Detection and identification of opportunistic Exophiala species using the rolling circle amplification of ribosomal internal transcribed spacers

Deep infections by melanized fungi deserve special attention because of a potentially fatal, cerebral or disseminated course of disease in otherwise healthy patients. Timely diagnostics are a major problem with these infections. Rolling circle amplification (RCA) is a sensitive, specific and reproducible isothermal DNA amplification technique for rapid molecular identification of microorganisms. RCA-based diagnostics are characterized by good reproducibility, with few amplification errors compared to PCR. The method is applied here to species of Exophiala known to cause systemic infections in humans. The ITS rDNA region of five Exophiala species (E. dermatitidis, E. oligosperma, E. spinifera, E. xenobiotica, and E. jeanselmei) was sequenced and aligned in view of designing specific padlock probes to be used for the detection of single nucleotide polymorphisms (SNPs) of the Exophiala species concerned. The assay proved to successfully amplify DNA of the target fungi at the level of species; while no cross-reactivity was observed. Amplification products were visualized on 1% agarose gels to verify the specificity of probe-template binding. Amounts of reagents were minimized to avoid the generation of false positive results. The sensitivity of RCA may help to improve early diagnostics of these difficult to diagnose infections.     

Report of chronic subcutaneous abscesses caused byExophiala spinifera

A case of phaeohyphomycosis with subcutaneous abscesses involving skin, muscle and bone was caused byExophiala spinifera. This is the first report ofE. spinifera causing bone degeneration in Recife-PE, Brazil.     

Essential krill species habitat resolved by seasonal upwelling and ocean circulation models within the large marine ecosystem of the California Current System

Due to their global distribution, high biomass and energy content, euphausiids (krill) are important prey for many mid and upper trophic level marine organisms. Understanding drivers of krill habitat is essential for forecasting range shifts, and to better understand the response of krill predators to climate change. We hypothesized that the distribution and abundance of krill species derived from ecosystem surveys in spring/summer relates to geomorphic features, coastal upwelling during the preceding winter, and spring mesoscale oceanographic conditions. To test this hypothesis, we used boosted regression trees with environmental data and ocean model output to quantify the habitat associations of two primary krill species (Euphausia pacifica and Thysanoessa spinifera) in the central California Current Ecosystem from 2002 to 2018. Models confirmed the neritic distribution of T. spinifera and pelagic, outer slope association of E. pacifica (deviance explained ~35%). Distribution of these species were influenced by depth and bottom rugosity; chlorophyll-a concentrations and increased winter upwelling conditions; and spring surface currents and wind stress. Thysanoessa spinifera and E. pacifica abundance responded negatively (positively) to warm (cold) climate events, confirming known relationships. As an independent evaluation of krill models, observations of krill predator (seabirds, marine mammals) distribution indicated they were present within habitats of predicted high krill species abundance. Our framework indicates species-specific habitat relationships for these foundational forage species and their negative response to large-scale climate variations, such as El Niño and marine heatwave conditions. The approach can be easily transferred to other ecosystems or krill species that respond to similar ocean and climate forcing.     

Devastating farmed abalone mortalities attributed to yessotoxin-producing dinoflagellates

A large dinoflagellate bloom in Walker Bay (South Africa) in January 2017 impacted 3 land-based abalone farms resulting in the death of several million animals. Satellite-derived images of Chl-a from the Ocean and Land Colour Imager (OLCI) on board the European Space Agency Sentinel-3 A showed bloom initiation in late December 2016 and dispersal in mid-February 2017. The bloom was dominated by two dinoflagellate species identified by light microscopy as Gonyaulax spinifera (Claparède & Lachmann) Diesing, 1866 and Lingulodinium polyedrum (Stein) Dodge, 1989. These morphologically based identifications were confirmed by phylogenetic analysis using partial sequences of the large subunit rDNA of both dinoflagellates. The appearance of yessotoxins (YTX) in abalone clearly coincided with increases in dinoflagellate concentrations. Yessotoxins in both the plankton and abalone were dominated by the two analogues homo-YTX and 45-hydroxy-YTX. The absence of toxins in a clonal culture of L. polyedrum implicated G. spinifera as the likely source of YTX. Toxin concentrations were found to be highest in the gills which showed the most significant pathology, including severe, generalized disruption of the gill epithelium characterized by degeneration and necrosis of epithelial cells accompanied by a modest inflammatory response. Some farms undertook pre-emptive or emergency harvesting to reduce financial losses.     

