Midgut Regions Research Articles

Nanoparticle Uptake in the Aging and Oncogenic Drosophila Midgut Measured with Surface-Enhanced Raman Spectroscopy.

Colorectal cancer remains a major global health concern. Colonoscopy, the gold-standard colorectal cancer diagnostic, relies on the visual detection of lesions and necessitates invasive biopsies for confirmation. Alternative diagnostic methods, based on nanomedicine, can facilitate early detection of malignancies. Here, we examine the uptake of surface-enhanced Raman scattering nanoparticles (SERS NPs) as a marker for intestinal tumor detection and imaging using an established Drosophila melanogaster model for gut disease. Young and old Oregon-R and w1118 flies were orally administered SERS NPs and scanned without and upon gut lumen clearance to assess nanoparticle retention as a function of aging. Neither young nor old flies showed significant NP retention in their body after gut lumen clearance. Moreover, tumorigenic flies of the esg-Gal4/UAS-RasV12 genotype were tested for SERS NP retention 2, 4 and 6 days after RasV12 oncogene induction in their midgut progenitor cells. Tumorigenic flies showed a statistically significant NP retention signal at 2 days, well before midgut epithelium impairment. The signal was then visualized in scans of dissected guts revealing areas of NP uptake in the posterior midgut region of high stem cell activity.

Hundreds of antimicrobial peptides create a selective barrier for insect gut symbionts

The spatial organization of gut microbiota is crucial for the functioning of the gut ecosystem, although the mechanisms that organize gut bacterial communities in microhabitats are only partially understood. The gut of the insect Riptortus pedestris has a characteristic microbiota biogeography with a multispecies community in the anterior midgut and a monospecific bacterial population in the posterior midgut. We show that the posterior midgut region produces massively hundreds of specific antimicrobial peptides (AMPs), the Crypt-specific Cysteine-Rich peptides (CCRs) that have membrane-damaging antimicrobial activity against diverse bacteria but posterior midgut symbionts have elevated resistance. We determined by transposon-sequencing the genetic repertoire in the symbiont Caballeronia insecticola to manage CCR stress, identifying different independent pathways, including AMP-resistance pathways unrelated to known membrane homeostasis functions as well as cell envelope functions. Mutants in the corresponding genes have reduced capacity to colonize the posterior midgut, demonstrating that CCRs create a selective barrier and resistance is crucial in gut symbionts. Moreover, once established in the gut, the bacteria differentiate into a CCR-sensitive state, suggesting a second function of the CCR peptide arsenal in protecting the gut epithelia or mediating metabolic exchanges between the host and the gut symbionts. Our study highlights the evolution of an extreme diverse AMP family that likely contributes to establish and control the gut microbiota.

Resistance to both aphids and nematodes in tobacco plants expressing a Bacillus thuringiensis crystal protein.

Bacillus thuringiensis (Bt) and its crystal toxin or δ-endotoxins (Cry) offer great potential for the efficient control of crop pests. A vast number of pests can potentially infect the same host plant, either simultaneously or sequentially. However, no effective Bt-Cry protein has been reported to control both aphids and plant parasitic nematodes due to its highly specific activity. Our study indicated that the Cry5Ba2 protein was toxic to the green peach aphid Myzus persicae, which had a median lethal concentration (LC50) of 9.7 ng μL-1 and fiducial limits of 3.1-34.6 ng μL-1. Immunohistochemical localization of Cry5Ba2 revealed that it could bind to the apical tip of microvilli in midgut regions. Moreover, transgenic tobacco plants expressing Cry5Ba2 exhibited significant resistance to Myzus persicae, as evidenced by reduced insect survival and impaired fecundity, and also intoxicated the Meloidogyne incognita as indicated by a decrease in galls and progeny reproduction. In sum, we identified a new aphicidal Bt toxin resource that could simultaneously control both aboveground and belowground pests, thus extending the application range of Bt-based strategy for crop protection. © 2024 Society of Chemical Industry.

