Contaminants and their ecological risk assessment in beach sediments and water along the Maharashtra coast of India: A comprehensive approach using microplastics, heavy metal(loid)s, pharmaceuticals, personal care products and plasticisers.

Emerging contaminants and their pervasive presence in freshwater ecosystems have been widely documented, but less is known about their prevalence and the harm they cause in marine ecosystems, particularly in developing countries. This study provides data on the prevalence and risk posed by microplastics, plasticisers, pharmaceuticals and personal care products (PPCPs), and heavy metal(loid)s (HMs) along the Maharashtra coast of India. The sediment and coastal water samples were collected from 17 sampling stations, processed, and subjected to FTIR-ATR, ICP-MS, SEM-EDX, LC-MS/MS, and GC-MS for further analysis. Higher MPs abundance, combined with the pollution load index, indicates that the northern zone is a high-impact zone with pollution concerns. Plasticisers in extracted MPs and HMs adsorption on MPs surface from surrounding waters reveal their roles as a source and vector for contaminants, respectively. The mean concentration of metoprolol (53.7-306 ng L-1), tramadol (16.6-198 ng L-1), venlafaxine (24.6-234 ng L-1), and triclosan (211-433 ng L-1) in Maharashtra's coastal waters were several folds higher than in other water systems, raising major health concerns. The hazard quotient (HQ) scores revealed that >70 % of study sites pose a high to medium (1 > HQ > 0.1) ecological risk to fish, crustaceans and algae, indicating serious concern. Fish and crustaceans (35.3 % each) show a higher level of risk than algae (29.5 %). Metoprolol and venlafaxine could represent greater ecological risks than tramadol. Similarly, HQ suggests that bisphenol A has larger ecological risks than bisphenol S along the Maharashtra coast. To the best of our knowledge, this is the first in-depth investigation into emerging pollutants in Indian coastal regions. This information is crucial for better policy formulation and coastal management in India in general, and Maharashtra in particular.

Contracaecum nematode parasites in hillstream loaches of the Western Ghats, India.

Nematode parasites of the family Anisakidae infect definitive hosts, such as fish-eating birds and mammals, through primary intermediate hosts like copepods and secondary intermediate hosts like fishes. However, consumption of raw or undercooked fish can lead to nematode infection called anisakidosis in humans. We observed the presence of nematode infection in hillstream loaches of families Cobitidae and Nemacheilidae available for human consumption in the local markets in the northern parts of Western Ghats, India. Scanning electron micrograph and genetic identification employing mitochondrial cytochrome oxidase subunit II, identified the nematode to the genus Contracaecum. Histology of infected host revealed the presence of the parasite in muscles. Antioxidant enzyme analysis of host liver suggested that infection leads to oxidative stress in the fish. We suspect that a gradual increase in parasite infection of the loaches in the last decade could be attributed to various anthropogenic stressors that are altering riverine habitats. Since loaches are consumed by tribal people who often prepare the fish without degutting and possibly undercooked, there is a potential threat of human infection.

A Fish of Multiple Faces, Which Show Us Enigmatic and Incredible Phenomena in Nature: Biology and Cytogenetics of the Genus Carassius.

Sexual vs. asexual reproduction-unisexual vs. bisexual populations-diploid vs. polyploid biotypes-genetic vs. environmental sex determination: all these natural phenomena are associated with the genus of teleost fish, Carassius. This review places emphasis on two Carassius entities with completely different biological characteristics: one globally widespread and invasive Carassius gibelio, and the other C. carassius with a decreasing trend of natural occurrence. Comprehensive biological and cytogenetic knowledge of both entities, including the physical interactions between them, can help to balance the advantages of highly invasive and disadvantages of threatened species. For example, the benefits of a wide-ranged colonization can lead to the extinction of native species or be compensated by parasitic enemies and lead to equilibrium. This review emphasizes the comprehensive biology and cytogenetic knowledge and the importance of the Carassius genus as one of the most useful experimental vertebrate models for evolutionary biology and genetics. Secondly, the review points out that effective molecular cytogenetics should be used for the identification of various species, ploidy levels, and hybrids. The proposed investigation of these hallmark characteristics in Carassius may be applied in conservation efforts to sustain threatened populations in their native ranges. Furthermore, the review focuses on the consequences of the co-occurrence of native and non-native species and outlines future perspectives of Carassius research.

