Dataset Information

Intestine proteome of Rhipicephalus microplus tick reveal key molecular features associated with resistant to ivermectin

ABSTRACT: Rhipicephalus microplus is among the critical tick parasite of cattle farming globally. However, the massive use of ivermectin last decades induced the generation of the tick-resistant population. Unfortunately, despite extensive research the mode of action of ivermectin remains unclear. Due to the limited molecular information related to the resistance of R. microplus to ivermectin, we carried out a midgut comparative proteomics study between susceptible and resistant tick. Our tissues specificity approach also included tandem mass tags (TMT) couple to synchronous precursor selection (SPS)-MS3. Our proteomics scrutiny exhibits key biological process associated with blood digestion and anticoagulation in midgut. In addition, we could observe the significant shutdown of the translation mechanism in susceptible thinks. At the same time, resistant thick exhibited the over-accumulation of cytochrome P450, glutathione-S-transferase, and ABC transporters. A similar trend was observed in proteins like glutathione peroxides glutaredoxin, sulfiredoxin, and disulfide-isomerase, which suggest a possible activation of the oxidative metabolism as a potential strategy to cope with ivermectin treatment.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Rhipicephalus Microplus

TISSUE(S): Intestine, Midgut

SUBMITTER: Miguel Elizalde

LAB HEAD: Ruiz-May Eliel

PROVIDER: PXD031231 | Pride | 2024-01-29

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	Garrapatas_Intestino_TMT.xlsx	Xlsx
	Garrapatas_Intestino_TMT_10.raw	Raw
	Garrapatas_Intestino_TMT_12.raw	Raw
	Garrapatas_Intestino_TMT_15.raw	Raw
	Garrapatas_Intestino_TMT_17.raw	Raw

Items per page:

1 - 5 of 11

Publications

Comparative proteome analysis of the midgut of Rhipicephalus microplus (Acari: Ixodidae) strains with contrasting resistance to ivermectin reveals the activation of proteins involved in the detoxification metabolism.

Ruiz-May Eliel E Álvarez-Sánchez María Elizbeth ME Aguilar-Tipacamú Gabriela G Elizalde-Contreras José M JM Bojórquez-Velázquez Esaú E Zamora-Briseño Jesús Alejandro JA Vázquez-Carrillo Laura I LI López-Esparza Areli A

Journal of proteomics 20220519

Rhipicephalus microplus is the most serious tick parasite for the livestock industry in tropical and subtropical regions. A cost-effective control method to manage the infestation of this parasite involves the use of chemicals such as ivermectin. However, massive overuse of ivermectin over recent decades has selected for ivermectin-resistant populations of R. microplus. Here, we carried out a comparative proteomic analysis of the midgut of ivermectin-susceptible versus ivermectin-resistant ticks ...[more]

PMID: 35598868

Similar Datasets

Project description:In addressing R. microplus - A. marginale interactions, we propose and test three linked hypotheses. The first is that the tick gene response is organ specific: the midgut gene regulation is unique during feeding and during acquisition of A. marginale as compared to the salivary gland. This distinction is relevant as the two organs serve very different roles in the transmission biology of A. marginale with early survival and replication within the midgut epithelium, composed of highly phagocytic cells, required for initial colonization while a second round of replication in the salivary gland acini, composed of highly secretory cells, is required for transmission of an infectious dose in the saliva. Importantly, both the midgut epithelium and salivary glands have been identified as separate and distinct barriers for transmission of A. marginale and thus represent two potential sites where transmission could be blocked. The second hypothesis to be tested is that the salivary gland transcriptome is temporally dynamic. Initiation of tick attachment and feeding involves secretion of a virtual pharmacopeia including lytic enzymes, anticoagulants, and inhibitors of the mammalian innate immune and nocioceptive systems. Concomitantly, the acini provide an environment where A. marginale replicates >100 fold and are secreted into the saliva. Prior studies show that duration of feeding is a critical component of transmission efficiency, with increased efficiency positively correlated with time of tick feeding. The third hypothesis to be tested is that A. marginale colonization does not significantly modulate the tick midgut and salivary gland transcriptome. This hypothesis is based on observations by ourselves and others that tick infection does not impart a significant fitness cost on the vector. This is in contrast to other bacterial and protozoal pathogens that have dramatic effects on success of tick attachment, engorgement, and survival. A. marginale, similar to other tick-borne pathogens in the Family Anaplasmataceeae, is believed to have evolved from an arthropod-specific bacterium with relatively late adaptation to specific niches in mammalian hosts. Consequently, we predict that A. marginale is well adapted to its tick vector and utilizes the normal signaling pathways of the feeding tick with few, if any, effects on the midgut and salivary gland transcriptome. In this manuscript, we report the testing of these three hypotheses and present the results in context of the vector-pathogen-mammalian host interaction at the time of transmission. A Roche NimbleGen high-density gene expression microarray was custom designed based on the expressed sequence tag (EST) database, B. microplus Gene Index Version 2 (BmiGI V2) for R. microplus. The expression level of 14,447 R. microplus genes was analyzed from total RNA extracted from 10 different tick tissue samples; 30 arrays were included since triplicates of each different sample were analyzed as follow: unfed (midgut and salivary glands), blood feeding (2 days midgut and 2, 6 and 9 days salivary glands), A. marginale-infected blood feeding (2 days midgut and 2, 6 and 9 days salivary glands).

