Network analysis uncovers putative genes affecting resistance to tick infestation in Braford cattle skin.

BACKGROUND: Genetic resistance in cattle is considered a suitable way to control tick burden and its consequent losses for livestock production. Exploring tick-resistant (R) and tick-susceptible (S) hosts, we investigated the genetic mechanisms underlying the variation of Braford resistance to tick infestation. Skin biopsies from four-times-artificially infested R (n = 20) and S (n = 19) hosts, obtained before the first and 24 h after the fourth tick infestation were submitted to RNA-Sequencing. Differential gene expression, functional enrichment, and network analysis were performed to identify genetic pathways and transcription factors (TFs) affecting host resistance. RESULTS: Intergroup comparisons of hosts before (Rpre vs. Spre) and after (Rpost vs. Spost) tick infestation found 51 differentially expressed genes (DEGs), of which almost all presented high variation (TopDEGs), and 38 were redundant genes. Gene expression was consistently different between R and S hosts, suggesting the existence of specific anti-tick mechanisms. In the intragroup comparisons, Rpost vs. Rpre and Spost vs. Spre, we found more than two thousand DEGs in response to tick infestation in both resistance groups. Redundant and non-redundant TopDEGs with potential anti-tick functions suggested a role in the development of different levels of resistance within the same breed. Leukocyte chemotaxis was over-represented in both hosts, whereas skin degradation and remodeling were only found in TopDEGs from R hosts. Also, these genes indicated the participation of cytokines, such as IL6 and IL22, and the activation of Wingless (WNT)-signaling pathway. A central gene of this pathway, WNT7A, was consistently modulated when hosts were compared. Moreover, the findings based on a genome-wide association study (GWAS) corroborate the prediction of the WNT-signaling pathway as a candidate mechanism of resistance. The regulation of immune response was the most relevant pathway predicted for S hosts. Members of Ap1 and NF-kB families were the most relevant TFs predicted for R and S, respectively. CONCLUSION: This work provides indications of genetic mechanisms presented by Braford cattle with different levels of resistance in response to tick infestation, contributing to the search of candidate genes for tick resistance in bovine.

Amblyomma americanum ticks utilizes countervailing pro and anti-inflammatory proteins to evade host defense.

Feeding and transmission of tick-borne disease (TBD) agents by ticks are facilitated by tick saliva proteins (TSP). Thus, defining functional roles of TSPs in tick evasion is expected to reveal potential targets in tick-antigen based vaccines to prevent TBD infections. This study describes two types of Amblyomma americanum TSPs: those that are similar to LPS activate macrophage (MΦ) to express pro-inflammation (PI) markers and another set that suppresses PI marker expression by activated MΦ. We show that similar to LPS, three recombinant (r) A. americanum insulin-like growth factor binding-related proteins (rAamIGFBP-rP1, rAamIGFBP-rP6S, and rAamIGFBP-rP6L), hereafter designated as PI-rTSPs, stimulated both PBMC -derived MΦ and mice RAW 267.4 MΦ to express PI co-stimulatory markers, CD40, CD80, and CD86 and cytokines, TNFα, IL-1, and IL-6. In contrast, two A. americanum tick saliva serine protease inhibitors (serpins), AAS27 and AAS41, hereafter designated as anti-inflammatory (AI) rTSPs, on their own did not affect MΦ function or suppress expression of PI markers, but enhanced expression of AI cytokines (IL-10 and TGFß) in MΦ that were pre-activated by LPS or PI-rTSPs. Mice paw edema test demonstrated that in vitro validated PI- and AI-rTSPs are functional in vivo since injection of HEK293-expressed PI-rTSPs (individually or as a cocktail) induced edema comparable to carrageenan-induced edema and was characterized by upregulation of CD40, CD80, CD86, TNF-α, IL-1, IL-6, and chemokines: CXCL1, CCL2, CCL3, CCL5, and CCL11, whereas the AI-rTSPs (individually and cocktail) were suppressive. We propose that the tick may utilize countervailing PI and AI TSPs to regulate evasion of host immune defenses whereby TSPs such as rAamIGFBP-rPs activate host immune cells and proteins such as AAS27 and AAS41 suppress the activated immune cells.