Micro-contaminant, but immense impact: Source and influence of diethyl phthalate plasticizer on bottom-dwelling fishes.

Sustainable plastic-waste management is becoming increasingly challenging as enormous loads of plastic debris regularly accumulate in susceptible ecosystems. The microplastic (MP) particles generated from these plastic wastes are imposing additional threats to these ecosystems due to their small size as well as their ability to adsorb and carry toxic chemicals. The current investigation deals with one such MP-originated toxicant, diethyl phthalate (DEP), and its impact on two species of freshwater loaches from the Western Ghats of India, Lepidocephalichthys thermalis and Indoreonectes evezardi. The MP samples were collected from the sediments of the Mula River and characterized using spectroscopic methods and scanning electron microscopy. Polymers, such as polyvinyl chloride and polypropylene, were identified in the collected MPs. GC-MS analysis of the MP extracts revealed the presence of DEP, confirming the MP waste as a potential source of DEP pollution. Further, to evaluate the effect of DEP on survival of selected loaches, L. thermalis and I. evezardi were exposed to DEP concentrations (18.75-300 mg L-1) and the lethal DEP dose (LC50) was estimated to be 44.53 mg L-1 for L. thermalis and 34.64 mg L-1 for I. evezardi. Fishes were further exposed to sub-lethal DEP concentration for one day (Short term exposure: STE) or eight days (Long term exposure: LTE) to analyze the histological condition and oxidative status of the liver in response to DEP treatment. Histology revealed congestion of sinusoids and vacuolization after the LTE. Higher lipid peroxidation levels were also measured in the livers of both species treated with DEP, which indicated DEP-mediated oxidative damage. The antioxidant enzymes including superoxide dismutase, catalase, peroxidase and glutathione-S-transferase displayed elevated activities in response to STE and LTE of DEP. Collectively, the results demonstrate that MPs in the Mula River are a potential source of DEP. The findings also show that DEP exposure can be fatal to freshwater fishes such as loaches, possibly by causing increased oxidative damage to the hepatic system.

Equilibrated evolution of the mixed auto-/allopolyploid haplotype-resolved genome of the invasive hexaploid Prussian carp.

Understanding genome evolution of polyploids requires dissection of their often highly similar subgenomes and haplotypes. Polyploid animal genome assemblies so far restricted homologous chromosomes to a 'collapsed' representation. Here, we sequenced the genome of the asexual Prussian carp, which is a close relative of the goldfish, and present a haplotype-resolved chromosome-scale assembly of a hexaploid animal. Genome-wide comparisons of the 150 chromosomes with those of two ancestral diploid cyprinids and the allotetraploid goldfish and common carp revealed the genomic structure, phylogeny and genome duplication history of its genome. It consists of 25 syntenic, homeologous chromosome groups and evolved by a recent autoploid addition to an allotetraploid ancestor. We show that de-polyploidization of the alloploid subgenomes on the individual gene level occurred in an equilibrated fashion. Analysis of the highly conserved actinopterygian gene set uncovered a subgenome dominance in duplicate gene loss of one ancestral chromosome set.

Big eyes can't see microplastics: Feeding selectivity and eco-morphological adaptations in oral cavity affect microplastic uptake in mud-dwelling amphibious mudskipper fish.

