DNA barcoding of Naididae (Annelida, Oligochaeta), based on cytochrome C oxidase gene and ITS2 region in China

Exploring the effectiveness of DNA barcoding in species identification is a prerequisite for biodiversity conservation and environmental monitoring. Aquatic oligochaetes could serve as excellent indicators in aquatic monitoring programmes. However, few studies have examined the effectiveness of DNA barcoding in these specific organisms. The mitochondrial cytochrome C oxidase (COI) gene of 83 specimens belonging to 40 species of 18 genera were sequenced in this study. The results showed that there was a barcode gap between species of Naididae and the intraspecific genetic distances of each species were smaller than interspecific genetic distances. The classification results of ABGD (Automatic Barcode Gap Discovery) were consistent with those of morphological identification, except for Tubifex tubifex and Lumbriculus variegatus. All species were successfully distinguished in the phylogenetic tree, based on the ITS2 region, which was coincident with the morphological result. Our results provided evidence that DNA barcoding can be used as an effective and convenient tool for species identification of the family Naididae and even for other aquatic oligochaetes.

Biodiversity of freshwater macroinvertebrates on Gökçeada Island (North Aegean Sea, Turkey)

Island ecosystems have attracted the attention of scientists since the early days of science because of their formation and biodiversity. Despite their abundant freshwater resources, data on freshwater diversity of Gökçeada are incomplete, especially for macroinvertebrates. This study was conducted in a wide range of freshwater ecosystems on Gökçeada between May 2016 and September 2018. Sampling of macroinvertebrates was performed using a hand net from 34 pre-selected sampling sites on the island. A total of 78 aquatic macroinvertebrate taxa were found, 46 of which are new records for the island. The most common species during the study were Gammarus komareki and Bythinella gokceadaensis with a frequency of 25.93%, followed by Tubifex tubifex, Gammarus sp. and Caenis sp. with a frequency of 22.22%. Endemic and invasive species on the island were assessed according to their presence and frequency values. Based on the results of the current study and previous data, it can be concluded that Gökçeada is characterized by a remarkable species diversity compared to aquatic macroinvertebrates on other Aegean islands.

Organochlorine Pesticides and Polychlorinated Biphenyls in High Mountain Lakes, Mexico

Pollution levels of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) were investigated in the El Sol and the La Luna alpine lakes. The lakes are located in central Mexico, in the crater of the Nevado de Toluca volcano. The El Sol and the La Luna lakes are extremely relevant in Mexico and in the world because they are recognized as pristine regions and environmental reservoirs. Samples of atmospheric aerosol, sediment, plankton, and Tubifex tubifex were collected at three different sample locations for three years (2017, 2018, and 2019) at three different times of year, meaning that the weather conditions at the time of sampling were different. Pollutants were analysed by gas chromatography–mass spectrometry with negative chemical ionisation (GC-MS/NCI). Endosulfan was the most frequent and abundant pollutant, showing the highest peaks of all. Atmospheric aerosol revealed Σ2 = 45 pg/m3, including α and β, while sediment lakes displayed α, β and endosulfan sulphate as Σ3 = 1963 pg/g, whereas plankton and Tubifex tubifex showed Σ2 = 576 pg/g and 540 pg/g for α and β respectively. Results of endosulfan ratios (α/β) and (α-β/endosulfan sulphate) suggest that both fresh and old discharges continue to arrive at the lakesThis study shows for the first time the pollution levels of OCP and PCB in high mountain lakes in Mexico. These results that must be considered by policy makers to mitigate their use in the various productive activities of the region.

In vitro study of anthelmintic effect of butterfly pea (Clitoria ternatea) flower aqueous extract on Tubifex tubifex

Clitoria ternatea (CT) or butterfly pea was one herbs used for Ayurvedic and other traditional medicines. Utilization of its roots, flowers, and leaves proposed several medical adventages with anthelmintic was one of it. Among current anthelmintic burden of resistance, this study aimed to examine the anthelmintic profile from CT flower aqueous extract on Tubifex tubifex. The study was done by conducting a pilot experimental study by extracting CT flowers to water and piping the extracts of different level of dilution to several group of pots containing Tubifex tubifex. Anthelmintic activity was determined with its paralyzing effect and was compared to negative and positive controls with levamisole. Phytochemicals substances were detected with alkaloids, flavonoids, glycosides, saponins, tannins, and triterpenoids detected. The extract exerts anthelmintic activity it 1:4, 1:40, and 1:400 dilutions and was comparable to 1mg/mL and 0.1mg/mL levamisole. Time to paralysis observed suggested a dose-response relationship of the extract on its anthelmintic activities. It was understood the phytochemicals influents the anthelmintic activity by paralysis worms and leads to death. It was concluded that CT flower aqueous extract presents anthelmintic activity, with further experimental study will needed to be conducted.