The midgut epithelium of mosquitoes adjusts cell proliferation and endoreplication to respond to physiological challenges

BackgroundHematophagous mosquitoes transmit many pathogens that cause human diseases. Pathogen acquisition and transmission occur when female mosquitoes blood feed to acquire nutrients for reproduction. The midgut epithelium of mosquitoes serves as the point of entry for transmissible viruses and parasites.ResultsWe studied midgut epithelial dynamics in five major mosquito vector species by quantifying PH3-positive cells (indicative of mitotic proliferation), the incorporation of nucleotide analogs (indicative of DNA synthesis accompanying proliferation and/or endoreplication), and the ploidy (by flow cytometry) of cell populations in the posterior midgut epithelium of adult females. Our results show that the epithelial dynamics of post-emergence maturation and of mature sugar-fed guts were similar in members of the Aedes, Culex, and Anopheles genera. In the first three days post-emergence, ~ 20% of cells in the posterior midgut region of interest incorporated nucleotide analogs, concurrent with both proliferative activity and a broad shift toward higher ploidy. In mature mosquitoes maintained on sugar, an average of 3.5% of cells in the posterior midgut region of interest incorporated nucleotide analogs from five to eight days post-emergence, with a consistent presence of mitotic cells indicating constant cell turnover. Oral bacterial infection triggered a sharp increase in mitosis and nucleotide analog incorporation, suggesting that the mosquito midgut undergoes accelerated cellular turnover in response to damage. Finally, blood feeding resulted in an increase in cell proliferation, but the nature and intensity of the response varied by mosquito species and by blood source (human, bovine, avian or artificial). In An. gambiae, enterocytes appeared to reenter the cell cycle to increase ploidy after consuming blood from all sources except avian.ConclusionsWe saw that epithelial proliferation, differentiation, and endoreplication reshape the blood-fed gut to increase ploidy, possibly to facilitate increased metabolic activity. Our results highlight the plasticity of the midgut epithelium in mosquitoes’ physiological responses to distinct challenges.Graphical

Bacterial biota composition in gut regions of black soldier fly larvae reared on industrial residual streams: revealing community dynamics along its intestinal tract.

Some insect species have gained attention as efficient bioconverters of low-value organic substrates (i.e., residual streams) into high-value biomass. Black soldier fly (BSF) (Hermetia illucens) larvae are particularly interesting for bioconversion due to their ability to grow on a wide range of substrates, including low-value industrial residual streams. This is in part due to the plasticity of the gut microbiota of polyphagous insects, like BSF. Gut microbiota composition varies depending on rearing substrates, via a mechanism that might support the recruitment of microorganisms that facilitate digestion of a specific substrate. At the same time, specific microbial genera do persist on different substrates via unknown mechanisms. This study aimed to offer insights on this microbial plasticity by investigating how the composition of the bacterial community present in the gut of BSF larvae responds to two industrial residual streams: swill (a mixture of catering and supermarket leftovers) and distiller's dried grains with solubles. The bacterial biota composition of substrates, whole larvae at the beginning of the rearing period and at harvest, rearing residues, and larval gut regions were investigated through 16S rRNA gene sequencing. It was observed that both substrate and insect development influenced the bacterial composition of the whole larvae. Zooming in on the gut regions, there was a clear shift in community composition from a higher to a lower diversity between the anterior/middle midgut and the posterior midgut/hindgut, indicating a selective pressure occurring in the middle midgut region. Additionally, the abundance of the bacterial biota was always high in the hindgut, while its diversity was relatively low. Even more, the bacterial community in the hindgut was found to be relatively more conserved over the different substrates, harboring members of the BSF core microbiota. We postulate a potential role of the hindgut as a reservoir for insect-associated microbes. This warrants further research on that underexplored region of the intestinal tract. Overall, these findings contribute to our understanding of the bacterial biota structure and dynamics along the intestinal tract, which can aid microbiome engineering efforts to enhance larval performance on (industrial) residual streams.

