella is described from hyperbenthic layers of the continental shelf of the eastern Weddell Sea. The genus Misophria is also reported from these waters. Misophriopsis australis sp. nov. is the fifth representative of this widely distributed genus, previously reported from the North Atlantic, the China Sea, and both NW and NE quadrants of the Pacific Ocean. Misophriella schminkei sp. nov. is the second representative of the genus, known to date only from a single female caught in North Atlantic bathyal depths. Traces of subdivisions on antennary exopodal segments of the latter species suggest that the first segment may be a triple segment, while the second and terminal segments may be double segments. In addition, the mandibular exopod of this taxon indicates that a six-segmented ramus with setal formula 0, 1, 1, 1, 1, 2 may represent the ground pattern in Copepoda. The presence of parts of a cyclopinid copepod in the gut of M. schminkei sp. nov. indicates that this species may be carnivorous. Received: 12 November 1998 / Received in revised form: 26 July 1999 / Accepted: 27 July 1999     

Chromosomal diversity in three species of electric fish (Apteronotidae,Gymnotiformes) from the Amazon Basin

Cytogenetic studies were carried out on samples of Parapteronotus hasemani, Sternarchogiton preto and Sternarchorhamphus muelleri (Apteronotidae, Gymnotiformes) from the Amazon basin. The first two species exhibited both a 2n = 52 karyotype, but differed in their karyotypic formulae, distribution of constitutive heterochromatin, and chromosomal location of the NOR. The third species, Sternarchorhamphus muelleri, was found to have a 2n = 32 karyotype. In all three species the DAPI and chromomycin A3 staining results were consistent with the C-banding results and nucleolar organizer region (NOR) localization. The 18S rDNA probe confirmed that there was only one pair of ribosomal DNA cistron bearers per species. The telomeric probe did not reveal interstitial telomeric sequences (ITS). The karyotypic differences among these species can be used for taxonomic identification. These data will be useful in future studies of these fishes and help understanding the phylogenetic relationships and chromosomal evolution of the Apteronotidae.     

Differential effects of Zn concentrations on Cr(VI) uptake by two Salvinia species: involvement of thiol compounds

Abstract

Floating ferns of the genus Salvinia have great potential for phytoremediation of heavy metals. To date, the effect of essential metals on the accumulation and transport of toxic metals by aquatic ferns has not been suitably established. The aim of this study was to compare the ability of floating leaves of Salvinia minima and Salvinia rotundifolia species to accumulate Cr from Cr(VI solutions containing very low (0.02?mg L^?1) and low (5?mg L^?1) Zn concentrations. After 7-day metal-exposure period, results showed that Zn increased Cr accumulation in S. minima leaves whereas in S. rotundifolia decreased significantly. Contrarily Zn accumulation did not show great differences between species. This fact may indicate that Zn interfere Cr(VI) uptake by S. rotundifolia. Bioconcentration factor (BCF) and translocation factor (TF) were affected differently by Zn in both Salvinia species. Membrane stability index (MSI) of both Salvinia species was decreased significantly by 5?mg L^?1 Zn concentration. Zn ions also increased hydrogen peroxide accumulation in fronds of Salvinia species. Total thiols (TT), non-protein thiols (NPT) and protein-bound thiols (PBT) were differentially affected by Cr(VI) and Zn ions. This study provides evidences on the involvement of different mechanisms against Cr(VI)/Zn toxicity in S. minima and S rotundifolia species.     

The Biology of Lophelia pertusa (Linnaeus 1758) and Other Deep-Water Reef-Forming Corals and Impacts from Human Activities.

