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.

First report on microplastics contamination in a meteorite impact Crater Lake from India.

Microplastic pollution is a worldwide concern affecting all environmental matrices, including pristine lakes. Lentic lakes operate as a sink for microplastics (MPs), which interfere with a biogeochemical cycle and, therefore, deserve immediate attention. We present a comprehensive assessment of MPs contamination in the sediment and surface water of a geo-heritage site, Lonar lake (India). It is the third largest natural saltwater lake and only basaltic crater in the world formed by meteoric impact around 52,000 years ago. Mean MPs abundance in lakeshore sediment and surface water was 14.44 particles/kg and 2.66 particles/L, respectively. Small-sized MPs dominate the hypersaline region of the lake. Transparent and green fragments and filaments morphotypes were abundant. Most of the MPs in Lonar lake were secondary in origin. FTIR-ATR analysis revealed 16 types of polymers in the lake, of which polypropylene, polyvinyl chloride, polyethylene, high-density polyethylene, low-density polyethylene, polystyrene, and polyester were the most common. The overall pollution load index (PLI) for Lonar lake sediment and water was 1.39 and 2.58, respectively. Although all sampling stations had significant MPs pollution (PLI > 1), there was noticeable station-specific variability, which could be linked to anthropogenic activities. Irresponsible tourist behavior and religious activities, coupled with poor waste management are the leading causes of MPs contamination in the lake. The current work fills a gap in the investigation of MP pollution in a crater lake formed by a meteorite impact by being the first to provide a precise estimate of the MPs contamination in the Lonar lake.

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.

Morpho-histochemical adaptations of the digestive tract in Gangetic mud-eel Ophichthys cuchia (Hamilton 1822) support utilization of mud-dwelling prey.

Freshwater mud-eel, Ophichthys cuchia is nocturnal, carnivorous and economically important fish, yet its digestive physiology is unknown. We therefore studied the gastrointestinal (GI) tract of O. cuchia using morphological, osteological, histological and histochemical approach to understand how the structural adequacy of GI tract helps in acquisition of bottom and mud-dwelling prey and supports utilization of uncommon food resources. Morphologically the GI tract showed typical features of carnivorous fishes in the form of sub-terminal mouth, short muscular esophagus, expendable stomach, short intestine and rectum. Osteological investigation clearly showed that the specialized arrangement of teeth in the oral cavity and pharyngeal region helps in digging and manipulation of bottom and mud-dwelling prey. Longitudinal mucosal folds, stratified squamous epithelium with numerous goblet cells of esophagus protect mucosa from mechanical harm and also allow easy transit of prey into the stomach. Large-sized rugae of stomach are complemented with numerous gastric glands which together increase the transit time of carnivorous food stuff and ensure its complete digestion. The pylorus specifically allows only digested and finely ground food stuff into the intestine and thus avoids the intestinal blockage due to entry of undigested food. The enterocytes and goblet cells of intestine are absorptive and lubricative in their functions respectively. A significant increase in the number of goblet cells and thickness of muscularis from the intestine to the rectum was evident which facilitates the easier transit of food, protection for the epithelium from abrasion, and helps in defecation. Histochemistry revealed that the mucins are secreted throughout the GI tract and thus catalyze faster digestion of carnivorous food stuff, providing protection to mucosal membrane from abrasion, acts as a co-factor to support digestion, absorption of proteins, ions, fluids and helps in defecation. The outcome of this study clearly supports the notion that structural adaptations in the digestive tract of fishes can be effectively used as a blueprint to understand why and how particular fish species feed and use unique food. Additionally, the structural and functional adequacy of the digestive tract helps the fishes to acquire and utilize novel habitats and food resources. The results presented in this study will serve as a reference point for future studies, which focus primarily on understanding the evolution of carnivory in Synbranchids.

Act now before its too late: Copper exposure drives chemo-ecology of predator-prey dynamics of freshwater common spiny loach, Lepidocephalichthys thermalis (Valenciennes, 1846).