Histopathology of chironomids exposed to fly ash and microplastics as a new biomarker of ecotoxicological assessment

In the last few decades, industrial pollution has gained extensive attention in terms of its effect on the aquatic environment. This imposes the need to develop sensitive biomarkers for early detection of pollutant toxicity in ecotoxicological assessment. The advantages of histopathological biomarkers are many, including quick reaction to the presence of contaminants, and the small number of individuals needed for efficient analysis. The present study analyzed the negative effect of lignite coal fly ash (LCFA) and microplastic particles (MPs) on Chironomus riparius, a suggested model organism by the Organization for Economic Cooperation and Development (OECD). This study aimed to perform histological analyses of larval tissues and target potential changes in treated groups that could serve as promising histopathological biomarkers of the contaminant's negative effects. Following that, other known sensitive sub-organismal biomarkers were analyzed and paired with the histopathological ones. Histological analysis of larvae showed a significantly decreased length of microvilli in midgut regions II and III in both treatments. Treatments with MPs affected oxidative stress parameters: thiobarbituric acid reactive substances (TBARS), advanced oxidation protein products (AOPP), superoxide dismutase (SOD), and hemoglobin levels, while LCFA significantly affected all tested sub-organismal biomarkers (DNA damage, levels of AOPP, SOD, and hemoglobin), except catalase (CAT) and TBARS. When observing histological slides, a significant shortage of brush border length in the posterior parts of the midgut was detected in all treatments. In the case of LCFA, the appearance of intensive vacuolization of digestive cells with inclusions resembling apoptotic bodies, in mentioned regions was also detected. This study demonstrated high sensitivity of brush border length to the MPs and LCFA exposure, complementary to other tested sub-organismal biomarkers. Revealing the great potential of this histopathological biomarker in ecotoxicological studies contributes to the international standard ecotoxicology assessment of emerging pollutants.

The larval midgut of Anopheles, Aedes, and Toxorhynchites mosquitoes (Diptera, Culicidae): a comparative approach in morphophysiology and evolution.

The mosquito larval midgut is responsible for acquiring and storing most of the nutrients that will sustain the events of metamorphosis and the insect's adult life. Despite its importance, the basic biology of this larval organ is poorly understood. To help fill this gap, we carried out a comparative morphophysiological investigation of three larval midgut regions (gastric caeca, anterior midgut, and posterior midgut) of phylogenetically distant mosquitoes: Anopheles gambiae (Anopheles albimanus was occasionally used as an alternate), Aedes aegypti, and Toxorhynchites theobaldi. Larvae of Toxorhynchites mosquitoes are predacious, in contrast to the other two species, that are detritivorous. In this work, we show that the larval gut of the three species shares basic histological characteristics, but differ in other aspects. The lipid and carbohydrate metabolism of the An. gambiae larval midgut is different compared with that of Ae. aegypti and Tx. theobaldi. The gastric caecum is the most variable region, with differences probably related to the chemical composition of the diet. The peritrophic matrix is morphologically similar in the three species, and processes involved in the post-embryonic development of the organ, such as cell differentiation and proliferation, were also similar. FMRF-positive enteroendocrine cells are grouped in the posterior midgut of Tx. theobaldi, but individualized in An. gambiae and Ae. aegypti. We hypothesize that Tx. theobaldi larval predation is an ancestral condition in mosquito evolution.

Trehalose Biosynthesis Gene otsA Protects against Stress in the Initial Infection Stage of Burkholderia-Bean Bug Symbiosis.

Trehalose, a nonreducing disaccharide, functions as a stress protectant in many organisms, including bacteria. In symbioses involving bacteria, the bacteria have to overcome various stressors to associate with their hosts; thus, trehalose biosynthesis may be important for symbiotic bacteria. Here, we investigated the role of trehalose biosynthesis in the Burkholderia-bean bug symbiosis. Expression levels of two trehalose biosynthesis genes, otsA and treS, were elevated in symbiotic Burkholderia insecticola cells, and hence mutant ΔotsA and ΔtreS strains were generated to examine the functions of these genes in symbiosis. An in vivo competition assay with the wild-type strain revealed that fewer ΔotsA cells, but not ΔtreS cells, colonized the host symbiotic organ, the M4 midgut, than wild-type cells. The ΔotsA strain was susceptible to osmotic pressure generated by high salt or high sucrose concentrations, suggesting that the reduced symbiotic competitiveness of the ΔotsA strain was due to the loss of stress resistance. We further demonstrated that fewer ΔotsA cells infected the M4 midgut initially but that fifth-instar nymphs exhibited similar symbiont population size as the wild-type strain. Together, these results demonstrated that the stress resistance role of otsA is important for B. insecticola to overcome the stresses it encounters during passage through the midgut regions to M4 in the initial infection stage but plays no role in resistance to stresses inside the M4 midgut in the persistent stage. IMPORTANCE Symbiotic bacteria have to overcome stressful conditions present in association with the host. In the Burkholderia-bean bug symbiosis, we speculated that a stress-resistant function of Burkholderia is important and that trehalose, known as a stress protectant, plays a role in the symbiotic association. Using otsA, the trehalose biosynthesis gene, and a mutant strain, we demonstrated that otsA confers Burkholderia with competitiveness when establishing a symbiotic association with bean bugs, especially playing a role in initial infection stage. In vitro assays revealed that otsA provides the resistance against osmotic stresses. Hemipteran insects, including bean bugs, feed on plant phloem sap, which may lead to high osmotic pressures in the midguts of hemipterans. Our results indicated that the stress-resistant role of otsA is important for Burkholderia to overcome the osmotic stresses present during the passage through midgut regions to reach the symbiotic organ.

