Effects of glyphosate-based herbicide on gut microbes and hepatopancreatic metabolism in Pomacea canaliculata.

Roundup®, a prominent glyphosate-based herbicide (GBH), holds a significant position in the global market. However, studies of its effects on aquatic invertebrates, including molluscs are limited. Pomacea canaliculata, a large freshwater snail naturally thrives in agricultural environments where GBH is extensively employed. Our investigation involved assessing the impact of two concentrations of GBH (at concentrations of 19.98 mg/L and 59.94 mg/L, corresponding to 6 mg/L and 18 mg/L glyphosate) during a 96 h exposure experiment on the intestinal bacterial composition and metabolites of P. canaliculata. Analysis of the 16 S rRNA gene demonstrated a notable reduction in the alpha diversity of intestinal bacteria due to GBH exposure. Higher GBH concentration caused a significant shift in the relative abundance of dominant bacteria, such as Bacteroides and Paludibacter. We employed widely-targeted metabolomics analysis to analyze alterations in the hepatopancreatic metabolic profile as a consequence of GBH exposure. The shifts in metabolites primarily affected lipid, amino acid, and glucose metabolism, resulting in compromised immune and adaptive capacities in P. canaliculata. These results suggested that exposure to varying GBH concentrations perpetuates adverse effects on intestinal and hepatopancreatic health of P. canaliculata. This study provides an understanding of the negative effects of GBH on P. canaliculata and may sheds light on its potential implications for other molluscs.

Screening the Efficacy and Safety of Molluscicides from Three Leaf Extracts of Chimonanthus against the Invasive Apple Snail, Pomacea canaliculata.

Pomacea canaliculata, the invasive snail, is a host of the parasitic nematode Angiostrongylus cantonensis, which has adverse effects on the agriculture system and human health. This work evaluated the molluscicidal activity of petroleum ether extracts (PEEs) from three species of Chimonanthus against the snail P. canaliculate. Pcp (PEE of C. praecox) showed the most effective molluscicide activity. Sixty-one compounds were identified by GC-MS and the main components were terpenoids and fatty acids. The half-lethal concentration (LC50) of Pcp at 24 h (0.27 mg/mL) and 48 h (0.19 mg/mL) was used to evaluate the biochemical alterations in snail tissue. These sublethal concentrations caused the levels of alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activity to increase, while acetylcholinesterase (AChE) activity decreased. Also, under LC50 treatment, several histological changes were observed in the hepatopancreas and foot of the snail compared with the control group. Moreover, the toxic test in rice demonstrated that Pcp has low toxicity. These results suggest that Pcp could be developed as an effective molluscicide for P. canaliculata control.

Sublethal effects of niclosamide on the aquatic snail Pomacea canaliculata.

Pomacea canaliculata is a malignant invasive aquatic snail found worldwide, and niclosamide (NS) is one of the primary agents used for its control. NS applied to water will exist in non-lethal concentrations for some time due to degradation or water exchange, thus resulting in sublethal effects on environmental organisms. To identify sublethal effects of NS on Pomacea canaliculata, we studied the aspects of histopathology, oxygen-nitrogen ratio (RO∶N), enzyme activity determination, and gene expression. After LC30 NS treatment (0.310 g/L), many muscle fibers of the feet degenerated and some acinar vesicles of the hepatopancreas collapsed and dissolved. The oxygen-nitrogen ratio (RO∶N) decreased significantly from 15.0494 to 11.5183, indicating that NS had changed the metabolic mode of Pomacea canaliculata and shifted it primarily to protein catabolism. Transcriptome analysis identified the sublethal effects of LC30 NS on the snails at the transcriptional level. 386, 322, and 583 differentially expressed genes (DEGs) were identified in the hepatopancreas, gills, and feet, respectively. GO (Gene Ontology) functional analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway annotations showed that DEGs in the hepatopancreas were mainly enriched for sugar metabolism, protein biosynthesis, immune response, and amino acid metabolism functional categories; DEGs in the gills were mainly enriched for ion transport and amino acid metabolism; DEGs in the feet were mainly enriched for transmembrane transport and inositol biosynthesis. In the future, we will perform functional validation of key genes to further explain the molecular mechanism of sublethal effects.