Due to the extensive use of copper (Cu) in various commercial products, its existence in aquatic bodies (freshwater and marine) is not unusual. Cu is well known for its effect on the olfactory physiology of fish. However, there are limited studies on the effect of Cu on important ecological functions in fish (predator-prey dynamics) that are primarily influenced by olfaction. In a series of experiments, we studied the effect of Cu exposure on the chemoreceptive behavior of the prey fish, Lepidocephalichthys thermalis. Prey fishes were exposed to an environmentally relevant concentration (5 µg/L) of Cu for 3 h and the anti-predator responses against native (Channa gachua) and alien predatory fish (tilapia) were quantified using an ethological assay. Cu exposed prey fishes did not recognize the native predator and had a lower survival rate than control (unexposed) fishes in predation trials. Cu exposed prey fishes have failed to learn associatively to detect a non-native predator resulting in higher mortality in prey population in direct encounters with tilapia. However, such a lack of predator recognition was found to be short-term and the treated prey fishes recovered anti-predator responses within 72 h. In addition, Cu inactivated the alarm cue which acts as a signal for the presence of predators and ensures associative learning and therefore it was considered to be an 'info-disruptor' in the present study. These outcomes together demonstrate that even at low concentration, Cu influences ecological decisions and survival against predators. Owing to the ubiquitous occurrence of Cu in water bodies, the present investigation will contribute to the knowledge of how environmental stressors alter the crucial ecological decisions of prey individuals in aquatic ecosystems. In addition, we suggest that freshwater reservoirs containing high levels of Cu could be unsuitable for the long-term survival of prey fishes and freshwater biodiversity.

Structural adequacy of the digestive tract supports dual feeding habit in catfish Pachypterus khavalchor (Siluriformes: Horabagridae).

Lepidophagy is comparatively rare amongst teleost fishes, yet our understanding of this specialization is lacking. Therefore we examined the digestive tract features of Pachypterus khavalchor using morphological, osteological, histological and histochemical techniques to comprehend and relate structural organization of digestive tract with scale eating habit. Morphologically, the alimentary canal is defined by a short and muscular esophagus, well-developed stomach and comparatively short intestine. Gut content analysis and intestinal coefficient value (0.53 ± 0.01) revealed that P. khavalchor exhibit both carnivory and lepidophagy. However, P. khavalchor primarily feeds on the scales (67.47%) and other chitin-rich material like aquatic insects (17.62%), aquatic larvae (8.66%) which affirms its solid association with chitinase producing endosymbionts in the gut. Lepidophagy is further supported by the osteological observations. The perfect segregation of the functions such as food capture, ingestion and processing amongst the different types of teeth located in the oral cavity and pharyngeal region thus could be taken as evolutionary adaptations in scale eaters to support lepidophagy. Specialized arrangement of the esophageal and stomach epithelial folds could be altogether taken as an adaptation with the end goal to frame the scale stacks and accordingly facilitate the handling and processing of chitin-rich bolus. The esophageal mucosa is simple squamous epithelium instead of stratified epithelium with numerous goblet cells to withstand the mechanical harm by hard-food stuff like scales. The cardiac and fundic regions exhibited large number tubular gastric glands with simple columnar epithelium. Surface cells of all three stomach regions stained positive for PAS staining. The intestine is without pyloric caeca and is divided into anterior and posterior region. Histologically it is characterized by simple columnar epithelium with brush border and numerous goblet cells throughout its length. Presence of large number microvilli on anterior and posterior intestine was noticeable. Intestinal goblet cells reacted positively to PAS, AB (pH 1) and AB (pH 2.5). Secretions of goblet cells are important for lubricating and protecting the epithelium. The results of present investigation improve the understanding of the digestive physiology of scale eaters in general and P. khavalchor in particular. Overall, our data indicates that though P. khavalchor predominantly feeds on scale, the digestive physiology is adapted to support dual feeding habit (lepidophagy and carnivory).

Impact, recovery and carryover effect of Roundup® on predator recognition in common spiny loach, Lepidocephalichthys thermalis.

Understanding the negative impact of a variety of environmental contaminants on aquatic animals is essential to curb biodiversity loss and stop degradation of ecological functions. Excessive and unrestricted use of pesticides is the most serious threat to aquatic animals including amphibians and fishes. Among the known pesticides, glyphosate based formulations have been shown to have lethal effects on many aquatic organisms. However, negative effects of pesticides on crucial ecological interactions such as prey-predator interactions are relatively unknown from tropics. In many aquatic organisms, recognition of predators is based on odor signatures; and therefore any anthropogenic alteration in water chemistry has the potential to impair recognition and learning of predators. Through a series of behavioral experiments we evaluated the effect of glyphosate based herbicide (Roundup®) on the antipredator behavior of common spiny loach, Lepidocephalichthys thermalis to understand the effects of pesticide-exposure on recognition of conspecific alarm cues, and associative learning to avoid predation. We exposed common spiny loach (for 3 h or 15 days) to sub-lethal concentration (0.5 mg a.e./L) of Roundup® and subsequently with conspecific alarm cues, signaling the proximity of a predator. Unexposed prey fish showed a significant reduction in activity level in response to conspecific alarm cues. Whereas such alarm response was not observed in prey fish that were exposed to Roundup® either for 3 h or 15 days. Such lack of response could be associated with alteration of olfactory function in prey individuals. However, this inability to detect the conspecific alarm cues was found to be transient and exposed fish recovered within 2 days. In subsequent experiments, we showed that Roundup® deactivates the conspecific alarm cues thus making them unavailable for prey to evoke the response. Furthermore, Roundup® mediated degradation of conspecific alarm cues and diminished the associative learning necessary for detection of the invasive/unknown/novel predators. Overall, due to the worldwide occurrence of glyphosate in water bodies, glyphosate mediated behavioral suppression exposes the prey animals to a considerable risk of predation, both by native and non-native predators.