Metabolic differentiation of early Lyme disease from southern tick-associated rash illness (STARI).

Lyme disease, the most commonly reported vector-borne disease in the United States, results from infection with Borrelia burgdorferi. Early clinical diagnosis of this disease is largely based on the presence of an erythematous skin lesion for individuals in high-risk regions. This, however, can be confused with other illnesses including southern tick-associated rash illness (STARI), an illness that lacks a defined etiological agent or laboratory diagnostic test, and is coprevalent with Lyme disease in portions of the eastern United States. By applying an unbiased metabolomics approach with sera retrospectively obtained from well-characterized patients, we defined biochemical and diagnostic differences between early Lyme disease and STARI. Specifically, a metabolic biosignature consisting of 261 molecular features (MFs) revealed that altered N-acyl ethanolamine and primary fatty acid amide metabolism discriminated early Lyme disease from STARI. Development of classification models with the 261-MF biosignature and testing against validation samples differentiated early Lyme disease from STARI with an accuracy of 85 to 98%. These findings revealed metabolic dissimilarity between early Lyme disease and STARI, and provide a powerful and new approach to inform patient management by objectively distinguishing early Lyme disease from an illness with nearly identical symptoms.

Espectro de las enfermedades transmitidas por garrapatas / Spectrum of thick-borne diseases

No disponible

Midgut proteome of an argasid tick, Ornithodoros erraticus: a comparison between unfed and engorged females.

BACKGROUND: The argasid tick Ornithodoros erraticus is the vector of African swine fever virus and of several Borrelia species that cause human relapsing fever in the Iberian Peninsula. The tick midgut is part of the ectoparasite-host interface and expresses proteins that are vital for the survival of the tick. Midgut proteins are therefore potential targets for drug and/or vaccine design aimed at the development of new strategies for tick control. Thus, the aim of this work was the characterization of the proteome of the O. erraticus midgut before and after a blood meal trying to elucidate the induced changes upon blood feeding. METHODS: Midgut tissues from unfed and engorged O. erraticus females were dissected and proteins were fractionated by centrifugation and SDS-PAGE, and the corresponding gel pieces analysed by LC-MS/MS. The identified proteins were classified according to their Protein Class and Molecular Function and the differences between fed and unfed specimens were analysed. RESULTS: Overall 555 tick proteins were identified: 414 in the midgut of the unfed specimens and 376 in the fed specimens, of which 235 were present in both groups. The proteins with catalytic, binding and structural functions were the most numerous and abundant, consistent with their role in the intracellular processing of the blood meal. The analysis of some groups of proteins putatively involved directly in blood meal digestion, including protein digestion (peptidase activity), iron metabolism, enzymes involved in oxidative stress and detoxification and membrane traffic and transport proteins, detected some differences between the fed and unfed ticks CONCLUSIONS: This work reports for the first time the collection and analysis of the midgut proteome of an argasid tick species and provides molecular information about the argasid machinery involved in blood digestion. This information represents a starting point for the identification and selection of new targets for the development of alternative control strategies.

Effects of parasites and antigenic challenge on metabolic rates and thermoregulation in northern red-backed voles (Myodes rutilus).

Perturbations in host energetics are considered to be an essential pathway for parasite impact on host fitness. However, direct estimations of parasite-induced variations in basal metabolic rates of vertebrate hosts have so far provided contradictory results. The energy requirements of immunity and other vital functions may be compromised in energy-demanding conditions in comparison to comfortable conditions; therefore, in our study performed on the wild red-backed vole, Myodes rutilus, we compared the values of indices that reflect metabolic and thermoregulatory responses to acute cooling in individuals that had been naturally infected by gut helminths or Ixodes persulcatus taiga ticks to individuals with no signs of infestation. To consider the possible effects of an acquired immune response on host energetics, we also injected some of the tested individuals with sheep red blood cells (SRBC). Red-backed voles infected by the nematode Heligmosomum mixtum injected with SRBC showed significantly lower cold-induced maximum oxygen consumption than the saline control. Additionally, individuals infected with H. mixtum showed significantly lower oxygen consumption during the final minute of the 15-min acute cooling period and a significantly greater decline in body temperature than individuals free from helminths. In individuals concurrently infected by H. mixtum and the cestodes Arostrilepis horrida, these indices did not differ from helminth-free individuals. The number of ticks simultaneously parasitizing the voles at the moment of capture correlated positively with their SMR. Our results suggest that even natural parasites produce deleterious effects on host aerobic capacity and thermoregulatory abilities, although the effects of different parasites might not be additive.

