Photoaged polystyrene microplastics result in neurotoxicity associated with neurotransmission and neurodevelopment in zebrafish larvae (Danio rerio).

Microplastics (MPs) are emerging pollutants widely distributed in the environment, inducing toxic effects in various organisms. However, the neurotoxicity and underlying mechanisms of simulated sunlight-aged MPs have rarely been investigated. In this study, zebrafish (Danio rerio) were exposed to environmentally relevant concentrations (0, 0.1, 1, 10, and 100 µg/L) of virgin polystyrene (V-PS) and aged polystyrene (A-PS) for 120 hpf to evaluate the neurotoxicity. The results demonstrated that simulated sunlight irradiation altered the physicochemical properties (morphology, functional groups, and chemical composition) of V-PS. Exposure to A-PS causes greater toxicity on locomotor ability in larval zebrafish than V-PS. Motor neuron development was disrupted by transgenic (hb9-GFP) zebrafish larvae exposed to A-PS, with significant alterations in neurotransmitter levels (ACh, DA, 5-HT, and GABA) and enzyme activity (AChE, ChAT, and ChE). Further investigation found that exposure to A-PS had a significantly impact on the expression of neurotransmission and neurodevelopment-related genes in zebrafish. These findings suggest that A-PS induces neurotoxicity by its effects on neurotransmission and neurodevelopment. This study highlights the neurotoxic effects and mechanisms of simulated sunlight irradiation of MPs, providing new insights for assessing the ecological risks of photoaged MPs in the environment.

A perspective on the impacts of microplastics on mosquito biology and their vectorial capacity.

Microplastics (plastic particles <5 mm) permeate aquatic and terrestrial ecosystems and constitute a hazard to animal life. Although much research has been conducted on the effects of microplastics on marine and benthic organisms, less consideration has been given to insects, especially those adapted to urban environments. Here, we provide a perspective on the potential consequences of exposure to microplastics within typical larval habitat on mosquito biology. Mosquitoes represent an ideal organism in which to explore the biological effects of microplastics on terrestrial insects, not least because of their importance as an infectious disease vector. Drawing on evidence from other organisms and knowledge of the mosquito life cycle, we summarise some of the more plausible impacts of microplastics including physiological, ecotoxicological and immunological responses. We conclude that although there remains little experimental evidence demonstrating any adverse effect on mosquito biology or pathogen transmission, significant knowledge gaps remain, and there is now a need to quantify the effects that microplastic pollution could have on such an important disease vector.

Early ontogenesis from embryo to juvenile in Senegalese sole Solea senegalensis under laboratory conditions.

Senegalese sole, Solea senegalensis, is a flatfish of high commercial value in the world. It has been identified as an interesting and promising species for marine commercial aquaculture diversification in Europe for at least four decades and was introduced to China in 2003. Early ontogenesis from embryo to juvenile stages in S. senegalensis was analysed under controlled laboratory conditions to provide morphological information for aquaculture. From 0 to 59 days post hatching (dph), 10-20 larvae were sampled and measured each day (0-17 dph) or every 2-6 days (17-59 dph). Morphological characteristics from the egg to the juvenile stage were described. The eggs were separate and spherical with multiple oil globules. After 3 dph, the yolk sac was completely absorbed, mouth and anus were open, a swim bladder appeared, and larvae began feeding on rotifers (Brachionus plicatilis). The larvae began metamorphosis as the notochord flexed upward and the left eye migrated upward after 10 dph. The left eye migrated to the dorsal midline at 15 dph. At 19 dph, the left eye was translocated to the right-ocular side, and the juveniles adopted a benthic lifestyle. The swim bladder degenerated, and the juveniles completed metamorphosis at 23 dph. The growth patterns of some parameters (TL, SL, BH, BW) during larval and juvenile development stages were identified. The inflection points, which are slopes of growth changes, were calculated in growth curves. Three inflection points occurring in the growth curves of larvae and juveniles were found to be associated with metamorphosis, weaning, and transitions in feeding habits. The basic information of embryo development and ontogenesis in this study represents a valuable contribution to the S. senegalensis industry, especially in artificial breeding and rearing techniques.

Behavioral toxicological tracking analysis of Drosophila larvae exposed to polystyrene microplastics based on machine learning.