Whirling disease in the Crowsnest River: an emerging threat to wild salmonids in Alberta

We provide the first documented case of whirling disease (WD) impacts to wild, self-sustaining rainbow trout (RNTR, Oncorynchus mykiss) populations in Canada. Myxobolus cerebralis (Mc), the causative agent of WD, was first confirmed in Alberta in 2016. However, evidence of disease in local fish populations was unknown. Using a weight-of-evidence approach, we examined multiple parasite life cycle stages in the Crowsnest River, Alberta. Percentage of infected Tubifex tubifex worms actively shedding triactinomyxons (TAMs) exceeded known thresholds of Mc establishment and TAM densities instream exceeded thresholds known to cause ≥90% declines in RNTR populations. Mc was detected at 5 of 6 study sites in water, fish, and worms. Disease severity was highest in the lower watershed where 100% of sentinel fish tested positive for Mc 7 to 14 days post-exposure; up to 85% of wild fingerling RNTR showed clinical signs of disease and yearling trout were largely absent from the river suggesting reduced survival. Our findings indicate conditions necessary for outbreak of WD exist in Alberta, highlighting the need to consider this disease as an emerging threat to wild salmonid populations.

Influence of Aging on Bioaccumulation and Toxicity of Copper Oxide Nanoparticles and Dissolved Copper in the Sediment-Dwelling Oligochaete Tubifex tubifex: A Long-Term Study Using a Stable Copper Isotope

For engineered metal nanoparticles (NPs), such as copper oxide (CuO) NPs, the sediment is recognized as a major compartment for NP accumulation. Sediment-dwelling organisms, such as the worm Tubifex tubifex, will be at particular risk of metal and metal NP exposure. However, a range of complex transformation processes in the sediment affects NP bioavailability and toxicity as the contamination ages. The objective of this study was to examine bioaccumulation and adverse effects of CuO NPs in T. tubifex compared to dissolved Cu (administered as CuCl2) and the influence of aging of spiked sediment. This was done in a 28-day exposure experiment with T. tubifex incubated in clean sediment or freshly spiked sediment with different concentrations of dissolved Cu (up to 230 μg g−1 dw) or CuO NPs (up to 40 μg g−1 dw). The experiment was repeated with the same sediments after it had been aged for 2 years. To obtain a distinct isotopic signature compared to background Cu, both Cu forms were based on the stable isotope 65Cu (>99%). The 28-day exposure to sediment-associated dissolved 65Cu and 65CuO NPs resulted in a clear concentration-dependent increase in the T. tubifex65Cu body burden. However, despite the elevated 65Cu body burdens in exposed worms, limited adverse effects were observed in either of the two experiments (e.g., above 80% survival in all treatments, low or no effects on the growth rate, feeding rate, and reproduction). Organisms exposed to aged sediments had lower body burdens of 65Cu than those exposed to freshly spiked sediments and we suggest that aging decreases the bioavailability of both 65Cu forms. In this study, the use of a stable isotope made it possible to use environmentally realistic Cu concentrations and, at the same time, differentiate between newly accumulated 65Cu and background Cu in experimental samples despite the high background Cu concentrations in sediment and T. tubifex tissue. Realistic exposure concentrations and aging of NPs should preferably be included in future studies to increase environmental realism to accurately predict the environmental risk of metal NPs.

DNA barcoding of Naididae (Annelida, Oligochaeta) based on COI and ITS2 genes in China

Exploring the effectiveness of DNA barcoding in species identification is prerequisite for biodiversity conservation and environmental monitoring. Aquatic oligochaete could serve as an excellent indicator in aquatic monitoring programs. However, few studies have examined the effectiveness of DNA barcoding in these specific organisms. The mitochondrial COI gene and nuclear ITS2 gene of 83 specimens belonging to 36 species of 18 genera were sequenced in this study. The results showed that there was a barcode gap between species of Naididae, and the intraspecific genetic distances of each species were smaller than interspecific genetic distances. The classification results of ABGD (Automatic Barcode Gap Discovery) were consistent with those of morphological identification except for Tubifex tubifex and Lumbriculus variegatus. All species were successfully distinguished in the phylogenetic tree based on ITS2 gene, which was coincident with morphological result. Our results provided evidence that DNA barcoding can be used as an effective and convenient tool for species identification of the family Naididae and even aquatic oligochaete.