scholarly journals Entomopathogenic Nematodes: Their Characterization, Bio-Control Properties and New Perspectives

2021 ◽  
Author(s):  
Himani Sharma ◽  
Aasha Rana ◽  
Aashaq H. Bhat ◽  
Ashok K. Chaubey

The insect parasitoid nematodes are a means boon to agronomy and serve as important bio-pesticides for controlling crop damaging insect pests. These nematodes inhabit moist soils and have been to exist in all the continents excluding Polar Regions. These nematodes have 3rd larval stage infective which is the only free living stage existing outside the host. These infective stages are mutually associated with bacteria which reside in their alimentary canal and duo are responsible for mortality of the insect host. These nematodes are currently given great attention by scientific community because of their insect killing properties and can be used to replace hazardous pesticides. These nematodes include various species belonging to genus Heterorhabditis and Steinernema, and members of insectivorous group of genus Oscheius. Before their use as bio-control agents, these nematodes need to be properly identified. Currently, these nematodes are characterized by using morphological and morphometrical parameters and advanced molecular tools including cross hybridization and scanning electron microscope studies. Their associated bacterial partners are studied through advanced molecular and biochemical techniques. The properly characterized nematodes having more entomopathogenic properties can be easily mass produced through in vitro and in vivo methods. They can be formulated in various carrier materials and supplied to farmers for effective control of damaging insect pests. Several countries have formulated various useful products of entomopathogenic nematodes which are available in markets for use by the farmer community and some have given very effective results. India is still at the early stage in the use of these nematodes for bio-control of insects in agronomy. More research in this field needs to be carried, especially in India to produce effective indigenous nematode products which may prove a boon for agriculture.


PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0242645
Author(s):  
Francois du Preez ◽  
Antoinette Paula Malan ◽  
Pia Addison

Entomopathogenic nematodes (EPNs) have been successfully applied as biological control agents against above ground and soil stages of insect pests. However, for commercial application, it is crucial to mass culture these nematodes using in vitro liquid culture technology, as it is not attainable when using susceptible insects as hosts. Lobesia vanillana (Lepidoptera: Tortricidae) is regarded a sporadic pest of wine grapes in South Africa. The in vivo- and in vitro-cultured South African EPNs, Steinernema yirgalemense and Steinernema jeffreyense (Rhabditida: Steinernematidae), were evaluated against larvae and pupae of L. vanillana in laboratory bioassays. For larvae, high mortality was observed for all treatments: In vitro-cultured S. yirgalemense (98%) performed better than S. jeffreyense (73%), while within in vivo cultures, there was no difference between nematode species (both 83%). No significant difference was detected between in vivo- and in vitro cultures of the same nematode species. The LD50 of the in vitro-cultured S. yirgalemense, was 7.33 nematodes per larva. Mortality by infection was established by dissecting L. vanillana cadavers and confirming the presence of nematodes, which was > 90% for all treatments. Within in vitro cultures, both S. yirgalemense and S. jeffreyense were able to produce a new cohort of infective juveniles from L. vanillana larvae. Pupae, however, were found to be considerably less susceptible to EPN infection. This is the first study on the use of EPNs to control L. vanillana. The relative success of in vitro-cultured EPN species in laboratory assays, without any loss in pathogenicity, is encouraging for further research and development of this technology.



2017 ◽  
Vol 2017 ◽  
pp. 1-8
Author(s):  
Lixia Ji ◽  
Lixia Cheng ◽  
Zhihong Yang

Objective.Lens osmotic expansion, provoked by overactivated aldose reductase (AR), is the most essential event of sugar cataract. Chloride channel 3 (Clcn3) is a volume-sensitive channel, mainly participating in the regulation of cell fundamental volume, and P-glycoprotein (P-gp) acts as its modulator. We aim to study whether P-gp and Clcn3 are involved in lens osmotic expansion of galactosemic cataract.Methods and Results.In vitro, lens epithelial cells (LECs) were primarily cultured in gradient galactose medium (10–60 mM), more and more vacuoles appeared in LEC cytoplasm, and mRNA and protein levels of AR, P-gp, and Clcn3 were synchronously upregulated along with the increase of galactose concentration. In vivo, we focused on the early stage of rat galactosemic cataract, amount of vacuoles arose from equatorial area and scattered to the whole anterior capsule of lenses from the 3rd day to the 9th day, and mRNA and protein levels of P-gp and Clcn3 reached the peak around the 9th or 12th day.Conclusion. Galactosemia caused the osmotic stress in lenses; it also markedly leads to the upregulations of AR, P-gp, and Clcn3 in LECs, together resulting in obvious osmotic expansion in vitro and in vivo.



2021 ◽  
Author(s):  
Yipu Wang ◽  
Dong Mei ◽  
Xinyi Zhang ◽  
Da-Hui Qu ◽  
Ju Mei ◽  
...  