Microplastics, as a pivotal concern within plastic pollution, have sparked widespread apprehension due to their ubiquitous presence. Recent research indicates that these minuscule plastic particles may exert discernible effects on the locomotor capabilities and behavior of insect larvae. This study focuses on the impact of polystyrene microplastics (PS-MPs) on the behavior of Drosophila melanogaster larvae, utilizing fruit flies as a model organism. Kinematic analysis methods were employed to assess and extrapolate the toxic effects of PS-MPs on the larvae. Drosophila larvae were exposed to varying concentrations (Control, 0.1 g/L, 1 g/L, 10 g/L, 20 g/L) of 5 µm PS-MPs during their developmental stages. The study involved calculating and evaluating parameters such as the proportion of larvae reaching the edge, distance covered, velocity, and angular velocity within a 5-min timeframe. Across different concentrations, Drosophila larvae exhibit differential degrees of impaired motor function and disrupted locomotor orientation. The proportion of larvae reaching the edge decreased, velocity significantly declined, and angular velocity exhibited a notable increase. These findings strongly suggest that when exposed to a PS-MPs environment, Drosophila larvae exhibit slower movement, increased angular rotation per unit time, leading to a reduction in the proportion of larvae reaching the edge. The altered behavior of Drosophila larvae implies potential damage of microplastics on insect larvae development and activity, consequently impacting the ecosystem and prompting heightened scrutiny regarding microplastics.

Mining tailings alter insects: revealing fluctuating asymmetry in the caddisfly Smicridea coronata.

The Samarco/Vale/BHP mine tailing dam breach that took place in Minas Gerais, southeastern Brazil, in 2015, deposited high concentrations of metals and metalloids in the Rio Doce basin, severely impacting freshwater and riverine forest ecosystems. To assess developmental instability of caddisflies in response to the environmental impacts of the dam breach, we investigated the fluctuating asymmetry (FA) in the species Smicridea (Rhyacophylax) coronata (Trichoptera: Hydropsychidae). FA was assessed at individual and populational scales using geometric morphometric methods in the cephalic capsule and mandibles of larvae and also on the forewings of adults, both collected under the impacted condition, and under the least disturbed condition. The levels of FA increased in response to stressors on the forewings at the populational scale, and on the mandibles, at individual scale. These morphological variations in the larval and adult stages may lead to detrimental effects and result in high mortality rates as well as lower adult fitness. Trichoptera forewings are revealed as suitable traits for assessing FA, holding potential for applications in biomonitoring programs. Directional asymmetry levels were higher than FA levels for all traits, and this correlation could be explained by a transition from fluctuating to directional asymmetry in the presence of heightened disturbance. Our results validate the relationship between the impacts from the dam breach and increased developmental instability in this species with likely cascade effects on the insect community.

Frog hatchlings use early environmental cues to produce an anticipatory resource-use phenotype.

Developmental plasticity can occur at any life stage, but plasticity that acts early in development may give individuals a competitive edge later in life. Here, we asked if early (pre-feeding) exposure to a nutrient-rich resource impacts hatchling morphology in Mexican spadefoot toad tadpoles, Spea multiplicata. A distinctive carnivore morph can be induced when tadpoles eat live fairy shrimp. We investigated whether cues from shrimp--detected before individuals are capable of feeding--alter hatchling morphology such that individuals could potentially take advantage of this nutritious resource once they begin feeding. We found that hatchlings with early developmental exposure to shrimp were larger and had larger jaw muscles--traits that, at later stages, increase a tadpole's competitive ability for shrimp. These results suggest that early developmental stages can assess and respond to environmental cues by producing resource-use phenotypes appropriate for future conditions. Such anticipatory plasticity may be an important but understudied form of developmental plasticity.

Pre-oral gut contributes to facial structures in non-teleost fishes.