Dose-dependent inhibitory effects of glyphosate on invasive Pomacea canaliculata reproductive and developmental growth under oxidative deposition.

Glyphosate (GLY) is the most widely used herbicide worldwide, and its effects on animals and plants have attracted increasing attention. In this study, we explored the following: (1) the effects of multigenerational chronic exposure to GLY and H2O2, alone or in combination, on the egg hatching rate and individual morphology of Pomacea canaliculata; and (2) the effects of short-term chronic exposure to GLY and H2O2, alone or in combination, on the reproductive system of P. canaliculata. The results showed that H2O2 and GLY exposure had distinct inhibitory effects on the hatching rate and individual growth indices with a substantial dose effect, and the F1 generation had the lowest resistance. In addition, with the prolongation of exposure time, the ovarian tissue was damaged, and the fecundity decreased; however, the snails could still lay eggs. In conclusion, these results suggest that P. canaliculata can tolerate low concentrations of pollution and in addition to drug dosage, the control should focus on two time points, the juvenile and early stage of spawning.

Transcriptome sequencing reveals the differentially expressed lncRNAs and mRNAs in response to cold acclimation and cold stress in Pomacea canaliculata.

BACKGROUND: Tolerance of low temperature has a significant impact on survival and expansion of invasive snail Pomacea canalicuata. Cold acclimation can enhance cold tolerance of Pomacea canalicuata. To elucidate the molecular mechanism of P. canaliculata's responses to cold acclimation and cold stress, a high-throughput transcriptome analysis of P. canaliculata was performed, and gene expression following artificial cold acclimation and then cold stress at 0 °C for 24 h was compared using RNA sequencing. RESULTS: Using the Illumina platform, we obtained 151.59 G subreads. A total of 5,416 novel lncRNAs were identified, and 3166 differentially expressed mRNAs and 211 differentially expressed lncRNAs were screened with stringent thresholds. The potential antisense, cis and trans targets of lncRNAs were predicted. Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that many target genes were involved in proteasome, linoleic acid metabolism and retinol metabolism under cold acclimation. The lncRNA of P. canaliculata could participate in cold acclimation by regulating the expression of E3 ubiquitin protein ligase, 26S proteasome non-ATPase dependent regulation subunit, glutathione S-transferase, sodium/glucose cotransporter and cytochrome P450. CONCLUSIONS: These results broaden our understanding of cold acclimation and cold stress associated lncRNAs and mRNAs, and provide new insights into lncRNA mediated regulation of P. canaliculata cold acclimation and cold stress response.

Research on the molluscicidal activity and molecular mechanisms of arecoline against Pomacea canaliculata.

Pomacea canaliculata, as an invasive snail in China, can adversely affect agricultural crop yields, ecological environment, and human health. In this paper, we studied the molluscicidal activity and mechanisms of arecoline against P. canaliculata. The molluscicidal activity tests showed that arecoline exhibits strong toxicity against P. canaliculata, and the LC50 value (72 h) was 1.05 mg/L (15 ± 2 mm shell diameter). Additionally, Molluscicidal toxicity were negatively correlated with the size of snails. Snails (25 ± 2 mm shell diameter) were choosed for mechanisms research and the result of microstructure and biochemistry showed that arecoline (4 mg/L, 20 â„ƒ) had strong toxic effect on the gill, and the main signs were the loss of cilia in the gill filaments. Moreover, arecoline significantly decreased the oxygen consumption rate, ammonia excretion rate and inhibited acetylcholinesterase (AChE). Then, the changes in protein expression were studied by iTRAQ, and 526 downregulated proteins were found. Among these, cilia and flagella-associated 157-like (PcCFP) and rootletin-like (PcRoo) were selected as candidate target proteins through bioinformatics analysis, and then RNA interference (RNAi) was adopted to verify the function of PcCFP and PcRoo. The results showed that after arecoline treated, the mortality and the cilia shedding rate of PcRoo RNAi treated group was significantly lower than control group. The above results indicate that arecoline can bind well with protein PcRoo, and then leads to the drop of gill cilia, affect respiratory metabolism, accelerate its entry into hemolymph, inhibit AChE and finally leads to the death of P. canaliculata.