Characterization of the tick-pathogen interface by quantitative proteomics.

Ticks are vectors of pathogens that affect human and animal health worldwide. Ticks and the pathogens they transmit have co-evolved molecular interactions involving genetic traits of both the tick and the pathogen that mediate their development and survival. Proteomics and genomics studies of infected ticks are required to understand tick-pathogen interactions and identify potential vaccine antigens to control tick infestations and pathogen transmission. In this paper, the application of quantitative proteomics to characterize differential protein expression in ticks and cultured tick cells in response to pathogen infection is reviewed. Analyses using (a) two-dimensional differential in gel electrophoresis (DIGE) labeling and (b) protein one-step in gel digestion, peptide iTRAQ labeling, and isoelectric focusing fractionation, both followed by peptide and protein identifications by mass spectrometry resulted in the identification of host, pathogen, and tick proteins differentially expressed in response to infection. Although at its infancy, these results showed that quantitative proteomics is a powerful approach to characterize the tick-pathogen interface and demonstrated pathogen and tick-specific differences in protein expression in ticks and cultured tick cells in response to pathogen infection.

Scavenger receptor mediates systemic RNA interference in ticks.

RNA interference is an efficient method to silence gene and protein expressions. Here, the class B scavenger receptor CD36 (SRB) mediated the uptake of exogenous dsRNAs in the induction of the RNAi responses in ticks. Unfed female Haemaphysalis longicornis ticks were injected with a single or a combination of H. longicornis SRB (HlSRB) dsRNA, vitellogenin-1 (HlVg-1) dsRNA, and vitellogenin receptor (HlVgR) dsRNA. We found that specific and systemic silencing of the HlSRB, HlVg-1, and HlVgR genes was achieved in ticks injected with a single dsRNA of HlSRB, HlVg-1, and HlVgR. In ticks injected first with HlVg-1 or HlVgR dsRNA followed 96 hours later with HlSRB dsRNA (HlVg-1/HlSRB or HlVgR/HlSRB), gene silencing of HlSRB was achieved in addition to first knockdown in HlVg-1 or HlVgR, and prominent phenotypic changes were observed in engorgement, mortality, and hatchability, indicating that a systemic and specific double knockdown of target genes had been simultaneously attained in these ticks. However, in ticks injected with HlSRB dsRNA followed 96 hours later with HlVg-1 or HlVgR dsRNAs, silencing of HlSRB was achieved, but no subsequent knockdown in HlVgR or HlVg-1 was observed. The Westernblot and immunohistochemical examinations revealed that the endogenous HlSRB protein was fully abolished in midguts of ticks injected with HlSRB/HlVg-1 dsRNAs but HlVg-1 was normally expressed in midguts, suggesting that HlVg-1 dsRNA-mediated RNAi was fully inhibited by the first knockdown of HlSRB. Similarly, the abolished localization of HlSRB protein was recognized in ovaries of ticks injected with HlSRB/HlVgR, while normal localization of HlVgR was observed in ovaries, suggesting that the failure to knock-down HlVgR could be attributed to the first knockdown of HlSRB. In summary, we demonstrated for the first time that SRB may not only mediate the effective knock-down of gene expression by RNAi but also play essential roles for systemic RNAi of ticks.

Expression of intracellular calcium signalling genes in cattle skin during tick infestation.