With increase of social aging, Alzheimer's disease (AD) has been one of the serious diseases threatening human health. The occurrence of A<i>β </i>fibrils<i> </i>or plaques is recognized as the hallmark of AD.<i> </i>Currently, optical imaging has stood out to be a promising technique for the imaging of A<i>β</i> fibrils/plaques and the diagnosis of AD. However, restricted by their poor blood-brain barrier (BBB) penetrability, short-wavelength excitation and emission, and aggregation-caused quenching (ACQ) effect, the clinically used gold-standard optical probes such as <a>thioflavin</a> T (ThT) and thioflavin S (ThS), are not effective enough in the early diagnosis of AD <i>in vivo</i>. Herein, we put forward an “all-in-one” design principle and demonstrate its feasibility in developing high-performance fluorescent probes which are specific to A<i>β</i> fibrils/plaques and promising for super-early <i>in</i>-<i>vivo</i> diagnosis of AD. As a proof of concept, a simple rod-like amphiphilic NIR fluorescent AIEgen, i.e., AIE-CNPy-AD, is developed by taking the specificity, BBB penetration ability, deep-tissue penetration capacity, high signal-to-noise ratio (SNR) into consideration. AIE-CNPy-AD is constituted by connecting the electron-donating and accepting moieties through single bonds and tagging with a propanesulfonate tail, giving rise to the NIR fluorescence, aggregation-induced emission (AIE) effect, amphiphilicity, and rod-like structure, which in turn result in high binding-affinity and excellent specificity to A<i>β</i> fibrils/plaques, satisfactory ability to penetrate BBB and deep tissues, ultrahigh SNR and sensitivity, and high-fidelity imaging capability. <i>In-vitro, ex-vivo,</i> and <i>in-vivo</i> <a>identifying of A<i>β</i> fibrils/plaques</a> in different strains of mice indicate that AIE-CNPy-AD holds the universality to the detection of A<i>β</i> fibrils/plaques. It is noteworthy that AIE-CNPy-AD is even able to trace the small and sparsely distributed A<i>β</i> fibrils/plaques in very young AD model mice such as 4-month-old APP/PS1 mice which are reported to be the youngest mice to have A<i>β</i> deposits in brains, suggesting its great potential in diagnosis and intervention of AD at a super-early stage.



2020 ◽  
Author(s):  
María A. Duque-Correa ◽  
David Goulding ◽  
Claire Cormie ◽  
Catherine Sharpe ◽  
Judit Gali Moya ◽  
...  

ABSTRACTHundreds of millions of people are infected with whipworms (Trichuris trichiura), large metazoan parasites that live in the caecum and proximal colon. Whipworms inhabit distinct multi-intracellular epithelial burrows that have been described as syncytial tunnels. However, the interactions between first-stage (L1) larvae and the host epithelia that determine parasite invasion and establishment in the syncytium remain unclear. In vivo experiments investigating these events have been severely hampered by the limited in situ accessibility to intracellular infective larvae at the bottom of the crypts of Lieberkühn, and the lack of genetic tools such as fluorescent organisms that are readily available for other pathogens but not parasitic nematodes. Moreover, cell lines, which do not mimic the complexity of the intestinal epithelium, have been unsuccessful in supporting infection by whipworm larvae. Here, we show that caecaloids grown in an open crypt-like conformation recapitulate the caecal epithelium. Using this system, we establish in vitro infections with T. muris L1 larvae for the first-time, and provide clear evidence that syncytial tunnels are formed at this early stage. We show that larval whipworms are completely intracellular but woven through multiple cells. Using the caecaloids, we are able to visualise the pathways taken by the larvae as they burrow through the epithelial cells. We also demonstrate that larvae degrade the mucus layers overlaying the epithelium, enabling them to access the cells below. We show that early syncytial tunnels are composed of enterocytes and goblet cells that are alive and actively interacting with the larvae during the first 24 h of the infection. Progression of infection results in damage to host cells and by 72 h post-infection, we show that desmosomes of cells from infected epithelium widen and some host cells appear to become liquified. Collectively, our work unravels processes mediating the intestinal epithelium invasion by whipworms and reveals new specific interactions between the host and the parasite that allow the whipworm to establish on its multi-intracellular niche. Our study demonstrates that caecaloids can be used as a relevant in vitro model to investigate the infection biology of T. muris during the early colonisation of its host.



2020 ◽  
Author(s):  
Hua Sang ◽  
Jiali Liu ◽  
Fang Zhou ◽  
Xiaofang Zhang ◽  
Jingwei Zhang ◽  
...  