Study on the relationship of Hsp70 with the temperature sensitivity of pedunsaponin A poisoning Pomacea canaliculata.

Previous studies have found that temperature influences molluscicidal the activity of pedunsaponin A (PA), which may be related to the expression of Hsp70, a cold-tolerance gene in Pomacea canaliculata. We determined the temperature effect of PA and the relationship between Hsp70 and temperature sensitivity of P. canaliculata poisoned by PA. Toxicity tests resulted in LC50 values of 17.7239 mg⋅L-1 at 10 °C, which decreased to 2.5774 mg⋅L-1 at 30 °C, implying a positive correlation between toxicity of PA and temperature. After Hsp70 being interfered, the mortality rate of P. canaliculata treated with PA for 72 h was 70%, which was significantly higher than that of snails treated with PA for 72 h without interfering (56.7%). Meanwhile, immune enzyme activities such as SOD, ACP and AKP were significantly increased in the interfered group and expression level of PcAdv in the gill was also significantly increased. These results suggest that deletion of Hsp70 promotes the activation of some immune enzymes of P. canaliculata and elevates the content of target proteins to cope with the dual stresses of low temperatures and molluscicides. These findings indicate that the Hsp70 plays an important role in influencing the temperature sensitivity of P. canaliculata when treated with PA.

Study on the Synergistic Molluscicidal Effect of Pedunsaponin A and Niclosamide.

Niclosamide (NI) is the main molluscicide used to control Pomacea canaliculata (Lamarck) (Architaenioglossa: Ampullariidae). However, NI failed to inhibit snail climbing during the treatment process. In this study, we examined the effect of NI combined with pedunsaponin A at an ineffective concentration. The molluscicidal effect of Pedunsaponin A on NI was evidently synergistic after 48 h, and the synergism ratio (SR) was 1.82 after treatment for 72 h at 0.8 mg·L-1. Examination of the climbing adhesion effect showed that a high concentration of Pedunsaponin A (0.4 mg·L-1 and 0.8 mg·L-1) combined with NI significantly inhibited the climbing of P. canaliculata. We further studied the synergism mechanism; the results of histopathological observation showed that the siphon appeared cavities, the muscle fibers of the ventricular were severely dissolved, and kidney tubule arrangement was distorted after NI adding Pedunsaponin A. In addition, the hemocyte survival rate and the content of hemocyanin decreased significantly. According to the results of our study, the synergism mechanism may hinder oxygen transport of P. canaliculata, influencing the supply of energy; the ability of immune defense and excretion and metabolic detoxification decreased, prolonging the action time of NI in the body.

A Novel Gelatin-Based Sustained-Release Molluscicide for Control of the Invasive Agricultural Pest and Disease Vector Pomacea canaliculata.

Pomacea canaliculata, one of the 100 most destructive invasive species in the world, and it is an important intermediate host of Angiostrongylus cantonensis. The molluscicides in current use are an effective method for controlling snails. However, most molluscicides have no slow-release effect and are toxic to nontarget organisms. Thus, these molluscicides cannot be used on a large scale to effectively act on snails. In this study, gelatin, a safe and nontoxic substance, was combined with sustained-release molluscicide and was found to reduce the toxicity of niclosamide to nontarget organisms. We assessed the effects of gelatin and molluscicide in controlling P. canaliculata snails and eggs. The results demonstrated that the niclosamide retention time with 1.0% and 1.5% gelatin sustained-release agents reached 20 days. Additionally, the mortality rate of P. canaliculata and their eggs increased as the concentration of the niclosamide sustained-release agents increased. The adult mortality rate of P. canaliculata reached 50% after the snails were exposed to gelatin with 0.1 mg/L niclosamide for 48 h. The hatching rate of P. canaliculata was only 28.5% of the normal group after the treatment was applied. The sustained-release molluscicide at this concentration was less toxic to zebrafish, which means that this molluscicide can increase the safety of niclosamide to control P. canaliculata in aquatic environments. In this study, we explored the safety of using niclosamide sustained-release agents with gelatin against P. canaliculata. The results suggest that gelatin is an ideal sustained-release agent that can provide a foundation for subsequent improvements in control of P. canaliculata.

