scholarly journals Production of germ-free mosquitoes via transient colonisation allows stage-specific investigation of host-microbiota interactions

Author(s):  
Ottavia Romoli ◽  
Johan Claes Schönbeck ◽  
Siegfried Hapfelmeier ◽  
Mathilde Gendrin

AbstractThe mosquito microbiota impacts the physiology of its host and is essential for normal larval development, thereby influencing transmission of vector-borne pathogens. Germ-free mosquitoes generated with current methods show larval stunting and developmental deficits. Therefore, functional studies of the mosquito microbiota have so far mostly been limited to antibiotic treatments of emerging adults. In this study, we developed a novel approach to produce germ-free Aedes aegypti mosquitoes. It is based on reversible colonisation with bacteria genetically modified to allow complete decolonisation at any developmental stage. We show that, unlike germ-free mosquitoes previously produced using sterile diets, reversibly colonised mosquitoes show no developmental retardation and reach the same size as control adults. This allowed us to uncouple the study of the microbiota in larvae and adults. In adults, we detected no impact of bacterial colonisation on mosquito fecundity or longevity. In larvae, we performed a transcriptome analysis and diet supplementation experiments following decolonisation during the third larval instar. Our data suggest that bacteria support larval development by contributing to folate biosynthesis and by enhancing energy storage. Our study establishes a novel tool to study the microbiota in insects and deepens our knowledge on the metabolic contribution of bacteria to mosquito development.

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ottavia Romoli ◽  
Johan Claes Schönbeck ◽  
Siegfried Hapfelmeier ◽  
Mathilde Gendrin

AbstractThe mosquito microbiota impacts the physiology of its host and is essential for normal larval development, thereby influencing transmission of vector-borne pathogens. Germ-free mosquitoes generated with current methods show larval stunting and developmental deficits. Therefore, functional studies of the mosquito microbiota have so far mostly been limited to antibiotic treatments of emerging adults. In this study, we introduce a method to produce germ-free Aedes aegypti mosquitoes. It is based on reversible colonisation with bacteria genetically modified to allow complete decolonisation at any developmental stage. We show that, unlike germ-free mosquitoes previously produced using sterile diets, reversibly colonised mosquitoes show no developmental retardation and reach the same size as control adults. This allows us to uncouple the study of the microbiota in larvae and adults. In adults, we detect no impact of bacterial colonisation on mosquito fecundity or longevity. In larvae, data from our transcriptome analysis and diet supplementation experiments following decolonisation suggest that bacteria support larval development by contributing to folate biosynthesis and by enhancing energy storage. Our study establishes a tool to study the microbiota in insects and deepens our knowledge on the metabolic contribution of bacteria to mosquito development.


2020 ◽  
Author(s):  
Fariha Azalea

University is relatively considered a stressful moment in the life of students due to numerous academic workloads and academic activities. The situation is further aggravated by the fact that some university students are in emerging adulthood, a developmental period which is psychologically fraught with uncertainty, instability and identity issues among others. Added to these, the context of most universities like Cameroon which is marred with political, economic and social turbulence common to other developing societies in the sub Saharan region makes life unbearable. Looking at the challenges that confront tertiary education students in the third decade of life, increases possibilities of fears that they will founder thus narrowing the route to a blossomed transition into adulthood and through the university from home into the world of work. However, observations reveal that some have remained hopeful as they continuously believe in themselves and their worth. As such, they have resiliently shrugged off the vast burden placed on them by the adult society as they struggle intentionally with continuous efforts to succeed. Being hopeful and self-efficacy beliefs are observed to be some of the effective drivers that pull emerging adults through the storms of university transition thus facilitating positive development into subsequent life stages. Unfortunately just a paucity of literature albeit theoretically actually narrates via scholarly corridors the monumental successes recorded by students as they sail flourishingly through university in the midst of storms an in the third decade of life. This paper examines and addresses the foregoing through the lenses of some theories.