It is widely acknowledged that changes in intracellular calcium ion (Ca(2+)) concentration provide dynamic signals that control a plethora of cellular processes, including triggering and mediating host defence mechanisms. In this study, quantitative real-time PCR was used to analyse gene expression of 14 Ca(2+) signalling proteins in skin obtained from high tick-resistant (HR) and low tick-resistant (LR) cattle following artificial challenge with cattle tick (Rhipicephalus (Boophilus) microplus). Up-regulation of numerous genes was observed in both HR and LR skin following tick challenge, however substantially higher transcription activation was found in HR tissue. The elevated expression in HR skin of specific Ca(2+) signalling genes such as AHNAK, CASQ, IL2, NFAT2CIP and PLCG1 may be related to host resistance. Our data suggest that Ca(2+) and its associated proteins might play an important role in host response to ticks and that further investigation is warranted.

Effects produced by the feeding of larvae of Ornithodoros aff. puertoricensis (Acari: Argasidae) on laboratory mice.

The clinical picture produced by the feeding of larvae of Ornithodoros aff. puertoricensis on laboratory mice, was studied using different larval infestation protocols that included 30, 40 or 50 larvae per mouse and control uninfested groups. Clinical effects appeared around 72 h of larval feeding, having a first stage characterized by hyperaemia in both nasal and ocular mucosa, followed by respiratory symptoms (96-120 h) and nervous incoordination (120-144 h). No one mouse evidenced paralysis, and nervous symptoms were never observed in animals infested with only 30 larvae. High mortality (commonly up to 70%) was observed in mice with respiratory symptoms, while 100% of animals in the nervous phase died between 168 and 192 h after the beginning of larval feeding. When some infested mice were treated with a solution of Amitraz the larvae were killed and reversion of symptoms was observed. These effects are ascribed to the presence of a toxin in the saliva of the feeding larvae.

[Toxic and allergic action of tick saliva components]. / Toksyczne i alergiczne dzialanie wydzieliny gruczolów slinowych kleszczy (Acari: Ixodida).

The paper presents the role of bioactive components secreted by ticks in pathogenesis local and general reactions in the host. The salivary glands of ticks produce cement, enzymes and their inhibitors, histamine agonists and antagonists, antihemostatic factors, prostaglandins, and toxins. Tick feeding induces host inflammatory lesion and host immune responses.

Cytokine production by lymph node cells from mice infested with Ixodes ricinus ticks and the effect of tick salivary gland extracts on IL-2 production.

In BALB/c mice repeatedly infested with nymphal Ixodes ricinus ticks, lymphocytes from axillary and brachial lymph nodes which drain the tick attachment site produced significant levels of IL-2, TNF-alpha and GM-CSF when stimulated in vitro with Con A or anti-CD3 antibodies. Cytokine production by cells from lymph nodes of the opposite flank was equivalent to that of cells from uninfested mice. Nine days after the first infestation and IL-2, GM-CSF were produced primarily by the CD4+ T cells, while some other cell types contributed also to the TNF-alpha production. In mice repeatedly infested, a gradual increase of lymph node cell production of IL-2 was observed. The IL-2 levels regularly increased from the first to the third infestation compared to TNF-alpha levels which gradually decreased. The in vitro production of GM-CSF was not affected by successive infestations. Spleen lymphocytes from naive mice produced higher levels of IL-2 than lymphocytes from axillary and brachial lymph nodes. Both tick salivary gland extracts and D-mannose inhibited IL-2 production by these lymphocytes.

Physiological and nutritional responses of steers infested with varying densities of Amblyomma americanum (Acari: Ixodidae).

The effects of varying densities of lone star tick, Amblyomma americanum (L.), on measured physiological parameters of beef cattle in a controlled environment was determined. Steers were infested with either 0, 20, 60, or 120 pairs of adult ticks. Heart rate, respiration, rectal temperature, fecal and urine excretions, and water and feed consumption were monitored daily. Blood samples were taken every 3rd d to measure cortisol, total proteins, urea nitrogen, and glucose levels. Hematocrits were also taken at each blood sampling. Results showed that A. americanum evoked elevated heart rates. The other measured physiological and nutritional responses of parasitized steers were similar to the control steers. These results suggest that tick densities were too low to cause physiological stress under the conditions used in this study. The methodology precluded detection of the parameters measured. A. americanum does not affect the parameters measured, or that fluctuating environmental parameters and varying host nutritional states may play major roles in modulating the effect of A. americanum infestations on cattle in nature.