Composition and diversity of gut microbiota in Pomacea canaliculata in sexes and between developmental stages.

BACKGROUND: The apple snail, Pomacea canaliculata, is one of the world's 100 worst invasive alien species and vector of some pathogens relevant to human health. METHODS: On account of the importance of gut microbiota to the host animals, we compared the communities of the intestinal microbiota from P. canaliculata collected at different developmental stages (juvenile and adult) and different sexes by using high-throughput sequencing. RESULTS: The core bacteria phyla of P. canaliculata gut microbiota included Tenericutes (at an average relative abundance of 45.7 %), Firmicutes (27.85 %), Proteobacteria (11.86 %), Actinobacteria (4.45 %), and Cyanobacteria (3.61 %). The female group possessed the highest richness values, whereas the male group possessed the lowest bacterial richness and diversity compared with the female and juvenile group. Both the developmental stages and sexes had important effects on the composition of the intestinal microbiota of P. canaliculata. By LEfSe analysis, microbes from the phyla Proteobacteria and Actinobacteria were enriched in the female group, phylum Bacteroidetes was enriched in the male group, family Mycoplasmataceae and genus Leuconostoc were enriched in the juvenile group. PICRUSt analysis predicted twenty-four metabolic functions in all samples, including general function prediction, amino acid transport and metabolism, transcription, replication, recombination and repair, carbohydrate transport and metabolism, etc. CONCLUSIONS: This study provided a general understanding of the diversity characteristics of intestinal microbial communities of P. canaliculata, and indicated that developmental stage and gender could both influence the intestinal microbes of P. canaliculata. Further study may focus on the interaction between the gut microbiota and their host.

Screening and functional verification of the target protein of pedunsaponin A in the killing of Pomacea canaliculata.

Previous study found that pedunsaponin A (PA) influenced the cytoskeleton of Pomacea canaliculata hemocytes, leading to depolarization and haemocyte destruction and eventually to snail death. In this study, we analysed the changes in protein expression by iTRAQ-mediated proteomics and identified 51 downregulated proteins. Among these, we focused on proteins related to cytoskeletal function and identified neural Wiskott-Aldrich syndrome isoform X1 (PcnWAS). The full-length PcnWAS gene contains 9791 bp and includes an open reading frame of 1401 bp that encodes 735 amino acids with a predicted molecular mass of 49.83 kD. PcnWAS exhibited a relatively distant genetic relationship with known species; the closest homologue is Biomphalaria glabrata (57%). RNA interference (RNAi) was adopted to verify the function of PcnWAS after screening the siRNA sequence with an efficiency of 97%. Interference with the gene expression of PcnWAS did not lead to snail death, but the depolarization level increased, which demonstrated that PcnWAS is an important depolarization-related protein. The results of PA treatment of snails subjected to RNAi proved that interfering with PcnWAS gene expression decreased the molluscicidal activity of PA toward P. canaliculata; snail mortality after RNAi was significantly lower (40%) than that in PA-treated snails without RNAi (54%), while the survival rate and depolarization level in haemocytes were not significant, indicating that PcnWAS is only one of the important target proteins of PA in P. canaliculata. This study lays the foundation for further exploration of the molecular mechanism by which PA kills this harmful snail.

Lantana camara Essential Oils from Vietnam: Chemical Composition, Molluscicidal, and Mosquito Larvicidal Activity.