<p></p><p>Key events including antibody-antigen affinity, ADC internalization, trafficking and lysosomal proteolysis-mediated payload release combinatorially determine the therapeutic efficacy and safety for ADCs. Nevertheless, a universal technology that efficiently and conveniently evaluates the involvement of these above elements to ADC payload release and hence the final therapeutic outcomes for mechanistic studies and quality assessment is lacking. Considering the plethora of ADC candidates under development owing to the ever-evolving linker and drug chemistry, we developed a TArget-Responsive Subcellular Catabolism (TARSC) approach that measures catabolites kinetics for given ADCs and elaborates how each individual step ranging from antigen binding to lysosomal proteolysis affects ADC catabolism by targeted interferences. Using a commercial and a biosimilar ado-trastuzumab emtansine (T-DM1) as model ADCs, we recorded unequivocal catabolites kinetics for the two T-DM1s in the presence and absence of the targeted interferences. Their negligible differences in TARSC profiles fitting with their undifferentiated therapeutic outcomes suggested by <i>in vitro</i> viability assays and <i>in vivo</i> tumor growth assays, highlighting TARSC analysis as a good indicator of ADC efficacy and bioequivalency. Lastly, we demonstrated the use of TARSC in assessing payload release efficiency for a new Trastuzumab-toxin conjugate. Collectively, we demonstrated the use of TARSC in characterizing ADC catabolism at (sub)cellular level, and in systematically depicting whether given target proteins affect ADC payload release and hence therapeutic efficacy. We anticipate its future use in high-throughput screening, quality assessment and mechanistic understanding of ADCs for drug R&D before proceeding to costly <i>in vivo</i> experiments.</p><br><p></p>



2020 ◽  
Author(s):  
Vera A van der Weijden ◽  
Meret Schmidhauser ◽  
Mayuko Kurome ◽  
Johannes Knubben ◽  
Veronika L Flöter ◽  
...  

Abstract Background: The transcriptional changes around the time of embryonic genome activation in pre-implantation embryos indicate that this process is highly dynamic. In vitro produced porcine blastocysts are known to be less competent than in vivo developed blastocysts. To understand the conditions that compromise developmental competence of in vitro embryos, it is crucial to evaluate the transcriptional profile of porcine embryos during pre-implantation stages. In this study, we investigated the transcriptome dynamics in in vivo developed and in vitro produced 4-cell embryos, morulae and hatched blastocysts.Results: In vivo developed and in vitro produced embryos displayed largely similar transcriptome profiles during development. Enriched canonical pathways from the 4-cell to the morula transition that were shared between in vivo developed and in vitro produced embryos included oxidative phosphorylation, tRNA charging, and EIF2 signaling. The shared canonical pathways from the morula to the hatched blastocyst transition were 14-3-3-mediated signaling, signaling of Rho family GTPases, and NRF2-mediated oxidative stress response. The in vivo developed and in vitro produced hatched blastocysts were compared to identify molecular signaling pathways indicative of lower developmental competence of in vitro produced hatched blastocysts. A higher metabolic rate and expression of the arginine transporter SLC7A1 were found in in vitro produced hatched blastocysts.Conclusions: Our findings suggest that embryos with compromised developmental potential are arrested at an early stage of development, while embryos developing to the hatched blastocyst stage display largely similar transcriptome profiles, irrespective of the embryo source. The hatched blastocysts derived from the in vitro fertilization-pipeline showed an enrichment in molecular signaling pathways associated with lower developmental competence, compared to the in vivo developed embryos.



2020 ◽  
Author(s):  
L Caló ◽  
E Hidari ◽  
M Wegrzynowicz ◽  
JW Dalley ◽  
BL Schneider ◽  
...  

AbstractαSynuclein aggregation at the synapse is an early event in Parkinson’s disease and is associated with impaired striatal synaptic function and dopaminergic neuronal death. The cysteine string protein (CSPα) and αsynuclein have partially overlapping roles in maintaining synaptic function and mutations in each cause neurodegenerative diseases. CSPα is a member of the DNAJ/HSP40 family of co-chaperones and like αsynuclein, chaperones the SNARE complex assembly and neurotransmitter release. αSynuclein can rescue neurodegeneration in CSPαKO mice. However, whether αsynuclein aggregation alters CSPα expression and function is unknown. Here we show that αsynuclein aggregation at the synapse induces a decrease in synaptic CSPα and a reduction in the complexes that CSPα forms with HSC70 and STGa. We further show that viral delivery of CSPα rescues in vitro the impaired vesicle recycling in PC12 cells with αsynuclein aggregates and in vivo reduces synaptic αsynuclein aggregates restoring normal dopamine release in 1-120hαsyn mice. These novel findings reveal a mechanism by which αsynuclein aggregation alters CSPα at the synapse, and show that CSPα rescues αsynuclein aggregation-related phenotype in 1-120hαsyn mice similar to the effect of αsynuclein in CSPαKO mice. These results implicate CSPα as a potential therapeutic target for the treatment of early-stage PD.