The effect of feeding immature Karoo paralysis ticks Ixodes rubicundus (Acari: Ixodidae) on the metabolic rate of the rock elephant shrew.

The effect of feeding immature Karoo paralysis ticks (Ixodes rubicundus) on the resting metabolic rate (RMR) of their principal natural host, the rock elephant shrew (Elephantulus myurus), was investigated under laboratory conditions. The elephant shrews were artificially infested with numbers of ticks simulating natural burdens. The RMR of the elephant shrews was quantified by measuring the oxygen consumption in an open through-flow system. The RMR of hosts infested only with larvae did not differ from that of the control group. Those infested with nymphs, or nymphs and larvae, had significantly lower RMR's compared to the control animals. There were no signs of paralysis in any of the infested hosts.

Some effects of tick infestations on juvenile northern brown bandicoot (Isoodon macrourus).

The effect of tick infestations on body weight and various blood parameters was monitored in juvenile northern brown bandicoots (Isoodon macrourus) after release into tick-infested or tick-free enclosures. Three species of ticks were observed in the enclosures, Haemaphysalis humerosa, Ixodes tasmani and Ixodes holocyclus. Bandicoots released into tick-infested enclosures showed a reduced growth rate (1.8 versus 2.5 g/day increase in body weight), a reduced haematocrit value (27.4 versus 40.0%) and an increased number of white blood cells when compared with bandicoots released into tick-free enclosures. These results suggest that tick infestations may influence the health of juvenile I. macrourus.

The effect of the brown ear-tick Rhipicephalus appendiculatus on the growth of Sanga and European breed cattle.

The effects of larvae, nymphs and adults of the brown ear-tick Rhipicephalus appendiculatus on the growth of Bos indicus X B. taurus (Sanga) steers and on B. taurus steers and heifers was measured by exposing groups, maintained in the same pastures, to different levels of tick challenge. Larvae and nymphs had no significant effect on liveweight gains, but adults had a large effect. Each engorging female tick counted was associated with a loss of 4 g. Their effect on the Sanga cattle was minimised by host resistance which severely limited the number of ticks feeding on them. The B. taurus animals were severely affected and suffered extensive ear damage. Both breeds were subjected to screw-worm fly (Chrysomya bezziana) strike in the lesions caused by heavy tick infestations, but the B. taurus animals were much more prone to attack. The results provide a basis for calculating losses in production caused by this species of tick which is widespread in Africa.

Metabolic changes in cattle due to the specific effect of the tick, Boophilus microplus.

1. An experiment was designed to provide information on the alterations in body metabolism which would account for the loss of body-weight in cattle due to the specific effect (factors other than reduced food intake) of the tick Boophilus microplus. 2. Two groups of British (Shorthorn x Hereford) and Africander x British calves, each approximately months olds, were used: one group (treated) of each breed was tick-infested and the other (control) was tick-free. Within breeds, calves in the control group were pair-fed to calves in the treated group. 3. In both breeds, the effect of ticks: (a) depressed packed cell volume, serum alkaline phosphatase (EC 3.1.3.1) and amylase (EC 3.2.1.1.) activities, plasma cholesterol and phospholipid levels, serum iron and albumin levels, (b) increased the plasma levels of urea-nitrogen and gamma-globulin (c) increased rectal temperature, water intake, urine volume, urinary and faecal total N, urinary urea-N and alpha-amino acids, the excretion of water, sodium and potassium in the faeces and (d) reduced N balance, N and dry-matter digestibilities. 4. In the British breed, ticks increased the excretion of K with a corresponding decrease in the excretion of Na in the urine and increased the plasma clearance of bromsulphthalein. 5. A second experiment showed that the specific effect of tick infestation increased the flow of organic matter (OM) from the abomasum and the fractional turnover of rumen fluid of Hereford steers. It was also shown that the decrease in OM digestibility in the gastrointestinal tract was largely due to a decrease in OM digestibility in the rumen and that the increased urinary urea excretion and plasma urea concentration was caused by higher production rates of urea despite a tendency for lowered urea degradation in the gastrointestinal tract.