Lantana camara is a troublesome invasive plant introduced to many tropical regions, including Southeast Asia. However, the plant does hold promise as a source of essential oils that may be explored for potential use. Fresh water snails such as Pomacea canaliculata, Gyraulus convexiusculus, and Tarebia granifera can be problematic agricultural pests as well as hosts for parasitic worms. Aedes and Culex mosquitoes are notorious vectors of numerous viral pathogens. Control of these vectors is of utmost importance. In this work, the essential oil compositions, molluscicidal, and mosquito larvicidal activities of four collections of L. camara from north-central Vietnam have been investigated. The sesquiterpene-rich L. camara essential oils showed wide variation in their compositions, not only compared to essential oils from other geographical locations (at least six possible chemotypes), but also between the four samples from Vietnam. L. camara essential oils showed molluscicidal activities comparable to the positive control, tea saponin, as well as other botanical agents. The median lethal concentrations (LC50 ) against the snails were 23.6-40.2 µg/mL (P. canaliculata), 7.9-29.6 µg/mL (G. convexiusculus), and 15.0-29.6 µg/mL (T. granifera). The essential oils showed good mosquito larvicidal activities with 24-h LC50 values of 15.1-29.0 µg/mL, 26.4-53.8 µg/mL, and 20.8-59.3 µg/mL against Ae. aegypti, Ae. albopictus, and Cx. quinquefasciatus, respectively. The essential oils were more toxic to snails and mosquito larvae than they were to the non-target water bug, Diplonychus rusticus (24-h LC50 =103.7-162.5 µg/mL). Sesquiterpene components of the essential oils may be acting as acetylcholinesterase (AChE) inhibitors. These results suggest that the invasive plant, L. camara, may be a renewable botanical pesticidal agent.

Ecotoxicological evaluation of fruit extracts from yerba mate progenies (Ilex paraguariensis a St-Hil.): a natural biopesticide.

This study evaluated the biocidal activity of aqueous fruit extracts from Ilex paraguariensis progenies at different ripening stages in two ecotoxicity assays using Pomacea canaliculata juveniles and Danio rerio larvae. Mutagenicity was verified by the Salmonella/microsome assay (TA 98 and TA 100) in the presence and absence of hepatic metabolism in vitro. The results showed that the snails were more sensitive than the larvae to the P.1 progeny extracts in all three evaluated ripening stages. P.3 progeny extracts were the most toxic to Danio rerio larvae after 96 hours of exposure. The toxicity results indicate that the progenies' selection influenced the metabolic contents present in the fruits in all ripening stages over time, indicating high molluscicidal activity for the P.1 progeny. In the Salmonella/microsome assays, the semi-ripe and ripe extracts from progenies P.1 and P.3 showed mainly base pair substitution mutation in assays with metabolism, but at higher doses than the toxic concentrations detected for Danio rerio or Pomacea canaliculata. Therefore, the toxicological investigation of the progenies' extracts can be interesting, given the selection of plant materials influenced the response of the bioassays.

Exaptation of two ancient immune proteins into a new dimeric pore-forming toxin in snails.

The Membrane Attack Complex-Perforin (MACPF) family is ubiquitously found in all kingdoms. They have diverse cellular roles, however MACPFs with pore-forming toxic function in venoms and poisons are very rare in animals. Here we present the structure of PmPV2, a MACPF toxin from the poisonous apple snail eggs, that can affect the digestive and nervous systems of potential predators. We report the three-dimensional structure of PmPV2, at 17.2 Å resolution determined by negative-stain electron microscopy and its solution structure by small angle X-ray scattering (SAXS). We found that PV2s differ from nearly all MACPFs in two respects: it is a dimer in solution and protomers combine two immune proteins into an AB toxin. The MACPF chain is linked by a single disulfide bond to a tachylectin chain, and two heterodimers are arranged head-to-tail by non-covalent forces in the native protein. MACPF domain is fused with a putative new Ct-accessory domain exclusive to invertebrates. The tachylectin is a six-bladed ß-propeller, similar to animal tectonins. We experimentally validated the predicted functions of both subunits and demonstrated for the first time that PV2s are true pore-forming toxins. The tachylectin "B" delivery subunit would bind to target membranes, and then the MACPF "A" toxic subunit would disrupt lipid bilayers forming large pores altering the plasma membrane conductance. These results indicate that PV2s toxicity evolved by linking two immune proteins where their combined preexisting functions gave rise to a new toxic entity with a novel role in defense against predation. This structure is an unparalleled example of protein exaptation.

