Development of microsatellite markers for the soft tick Ornithodoros phacochoerus.

BACKGROUND: Soft ticks of the genus Ornithodoros are responsible for the maintenance and transmission of the African swine fever (ASF) virus in the sylvatic and domestic viral cycles in Southern Africa. They are also the main vectors of the Borrelia species causing relapsing fevers. Currently, no genetic markers are available for Afrotropical Ornithodoros ticks. As ASF spreads globally, such markers are needed to assess the role of ticks in the emergence of new outbreaks. The aim of this study is to design microsatellite markers that could be used for ticks of the Ornithodoros moubata complex, particularly Ornithodoros phacochoerus, to assess population structure and tick movements in ASF endemic areas. METHODS: A total of 151 markers were designed using the O. moubata and O. porcinus genomes after elimination of repeated sequences in the genomes. All designed markers were tested on O. phacochoerus and O. porcinus DNA to select the best markers. RESULTS: A total of 24 microsatellite markers were genotyped on two populations of O. phacochoerus and on individuals from four other Ornithodoros species. Nineteen markers were selected to be as robust as possible for population genetic studies on O. phacochoerus. CONCLUSIONS: The microsatellite markers developed here represent the first genetic tool to study nidicolous populations of O. phacochoerus.

Nation-wide surveillance of ticks (Acari: Argasidae) on bats (Chiroptera) in Singapore.

Bats and ticks are important sources of zoonotic pathogens. Therefore, understanding the diversity, distribution, and ecology of both groups is crucial for public health preparedness. Soft ticks (Argasidae) are a major group of ectoparasites commonly associated with bats. The multi-host life cycle of many argasids make them important vectors of pathogens. Over nine years (2011-2020), surveillance was undertaken to identify the ticks associated with common bats in Singapore. During this period, the bat tick Ornithodoros batuensis was detected within populations of two cave roosting bat species: Eonycteris spelaea and Penthetor lucasi. We examined the relationship between bat species, roosting behaviour, and probability of O. batuensis infestation. We also estimated the relationship between bat life history variables (body condition index, sex, and age) on the probability of infestation and tick count. This represents the first detection of O. batuensis and the genus Ornithodoros within Singapore. We also provide evidence of the continued persistence of Argas pusillus in Singapore with the second local record.

Experimental transmission of a novel relapsing fever group Borrelia harbored by Ornithodoros octodontus (Ixodida: Argasidae) in Chile.

Tick-borne relapsing fever spirochetes of genus Borrelia thrive in enzootic cycles involving Ornithodoros spp. (Argasidae) mainly, and rodents. The isolation of these spirochetes usually involves a murine model in which ticks are fed and the spirochetes detected in blood several days later. Such an experiment also demonstrates that a given species of tick is competent in the transmission of the bacteria. Here, soft ticks Ornithodoros octodontus were collected in Northern Chile with the objective to experimentally determine its capacity to transmit a Borrelia sp. detected in a previous study. Two Guinea pigs (Cavia porcellus) were used to feed nymphs and adults of O. octodontus and the spirochetes in blood were inspected by dark-field microscopy and nested PCR. Although spirochetes were not seen in blood, DNA was detected in only one animal 11 days after the ticks were fed. Genetic sequences of Borrelia flaB, clpX, pepX, recG, rplB, and uvrA genes retrieved from DNA extraction of positive blood were employed to construct two phylogenetic analyses. On the one hand, the flaB tree showed the Borrelia sp. transmitted by O. octodontus clustering with Borrelia sp. Alcohuaz, which was previously detected in that same tick species. On the other hand, concatenated clpX-pepX-recG-rplB-uvrA demonstrated that the characterized spirochete branches together with "Candidatus Borrelia caatinga", a recently discovered species from Brazil. Based on the genetic profile presented in this study, the name "Candidatus Borrelia octodonta" is proposed for the species transmitted by O. octodontus. The fact that spirochetes were not observed in blood of guinea pigs, may reflect the occurrence of low spirochetemia, which could be explained because the susceptibility of infection varies depending on the rodent species that is used in experimental models. Although the vertebrate reservoir of "Ca. Borrelia octodonta" is still unknown, Octodon degus, a rodent species that is commonly parasitized by O. octodontus, should be a future target to elucidate this issue.

DNA barcoding and new records of Ornithodoros yumatensis from Central Mexico.

Bats represent the second order of mammals with the highest number of species worldwide with over 1,616 species, and almost 10% of them are recorded in Mexico. These mammals have a great diversity of ectoparasites, in particular soft ticks of the genus Ornithodoros. Desmodus rotundus is one of the bat species that has scarcely been studied in terms of tick species richness in Mexico, with three tick species reported in five of the 32 Mexican states. For this reason, the aim of the present work was to identify ticks associated with D. rotundus from Central Mexico. Fieldwork was undertaken in the municipality El Marqués, Ejido Atongo A, Querétaro, Mexico. Bats were captured using mist nets and were visually inspected for tick presence. The ectoparasites were identified morphologically and molecularly with the use of mitochondrial markers 16SrDNA and cytochrome oxidase subunit I (COI). A total of 30 D. rotundus (1 female, 29 males) were captured, from which 20 larvae identified as Ornithodoros yumatensis were recovered. Molecular analysis confirmed the presence of this species with identity values of 99-100% with sequences of this species from the southwestern US, and the Yucatán Peninsula, Mexico. This is the first report of ticks associated with bats for the state of Querétaro, providing the first sequences of the COI gene from Mexican populations of O. yumatensis and shows an increase in the distribution of this soft tick across Central Mexico.

Description of a new Pavlovskyella species (Acari: Argasidae) from Chile.