Insecticidal activity and changes in midgut histology of the generalist herbivore, Spodoptera litura F. (Lepidoptera: Noctuidae) in response to seed extract of Annona squamosa Linn.

The tobacco caterpillar, Spodoptera litura F., is one of the most devastating, cosmopolitan polyphagous pests affecting major crops that significantly impact agricultural productivity. The present study aimed to evaluate the toxicological effect of seed extract of the medicinal plant, Annona squamosa L. and the histological effect on the midgut of Spodoptera litura F., under laboratory conditions. The crude methanolic extract of A. squamosa seed was tested against the third instar larvae of S. litura at five different concentrations viz., 0.5%, 1.0%, 1.5%, 2.0% and 2.5 % by leaf dip bioassay method. The seed extract exhibited larval mortality of 96.67%, 83.37 %,70%, 53.33% and 40 % at the concentrations 2.5 %, 2.0%, 1.5%, 1.0% and 0.5%, respectively.The results indicated that the response of larval mortality to the seed extract was dose-dependent. The dose of 0.5% showed the lowest mortality (40.00%), while the dose of 2.5 % showed maximum larval mortality (96.67%). Hence they were subjected to histological analysis. The anatomical sectioning of S. litura larval midgut treated with 0.5% concentration showed disruption in the peritrophic membrane and striated border of epithelial cells. The midgut of larvae treated with 2.5 % concentration showed an irregular epithelium and high vacuolization in the cytoplasmic cells. The results indicated that methanol extract of A. squamosa seed extract has the ability to cause changes in the midgut region, thereby affecting the digestion and nutrient absorption of S. litura larvae, which will lead to a debilitating effect on the development of larvae. Based on the present study A. squamosa seed extract will be a potential biopesticide for managing S. litura.

Symbiont coordinates stem cell proliferation, apoptosis, and morphogenesis of gut symbiotic organ in the stinkbug-Caballeronia symbiosis.

The bean bug Riptortus pedestris obtains a specific bacterial symbiont, Caballeronia insecticola (Burkholderia insecticola), from the environmental soil and harbors it in the posterior midgut region that is composed of hundreds of crypts. While newly hatched aposymbiotic insects possess primordial midgut crypts with little or no lumen, colonization of C. insecticola triggers swift development of the symbiotic organ, forming enlarged and opened crypts, and the symbiont subsequently fills the luminal cavities of those mature crypts. The cellular processes of crypt development triggered by C. insecticola colonization are poorly understood. Here we identified a fundamental mechanism of the symbiont-mediated midgut development by investigating cell cycles of intestinal epithelial cells. Intestinal stem cells of the bean bug are located and proliferate at the crypt base. Differentiated enterocytes migrate upward along the epithelial cell layer of the crypt as the midgut develops, induction of apoptosis in enterocytes primarily occurred on the tip side of the crypts, and apoptotic cells then eventually were shed from the crypts into the hemolymph. The proliferation rate of the stem cells at the base of the crypts was low while a high apoptotic rate was observed at the crypt tip in aposymbiotic insects, resulting in undeveloped short crypts. On the contrary, the gut-colonizing C. insecticola promoted the proliferation of the stem cells at the base of crypts and simultaneously inhibited apoptosis at the tip of crypts, resulting in a net growth of the crypts and the generation of a crypt lumen that becomes colonized by the bacterial symbiont. These results demonstrated that the Caballeronia symbiont colonization induces the development of the midgut crypts via finely regulating the enterocyte cell cycles, enabling it to stably and abundantly colonize the generated spacious crypts of the bean bug host.