Despite the wide variety of adaptive modifications in the oral and facial regions of vertebrates, their early oropharyngeal development is considered strictly uniform. It involves sequential formation of the mouth and pharyngeal pouches, with ectoderm outlining the outer surface and endoderm the inner surface, as a rule. At the extreme anterior domain of vertebrate embryos, the ectoderm and endoderm directly juxtapose and initial development of this earliest ecto-endoderm interface, the primary mouth, typically involves ectodermal stomodeal invagination that limits the anterior expansion of the foregut endoderm. Here we present evidence that in embryos of extant non-teleost fishes, oral (stomodeal) formation is preceded by the development of prominent pre-oral gut diverticula (POGD) between the forebrain and roof of the forming mouth. Micro-computed tomography (micro-CT) imaging of bichir, sturgeon and gar embryos revealed that foregut outpocketing at the pre-oral domain begins even before the sequential formation of pharyngeal pouches. The presence of foregut-derived cells in the front of the mouth was further confirmed by in vivo experiments that allowed specific tracing of the early endodermal lining. We show that POGD in sturgeons contribute to the orofacial surface of their larvae, comprising oral teeth, lips, and sensory barbels. To our knowledge, this is the first thorough evidence for endodermal origin of external craniofacial structures in any vertebrate. In bichir and gar embryos, POGD form prominent cranial adhesive organs that are characteristic of the ancient bauplan of free-living chordate larvae. POGD hence seem arguably to be ancestral for all ray-finned fishes, and their topology, pharyngeal-like morphogenesis and gene expression suggest that they are evolutionarily related to the foregut-derived diverticula of early chordate and hemichordate embryos. The formation of POGD might thus represent an ancestral developmental module with deep deuterostome origins.

Controlled decoration of nanoceria on the surface of MoS2nanoflowers to improve the biodegradability and biocompatibility inDrosophila melanogastermodel.

In recent years, nanozymes based on two-dimensional (2D) nanomaterials have been receiving great interest for cancer photothermal therapy. 2D materials decorated with nanoparticles (NPs) on their surface are advantageous over conventional NPs and 2D material based systems because of their ability to synergistically improve the unique properties of both NPs and 2D materials. In this work, we report a nanozyme based on flower-like MoS2nanoflakes (NFs) by decorating their flower petals with NCeO2using polyethylenimine (PEI) as a linker molecule. A detailed investigation on toxicity, biocompatibility and degradation behavior of fabricated nanozymes in wild-typeDrosophila melanogastermodel revealed that there were no significant effects on the larval size, morphology, larval length, breadth and no time delay in changing larvae to the third instar stage at 7-10 d for MoS2NFs before and after NCeO2decoration. The muscle contraction and locomotion behavior of third instar larvae exhibited high distance coverage for NCeO2decorated MoS2NFs when compared to bare MoS2NFs and control groups. Notably, the MoS2and NCeO2-PEI-MoS2NFs treated groups at 100µg ml-1covered a distance of 38.2 mm (19.4% increase when compared with control) and 49.88 mm (no change when compared with control), respectively. High-resolution transmission electron microscopy investigations on the new born fly gut showed that the NCeO2decoration improved the degradation rate of MoS2NFs. Hence, nanozymes reported here have huge potential in various fields ranging from biosensing, cancer therapy and theranostics to tissue engineering and the treatment of Alzheimer's disease and retinal therapy.

BMP controls dorsoventral and neural patterning in indirect-developing hemichordates providing insight into a possible origin of chordates.

A defining feature of chordates is the unique presence of a dorsal hollow neural tube that forms by internalization of the ectodermal neural plate specified via inhibition of BMP signaling during gastrulation. While BMP controls dorsoventral (DV) patterning across diverse bilaterians, the BMP-active side is ventral in chordates and dorsal in many other bilaterians. How this phylum-specific DV inversion occurs and whether it is coupled to the emergence of the dorsal neural plate are unknown. Here we explore these questions by investigating an indirect-developing enteropneust from the hemichordate phylum, which together with echinoderms form a sister group of the chordates. We found that in the hemichordate larva, BMP signaling is required for DV patterning and is sufficient to repress neurogenesis. We also found that transient overactivation of BMP signaling during gastrulation concomitantly blocked mouth formation and centralized the nervous system to the ventral ectoderm in both hemichordate and sea urchin larvae. Moreover, this mouthless, neurogenic ventral ectoderm displayed a medial-to-lateral organization similar to that of the chordate neural plate. Thus, indirect-developing deuterostomes use BMP signaling in DV and neural patterning, and an elevated BMP level during gastrulation drives pronounced morphological changes reminiscent of a DV inversion. These findings provide a mechanistic basis to support the hypothesis that an inverse chordate body plan emerged from an indirect-developing ancestor by tinkering with BMP signaling.