Microplastic contamination is a widespread global problem. Plastic pollution in the oceans has received a lot of news coverage, but there is a significant gap in our knowledge about its effect in estuarine areas and a profound regional bias in available information. Here, we estimated the degree of microplastic pollution, its impact on a selected fish, and its function as a vector for heavy metals in the Ulhas River estuary, which is one of the most fragile, polluted, and anthropogenically impacted estuaries in India. Using mudskipper fish, we have also assessed how the feeding guild and ecomorphological adaptations in the feeding apparatus affected the microplastic intake and life history traits of the fish. Sediment, water and fish samples were collected from three sampling localities (S1, S2 and S3) in the Ulhas River estuary and analysed. Findings showed an increase in microplastic abundance from S1 (suburban) to S3 (urban industrial belt) in sediment (96.67-130.0 particles kg-1), water (0.28-0.41 particles L-1) and fish (3.75-6.11 particles per fish). Fragments, followed by pellets and filaments largely contribute to the plastic morphotypes in sediment and water. FTIR analysis revealed polymers of anthropogenic and industrial origin such as polypropylene, Surlyn ionomer, low-density polyethylene, and polyethylene or polybutylene terephthalate. Only filaments were found in the guts of 74% of the mudskippers examined, which may be due to their filter-feeding habit and unique anatomical arrangement of oral structures that effectively filter large microplastic particles. Microplastic abundance showed a strong negative correlation with condition factor, fullness index and hepatosomatic index of fish. SEM-EDS analysis revealed that the microplastic surface topography played an important part in adsorbing heavy metals from a water body containing these contaminants. Results highlight the contamination of vulnerable estuarine habitats, harmful effects on resident biota, and health threats to dependent populations.

Expanding conservation culturomics and iEcology from terrestrial to aquatic realms.

The ongoing digital revolution in the age of big data is opening new research opportunities. Culturomics and iEcology, two emerging research areas based on the analysis of online data resources, can provide novel scientific insights and inform conservation and management efforts. To date, culturomics and iEcology have been applied primarily in the terrestrial realm. Here, we advocate for expanding such applications to the aquatic realm by providing a brief overview of these new approaches and outlining key areas in which culturomics and iEcology are likely to have the highest impact, including the management of protected areas; fisheries; flagship species identification; detection and distribution of threatened, rare, and alien species; assessment of ecosystem status and anthropogenic impacts; and social impact assessment. When deployed in the right context with awareness of potential biases, culturomics and iEcology are ripe for rapid development as low-cost research approaches based on data available from digital sources, with increasingly diverse applications for aquatic ecosystems.

Early ontogeny social deprivation modifies future agonistic behaviour in crayfish.

Social deprivation early in life affects further individual development and leads to irreversible behavioural alterations later in life. Although the syndrome is well-studied in vertebrates including humans, its presence in invertebrates has been described only in eusocial insects and cockroaches. Here we show the first evidence of social deprivation in subsocial decapod crustaceans, based on analysis of video-recorded agonistic encounters of juvenile red swamp crayfish (Procambarus clarkii, Girard). In comparison with maternally incubated juveniles, isolated crayfish had altered repertoires, numbers and frequency of agonistic interactions similar to those described in vertebrates. Our results support the view on the syndrome of social deprivation as a ubiquitous trait in species with developed maternal care across diverse taxa.

Morphologically indistinguishable hybrid Carassius female with 156 chromosomes: A threat for the threatened crucian carp, C. carassius, L.