1957 ◽  
Vol s3-98 (41) ◽  
pp. 123-150
Author(s):  
JOAN M. WHITTEN

The fate of the tracheal system is traced from the first larval instar to the adult stage. The basic larval pattern conforms to that shown for other Diptera Cyclorrhapha (Whitten, 1955), and is identical in all three instars. According to previous accounts the adult system directly replaces the larval: the larval system is partly shed, partly histolysed, and the adult system arises from imaginal cell clusters independently of the preceding larval system. In contrast, it is shown here that in the cephalic, thoracic, and anterior abdominal region there is a definite continuity in the tracheal system, from larval, through pupal to the adult stage, whereas in the posterior abdominal region the larval system is histolysed, and the adult system is independent of it in origin. Moreover, in the pupal stage this region is tracheated by tracheae arising from the anterior abdominal region and belonging to a distinct pupal system. Moulting of the tracheal linings is complete at the first and second larval ecdyses, but incomplete at the third larval-pupal and pupal-adult ecdyses. In consequence, in both pupal and adult systems there are tracheae which are secreted around preexisting tracheae, others formed as new ‘branch’ tracheae, and those which have been carried over from the previous instar. In the adult the newly formed tracheae of the posterior abdominal region fall into a fourth category. Most of the adult thoracic air sacs correspond to new ‘branch’ tracheae of other instars. The pre-pupal moult and instar are discussed with reference to the tracheal system and tentative suggestions are made concerning the true nature of the pre-pupal cuticle. There is no pre-pupal tracheal system. Events traced for Drosophila would seem to be general for Cyclorrhapha, both Acalypterae and Calypterae. The separate fates of the anterior and posterior abdom inal systems, in contrast with the straightforward development in Dipterc Nematocera, would appear to mark a distinct step in the evolution of the system in Diptera.


2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Naz Fatima ◽  
Tasleem Akhtar ◽  
Nadeem Sheikh

Hepatocellular carcinoma is one of the fatal malignancies and is considered as the third leading cause of death. Mutations, genetic modifications, dietary aflatoxins, or impairments in the regulation of oncogenic pathways may bring about liver cancer. An effective barrier against hepatotoxins is offered by gut-liver axis as a change in gut permeability and expanded translocation of lipopolysaccharides triggers the activation of Toll-like receptors which stimulate the process of hepatocarcinogenesis. Prebiotics, nondigestible oligosaccharides, have a pivotal role to play when it comes to inducing an antitumor effect. A healthy gut flora balance is imperative to downregulation of inflammatory cytokines and reducing lipopolysaccharides induced endotoxemia, thus inducing the antitumor effect.


2013 ◽  
Vol 47 (1) ◽  
pp. 24-34
Author(s):  
I. V. Dolinskaya

Abstract Larval cranial setae of each larval instar of 66 species belonging to 35 genera of Palaeartic Notodontid moths from Ukraine and Far East of Russia (Primorskii krai) was examined with the use of a scanning electron microscope. A comparison with outgroup species - Lasiocampoidea (Lasiocampidae), Sphingoidea (Sphingidae) and Noctuoidea (Erebidae: Lymantriinae, Arctiinae; Noctuidae) is conducted. Main kinds of setae during larval development and their transformation are discussed. Possible apomorphic and plesiomorphic states of the different characters are discussed in relation to the different taxa.


2014 ◽  
Vol 11 (2) ◽  
pp. 943-951
Author(s):  
Baghdad Science Journal

The following dilution 5×10-1, 10-1, 10?2 , 10-3 gm/L for the indigenous isolate of Bacillus thuringiensis bacteria and the commercially isalate were used for experiments against the different stages of fig moth of E.cautella which exposed by filter paper method. The results showed that mortality of larval stages was increased with the increasing concentration of the biocide, in addition to increase in the mortality of the larval stages reached to the highest percentage in the third days of treatment of the larval stage in comparison with the first and second days of exposure. The results also showed that the sensitivity of larval stages was increased in first and second instars while reduced in the last instars .The high percentage of first instar mortality for the indigenous isolate in the concentration of 5×10-1 was 72.8% , while the low percentage of mortality showed in the concentration of 5×10-1 for the fifth instar larvae which was 13.3% in third days of treatment while a high percentage of mortality was showed for the first instar larvae for the commercially isulate in the concentration of 5×10-1 was 59.4% Furthermore, low percentage of mortality was shown in the concentration of 5×10-1 in fifth instar larval which was 8.3% in the third days of treatment. The results also showed that the indigenous isolated was more effective than the commercially produced bacteria for killing larval instars of fig moth E.cautella .The total percentage of larval instar mortality reached to 44.5 % after the third days of treatment in concentration 5×10-1 in the indigenous isolate , and it was 33.8 % in the commercially produced bacteria .


