Methane emitted by insects is considered to be an important source of atmospheric methane. Here we report the stimulation of methane emission in the cockroach Periplaneta americana and termite Coptotermes chaohuensis, insects with abundant methanogens, by neonicotinoids, insecticides widely used to control insect pests. Cycloxaprid (CYC) and imidacloprid (IMI) caused foregut expansion in P. americana, and increased the methane emission. Antibiotics mostly eliminated the effects. In P. americana guts, hydrogen levels increased and pH values decreased, which could be significantly explained by the gut bacterium community change. The proportion of several bacterium genera increased in guts following CYC treatment, and two genera from four could generate hydrogen. Hydrogen is a central intermediate in methanogenesis. All increased methanogens in both foregut and hindgut used hydrogen as electron donor to produce methane. Besides, the up-regulation of mcrA, encoding the enzyme for the final step of methanogenesis suggested the enhanced methane production ability in present methanogens. In the termite, hydrogen levels in gut and methane emission also significantly increased after neonicotinoid treatment, which was similar to the results in P. americana. In summary, neonicotinoids changed bacterium community in P. americana gut to generate more hydrogen, which then stimulated gut methanogens to produce and emit more methane. The finding raised a new concern over neonicotinoid applications, and might be a potential environmental risk associated with atmospheric methane.

The effects of microplastic exposure in aquatic organisms have been widely reported. Nonetheless, there is limited evidence of the effects of exposure in soil systems. Thus, the objective of this study was to evaluate the effects of microplastic exposure using as a bioindicator the species Lumbricus terrestris, exposed to different concentrations of microplastic (2.5%; 5%, and 7% w/w). Avoidance bioassays were carried out for 48 h in soil with and without microplastic; the gastrointestinal tract—crop/gizzard, foregut, and midgut—was dissected and acetylcholinesterase (AChE) was used as a biomarker of neurotoxicity stress. In parallel, bioassays of microplastic ingestion were carried out, and after 48 h of initiating the ingestion assay, using a stereo and fluorescence microscope, the microplastic distribution was observed in the different earthworm segments. The results obtained in the avoidance assay indicated a lack of preference for either soil type; however, upon moving, the earthworms lost surface mucus, resulting in burns and lesions on their bodies, which were reflected in the increase in AChE enzyme levels, which was not directly related to microplastic ingestion, but rather likely acts as an external physical stress agent. The results of the ingestion bioassay showed that microplastic was present in all the earthworm segments, with a higher number of particles in the hindgut. The Lumbricus terrestris did not distinguish microplastics from soil particles, and given the high exposure concentrations, microplastics produced physical lesions on the mucus membranes of earthworms. Lumbricus terrestris showed to be a suitable bioindicator for testing the exposure to microplastic contamination in soil.

Earthworms are commonly referred as environmental engineers and their guts are often compared with chemical reactors. However, modeling experiments to substantiate it are lacking. The aim of this study was to use established reactor models, particularly PFR, on the gut of the vermicomposting earthworm Eudrilus eugeniae to understand more on its digestion. To achieve the objective, a mathematical model based on first-order kinetics was framed and used to determine the pattern of digestion rates of nutrient indicators, namely total carbon (%), total nitrogen (%), C/N ratio, ¹³C (‰), and ¹⁵N (‰) at five intersections (pre-intestine, foregut, midgut A, midgut B, and hindgut) along the gut of E. eugeniae. The experimental results revealed that the concentrations of TC, TN, ¹³C, and ¹⁵N decreased during gut transit, whereas C/N ratio increased. The first-order model demonstrated that all the nutrients exhibit a linear pattern of digestion during gut transit, which supports the PFR model. On this basis, the present study concludes that the gut of E. eugeniae functions as PFR.

To study the responses of digestive system of the freshwater crab Sinopotamon henanense to the exposure with cadmium (Cd), crabs were acutely exposed to 7.25, 14.50, and 29.00 mg/l Cd for 96 h and subchronically exposed to 0.725, 1.450, and 2.900 mg/l for 21 days. Cd bioaccumulation in the hepatopancreas and digestive tract (esophagus and intestine) was examined. Furthermore, histopathological alterations of the esophagus, midgut, hindgut, and hepatopancreas were assessed in animals from the 29.0 and 2.90 mg/l Cd treatment groups, and expression of metallothionein messenger RNA (MT mRNA) in the hepatopancreas and intestine was measured in all treatment groups. The results showed difference in the middle and high concentrations between acute and subchronic treatment groups. Cd content in digestive tract after acute 14.5 and 29.0 mg/l Cd exposure was significantly higher than that at subchronic 1.45 and 2.90 mg/l exposure, but Cd levels in hepatopancreas were not significantly different under the same condition. Acute exposure to Cd induced greater morphological damage than subchronic exposure: large areas of epithelial cells were necrotic in hepatopancreas and midgut, which detached from the basal lamina. Vacuolated muscle cells were observed in the hindgut of animals from the acute exposure group, but the changes of esophageal morphology were not obvious after acute or subchronic treatments. The expression of MT mRNA increased with increasing Cd concentration, and MT mRNA level in acute exposure groups was significantly lower when compared to the subchronic exposure groups. Higher Cd content and lower MT mRNA expression in the acutely exposed groups may be responsible for more severe damage of digestive system in these exposure groups.

Vermi-composting is considered to be a feasible method for reducing tetracycline resistance genes (TRGs) in the sludge. Nevertheless, the way different gut passages of earthworm might affect the fates of TRGs and whether this process is affected by tetracycline (TC) concentrations need to be further investigated. In this study, we examined the effects of TC concentrations on changes in TRGs and bacterial communities in gut passages of earthworm were determined by using quantitative PCR and Illumina high-throughput sequencing. TRGs and intI1 were mainly reduced in the hindgut under the TC concentrations ranging from 0 to 25 mg/kg, while they were enriched under higher TC stress exposure. Consequently, we suggest the TC limitation of 25 mg/kg in the domestic sludge (DS) for vermi-composting. Although the predominant genera were TC sensitive under TC stress, many bacterial hosts harboring multiple TRGs (especially those in the hindgut) should be paid further attention to. In the foregut, five genera with abundant tetracycline-resistant bacteria (TRB) were specialized taxa. Among these genera, Unclassified_Solirubrobacterales and Pirellulaceae were probably related to the digestion processes. Other unclassified taxa related to the TRGs were probably derived from the DS. Five genera with abundant TRB were shared in the gut passages, and three specialized genera in the hindgut. These genera could spread TRGs and intI1 to the environment. These results suggest that vermi-composting is a feasible approach for TRG control in the DS containing TC concentration that does not exceed 25 mg/kg. Fates of TRGs and intI1 widely differ in the gut passages, showing inevitable connections with bacterial communities.