Fine structure of the posterior midgut in the mite Anystis baccarum (L.)

Homology of the posterior midgut regions (PMG) in different phylogenetic lineages of acariform mites (superorder Acariformes) remains unresolved. In the order Trombidiformes, the ultrastructure of the PMG is known primarily in derived groups; thus this study focuses on species belonging to a relatively basal trombidiform family.PMG of Anystis baccarum consists of the colon and postcolon separated by a small intercolon. The fine structure of the colon and postcolon is close to that of the corresponding organs of sarcoptiform mites with the epithelium showing absorptive and endocytotic activity. The epithelial cells produce a variety of excretory vacuoles and a peritrophic matrix around the feces. Morover, the epithelium of the postcolon is characterized by the highest apical brush border and especially numerous mitochondria suggesting involvement in water and ion absorption. The intercolon functions as a sphincter lined with an epithelium capable of producing excretory granules. A pair of short blind extensions arises assimmetrically from the intercolon into the body cavity. Ultrastructurally, these extensions are similar to the arachnid Malpighian tubules and may be their reduced version. Rare endocrine-like cells have been observed in the colon and postcolon.

Eco-friendly biosynthesis of TiO2 nanoparticles using Desmostachya bipinnata extract: Larvicidal and pupicidal potential against Aedes aegypti and Spodoptera litura and acute toxicity in non-target organisms

The resistance to insecticides among insects, including mosquitoes and agricultural pests, and the impact of these compounds' environmental risks and health issues have motivated the proposition of eco-friendly alternatives. Thus, we aimed to explore the potential use of Desmostachya bipinnata for the biosynthesis of TiO2NPs and evaluate their larvicidal and pupicidal activity of target (Aedes aegypti and Spodoptera litura) and acute toxicity in non-target organisms (Toxorhynchites splendens and Eisenia fetida), at distinct concentrations, after 24 h of exposure. The characterization of the biosynthesized TiO2NPs was carried out by FT-IR, XRD, SEM, and EDX analysis. Under the UV–vis spectrum analysis, a sharp peak was recorded at 200 to 800 nm, which indicated the production of TiO2NPs by the plant extract. The SEM analysis revealed that the synthesized TiO2NPs were spherical with a diameter of 36.4 nm and were detected in the XRD spectrum analysis related to the TiO2NPs. The highest percentage of mortality recorded at 900 μg/mL was 96 % and 94 % in the 2nd instar of A. aegypti and S. litura larvae, respectively, and exhibited the LC50 and LC90 values 5 of 458.79 and 531.01 μg/mL, respectively. The biosynthesized TiO2NPs showed concentration-dependent increased pupal lethality for both A. aegypti and S. litura. We also observed increased detoxification enzyme activity (α esterase, β esterase, and glutathione-S-transferase) of A. aegypti and S. litura exposed to different concentrations of biosynthesized TiO2NPs as histopathological changes in the midgut region of these animals. On the other hand, the mortality rate of non-target organisms (T. splendens and E. fetida) was lower when exposed to TiO2NPs, compared to the high lethality induced by synthetic pesticides (cypermethrin and monocrotophos for E. fetida; and cypermethrin and temphos for T. splendens). Thus, our study provides pioneering evidence on the potential use of D. bipinnata-mediated TiO2NPs for controlling mosquito vectors and agricultural pest management.