Preliminary investigation demonstrating the GHITM gene probably involved in apoptosis and growth of the golden apple snail (Pomacea canaliculata).

BACKGROUND: Growth hormone inducible transmembrane protein (GHITM) is a highly conserved transmembrane protein. This study was conducted to investigate the role of GHITM gene in the apoptosis and growth of the golden apple snail Pomacea canaliculate. RESULTS: The complete cDNA of this gene was cloned using the rapid amplification of cDNA ends (RACE) method and subjected to bioinformatics analysis. The full-length cDNA was 2242 bp, including an open reading frame of 1021 bp that encoded a protein of 342 amino acid residues. The mRNA expression profiles of GHITM gene in different tissues (liver, kidney, gonad and foot) and different growth phases (6-months old and 2-years old) showed that it was expressed in various tissues and different growth phases. Silencing of the GHITM gene by RNAi (RNA interference) experiments revealed that the GHITM gene possibly plays a role in inhibiting apoptosis through detecting the Caspase (Cysteine-requiring Aspartate Protease)-3 activity. In addition, the aperture width and body whorl length of the snail was significantly affected by RNAi, suggesting that this gene plays a significant role in promoting the growth of the organism. CONCLUSIONS: These results demonstrated that the GHITM gene was involved in apoptosis and growth in golden apple snail.

Molluscicidal activity of fatty acids in the kernel of Chimonanthus praecox cv. Luteus against the golden apple snail Pomacea canaliculata.

The fatty acid composition of the kernel of Chimonanthus praecox cv. Luteus (FKC) was analyzed by gas chromatography-mass spectrometry (GC-MS), its ability to kill Pomacea canaliculata was detected, and the degree of damage and physiological and biochemical effects of an FKC soaking treatment on the hepatopancreas tissue of P. canaliculata were evaluated. In total, 16 fatty acids were detected in FKC, among which 13 were qualitatively identified; octadecadienoic acid (56.76%) and palmitic acid (17.03%) had the highest contents. After 48 h of treatment with FKC, the hepatopancreas of P. canaliculata had a large area of necrosis. The contents of soluble sugar, soluble protein, and albumin (Alb) in the hepatopancreas of P. canaliculata decreased with increasing FKC concentration. The content of malondialdehyde (MDA) and the activities of cereal third transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (AKP), and acetylcholinesterase (AChE) increased with increasing FKC concentration. The results showed that FKC has an obvious negative effect on the hepatopancreas cell structure and physiological function of P. canaliculata, i.e., has strong molluscicidal activity.

Two new compounds from the roots of Pueraria peduncularis and their molluscicidal effects on Pomacea canaliculata.

Two oleanane-type triterpenoid saponins named pedunsaponin D (1) and pedunsaponin E (2) were isolated from the roots of Pueraria peduncularis. The structures of the new compounds were elucidated based on chemical and physicochemical evidence as follows: pedunsaponin D, 3-O-ß-glucopyranosyl-(1-3)-ß-glucuronopyranosyl-3ß,15α,23α-trihydroxy-11,13(18)-oleanadien-16-one (1); pedunsaponin E, 3-O-ß-glucopyranosyl-(1-2)-ß-glucopy ranosyl(1-2)[ß-glucopyranosyl(1-3)-ß-glucuronopyranosyl]-3ß-hydroxy-16-oxoolean-12-en-30-oic acid (2). The two compounds showed moderate molluscicidal activity.[Formula: see text].

Two Unconventional Metallothioneins in the Apple Snail Pomacea bridgesii Have Lost Their Metal Specificity during Adaptation to Freshwater Habitats.