Interplay between behavior, morphology and physiology supports lepidophagy in the catfish Pachypterus khavalchor (Siluriformes: Horabagridae).

The present study demonstrates the scale-eating behavior of a siluroid catfish, Pachypterus khavalchor, and the role of its oral structure and bacterial endosymbionts in shaping this lepidophagous habit. Scale-eating behavior in P. khavalchor was studied using a series of behavioral experiments. P. khavalchor was found to feed only on the scales of live fish and never of dead fish, even after 72h of starvation. It was nocturnal in habit and attacked all species that were used as prey. Attacking behavior showed extensive chasing of prey species followed by a powerful random strike at the flank or close to the caudal region in posterior oblique position. After a strike, P. khavalchor was found to immediately turn back and pick up the falling dislodged scales. SEM analysis of oral structures of P. khavalchor revealed three different types of teeth arranged in specific order on the upper jaw, lower jaw and pharyngeal region that could facilitate their lepidophagous habit. Teeth of upper and lower jaws are likely to help in dislodging the scales and pharyngeal teeth may help to engulf the scales. Gut microflora analysis and enzyme assay revealed two isolates, namely Bacillus pumilus and Bacillus licheniformis, which were positive for chitinase production. These two isolates were found to be capable of producing chitinase indicating that they are likely to be involved in the digestion of chitin-rich scales in the host fish gut. An in vitro scale degradation assay further strengthens the results since both isolates were found to be efficient in chitinase production and degradation of scales.

Determining sensitive stages for learning to detect predators in larval bronzed frogs: importance of alarm cues in learning.

Successful survival and reproduction of prey organisms depend on their ability to detect their potential predators accurately and respond effectively with suitable defences. Predator detection can be innate or can be acquired through learning.We studied prey-predator interactions in the larval bronzed frogs (Sylvirana temporalis), which have the innate ability to detect certain predators. We conducted a series of experiments to determine if the larval S. temporalis rely solely on innate predator detection mechanisms or can also learn to use more specific cues such as conspecific alarm cues for the purpose. The results of our study clearly indicate that larval S. temporalis use both innate and learned mechanisms for predator detection. Predator-naive tadpoles could detect kairomones alone as a potential threat and responded by reducing activity, suggesting an innate predator detection mechanism. Surprisingly, predator-naive tadpoles failed to detect conspecific alarm cues as a potential threat, but learned to do so through experience. After acquiring the ability to detect conspecific alarm cues, they could associate novel predator cues with conspecific alarm cues. Further, post feeding stages of larval S. temporalis are sensitive for learning to detect conspecific alarm cues to label novel predators.

Occurrence of parotoid glands in tadpoles of the tropical frog, Clinotarsus curtipes and their role in predator deterrence.

Tadpoles of the tropical bicolored frog, Clinotarsus curtipes are unique in having parotoid glands secreting a white viscous fluid and are structurally similar to granular glands from other amphibians. To ascertain the involvement of these glands and their secretion in predator deterrence, it was tested against a predatory fish, Clarias gariepinus, using a paired choice behavioral assay. The results showed that the fish avoid eating C. curtipes tadpoles when paired with tadpoles of a sympatric species, Sylvirana temporalis. While the fish fed on C. curtipes tadpoles whose parotoid glands were surgically removed, did not touch those with intact glands, suggesting a role for the parotoid gland secretion in predator deterrence. Histochemical and biochemical analyses of the gland secretion revealed the presence of high concentrations of proteins, lipids, and alkaloids. SDS-PAGE showed the presence of proteins with prominent bands at 17 and 50kDa. The presence of other small molecules (950-2000amu) as detected by LC-MS showed the presence of five major peaks. Peaks 1 and 2 are probably tetrodotoxin and/or its analogs. Peaks 3 and 5 are possibly bufalin and argininosuccinic acid, respectively while peak 4 remains unidentified. Thus, secretion of parotoid glands of larval C. curtipes contains chemicals which, either alone or in combination, might be responsible for deterring predators.