Histological changes in the Dengue vector, Aedes aegypti (Diptera: Culicidae) larvae treated with neem oil loaded niosomes

Mosquitoes are one of the greatest threats to human health around the globe. They act as vectors for common diseases like Malaria, Dengue, Chikungunya, etc. Niosomes encapsulated with neem oil showed a significant mortality rate against Aedes aegypti larvae when treated for 24 h. In this study, the histological changes that led to the mortality of the Aedes aegypti mosquito larvae were studied. Organs of the late III-instar stage larvae such as head, optic lobes, cuticle, adipose tissue, midgut region, haemolymph were investigated. Several histological alterations such as disorientation of the brain and antenna in the head part, damage in the optic lobe and microvilli were observed. Total disruption was seen in the inner and outer retractor muscles of the larval body. The midgut and hindgut regions were disintegrated due to the damage to the fat bodies in the region. A Series of such histological changes in the body of mosquito larvae compared to the control larvae hindered metabolic functions leading to death. The results suggested that the neem oil loaded niosomes could be used as a biocontrol agent against the Dengue vector, Aedes aegypti larvae.

Comparative Study on Bacterial Population Dynamics of Foregut, Midgut, and Hindgut Content of Perionyx excavatus (Perrier) Isolated from Eco-friendly, Non-hazardous Vermicompost.

The ideal condition of earthworm gut promotes growth and multiplication of beneficial soil microorganisms eliminating pathogens and converts organic wastes into nutrients rich compost. The present study has been carried out to determine the population dynamics of earthworm gut bacteria and to find out relative abundance of different functional bacterial groups in the foregut, midgut, and hindgut of earthworm Perionyx excavatus. To assess bacterial diversity, a viable plate count method was adopted. In the different gut region of earthworm, aerobic heterotrophic, amylolytic, Bacillus, Gram-negative, proteolytic, fat hydrolyzing, nitrate-reducing, nitrifying, asymbiotic nitrogen-fixing, Azotobacter, and phosphate solubilizing bacterial populations ranged from 22.2 to 241.6 × 106, 8.0 to 171.60 × 106, 1.83 to 2.79 × 106, 10.68 to 23.04 × 104, 3.70 to 5.52 × 104, 59.60 to 208.40 × 104, 1.86 to 7.34 × 104, 10.94 to 19.78 × 104, 0.80 to 3.42 × 104, 7.83 to 13.70 × 104, 1.31 to 2.67 × 104cfu/ml gut suspension, respectively. The results of the one-way ANOVA revealed that the bacterial load of most of the bacterial groups was significantly higher (p < 0.05) in the hindgut region, followed by midgut and foregut. Only the density of the proteolytic group was significantly higher (p < 0.05) in the midgut region followed by foregut and hindgut. Starch hydrolyzing bacteria constitute the largest group of bacteria in the gut content. From principal component analysis, two components were extracted with the eigenvalues of 8.485 and 1.132. Agglomerative hierarchical cluster analysis revealed that the bacterial populations were clustered into four different groups. Quantitative variation among bacterial groups in earthworm's gut seems to determine the soil health and composting efficiency; from this point of view, the present study will provide a better understanding about different functional bacterial groups of earthworm's guts and might be helpful in sustainable agriculture and waste management.

Bacterial Competition Influences the Ability of Symbiotic Bacteria to Colonize Western Flower Thrips.

Symbiont mediated RNAi (SMR) is a promising method for precision control of pest insect species such as Western Flower Thrips (WFT). Two species of bacteria are known to be dominant symbiotic bacteria in WFT, namely BFo1 and BFo2 (Bacteria from Frankliniella occidentalis 1 and 2), as we here confirm by analysis of next-generation sequence data derived to obtain a reference WFT genome sequence. Our first demonstration of SMR in WFT used BFo2, related to Pantoea, isolated from a domesticated Dutch thrips population. However, for successful use of SMR as a thrips control measure, these bacteria need to successfully colonize different environmental thrips populations. Here, we describe a United Kingdom thrips population that does not harbour BFo2, but does contain BFo1, a species related to Erwinia. Attempts to introduce BFo2 indicate that this bacterium is unable to establish itself in the United Kingdom thrips, in contrast to successful colonization by a strain of BFo1 expressing green fluorescent protein. Fluorescence microscopy indicates that BFo1 occupies similar regions of the thrips posterior midgut and hindgut as BFo2. Bacterial competition assays revealed that a barrier to BFo2 establishing itself in thrips is the identity of the resident BFo1; BFo1 isolated from the United Kingdom thrips suppresses growth of BFo2 to a greater extent than BFo1 from the Dutch thrips that is permissive for BFo2 colonization. The ability of the latter strain of BFo1 to colonize the United Kingdom thrips is also likely attributable to its ability to out-compete the resident BFo1. Lastly, we observed that United Kingdom thrips pre-exposed to the Dutch BFo1 could then be successfully colonized by BFo2. These results indicate, for the first time, that microbial competition and strain differences can have a large influence on how symbiotic bacteria can colonize different populations of an insect species.

