Preparation of Red Palm Weevil Rhynchophorus Ferrugineus (Olivier) (Coleoptera: Dryophthoridae) Germ-free Larvae for Host-gut Microbes Interaction Studies

Muhammad, Aliyu Idris; Habineza, Prosper; Hou, Youming; Shi, Zhanghong

doi:10.21769/bioprotoc.3456

Cited by 5 publications

(4 citation statements)

References 24 publications

(3 reference statements)

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“…2018; Muhammad et al . 2019). The other half of the non‐axenic eggs served as the conventionally reared group (CR) which was treated with only sterilized water.…”

Section: Methodsmentioning

confidence: 99%

“…Fresh eggs were collected after oviposition and placed on moist absorbent paper to avoid water loss. At 60 h after oviposition, half of the collected eggs were surface sterilized with 75% ethanol for 8 min and washed with sterilized water 3 times to obtain germ-free (GF) larvae (Kietz et al 2018;Muhammad et al 2019). The other half of the non-axenic eggs served as the conventionally reared group (CR) which was treated with only sterilized water.…”

Section: P Versicolora Rearing and Preparation Of Conventional And Germ-free Larvaementioning

confidence: 99%

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Metabolic and immunological effects of gut microbiota in leaf beetles at the local and systemic levels

Luo

et al. 2021

Integr. Zool.

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Insects' intestinal microbes have profound effects on the host's physiological traits, which can impact their physiology at both the local (gut) and systemic (body) levels. Nevertheless, the molecular mechanisms underlying hostmicrobiota interactions, especially in non-model insects, remain elusive. Recently, tissue-specific transcriptomic analysis has been highlighted as a robust tool in studying host-microbe interactions. Plagiodera versicolora is a worldwide leaf-eating pest that primarily feeds on willows and poplar. The interplay between gut microflora and this host beetle has yet to be studied. Herein, we investigate the effects of the gut microbiota on the body mass of P. versicolora larvae, compare the nutrition status of larvae in absence and presence of gut microbiota, and profile gut bacterial loads throughout its developmental larval stages. We then perform comparative transcriptomic analysis of gut and body tissues in axenic and non-axenic larvae. Finally, we confirm the expression patterns of representative genes in nutritional metabolism and immunity. Results show that weight growth is retarded in conventional larvae, with a concomitant increase of total bacterial load by the 5 th development day, and germ-free larvae have a higher glucose content than conventional-reared larvae. Both nutritional and immunological analyses indicate that gut bacteria are a burden in the beetle's larval development. These findings elucidate the impacts of gut microbiota on P. versicolora, and provide insight into tissue-specific responses to gut microflora in this pest at the genetic level, boosting our understanding of the molecular mechanisms underlying host-microbe interactions in leaf beetles and beyond.

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“…2018; Muhammad et al . 2019). The other half of the non‐axenic eggs served as the conventionally reared group (CR) which was treated with only sterilized water.…”

Section: Methodsmentioning

confidence: 99%

Section: P Versicolora Rearing and Preparation Of Conventional And Germ-free Larvaementioning

confidence: 99%

Metabolic and immunological effects of gut microbiota in leaf beetles at the local and systemic levels

Luo

et al. 2021

Integr. Zool.

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“…In addition to the RPW’s strong mouthpart, this economically important pest also possesses an alimentary gut system that can digest the high-fiber food that it ingests. The gut can be divided into three different parts ( Figure 2 ): foregut, midgut and hindgut [ 13 , 15 , 24 ]. Mainly, the foregut is used to eat the food, and the hindgut is for the absorption of water.…”

Section: Anatomy Of Rpw’s Digestive Systemmentioning

confidence: 99%

A Review on Digestive System of Rhynchophorus ferrugineus as Potential Target to Develop Control Strategies

Seman-Kamarulzaman,

Pariamiskal,

Azidi

et al. 2023

Insects

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Rhynchophorus ferrugineus, commonly known as red palm weevil (RPW), is a high-risk insect pest that has become a threat to many important palm species. There are several dominant factors that lead to the successful infestation of RPW, including its stealthy lifestyle, highly chitinized mouthpart, and high fecundity rate. Due to that, millions of dollars of losses have been suffered by many countries invaded by RPW. Several methods have been designed to control its invasion, including the usage of insecticides, but many cause resistance and environmental pollution. Therefore, an environmentally friendly insecticide that targets specific systems or pathways in RPW is urgently needed. One of the potential targets is the digestive system of RPW, as it is the major interface between the insect and its plant host. The related knowledge of RPW’s digestive system, such as the anatomy, microflora, transcriptomic analysis, and proteomic analysis, is important to understand its effects on RPW’s survival. Several data from different omics regarding the digestive systems of RPW have been published in separate reports. Some of the potential targets have been reported to be inhibited by certain potential insecticides, while other targets have not yet been tested with any inhibitors. Hence, this review may lead to a better understanding on managing infestations of RPW using the system biology approach for its digestive system.

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“…In 1969, Marion Bakula first described the use of disinfectants to remove pigment from Drosophila embryos to produce germ‐free Drosophila (Blum et al, 2013). Currently, large numbers of axenic hosts such as Caenorhabditis elegans (Maupas) (Szewczyk et al, 2003), zebrafish (Melancon et al, 2017), Aedes aegypti (L.) (Correa et al, 2018), Rhynchophorus ferrugineus (Olivier) (Muhammad et al, 2019), and Bombyx mori (L.) (Chen et al, 2020) can be generated by surface‐sterilizing eggs, typically using bleach, and raising the resultant animals in sterile dishes or tubes.…”

Section: Introductionmentioning

confidence: 99%

Generating Ectropis obliqua axenic larvae in support of functional microbiome studies

Zhao

Zhou

et al. 2022

Entomologia Exp Applicata

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Ectropis obliqua Prout (Lepidoptera: Geometridae) is a serious leaf‐eating pest in tea gardens throughout China, with rich bacterial communities in the larval gut. Few studies have investigated whether these bacteria play an important role in growth, development, and adaptability of E. obliqua. Axenic larval models are a useful tool for the study of interactions between gut bacteria and host insects. At present, artificial diet and antibiotics are typically used to remove larval gut bacteria for study; however, this method may change the natural food of insects and the habitat provided by their host plants, and cannot completely eliminate insect gut bacteria. Therefore, in order to establish an axenic insect feeding model, we first investigated the effects of egg age, different disinfectant concentration, and treatment time on the eggs of E. obliqua through orthogonal experiments. Next, we fed E. obliqua larvae tissue‐cultured tea seedlings under aseptic conditions. Thus, we successfully generated a feeding model of gut axenic larvae in the laboratory. Based on it, we then compared the mortality of axenic and non‐axenic E. obliqua larvae infected with the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuill. These results demonstrated the feasibility of this axenic larval model and that B. bassiana is a broad‐spectrum entomogenous fungus with a toxic effect on E. obliqua.

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Preparation of Red Palm Weevil Rhynchophorus Ferrugineus (Olivier) (Coleoptera: Dryophthoridae) Germ-free Larvae for Host-gut Microbes Interaction Studies

Cited by 5 publications

References 24 publications

Metabolic and immunological effects of gut microbiota in leaf beetles at the local and systemic levels

Metabolic and immunological effects of gut microbiota in leaf beetles at the local and systemic levels

A Review on Digestive System of Rhynchophorus ferrugineus as Potential Target to Develop Control Strategies

Generating Ectropis obliqua axenic larvae in support of functional microbiome studies

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