1972 ◽  
Vol 50 (6) ◽  
pp. 713-719 ◽  
Author(s):  
William E. Bradshaw ◽  
L. Philip Lounibos

Wyeomyia smithii diapause in the third larval instar. Long days avert or terminate and short days promote or maintain diapause. Diapause occurs early in the third instar and may be terminated by photoperiodic stimuli without the intervention of chilling or other factors. Fifty percent termination of diapause requires about 3 long days and another [Formula: see text] days are consumed in the third instar for postdiapause development. The critical daylength is identical for both the initiation and termination of diapause, 14.75 h of light per day. But, the photoperiodic clock monitoring diapause decisions is several times as accurate during initiation as in termination, reflecting the more drastic environmental consequences of development misdirection in the fall than in the spring. This accuracy is further enhanced by a prolongation of the second instar under short-day conditions. The doubling in the duration of the second instar exhibits the same critical daylength properties as diapause determination.The third instar is divisible into four distinct developmental periods: prediapause, diapause, termination of diapause, and postdiapause. Methods for quantifying these periods are presented. Similar manipulations could be employed for other diapausing arthropods, regardless of the stage at which dormancy occurs or the cues used in its regulation.


Development ◽  
1975 ◽  
Vol 33 (2) ◽  
pp. 487-498
Author(s):  
Andreas Dübendorfer ◽  
Glen Shields ◽  
James H. Sang

Embryos of Drosophila melanogaster, 6–8 h after oviposition, were dissociated and the cells cultured in vitro. Besides larval cell types, imaginal disc cells, assembled and growing in bloated monolayered vesicles, were obtained. The cells of these vesicles become competent to differentiate adult structures when treated with α-ecdysone or ecdysterone in vitro. Recognizable patterns of the adult fly are not formed though. If metamorphosis of imaginal cell vesicles from in vitro-cultures is induced in vivo by transplantation into host larvae of various ages within the third larval instar, recognizable patterns can differentiate provided the host larva does not metamorphose prior to 2 days after transplantation. The frequency of specific patterns in the implants can be increased by providing 9 days of culture in vivo (adult host flies) before metamorphosis. Passage through the third larval instar is not essential for these cells to produce identifiable patterns since culture in adult flies alone can achieve this. The quality of the differentiated pattern is not correlated with the extent of cell proliferation in the cultured tissues. The problem of pattern specification in vitro and in vivo is discussed.


Author(s):  
Poulami Sarkar ◽  
Svetlana Kontsedalov ◽  
Galina Lebedev ◽  
Murad Ghanim

Several vector-borne plant pathogens have evolved mechanisms to exploit and hijack vector host cellular, molecular and defense mechanisms for their transmission. Over the past few years, Liberibacter species, which are transmitted by several psyllid vectors, have become an economically important group of pathogens that devastated the citrus industry and caused tremendous losses to many other important crops worldwide. The molecular mechanisms underlying the interactions of Liberibacter species with their psyllid vectors are poorly studied. Candidatus Liberibacter solanacearum (CLso) associated with important vegetable diseases is transmitted by the carrot psyllid, Bactericera trigonica in a persistent manner. Here, we elucidated the role of B. trigonica Arp2/3 protein complex, which plays a major role in the regulation of the actin cytoskeleton, in the transmission of CLso. CLso co-localized with ArpC2, a key protein in this complex, and this co-localization strongly associated with actin filaments. Silencing the psyllid ArpC2 disrupted the co-localization and the dynamics of F-actin. Silencing RhoGAP21 and Cdc42, which act in the signaling cascade leading to upregulation of Arp2/3 and F-actin bundling, also showed similar results. On the other hand, silencing ArpC5, another component of the complex, did not induce any significant effects on F-actin formation. Finally, ArpC2 silencing caused 73.4% reduction in CLso transmission by psyllids, strongly suggesting that its transmission by B. trigonica is cytoskeleton-dependent and it interacts with ArpC2 to exploit the intracellular actin nucleation process for transmission. Targeting this unique interaction could lead to developing a novel strategy for the management of Liberibacter-associated diseases. IMPORTANCE Plant diseases caused by vector-borne pathogens are responsible for tremendous losses and threaten some of the most important agricultural crops. A good example is the citrus greening disease caused by bacteria of the genus Liberibacter and transmitted by psyllids, and has devastated the citrus industry in the US, China and Brazil. Here we show that the psyllid-transmitted Candidatus Liberibacter solanacearum (CLso) employs the actin cytoskeleton of psyllid gut cells, specifically the ArpC2 protein in the Arp2/3 complex of this system, for movement and transmission in the vector. Silencing ArpC2 dramatically influenced interaction of CLso with the cytoskeleton and decreased the bacteria transmission to plants. This system could be targeted for developing a novel approach for the control of Liberibacter- associated diseases.


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