Larvicidal Effects Of Pongamiaglabra and Syzygiumaromaticum On Aedes Aegypti Mosquito Larvae

Mosquitoes are considered as the most nuisance causing vectors as they are responsible for a widespread of diseases. Malaria-borne mosquitoes are responsible of killing two to three million people and infect an estimated two hundred million or more annually. A host of other mosquito-borne diseases including filariasis, yellow fever, dengue, and encephalitis kill and debilitate hundreds of millions more. The present study investigates the bio efficacy of two commercially available plant oils from Pongamia glabra (Karanja) and Syzygium aromaticum (Clove). These two plant oils were tested against Aedes aegypti (Dengue fever mosquito) to determine their lethal concentration doses (LC50). A larval toxicity assay was performed where a stock solution of 10% was prepared for both the oils at a concentration range of 100 to 900 ppm and 90 to 1300 ppm was set for Pongamia glabra and Syzygium aromaticum respectively. A batch of each 10 IVth instar larvae was subjected to the assay with test, combination and a control set for a period of 24 and 48 hrs. LC50 of two oils were determined while the combination of two oils showed a synergistic effect. The effect of P. glabra and S. aromaticum on the anterior and posterior regions of the midgut region of A. aegypti larvae were found damage to epithelial cells, basement membrane, peritrophic membrane, fat body cells and nuclei were observed. The results of the study showed that the synergistic method of using common eco-safe combination of plant oils were effective to handle and monitor A. aegypti larva. Pongamia glabra and Syzygium aromaticum are among the successful mosquito control agents.

Insecticidal Serralysin of Serratia marcescens Is Detoxified in M3 Midgut Region of Riptortus pedestris.

Riptortus pedestris insect indiscriminately acquires not only the symbiotic bacterium Burkholderia insecticola, but also entomopathogens that are abundant in the soil via feeding. However, it is unclear how the host insect survives oral infections of entomopathogens. A previous study suggested that serralysin, a potent virulence factor produced by Serratia marcescens, suppresses cellular immunity by degrading adhesion molecules, thereby contributing to bacterial pathogenesis. Here, we observed that S. marcescens orally administered to R. pedestris stably colonized the insect midgut, while not exhibiting insecticidal activity. Additionally, oral infection with S. marcescens did not affect the host growth or fitness. When co-incubated with the midgut lysates of R. pedestris, serralysin was remarkably degraded. The detoxification activity against serralysin was enhanced in the midgut extract of gut symbiont-colonizing insects. The mRNA expression levels of serralysin genes were negligible in M3-colonizing S. marcescens. M3-colonizing S. marcescens did not produce serralysin toxin. Immunoblot analyses revealed that serralysin was not detected in the M3 midgut region. The findings of our study suggest that orally infected S. marcescens lose entomopathogenicity through host-derived degrading factors and suppression of serralysin.

Biomimetically synthesized Physalis minima fruit extract-based zinc oxide nanoparticles as eco-friendly biomaterials for biological applications