Over the last twenty years, human exploitation has begun to have an impact in the deep sea, especially in the upper bathyal zone. This has mainly taken the form of deep-sea fishing but more recently oil exploration has extended beyond the continental shelf. Deep-water coral reefs occur in the upper bathyal zone throughout the world. These structures, however, are poorly studied with respect to their occurrence, biology and the diversity of the communities associated with them. In the North-East Atlantic the coral Lophelia pertusa has frequently been recorded. The present review examines the current knowledge on L. pertusa and discusses similarities between its biology and that of other deep-water, reef-forming, corals. It is concluded that L. pertusa is a reef-forming coral that has a highly diverse associated fauna. Associated diversity is compared with that of tropical shallow-water reefs. Such a highly diverse fauna may be shared with other deep-water, reef-forming, corals though as yet many of these are poorly studied. The main potential threats to L. pertusa in the North-East Atlantic are considered to be natural phenomena, such as slope failures and changes in ocean circulation and anthropogenic impacts such as deep-sea fishing and oil exploration. The existing and potential impacts of these activities on L. pertusa are discussed. Deep-sea fishing is also known to have had a significant impact on deep-water reefs in other parts of the world.     

Genetic diversity and conservation of Ipomoea microdactyla (Convolvulaceae): an endemic vine from the Bahamas,Cuba, and southeastern Florida

Ipomoea microdactyla Griseb. (Convolvulaceae) is restricted to the Bahamian archipelago, Cuba, and southeastern Florida. The species is listed as a state endangered species in Florida, where it is mostly restricted to the hyperfragmented pine rockland of Miami‐Dade County. Using seven DNA microsatellite loci, we assessed levels of genetic diversity for 12 populations of this species from Andros Island in the Bahamas (six sites), Cuba (one site), and Florida (five sites). We found significantly greater mean numbers of alleles, and higher mean values for both observed and expected heterozygosity in populations from the continuous forest on Andros than those from the habitat fragments in Florida. It is unknown if these patterns of genetic diversity in the Florida populations are the result of habitat fragmentation or founder effects. The population from Cuba exhibited relatively high levels of genetic variation, suggesting that this island is a major center of diversity and dispersal for this species. It appears that hybrid introgression for I. carolina alleles within I. microdactyla individuals occurred at a single site on Andros Island. Overall, the mean inbreeding coefficient value was 0.089, suggesting low levels of inbreeding. The highest inbreeding coefficient values were mostly recorded in Florida. Two groups were revealed, one containing the populations from Florida, and the second one encompassing those from the Bahamas and Cuba. Our results highlight the negative genetic consequences of habitat fragmentation and support initiatives recently established to establish corridors to connect the remnants of the pine forest of the Miami‐Dade County.     

Synergistic interactions in two-drug and three-drug combinations (thymol,EDTA and vancomycin) against multi drug resistant bacteria including E. coli

Combinations of two or more drugs, which affect different targets, have frequently been used as a new approach against resistant bacteria. In our work we studied the antimicrobial activity (MIC, MBC) of individual drugs (the phenolic monoterpene thymol, EDTA and vancomycin), of two-drug interactions between thymol and EDTA in comparison with three-drug interactions with vancomycin against sensitive and resistant bacteria. Thymol demonstrated moderate bactericidal activity (MBC between 60 and 4000 μg/ml) while EDTA only exhibited bacteriostatic activity over a range of 60–4000 μg/ml. MICs of vancomycin were between 0.125 and 16 μg/ml against Gram-positive and between 32 and 128 μg/ml against Gram-negative bacteria. Checkerboard dilution and time-kill curve assays were performed to evaluate the mode of interaction of several combinations against Methicillin-resistant Staphylococcus aureus (MRSA NCTC 10442) and Escherichia coli (ATCC 25922). Checkerboard data indicate indifferent interaction against Gram-positive (FICI = 1–1.3) and synergy against Gram-negative bacteria (FICI ≈ 0.4), while time kill analyses suggest synergistic effect in different combinations against both types of bacteria. It is remarkable that the combinations could enhance the sensitivity of E. coli to vancomycin 16-fold to which it is normally insensitive. We have provided proof for the concept, that combinations of known antibiotics with modern phytotherapeutics can expand the spectrum of useful therapeutics.