Effect of acclimation temperature and substrate type on selected temperature,movement and activity of juvenile spiny softshell turtles (Apalone spinifera) in an aquatic thermal gradient

In some turtle species, temperature selection may be influenced by environmental conditions, including acclimation temperature and substrate quality. These factors may be particularly important for softshell turtles that are highly aquatic and often thermoregulate by burying in the substrate in shallow water microhabitats. We tested for effects of acclimation temperature (22 °C or 27 °C) and substrate type (sand or gravel) on the selected temperature and movement patterns of 20 juvenile spiny softhshell turtles (Apalone spinifera; Reptilia: Trionychidae) in an aquatic thermal gradient of 14–34 °C. Among 7–11 month old juvenile softshell turtles, acclimation temperature and substrate type did not influence temperature selection, nor alter activity and movement patterns. During thermal gradient tests, both 22- and 27 °C-acclimated turtles selected the warmest temperature (34 °C) available most frequently, regardless of substrate type (sand or gravel). Similarly, acclimation temperature and substrate type did not influence movement patterns of turtles, nor the number of chambers used in the gradient tests. These results suggest that juvenile Apalone spinifera are capable of detecting small temperature increments and prefer warm temperatures that may positively influence growth and metabolism, and that thermal factors more significantly influence aquatic thermoregulation in this species than does substrate type.     

Relationship between the dinoflagellate cyst Spiniferites pachydermus and Gonyaulax ellegaardiae sp. nov. from Izmir Bay,Turkey, and molecular characterization

Here, we established the cyst‐motile stage relation‐ship for Spiniferites pachydermus through incubation of cysts with a characteristically microreticulate/perforate surface isolated from Izmir Bay in the eastern Aegean Sea of the eastern Mediterranean. The morphology of the motile stage was similar to Gonyaulax spinifera but had a different size, overhang, displacement and reticulations. Based on the distinct morphology of the cyst and morphological differences in motile cells, we assigned S. pachydermus from Izmir Bay to the new species Gonyaulax ellegaardiae. We elucidate the phylogenetic relationship of G. ellegaardiae through large and small subunit ribosomal DNA and show that it forms a clade with other species that belong to the G. spinifera complex.     

Redescription of Scapholeberis echinulata Sars 1903 (Crustacea,Cladocera)

Scapholeberis echinulata Sars is rediscribed, based on type material of Sars (S. echinulata), Daday (s. erinaceus) and the author's own samples. A differential diagnosis with Scapholeberis spinifera Nicolet and S. mucronata O. F. Müller, S. microcephala Sars, S. kingi Sars and S. aurita Fischer is given.     

Further studies on the phylogenesis of the genus Exophiala and Hortaea

The conidial ontogenesis of the pathogenic black yeasts is studied at an ultrastructural level and their phylogenesis is discussed. Five cultures of Exophiala dermatitidis, four of E. jeanselmei, one of E. moniliae, one of E. spinifera and six of H. werneckii were observed using a scanning electron microscope.The conidial ontogenesis of the Exophiala species is not pleomorphic but only annellidic. There are definite differences in morphology of annellated tips among the Exophiala species. The ontogenesis of Hortaea werneckii consists of a combination of sympodial and annellidic conidiogenesis. Its sympodial anamorph is unique and the annellidic anamorph is considered to be a homology of the sympodial one.     