The biological regulation of sea urchin larval skeletogenesis - From genes to biomineralized tissue.

Biomineralization is the process in which soft organic tissues use minerals to produce shells, skeletons and teeth for various functions such as protection and physical support. The ability of the cells to control the time and place of crystal nucleation as well as crystal orientation and stiffness is far beyond the state-of-the art of human technologies. Thus, understanding the biological control of biomineralization will promote our understanding of embryo development as well as provide novel approaches for material engineering. Sea urchin larval skeletogenesis offers an excellent platform for functional analyses of both the molecular control system and mineral uptake and deposition. Here we describe the current understanding of the genetic, molecular and cellular processes that underlie sea urchin larval skeletogenesis. We portray the regulatory genes that define the specification of the skeletogenic cells and drive the various morphogenetic processes that occur in the skeletogenic lineage, including: epithelial to mesenchymal transition, cell migration, spicule cavity formation and mineral deposition into the spicule cavity. We describe recent characterizations of the size, motion and mineral concentration of the calcium-bearing vesicles in the skeletogenic cells. We review the distinct specification states within the skeletogenic lineage that drive localized skeletal growth at the tips of the spicules. Finally, we discuss the surprising similarity between the regulatory network and cellular processes that drive sea urchin skeletogenesis and those that control vertebrate vascularization. Overall, we illustrate the novel insights on the biological regulation and evolution of biomineralization, gained from studies of the sea urchin larval skeletogenesis.

A pair-rule function of odd-skipped in germband stages of Tribolium development.

While pair-rule patterning has been observed in most insects examined, the orthologs of Drosophila pair-rule genes have shown divergent roles in insect segmentation. In the beetle Tribolium castaneum, while odd-skipped (Tc-odd) was expressed as a series of pair-rule stripes, RNAi-mediated knockdown of Tc-odd (Tc-oddRNAi) resulted in severely truncated, almost asegmental phenotypes rather than the classical pair-rule phenotypes observed in germbands and larval cuticles. However, considering that most segments arise later in germband stages of Tribolium development, the roles of Tc-odd in segmentation of growing germbands could not be analyzed properly in the truncated Tc-oddRNAi germbands. Here, we investigated the segmentation function of Tc-odd in germband stages of Tribolium development by analyzing Tc-oddRNAi embryos that resumed germband extension. In the larval cuticles of Tc-oddRNAi embryos, normal mandibular and maxillary and loss of the labial segments were consistent in the head, whereas a broad range of segmentation defects including loss or fusion of thoracic and/or abdominal segments was observed in the trunk. Interestingly, a group of Tc-oddRNAi germbands showed pair-rule-like defects in the segmental stripes of the segment-polarity genes, engrailed, hedgehog, or wingless, in the abdominal regions. While the pair-rule genes even-skipped, runt, odd, and paired were misregulated in the growing Tc-oddRNAi germbands, paired expression required for odd-numbered segment formation was largely abolished, which might cause the pair-rule-like defects. Taken together, these findings suggest that Tc-odd can function as a pair-rule gene in the germband stages of Tribolium development.

Adverse biological effects of ingested polystyrene microplastics using Drosophila melanogaster as a model in vivo organism.

The abundant presence and extensive use of polystyrene microplastics (PSMPs) has recently become a serious environmental concern, as impact of exposure to these substances on human health remains unknown. While in vitro studies yield data on adverse effect of PSMPs, in vivo approaches are more relevant for risk assessment. Drosophila melanogaster is one of the most genetically and experimentally accessible model organisms used in biology as an in vivo model. D. melanogaster was selected as a representative in vivo model organism to examine the genotoxic potential of PSMPs at 5 concentrations of three different sizes namely 4, 10, or 20 µm. In particular, the wing somatic mutation and recombination test (SMART), a scalable, time-efficient in vivo assay developed to study genotoxicity of various compounds in a rapid manner at low costs was used. The third-instar Drosophila larvae were exposed to PSMPs through food at 5 concentrations ranging from 0.01-10 mM. Viability (lethality), larval length, morphological deformations, locomotor activity (climbing behavior), and genotoxic effects were the end-points measured. Exposure to PSMPs at 4, 10, or 20 µm produced significant morphological defects, impaired climbing behavior, and genotoxicity as evidenced by the SMART test demonstrating induction of somatic recombination. Significant increases were observed in the frequency of total spots, suggesting that PSMPs might induce genotoxic activity predominantly via initiation of somatic DNA recombination in a concentration-dependent manner.