Project description:Ticks are among the most economically important arthropod parasites due to the heavy infestations they provoke, affecting human and animal health and food production. Ticks are also important vectors of pathogens, mediating the dispersion of livestock and zoonotic diseases. Some examples of livestock diseases vectored by ticks include bovine babesiosis, anaplasmosis, theileriosis, and/or heartwater disease, all of which are enlisted by the World Organization for Animals as possible threats to animals and public health. The family Ixodidae (hard ticks) is of veterinary relevance, and many ixodid tick species within the genera Ixodes, Dermacentor, Amblyomma, Haemophysalis, Hyalomma, and Rhipicephalus, among others, impact farming production worldwide. However, there is no doubt that most pernicious tick species is Rhipicephalus microplus, commonly known as the southern cattle fever tick. This specie is not only the major economically important tick parasite for livestock production, but it is also the vector of Babesia bovis and B. bigemina, the causal agents of babesiosis, a severe disease that affects cattle from tropical and subtropical regions. Currently, global economic losses of the cattle industry attributable to R. microplus are estimated at around $18.7 US billion dollars per year. The economic impact is the leading reason to develop different strategies to constrains R. microplus and its vector-borne pathogens, including chemical, mechanical or environmental treatments, biological control, and/or genetic methods. Chemical, among other methods, are the most frequently applied, but the intense use worldwide, has led to the selection of acaricidal-resistant ticks populations. The molecular adaptations of ticks to overcome the effects of acaricides (e.g., deltamethrin, cypermethrin, amitraz, ivermectin) includes metabolic detoxiﬁcation mechanisms and/or the selection of mutated molecular targets. Recently, a couple of systematic reviews pointed out a significant increment in the number of studies intended to understand the acaricidal resistance in ticks. These reports highlighted the acaricide resistance observed in R. microplus and the wide geographical dispersion of this problem. Whereas many acaricide-resistant R. microplus populations have been reported in Brazil, India, and Mexico, the lack of strict regulatory policies on tick control, leaving the application of acaricides at discretion of farmers. Therefore, the correct implementation of existing strategies together with novel sustainable control methods may be crucial to safeguard livestock production in the context of growing population and global warming. However, until the implementation of such sanitary measures becomes the rule, the main control strategy will still rely on the use of chemical acaricides, such as ivermectin (IVM). Ivermectin is an antiparasitic macrocyclic lactone used to control the populations of R. microplus and other ticks in livestock farming. As occurs in other countries, in Mexico ivermectin is the primary synthetic antiparasitic drug, and its abuse during the last three decades represents an important selection pressure that boosted the development of ivermectin-resistant populations of R. microplus. Therefore, the understanding of the molecular basis of the R. microplus resistance to ivermectin, especially in those cases occurring in tropical and subtropical areas of developing countries, can help to underpin the development of novel strategies to tackle this problem. In this context, omics tools can provide helpful information for the discovery of the molecular mechanisms behind the ivermectin resistance in R. microplus and other related tick species. Previous studies identified the main proteomic components of ovaries of R. microplus, including proteins involved in protein synthesis, post-translational modification, nuclear regulation, cytoskeleton, proteasome, transcriptional regulation, energetic metabolism, detoxification metabolism, or energy storage. Recently, the vitellogenin receptor (VgR) was suggested as a target for tick control because of its essential role in egg formation and maturation. However, molecular information on tick ovaries currently available limits explores the feasibility of VgR or other reproduction-related molecules as promissory targets for tick control.

Project description:Ticks are obligate hematophagous arthropods that transmit a wide range of pathogens to humans as well as wild and domestic animals. They also harbor a non-pathogenic microbiota, although our previous study has shown that the diverse bacterial microbiome in the midgut of Ixodes ricinus is quantitatively poor and lacks a core microbe. In artificial infections by capillary feeding of ticks with two model bacteria (Gram-positive Micrococcus luteus and Gram-negative Pantoea sp.), rapid clearance of these microbes from the midgut was observed, indicating the presence of active immune mechanisms in this organ. In the current study, RNA-seq analysis was performed on the midgut of I. ricinus females inoculated with either M. luteus or Pantoea sp. or with sterile water as a control. While no immune-related transcripts were upregulated by microbial inoculation compared to the sterile control, capillary feeding itself triggered dramatic transcriptional changes in the tick midgut. Manual curation of the transcriptome from the midgut of unfed I. ricinus females, complemented by proteomic analysis, revealed the presence of several constitutively expressed putative antimicrobial peptides (AMPs) that are independent of microbial stimulation and are referred to here as ‘guard’ AMPs. These included two types of midgut-specific defensins, two different domesticated amidase effector 2 (Dae2), microplusin/ricinusin-related molecules, two lysozymes and two gamma interferon-inducible lysosomal thiol reductases (GILTs). The in vitro antimicrobial activity assays of two synthetic mature defensins, defensin 1 and defensin 8, confirmed their specificity against Gram-positive bacteria showing exceptional potency to inhibit the growth of M. luteus at nanomolar concentrations. The antimicrobial activity of midgut defensins is likely part of a multicomponent system responsible for the rapid clearance of bacteria in the tick midgut. Further studies are needed to evaluate the role of other identified ‘guard‘ AMPs in controlling microorganisms entering the tick midgut.