Infection with Haemoproteus iwa affects vector movement in a hippoboscid fly—frigatebird system

Haemosporidian parasites, which require both a vertebrate and invertebrate host, are most commonly studied in the life stages occurring in the vertebrate. However, aspects of the vector's behaviour and biology can have profound effects on parasite dynamics. We explored the effects of a haemosporidian parasite, Haemoproteus iwa, on a hippoboscid fly vector, Olfersia spinifera. Olfersia spinifera is an obligate ectoparasite of the great frigatebird, Fregata minor, living among bird feathers for all of its adult life. This study examined the movements of O. spinifera between great frigatebird hosts. Movement, or host switching, was inferred by identifying host (frigatebird) microsatellite genotypes from fly bloodmeals that did not match the host from which the fly was collected. Such host switches were analysed using a logistic regression model, and the best‐fit model included the H. iwa infection status of the fly and the bird host sex. Uninfected flies were more likely to have a bird genotype in their bloodmeal that was different from their current host's genotype (i.e. to have switched hosts) than infected flies. Flies collected from female birds were more likely to have switched hosts than those collected on males. Reduced movement of infected flies suggests that there may be a cost of parasitism for the fly. The effect of host sex is probably driven by differences in the sex ratio of bird hosts available to moving flies.     

Daily variation of marine cladoceran densities in a tropical bay – Brazil

This contribution presents the daily variation of marine cladoceran densities in Guanabara Bay. The samples were obtained by vertical tows with a 200 m mesh-size conical net at different times over a 4-day period in September, 1995. Penilia avirostris was the most common species, followed by Evadne tergestina and E. spinifera. These organisms presented higher densities at nightfall. Physical factors like tide and light are responsible for the highest densities. High tide and nightfall time are requested conditions for the P. avirostris and E. tergestina abundance. Temperature and salinity appear to have no relation with cladoceran densities.     

Three new colorless Euglenophyceae from marine situations

Summary Investigation of the colorless Euglenophyceae of sandy beaches and marine sediment-water interfaces on the east and west coasts of the U.S.A. has resulted in the discovery of two new species, Sphenomonas elongata and Entosiphon cuneatiformis, and one new genus and species, Pentamonas spinifera, herein described and figured.This paper is dedicated to Professor Dr. E. G. Pringsheim on the occasion of his 80 th birthday.     

In situ measurements of bioluminescence response of Gonyaulax spinifera to various mechanical stimuli

A conspicuous bioluminescence during nighttime was reported in an aquaculture farm in the Cochin estuary due to Gonyaulax spinifera bloom on March 20, 2020. In situ measurements on bioluminescence was carried out during nighttime to quantify the response of G. spinifera to various mechanical stimuli. The bioluminescence intensity (BI) was measured using Glowtracka, an advanced single channel sensor, attached to a Conductivity–Temperature–Depth Profiler. In steady environment, without any external stimuli, the bioluminescence generated due to the movement of fishes and shrimps in the water column was not detected by the sensor. However, stimuli such as a hand splash, oar and swimming movements, and a mixer could generate measurable bioluminescence responses. An abundance of?~?2.7?×?10⁶ cells L^?1 of G. spinifera with exceptionally high chlorophyll a of 25 mg m^?3 was recorded. The BI in response to hand splash was recorded as high as 1.6?×?10¹¹ photons cm^?2 s^?1. Similarly, BI of?~?1–6?×?10¹⁰ photons cm^?2 s^?1 with a cumulative bioluminescence of?~?2.51?×?10¹² photons cm^?2 (for 35 s) was recorded when there is a mixer with a constant force of 494 N/800 rpm min^?1. The response of G. spinifera was spontaneous with no time lapse between application of stimuli and the bioluminescence response. Interestingly, in natural environment, application of stimulus for longer time periods (10 min) does not lower the bioluminescence intensity due to the replenishment of water thrusted in by the mixer from surrounding areas. We also demonstrated that the bioluminescence intensity decreases with increase in distance from the source of stimuli (mixer) (av. 1.84?×?10¹⁰ photons cm^?2 s^?1 at 0.2 m to av. 0.05?×?10¹⁰ photons cm^?2 s^?1 at 1 m). The BI was highest in the periphery of the turbulent wake generated by the stimuli (av. 3.1?×?10¹⁰ photons cm^?2 s^?1) compared to the center (av. 1.8?×?10¹⁰ photons cm^?2 s^?1). When the stimuli was applied vertically down, the BI decreased from 0.2 m (0.3?×?10¹⁰ photons cm^?2 s^?1) to 0.5 m (0.10?×?10¹⁰ photons cm^?2 s^?1). Our study demonstrates that the BI of G. spinifera increases with increase in mechanical stimuli and decreases with increase in distance from the stimuli.