Increasing intensities of Anisakis simplex (Rudolphi, 1809 det. Krabbe, 1878) larvae with weight and sea age in returning adult Atlantic salmon, Salmo salar L., of coastal waters of Norway.

Ninety wild Atlantic salmon, Salmo salar L., (1.5-10.3 kg) were caught in the Namsen Fjord near the mouth of River Namsen, mid-Norway, and examined for the presence and distribution of Anisakis simplex (Rudolphi, 1809 det. Krabbe, 1878) larvae by digestion of the viscera and muscles in a pepsin/HCl solution. All salmon were migrating spawners after 1-4 years of feeding in the Atlantic Ocean. All 90 Atlantic salmon had A. simplex larvae in the viscera, and all, except two, had A. simplex larvae in the musculature. The number of A. simplex larvae in each fish varied between 3 and 181, and the total mean number of nematode larvae was 44.5. The intensity of A. simplex larvae was positively correlated with increasing weight and sea age of the host. However, the proportion of larvae in the muscle fillets decreased with increasing host weight and sea age. Atlantic salmon females had more A. simplex larvae than males. In all the fish examined, 70.2% of the A. simplex larvae were found in the viscera and 29.8% in the musculature. The majority (93%) of the larvae in the musculature occurred in the hypaxial sections anterior to the anus. As A. simplex larvae commonly occur in the musculature of wild Atlantic salmon, consumption of unfrozen, raw or semi-raw musculature represents a risk for humans developing anisakiasis.

Onset of nutrient consumption during early life stage digestive system development of two tuna species (Thunnus orientalis and Thunnus albacares).

To specify the timing of exogenous nutrient consumption in the larvae of two commercially important tuna species, the Pacific bluefin tuna (PBF) Thunnus orientalis and the yellowfin tuna (YFT) Thunnus albacares, the gene expressions of peptide transporter 1 (PEPT1) were examined. The mRNA expressions of PEPT1 first occurred at 2 days post hatching (dph) in PBF larvae and 3 dph for the YFT, and PEPT1 was found to only be expressed in the intestinal tract. The histological changes of the digestive tract of the YFT larvae were observed and compared to PBF larvae from a previous study. The intestines were developed at the hatching day for both species. It was found that the developmental timing of internal organs differed between the species, with the YFT showing an approximately one-day delay. The major organs such as liver, pancreas and gall bladder that excrete digestive enzymes appeared at 1 dph for PBF and 2 dph for YFT. The development of external morphological features was similar to organ development timings, with mouth-opening and first feeding starting at 2 dph for PBF, and 3 dph for YFT. Growth during the first month is rapid and variable for both species, ranging from 1.06 to 1.56 mm/d. Our findings provide new information about the early onset of feeding and larval development for the two species which would contribute to future aquaculture.

Ontogeny and development of the gastrointestinal system in Indian walking catfish (Clarias magur) during its early development.

Ontogeny of the digestive tract and its accessory organs and their further development in the Indian walking catfish (Clarias magur) were examined in larvae, starting from the day of hatching until 35 days post-hatching (dph) reared at 28-29 °C. Many organs at their primordial stage were seen on the day of hatching. These include opened oral cavity with monolayered epithelial lining and very few newly emerging taste buds and goblet cells, primordial pharyngeal teeth on slightly stratified epithelia of the pharyngeal plate, stomach anlage with some degree of the mucosal fold, and a few newly forming gastric glands embedded under its mucosa, primordial anterior and posterior intestine with the smooth mucosal surface, anal opening, and primordial liver and pancreas. At 1 dph, the stomach appeared to be bilobed with the first evidence of food particle in it, and the intestine had some initial folding. On the day of hatching, goblet cells appeared in all lengths of the gut, but not densely, except in the stomach; on it, they appeared at 2 dph. Pancreatic zymogen granules also appeared on this day. Supranuclear vesicles first appeared on 4-5 dph (7.9 ± 0.5-8.6 ± 0.8 mm TL), and they continue to exist until 35 dph. The developmental sequence in this fish confirmed it as an altricial species with some major histomorphological events after the onset of feeding; these include-the appearance of fully developed-pharyngeal teeth at 4 dph and onwards, pyloric sphincter, anterior to posterior intestinal sphincter at 6 dph, and the continuous development of buccopharyngeal cavity and stomach in their shape, size, and functionality until the completion of metamorphosis. Overall, the information on gastrointestinal development in the early life stage of C. magur will be useful for understanding its larval digestive physiology, and this, in turn, will help in designing effective larval feed for growth and survival.