Metallothioneins (MTs) are a diverse group of proteins responsible for the control of metal homeostasis and detoxification. To investigate the impact that environmental conditions might have had on the metal-binding abilities of these proteins, we have characterized the MTs from the apple snail Pomacea bridgesii, a gastropod species belonging to the class of Caenogastropoda with an amphibious lifestyle facing diverse situations of metal bioavailability. P. bridgesii has two structurally divergent MTs, named PbrMT1 and PbrMT2, that are longer than other gastropod MTs due to the presence of extra sequence motifs and metal-binding domains. We have characterized the Zn(II), Cd(II), and Cu(I) binding abilities of these two MTs after their heterologous expression in E. coli. Our results have revealed that despite their structural differences, both MTs share an unspecific metal-binding character, and a great ability to cope with elevated amounts of different metal ions. Our analyses have also revealed slight divergences in their metal-binding features: PbrMT1 shows a more pronounced Zn(II)-thionein character than PbrMT2, while the latter has a stronger Cu(I)-thionein character. The characterization of these two unconventional PbrMTs supports the loss of the metal-binding specificity during the evolution of the MTs of the Ampullariid family, and further suggests an evolutionary link of this loss with the adaptation of these gastropod lineages to metal-poor freshwater habitats.

Spatial structure of the microbiome in the gut of Pomacea canaliculata.

BACKGROUND: Gut microbes can contribute to their hosts in food digestion, nutrient absorption, and inhibiting the growth of pathogens. However, only limited studies have focused on the gut microbiota of freshwater snails. Pomacea canaliculata is considered one of the worst invasive alien species in the world. Elucidating the diversity and composition of the microbiota in the gut of P. canaliculata snails may be helpful for better understanding the widespread invasion of this snail species. In this study, the buccal masses, stomachs, and intestines were isolated from seven P. canaliculata snails. The diversity and composition of the microbiota in the three gut sections were then investigated based on high-throughput Illumina sequencing targeting the V3-V4 regions of the 16S rRNA gene. RESULTS: The diversity of the microbiota was highest in the intestine but lowest in the buccal mass. A total of 29 phyla and 111 genera of bacteria were identified in all of the samples. In general, Ochrobactrum, a genus of putative cellulose-degrading bacteria, was the most abundant (overall relative abundance: 13.6%), followed by Sediminibacterium (9.7%), Desulfovibrio (7.8%), an unclassified genus in the family Aeromonadaceae (5.4%), and Cloacibacterium (5.4%). The composition of the microbiota was diverse among the different gut sections. Ochrobactrum (relative abundance: 23.15% ± 7.92%) and Sediminibacterium (16.95 ± 5.70%) were most abundant in the stomach, an unclassified genus in the family Porphyromonadaceae (14.28 ± 7.29%) and Leptotrichia (8.70 ± 4.46%) were highest in the buccal mass, and two genera in the families Aeromonadaceae (7.55 ± 4.53%) and Mollicutes (13.47 ± 13.03%) were highest in the intestine. CONCLUSIONS: The diversity and composition of the microbiome vary among different gut sections of P. canaliculata snails. Putative cellulose-degrading bacteria are enriched in the gut of P. canaliculata.

Toxicity of Chimonanthus nitens flower extracts to the golden apple snail, Pomacea canaliculata.

We studied the molluscicidal activity of Chimonanthus nitens extracts on Pomacea canaliculata (Ampullariidae). The degree of hepatopancreatic tissue damage, and its physiological and biochemical effects, was evaluated on individuals exposed to petroleum ether extracts (PEEEs). The PEEEs, ethyl acetate extract (EAEE) and water saturated n-butyl extract (SBEE) of C. nitens also had toxic effects on P. canaliculata but PEEE had the greatest molluscicidal activity. After exposure to PEEE for 24 h, the hepatopancreas of P. canaliculata had a large necrotic area. The levels of soluble sugar, soluble protein and albumin (Alb) in the hepatopancreas of P. canaliculata decreased with increasing PEEE concentration, while the activities of glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT) and acetylcholinesterase (AchE) increased with increasing PEEE concentration. A total of 29 compounds were identified from the PEEE of C. nitens by gas chromatography-mass spectrometry analysis. The main components were esters (48.13%), alcohols (18.43%) and the compound Chimonanthine (14.70%). The results of the molluscicidal assay, histological experiments and the physiological and biochemical experiments show that the PEEE of C. nitens could potentially be used for P. canaliculata management.