     

Differential use of zooplankton prey by Ancient murrelets and Cassin's auklets in the Queen Charlotte Islands

The distribution at sea and the food of two similar sized plankton-feedingalcids were examined during the 1981 breeding seasons in thenorthwestern Queen Charlotte Islands, British Columbia. Thetwo alcids, the Ancient murrelet (Synthliboramphus antiquus)and the Cassin's auklet (Prychoramphus aleuticus) have differentchick-rearing strategies. Both species fed predominantly atthe shelf break, although the Cassin's auklet also foraged overseamounts. The feeding distributions of the species appear tobe related to those of their prey. Zooplankton sampling indicatedthat each alcid selects a small and different portion of thezooplankton available in surface waters. The Ancient murrelet'smain foods were euphausiids (Thysanoessa spinifera and Euphausiapacifica) and larval and juvenile fishes. The Cassin's aukletchicks fed chiefly on calanoid copepods (Neocalanus cristatus).euphausiids (mostly Thysanoessa longipes in 1981, but in otheryears also Thysanoessa spinifera), and larval and juvenile fishes.The Cassin's auklets took smaller prey than the Ancient murrelet.Differences in the diets of the two alcid species were associatedwith differences in morphology and chick-rearing strategies.     

Late Darriwilian–early Katian (Ordovician) ostracods of northeastern Yunnan,Southwest China

Ordovician silicified ostracods of the western Yangtze Platform, South China, are documented for the first time from the Huadan Formation of Qiaojia County, northeastern Yunnan Province, Southwest China. Twenty-one species of 17 genera are identified with new taxa including Vendona spinifera n. sp. and Nikitinella songliangensis n. sp. These two species, together with Medianella gigantea are the most common constituents of the fauna. Faunal comparison shows close connections between South China, Baltica, the Argentine Precordillera, Kazakhstan, Tarim, and Sibumasu during this time interval. This fauna is considered to be related to a nearshore carbonate environment, and biostratigraphical analysis of it supports a late Darriwilian to early Katian age for the Huadan Formation.     

Frequency and molecular characterisation of Entamoeba histolytica,Entamoeba dispar,Entamoeba moshkovskii,and Entamoeba hartmanni in the context of water scarcity in northeastern Brazil

This study aimed to estimate the frequency, associated factors, and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, andEntamoeba hartmanni infections. We performed a survey (n = 213 subjects) to obtain parasitological, sanitation, and sociodemographic data. Faecal samples were processed through flotation and centrifugation methods.E. histolytica, E. dispar, E. moshkovskii, and E. hartmanni were identified by nested-polymerase chain reaction (PCR). The overall prevalence of infection was 22/213 (10.3%). The infection rate among subjects who drink rainwater collected from roofs in tanks was higher than the rate in subjects who drink desalinated water pumped from wells; similarly, the infection rate among subjects who practice open defecation was significantly higher than that of subjects with latrines. Out of the 22 samples positive for morphologically indistinguishableEntamoeba species, the differentiation by PCR was successful for 21. The species distribution was as follows: 57.1% to E. dispar, 23.8% to E. histolytica, 14.3% toE. histolytica and E. dispar, and 4.8% E. dispar and E. hartmanni. These data suggest a high prevalence of asymptomatic infection by the group of morphologically indistinguishable Entamoeba histolytica/dispar/moshkovskiicomplex and E. hartmanni species. In this context of water scarcity, the sanitary and socioenvironmental characteristics of the region appear to favour transmission.