Transcriptome analysis reveals the importance of exogenous nutrition in regulating antioxidant defenses during the mouth-opening stage in oviparous fish.

Antioxidant system is crucial for protecting against environmental oxidative stress in fish life cycle. Although the effects of starvation on the antioxidant defenses in several adult fish have been defined, no relevant researches have been reported in the larval stage, particularly during the transition from endogenous to exogenous feeding. To clarify the molecular response of antioxidant system that occurs during the mouth-opening stage under starvation stress and explore its association with energy metabolism, we employed RNA-seq to analyze the gene expression profiles in zebrafish larvae that received a delayed first feeding for 3 days. Our data showed that delayed feeding resulted in downregulation of 7078 genes and upregulation of 497 genes. These differentially expressed genes are mainly involved in growth regulation (i.e., DNA replication and cell cycle), energy metabolism (i.e., glycolysis/gluconeogenesis and fatty acid metabolism), and antioxidant defenses. We demonstrated that the starved larvae are in an extremely malnourished state in the absence of exogenous nutrition, and the consequence is that numerous antioxidant genes are downregulated. Meanwhile, the antioxidant defenses also respond negatively to oxidative stress. After nutritional supply, the expression of these inhibited antioxidant genes was restored. These results suggest that the establishment of antioxidant defenses during the mouth-opening stage depends highly on exogenous nutrition. Our findings would contribute to comprehending the nutritional stress and metabolic switches during the mouth-opening stage and are essential for reducing high mortality in commercial fish farming.

Mouth opening is mediated by separation of dorsal and ventral daughter cells of the lip precursor cells in the larvacean, Oikopleura dioica.

Mouth formation involves the processes of mouth opening, formation of the oral cavity, and the development of associated sensory organs. In deuterostomes, the surface ectoderm and the anterior part of the archenteron are reconfigured and reconnected to make a mouth opening. This study of the larval development of the larvacean, Oikopleura dioica, investigates the cellular organization of the oral region, the developmental processes of the mouth, and the formation of associated sensory cells. O. dioica is a simple chordate whose larvae are transparent and have a small number of constituent cells. It completes organ morphogenesis in 7 h, between hatching 3 h after fertilization and the juvenile stage at 10 h, when it attains adult form and starts to feed. It has two types of mechanosensory cell embedded in the oral epithelium, which is a single layer of cells. There are twenty coronal sensory cells in the circumoral nerve ring and two dorsal sensory organ cells. Two bilateral lip precursor cells (LPCs), facing the anterior surface, divide dorsoventrally and make a wedge-shaped cleft between the two daughter cells named the dorsal lip cell (DLC) and the ventral lip cell (VLC). Eventually, the DLC and VLC become detached and separated into dorsal and ventral lips, triggering mouth opening. This is an intriguing example of cell division itself contributing to morphogenesis. The boundary between the ectoderm and endoderm is present between the lip cells and coronal sensory cells. All oral sensory cells, including dorsal sensory organ cells, were of endodermal origin and were not derived from the ectodermal placode. These observations on mouth formation provide a cellular basis for further studies at a molecular level, in this simple chordate.

Fast and Facile Biodegradation of Polystyrene by the Gut Microbial Flora of Plesiophthalmus davidis Larvae.