The crucian carp Carassius carassius (Linnaeus, 1758), is native to many European freshwaters. Despite its wide distribution, the crucian carp is declining in both the number and sizes of populations across much of its range. Here we studied 30 individuals of a putative pure population from Helsinki, Finland. Despite clear external morphological features of C. carassius, an individual was of a higher ploidy level than the others. We therefore applied a set of molecular genetic (S7 nuclear and cytochrome b mitochondrial genes) and cytogenetic tools (sequential fluorescent 4', 6-diamidino-2-phenylindole [DAPI], Chromomycin A3 [CMA3], C-banding and in situ hybridization [FISH] with both 5S and 28S ribosomal DNA probes) to determine its origin. While all examined characteristics of a diploid representative male (CCAHe2Fi) clearly corresponded to those of C. carassius, a triploid individual (CCAHe1Fi) was more complex. Phylogenetic analysis revealed that the nuclear genome of CCAHe1Fi contained three haploid sets: two C. gibelio and one C. carassius. However the mitochondrial DNA was that of C. gibelio, demonstrating its hybrid origin. The FISH revealed three strong (more intensive) 5S rDNA loci, confirming the triploid status, and an additional 24 weak (less intensive) signals were observed in the chromosome complement of CCAHe1Fi. On the other hand, only two strong and 16 weak 5S rDNA signals were visible on the chromosomes of the CCAHe2Fi male. 28S rDNA FISH revealed four strong signals in both CCAHe1Fi and CCAHe2Fi individuals. CMA3 staining revealed four to six CMA3-positive bands of CCAHe1Fi, while that of diploids contained only two to four. The fact that a polyploid hybrid Carassius female with a strong invasive potential may share morphological characters typical for endangered C. carassius highlights a need to combine genetic investigations of Carassius cryptic diversity with conservation measures of C. carassius in Europe.

Potential pest transfer mediated by international ornamental plant trade.

In recent years, the keeping of ornamental freshwater animals and plants in garden ponds has been growing in popularity. Water hyacinth (Eichhornia crassipes) is one of the preferred macrophytes seasonally imported mainly from South-eastern Asia throughout the world. This constitutes a secondary introduction inasmuch as the species is native to South America. Although many assemblages of aquatic invertebrates have been described as associated with this plant in the wild, there has been no research focused on their potential introduction via the international plant trade. We examined 216 specimens of water hyacinths imported for ornamental purposes from Indonesia into the Czech Republic. Numerous meio- and macroinvertebrates belonging to at least 39 species were captured. On the total number of individuals, the highest prevalence was of Tubulinea and Rotifera. Most of these were still alive and vital, including a caterpillar of the Indo-Australian invasive moth Spodoptera litura. Water hyacinths are usually placed into outdoor ponds immediately after import, which facilitates the release of non-target alien species. The present paper aims to draw attention to "hitchhikers" associated with the ornamental trade.

Comparative cytogenetics of Neotropical cichlid fishes (Nannacara, Ivanacara and Cleithracara) indicates evolutionary reduction of diploid chromosome numbers.

A comparative cytogenetic analysis was carried out in five species of a monophyletic clade of neotropical Cichlasomatine cichlids, namely Cleithracara maronii Steindachner, 1881, Ivanacara adoketa (Kullander & Prada-Pedreros, 1993), Nannacara anomala Regan, 1905, N. aureocephalus Allgayer, 1983 and N. taenia Regan, 1912. Karyotypes and other chromosomal characteristics were revealed by CDD banding and mapped onto the phylogenetic hypothesis based on molecular analyses of four genes, namely cyt b, 16S rRNA, S7 and RAG1. The diploid numbers of chromosomes ranged from 44 to 50, karyotypes were composed predominantly of monoarmed chromosomes and one to three pairs of CMA3 signal were observed. The results showed evolutionary reduction in this monophyletic clade and the cytogenetic mechanisms (fissions/fusions) were hypothesized and discussed.

Importance of fish gender as a factor in environmental monitoring of mercury.

Total mercury concentrations were determined in the gonadal tissues of 15 female and 10 male European perch (Perca fluviatilis) from one location of the stream "Jevanský potok" located about 30 km from Prague (Czech Republic). Tissue samples were frozen at -26 °C in polypropylene containers until further processing, which was carried out using an Advance Mercury Analyser (single purpose atomic absorption spectrometer). Mercury concentrations were present in all analysed gonad samples, and ranged from 2.3 to 12.7 µg/kg wet weight. However, we determined a mean Hg concentration (9.45 µg/kg) in male gonads that was 2.4 times greater than that of female gonads (3.9 µg/kg). This is an important finding when taking into account fish sex in environmental pollution monitoring (especially for mercury contamination).

Karyotype and chromosome banding of endangered crucian carp, Carassius carassius (Linnaeus, 1758) (Teleostei, Cyprinidae).

