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Complete mitochondrial genome of Asian longhorned tick, Haemaphysalis longicornis, Neumann, 1901 (Acari: Ixodida: Ixodidae) identified in the United States.

ABSTRACT: Haemaphysalis longicornis (Ixodida: Ixodidae), the Asian longhorned tick, which is native to temperate East Asia, has been recently detected in the northeastern region of the United States, drawing concerns about its potential impact on the US animal and public health sectors. Knowledge about the genetic features of H. longicornis found in the US is limited. Therefore, we sequenced the complete mitochondrial genome (mt-genome) from two H. longicornis ticks recently collected in the State of New York, USA, in 2020. These ticks were morphologically identified and tested for tick-borne pathogens at the Connecticut Veterinary Medical Diagnostic Laboratory (Storrs, CT). The mt-genome was 14,694 bp in length and encoded 37 genes, including 13 protein-coding genes, 22 transfer RNAs, and two ribosomal RNAs. Phylogenetic analysis showed that the mt-genome clustered with those of other H. longicornis identified in China. The mt-genome sequence was 99.7% identical to a H. longicornis mt-genome (GenBank: MK439888) collected in China. The cox1 gene haplotype in these ticks belonged to the H1 type, which is the dominant haplotype present in central NJ and Staten Island, NY. The complete mt-genome data are needed to provide insights into genetic changes and phylogenetic studies of H. longicornis ticks.

SUBMITTER: Hyeon JY

PROVIDER: S-EPMC8291064 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND:Close relationships between ticks and microbial communities are important for tick fitness and pathogen colonization and transmission. Haemaphysalis longicornis, distributed widely in China, can carry and transmit various pathogens and pose serious damages to public health and economics. However, little is known about the broader array of microbial communities and symbionts in H. longicornis under natural conditions. In the present study, we investigated the composition of bacterial communities associated with H. longicornis and evaluated the putative symbionts. METHODS:The eubacterial 16S rRNA gene clone libraries of H. longicornis were constructed and analyzed by restriction fragment length polymorphism (RFLP) and DNA sequencing. In addition, diagnostic PCR was performed to assess the prevalence, vertical transmission and infection sites of the symbionts in H. longicornis. RESULTS:Vertically-transmitted symbionts, potential pathogens and allochthonous nonpathogenic bacteria were identified from the field-collected H. longicornis. Three types of symbionts (Coxiella-like, Arsenophonus-like and Rickettsia-like symbionts) were identified in a single host simultaneously. A series of analyses revealed the vertical transmission, prevalence, and infection sites of these symbionts. However, only Coxiella-like bacteria were transmitted stably in the laboratory-reared ticks. In addition, we identified a novel Coxiella-like agent with 95.31% sequence similarity to the taxon described previously. CONCLUSIONS:The present study demonstrated that natural H. longicornis harboured a diverse array of microbial communities. Three types of symbionts were identified in a single host simultaneously. Moreover, high prevalence, vertical transmission and the infection sites supported an obligate symbiotic association between Coxiella symbiont and its host. The role of Coxiella symbiont in the host fitness and the interaction among microbial communities remained to be elucidated. Our investigation of microbial communities in the ticks revealed the complexity of ecological interactions between host and microbe and provided insight for the biological control of ticks.

Project description:Tick saliva contains many bioactive molecules that are involved in attachment to the host, blood feeding and transmission of pathogens. MicroRNAs (miRNAs) are a class of short non-coding RNAs with a length of 19-24 nucleotides. They act as regulators of gene expression by binding to their target mRNA at the post-transcriptional level and control a variety of cellular functions, including regulation of growth, metabolism, and development. The detection and characterizations of miRNAs from tick saliva may help explain the molecular mechanisms involved in the interaction between ticks, pathogens, and hosts. They may also contribute to the discovery of vaccines, which can control ticks and the pathogens they transmit. An RNA library was generated from the saliva of fed adult Haemaphysalis longicornis ticks, and it contained 17.4 million clean reads of 18-30 nucleotides. Overall, 319 known miRNAs and 1 novel miRNA were found. The 10 most abundantly expressed miRNAs present in tick saliva were miR-100_2, miR-315, miR-184_1, miR-100-5p_2, miR-5307, miR-184-3p_3, Let-7-5p_6, miR-71_5, miR-1-3p_6, and miR-10-5p_2. The miR-375, one of the abundantly expressed, was subjected to Quantitative Real-Time PCR analysis (qRT-PCR) in various tick developmental stages, as well as in different tissues isolated from adult ticks. The expression of miR-375 in different tick development stages was highest in unfed nymphs and lowest in the egg stage. In the tissues of adult ticks, miR-375 was most highly expressed in the salivary gland. To investigate the possible role of miR-375, Ant-375 was used to inhibit the miR-375. Treated group (Ant-375) had a reduced number of eggs (t(10)= 2.652, P=0.0242), eggs that were partially desiccated, and reduced egg hatchability (t(10)=2.272, P=0.044) compared to Ms-Ant and the non-injected control. This is the first study to investigate the miRNAs profile in tick saliva and the role of miR-375 in H. longicornis. The identification and characterization of miRNA in tick saliva may help to reveal the molecular mechanisms of interactions among ticks, pathogens, and hosts and suggest new vaccine strategies to control tick borne diseases.

Project description:BackgroundCharacterization of the microbial diversity and symbiont dynamics of ticks may help to understand the development of ticks and reveal new strategies to control tick-transmitted pathogens, which has not yet been explored in the Tibetan tick Haemaphysalis tibetensis. This tick species is widely distributed in the Tibetan Plateau, and is recognized as one of the primary parasites affecting domestic and wild animals.MethodsIn the present study, the endosymbionts of H. tibetensis were characterized using diagnostic polymerase chain reaction (diagnostic PCR), and further evaluated for tissue distribution and population dynamics at each developmental stage of ticks and in tissues at different reproductive statuses by real-time quantitative polymerase chain reaction (RT-qPCR).ResultsTwo symbionts were found in H. tibetensis, and named as CLS-Ht (Coxiella-like symbiont in H. tibetensis) and RLS-Ht (Rickettsia-like symbiont in H. tibetensis). They showed 100% infection rate in both females and males of H. tibetensis. CLS-Ht and RLS-Ht can be observed within eggs, larvae, nymphs and adults, which indicates vertical transmission in H. tibetensis. CLS-Ht was specifically distributed in the female ovaries and Malpighian tubules, whereas RLS-Ht was detected within ovaries, Malpighian tubules, salivary glands and midguts of the ticks. Real-time qPCR suggested that adult ticks carried the largest amount of CLS-Ht and RLS-Ht with CLS-Ht having a significantly higher presence in females than in males (P < 0.05), whereas the presence of RLS-Ht showed no significant differences between sexes. In the ovaries, CLS-Ht distribution reached a peak at one day post-engorgement, and then gradually declined to a lower level, whereas no change was observed in RLS-Ht. In Malpighian tubules, the amount of both symbionts displayed an increasing trend with time post-engorgement. In midguts and salivary glands, the amount of RLS-Ht showed no significant differences.ConclusionTwo novel endosymbionts (CLS-Ht and RLS-Ht) were characterized in H. tibetensis both showing a high prevalence and stable vertical transmission. The described tissue distribution and population dynamics might imply the important functions of these symbionts during the development and reproduction of ticks.

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Complete mitochondrial genome of Asian longhorned tick, Haemaphysalis longicornis, Neumann, 1901 (Acari: Ixodida: Ixodidae) identified in the United States.

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