Polystyrene (PS), which accounts for a significant fraction of plastic wastes, is difficult to biodegrade due to its unique molecular structure. Therefore, biodegradation and chemical modification of PS are limited. In this study, we report PS biodegradation by the larvae of the darkling beetle Plesiophthalmus davidis (Coleoptera: Tenebrionidae). In 14 days, P. davidis ingested 34.27 ± 4.04 mg of Styrofoam (PS foam) per larva and survived by feeding only on Styrofoam. Fourier transform infrared spectroscopy confirmed that the ingested Styrofoam was oxidized. Gel permeation chromatography analysis indicated the decrease in average molecular weight of the residual PS in the frass compared with the feed Styrofoam. When the extracted gut flora was cultured for 20 days with PS films, biofilm and cavities were observed by scanning electron microscopy and atomic force microscopy. X-ray photoelectron spectroscopy (XPS) studies revealed that C-O bonding was introduced into the biodegraded PS film. Serratia sp. strain WSW (KCTC 82146), which was isolated from the gut flora, also formed a biofilm and cavities on the PS film in 20 days, but its degradation was less prominent than the gut flora. XPS confirmed that C-O and C=O bonds were introduced into the biodegraded PS film by Serratia sp. WSW. Microbial community analysis revealed that Serratia was in the gut flora in significant amounts and increased sixfold when the larvae were fed Styrofoam for 2 weeks. This suggests that P. davidis larvae and its gut bacteria could be used to chemically modify and rapidly degrade PS.IMPORTANCE PS is widely produced in the modern world, but it is robust against biodegradation. A few studies reported the biodegradation of PS, but most of them merely observed its weight loss; fewer were able to find its chemical modifications, which are rather direct evidence of biodegradation, by using limited organisms. Therefore, it is required to find an effective way to decompose PS using various kinds of organisms. Herein, we discovered a new PS-degrading insect species and bacterial strain, and we found that the genus that includes the PS-degrading bacterial strain occurs in significant amounts in the larval gut flora, and the proportion of this genus increased as the larvae were fed Styrofoam. Our research offers a wider selection of PS-degrading insects and the possibility of using a certain mixture of bacteria that resemble the gut flora of a PS-degrading insect to biodegrade PS, and thus could contribute to solving the global plastic crisis.

Evaluation of hydrolytic enzyme activities from digestive fluids of Anthonomus grandis (Coleoptera: Curculionidae).

The cotton boll weevil, Anthonomus grandis, is a major pest of cotton crops in South America. In this work, partial biochemical characterizations of (hemi) cellulases and pectinases activities in the digestive system (head- and gut- extracts) of A. grandis were evaluated. Gut extract section from third instar larvae exhibited endoglucanase, xylanase, ß-glucosidase, and pectinase activities. The endoglucanase and xylanase activities were localized in the foregut, whereas ß-glucosidase activity was mainly detected in the hindgut. In addition, no difference in pectinase activity was observed across the gut sections. Thus, A. grandis digestive system is a potentially interesting reservoir for further lignocellulolytic enzymes research.

Morphological and molecular description of Armillifer moniliformis larvae isolated from Sri Lankan brown palm civet (Paradoxurus montanus).

We report Armillifer moniliformis species infecting the endemic Sri Lankan brown palm civet (Paradoxurus montanus) from the Knuckles Range Forest Conservation Area, Sri Lanka. Larval stages of A. moniliformis were found during the postmortem of three civet cats found dead. Morphological studies were done by a light microscope and a scanning electron microscope (SEM). Histopathological examination was conducted using tissue samples obtained from the liver. For the molecular analysis, DNA was extracted from the isolated third-stage larvae. The NADH dehydrogenase subunit 5 (ND5) and the second internal transcribed spacer region (ITS-2), a portion of the large subunit nuclear ribosomal DNA (28S), a portion of 18S ribosomal rRNA gene (18S), and cytochrome c oxidase subunit 1 gene (COX1) were amplified using polymerase chain reaction (PCR). Excysted third-stage larvae were observed in the lungs, omentum, the pleural cavity, the abdominal cavity, and the surface of the spleen and the pericardium. Around 88 third-stage larvae were isolated from three civet cats. First-stage larvae in the liver were surrounded by outer fibrous layer over the inner germinal layer and filled with clear fluid. Slight hemorrhage, leukocyte infiltration, and mild hepatocellular degeneration in the liver were observed. The SEM examination indicated the unique oral apparatus comprises the oval-shaped mouth opening in between two pairs of curved, retractile hamuli. The sequences obtained for ND5, ITS-2, 28S, 18S, and COX1 were 301, 382, 325, 414, and 644 bp in length respectively. Morphology, sequence similarity search, sequence alignment, and phylogenetic analysis identified this parasite as A. moniliformis.