2018 ◽  
Vol 62 (4) ◽  
pp. e01505-17 ◽  
Author(s):  
R. S. Jumani ◽  
K. Bessoff ◽  
M. S. Love ◽  
P. Miller ◽  
E. E. Stebbins ◽  
...  

ABSTRACTCryptosporidiosis causes life-threatening diarrhea in children under the age of 5 years and prolonged diarrhea in immunodeficient people, especially AIDS patients. The standard of care, nitazoxanide, is modestly effective in children and ineffective in immunocompromised individuals. In addition to the need for new drugs, better knowledge of drug properties that drivein vivoefficacy is needed to facilitate drug development. We report the identification of a piperazine-based lead compound forCryptosporidiumdrug development, MMV665917, and a new pharmacodynamic method used for its characterization. The identification of MMV665917 from the Medicines for Malaria Venture Malaria Box was followed by dose-response studies,in vitrotoxicity studies, and structure-activity relationship studies using commercial analogues. The potency of this compound againstCryptosporidium parvumIowa and field isolates was comparable to that againstCryptosporidium hominis. Furthermore, unlike nitazoxanide, clofazimine, and paromomycin, MMV665917 appeared to be curative in a NOD SCID gamma mouse model of chronic cryptosporidiosis. MMV665917 was also efficacious in a gamma interferon knockout mouse model of acute cryptosporidiosis. To determine if efficacy in this mouse model of chronic infection might relate to whether compounds are parasiticidal or parasitistatic forC. parvum, we developed a novelin vitroparasite persistence assay. This assay suggested that MMV665917 was parasiticidal, unlike nitazoxanide, clofazimine, and paromomycin. The assay also enabled determination of the concentration of the compound required to maximize the rate of parasite elimination. This time-kill assay can be used to prioritize early-stageCryptosporidiumdrug leads and may aid in planningin vivoefficacy experiments. Collectively, these results identify MMV665917 as a promising lead and establish a new method for characterizing potential anticryptosporidial agents.



Plant Disease ◽  
2021 ◽  
Author(s):  
Phinda Magagula ◽  
Nicky Taylor ◽  
Velushka Swart ◽  
Noëlani van den Berg

Rosellinia necatrix is the causal agent of white root rot (WRR), a fatal disease affecting many woody plants, including avocado (Persea americana). As with other root diseases, an integrated approach is required to control WRR. No fully effective control methods are available, and no chemical or biological agents against R. necatrix have been registered for use on avocado in South Africa. Fluazinam has shown promising results in the greenhouse and field in other countries, including Spain. The current study aimed to investigate the potential of a fumigant, chloropicrin, and biological control agents (B-Rus, Beta-Bak, Mity-Gro, and Trichoderma) against R. necatrix both in vitro and in vivo as compared with fluazinam. In a greenhouse trial, results showed that Trichoderma and B-Rus were as effective as fluazinam at inhibiting R. necatrix in vitro and suppressed WRR symptoms when applied before inoculation with R. necatrix. In contrast, Mity-Gro and Beta-Bak failed to inhibit the pathogen in vitro and in the greenhouse trial, despite application of the products to plants before R. necatrix infection. Fluazinam suppressed WRR symptoms in plants when applied at the early stages of infection, whereas chloropicrin rendered the pathogen nonviable when used as a preplant treatment. Plants treated with Trichoderma, B-Rus, and fluazinam sustained dry mass production and net CO2 assimilation by maintaining the green leaf tissues despite being infected with the pathogen. This study has important implications for the integrated management of WRR.



2021 ◽  
Vol 17 (9) ◽  
pp. 1812-1823
Author(s):  
Fei Liu ◽  
Xinyu Wang ◽  
Shujun Li ◽  
Yiheng Liao ◽  
Xinxin Zhan ◽  
...  

Ti–24Nb–4Zr–8Sn (Ti2448) alloys, with a relatively low elastic modulus and unique mechanical properties, are desirable materials for oral implantation. In the current study, a multifaceted strontium-incorporating nanotube coating was fabricated on a Ti2448 alloy (Ti2-NTSr) through anodization and hydrothermal procedures. In vitro, the Ti2-NTSr specimens demonstrated better osteogenic properties and more favorable osteoimmunomodulatory abilities. Moreover, macrophages on Ti2-NTSr specimens could improve the recruitment and osteogenic differentiation of osteoblasts. In vivo, dense clots with highly branched, thin fibrins and small pores existed on the Ti2-NTSr implant in the early stage after surgery. Analysis of the deposition of Ca and P elements, hard tissue slices and the bone-implant contact rate (BIC%) of the Ti2-NTSr implants also showed superior osseointegration. Taken together, these results demonstrate that the Ti2-NTSr coating may maximize the clinical outcomes of Ti2448 alloys for implantation applications.



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