In the present study, the fruit extract of Physalis minima was used to synthesize zinc oxide nanoparticles (Pm-ZnO NPs). The first validation of the Pm-ZnO NPs production was obtained using a UV–Vis spectrophotometer at 327 nm. Pm-ZnO NPs crystalline nature was determined as hexagonal, relating to a joint corporation of powder diffraction standards (JCPDS) 75-1526 examined in XRD analysis. FTIR spectroscopy was used to identify functional groups associated with Pm-ZnO NPs, i.e., O–H and C=N. Pm-ZnO NPs were measured to be 50-150 nm in size and spherical in form using a TEM. Arrested augmentation of Gram-positive (Enterococci faecalis) and Gram-negative bacteria (Pseudomonas aeruginosa) populations was recorded at a 50 μg mL-1 concentration of Pm-ZnO NPs, where the zones of inhibition were measured as 31.4 ± 2.8 mm & 33.5 ± 0.8 mm, respectively. Pm-ZnO NPs inhibited the growth of bacteria E. faecalis and P. aeruginosa at a minimum concentration of 30 μg mL-1. Inverted microscopic images revealed that Pm-ZnO NPs diminished bacterial biofilm thickness at 50 and 100 μg mL−1. The biocompatibility of Pm-ZnO NPs was examined by potential antioxidant properties (DPPH assay). Bio-toxicity values of Pm-ZnO NPs were determined as LC50 – 76.5 (68.7–86.4) μg mL-1 and LC90 – 153.4 (132.4–189.6) μg mL-1 on 48 h treatment with Artemia salina nauplii and damages in the mid-gut region were examined under a stereomicroscope. Pm-ZnO NPs exhibited photocatalytic activity at 48 h duration against methylene blue (MB) dye. Concludingly, all the reported findings suggest the utilization of these generated nanoparticles as nanomedicine against microbial infection.

Pupation inhibition and larvicidal activity of tyrosinase on Culex pipiens third-instar larvae

BackgroundMosquitoes are considered to be the main vector of a variety of diseases in both humans and domesticated animals. The development of insecticide resistance and the limitations of traditional insecticide-based strategies have resulted in significant efforts to develop eco-friendly, alternative methods.Main bodyIn this study, nine fungi species were screened to produce tyrosinase enzyme. This was done in order to evaluate its inhibitory activity against Culex pipiens third-instar larvae. The extracellular tyrosinase was produced by five strains. Aspergillus tamarii NRC3 was found to possess the highest tyrosinase activity and was therefore used in this study. Some factors were studied to enhance the production of the enzyme. The enzyme was partially purified using ammonium sulfate at a 70% saturation, giving 1.861 purification fold. The toxicity on Culex pipiens varied, depending upon the tyrosinase concentration and the period of exposure. In addition, notable histological effects were seen in the midgut region.ConclusionsA concentration of 80% on third-stage larvae showed 90% inhibition in the formation of pupae at 72 h post-treatment. In addition, a significant cellular microvillus disruption was seen in the midgut region at 24 h post-treatment.

Exploring the region-wise diversity and functions of symbiotic bacteria in the gut system of wood-feeding termite, Coptotermes formosanus, toward the degradation of cellulose, hemicellulose, and organic dyes.

The wood-feeding termite Coptotermes formosanus represents a unique and impressive system for lignocellulose degradation. The highly efficient digestion of lignocellulose is achieved through symbiosis with gut symbionts like bacteria. Despite extensive research during the last three decades, diversity of bacterial symbionts residing in individual gut regions of the termite and their associated functions is still lacking. To this end, cellulose, xylan, and dye-decolorization bacteria residing in foregut, midgut, and hindgut regions of C. formosanus were enlisted by using enrichment and culture-dependent molecular methods. A total of 87 bacterial strains were successfully isolated from different gut regions of C. formosanus which belonged to 27 different species of 10 genera, majorly affiliated with Proteobacteria (80%) and Firmicutes (18.3%). Among the gut regions, 37.9% of the total bacterial isolates were observed in the hindgut that demonstrated predominance of cellulolytic bacteria (47.6%). The majority of the xylanolytic and dye-decolorization bacteria (50%) were obtained from the foregut and midgut, respectively. Actinobacteria represented by Dietza sp. was observed in the hindgut only. Based on species richness, the highest diversity was observed in midgut and hindgut regions each of which harbored seven unique bacterial species. The members of Enterobacter, Klebsiella, and Pseudomonas were common among the gut regions. The lignocellulolytic activities of the selected potential bacteria signpost their assistance to the host for lignocellulose digestion. The overall results indicate that C. formosanus harbors diverse communities of lignocellulolytic bacteria in different regions of the gut system. These observations will significantly advance our understanding of the termite-bacteria symbiosis and their microbial ecology uniquely existed in different gut regions of C. formosanus, which may further shed a light on its potential values at termite-modeled biotechnology.