The karyotype and other chromosomal characteristics the crucian carp (Carassius carassius (Linnaeus, 1758)) were revealed by means of conventional banding protocols (C, CMA3, AgNOR). The diploid chromosome number (2n) in this species was 100. Its karyotype was composed of 10 pairs of metacentric, 18 pairs of submetacentric and 22 pairs of subtelo- to acrocentric chromosomes without any microchromosomes. C-banding identified blocks of telomeric heterochromatin on seven chromosome pairs. The NORs were situated on the p arms of the 14(th) pair of submetacentric chromosomes and on the p arms of the 32(nd) pair of subtelo-acrocentric chromosomes; AgNOR-positive signals corresponded to the CMA3-positive signals. These chromosome characteristics may suggest a paleo-allotetraploid origin of Carassius carassius genome.

Cichlid fishes in the Angolan headwaters region: molecular evidence of the ichthyofaunal contact between the Cuanza and Okavango-Zambezi systems.

The headwaters of five large African river basins flow through the Bié Plateau in Angola and still remain faunistically largely unexplored. We investigated fish fauna from the Cuanza and Okavango-Zambezi river systems from central Angola. We reconstructed molecular phylogenies of the most common cichlid species from the region, Tilapia sparrmanii and Serranochromis macrocephalus, using both mitochondrial and nuclear markers. We found evidence for ichthyofaunal contact and gene flow between the Cuanza and Okavango-Zambezi watersheds in the Bié Plateau in central Angola. Waterfalls and rapids also appeared to restrict genetic exchange among populations within the Cuanza basin. Further, we found that the Angolan Serranochromis cichlid fishes represent a monophyletic lineage with respect to other haplochromines, including the serranochromines from the Congo and Zambezi rivers. This study represents an important initial step in a biodiversity survey of this extremely poorly explored region, as well as giving further understanding to species distributions and gene flow both between and within river basins.

Effect of Acanthocephalus lucii infection on total mercury concentrations in muscle and gonads of fish host (Perca fluviatilis).

The samples from 13 perches (Perca fluviatilis) - muscle with skin and bones; fish gonads; and acanthocephalan parasites were analysed for mercury (Hg). Hg concentrations were present in all analysed samples. There were found no statistically significant difference in Hg concentration in fish tissues between perches either with or without infection by the acanthocephalan parasite, Acanthocephalus lucii). In this study there was no evidence that acanthocephalan worms accumulate mercury from hosts. For this reason, A. lucii is not a suitable bioindicator for mercury pollution.

Concentrations of Zn, Mn, Cu and Cd in different tissues of perch (Perca fluviatilis) and in perch intestinal parasite (Acanthocephalus lucii) from the stream near Prague (Czech Republic).

We monitored concentrations of Cd, Cu, Mn and Zn in acantocephalan parasites (Acanthocephalus lucii) and its final host (Perca fluviatilis). The concentrations in parasites were found to be significantly higher than those found in the muscle, gonads and liver of fish host. The bioaccumulation factor values (BF=C(parasite)/C(host muscle)) were 194, 24.4, 2.2 and 4.7 for Cd, Cu, Mn and Zn, respectively. This suggests a benefit for the host due to the high accumulation of toxic cadmium.

Intestinal parasite Acanthocephalus lucii (Acanthocephala) from European perch (Perca fluviatilis) as a bioindicator for lead pollution in the stream "Jevanský potok" near Prague, Czech Republic.

Lead concentrations in the tissues of perch and its parasites were determined as mg/kg dw. Lead was found at higher concentrations in the acanthocephalans (11.56) than in different tissues (liver, gonads and muscle with skin and bone) of perch. With respect to fish tissues, the highest concentrations of lead were present in the liver (1.24), followed by the gonads (0.57) whereas the lowest concentrations were in the muscle with skin and bone (0.21). The bioconcentration factors for lead indicated that parasites accumulate metals to a higher degree than fish tissues--lead concentrations in acanthocephalans were 9.32, 19.27 and 55.05 higher than in liver, gonads and muscles of host, respectively.