Soft ticks (Argasidae) of the Pavlovskyella Pospelova-Shtrom subgenus are important vectors of relapsing fever spirochetes, which are agents of disease globally. South American representatives of the Pavlovskyella subgenus include 3 species: Ornithodoros (Pavlovskyella) brasiliensis Aragão, Ornithodoros (Pavlovskyella) furcosus Neumann, and Ornithodoros (Pavlovskyella) rostratus Aragão. Here, we describe a fourth species based on morphological and mitogenomic evidence of ticks collected in burrows of unknown hosts in central Chile. The larva of the new species separates from other South American soft ticks by the following combination of characters: 13 pairs of dorsolateral setae, dorsal plate hexagonal, hypostome blunt with denticles from apex almost to the base. Adults of this new species lack cheeks, possess a dorsoventral groove, and have humps, similar to O. (P.) brasiliensis; however, they lack bulging structures on the flanks of idiosoma. Moreover, females and males differ from O. (P.) rostratus by having 3 humps instead of spurs in tarsi I and from O. (P.) furcosus because of their smaller size and thinner anterior lip of the genital aperture in females. The phylogenetic analysis performed with mitogenomes of the Argasidae family depicts the new Pavlovskyella species from Chile in a monophyletic clade with other South American species in the subgenus, confirming a regional group.

Confirmation of the presence of Rickettsia felis infecting Ornithodoros puertoricensis in Mexico.

Soft ticks are neglected competent vectors of a wide range of pathogenic microorganisms, among which bacteria of the genera Rickettsia and Borrelia stand out. In Mexico, previous studies have shown the presence of a member of the Ornithodoros talaje complex in the Virginia opossum (Didelphis virginiana Didelphimorphia: Didelphidae Kerr) from southeastern Mexico. However, its specific identification has not been achieved. Two D. virginiana were treated in a private clinic during the period of April-May 2022. Tick larvae were manually removed, DNA extraction was performed, and some genes from various bacterial and parasitic pathogens were amplified and sequenced. A total of 96 larvae were recovered, which were morphologically identified as Ornithodoros puertoricensis (Ixodida: Argasidae Fox); the 16 S sequences showed a similarity of 96.79%-99.51% with sequences of O. puertoricensis from Panama and Colombia. The presence of Rickettsia felis (Rickettsiales: Rickettsiaceae Bouyer et al.) was detected in 15 specimens from one host. The soft tick O. puertoricensis is recorded for the first time as an ectoparasite of the Virginia opossum in America and represents the second report for this soft tick in Mexico since 1963. This represents the most northern record of this tick species in its geographic distribution and brings a new soft tick-Rickettsia association.

Ornithodoros mimon colonizing a residence in Campinas, state of São Paulo, associated with human parasitism / Ornithodoros mimon colonizando uma residência em Campinas, estado de São Paulo, associado a parasitismo humano

Ornithodoros mimon is an argasid tick species usually associated with bats and marsupials and occasionally parasitizes humans inside their homes. This paper reports a tick infestation in a residence in the municipality of Campinas, located in the interior of the state of São Paulo (SP). This report increases O. mimon occurrence in SP and corroborates its anthropophilic activity. Further studies are needed to clarify its role as a vector of pathogens. We highlighted the presence of O. mimon in an area with a large human population (Campinas) associated with synanthropic animals.(AU)

Ornithodoros mimon é uma espécie de carrapato argasídeo, geralmente associada a morcegos e marsupiais, sendo ocasionalmente relatada parasitando humanos dentro de seus domicílios. Este trabalho relata a infestação por carrapatos em uma residência no município de Campinas, interior do estado de São Paulo (SP). O presente relato amplia a ocorrência de O. mimon no estado de SP, corroborando sua atividade antropofílica, sendo necessários mais estudos para esclarecer o seu possível papel como vetor de patógenos. Destaca-se a presença de O. mimon numa área de grande contingente humano (Campinas), associado a animais sinantrópicos.(AU)

Bacteria related to tick-borne pathogen assemblages in Ornithodoros cf. hasei (Acari: Argasidae) and blood of the wild mammal hosts in the Orinoquia region, Colombia.

Interest in research on soft ticks has increased in recent decades, leading to valuable insight into their role as disease vectors. The use of metagenomics-based analyses have helped to elucidate ecological factors involved in pathogen, vector, and host dynamics. To understand the main bacterial assemblages present in Ornithodoros cf. hasei and its mammalian hosts, 84 ticks and 13 blood samples from bat hosts (Chiroptera) were selected, and the 16S rRNA gene V4 region was sequenced in five pools (each one related to each host-tick pairing). Bacterial taxonomic assignment analyses were performed by comparing operational taxonomic units (OTUs) shared between ticks and their host blood. This analysis showed the presence of Proteobacteria (38.8%), Enterobacteriaceae (25%), Firmicutes (12.3%), and Actinobacteria (10.9%) within blood samples, and Rickettsiaceae (39%), Firmicutes (25%), Actinobacteria (13.1%), and Proteobacteria (9%) within ticks. Species related to potentially pathogenic genera were detected in ticks, such as Borrelia sp., Bartonella tamiae, Ehrlichia sp. and Rickettsia-like endosymbiont, and the presence of these organisms was found in all analyzed bat species (Cynomops planirostris, Molossus pretiosus, Noctilio albiventris), and O. cf. hasei. About 41-48.6% of bacterial OTUs (genera and species) were shared between ticks and the blood of bat hosts. Targeted metagenomic screening techniques allowed the detection of tick-associated pathogens for O. cf. hasei and small mammals for the first time, enabling future research on many of these pathogens.

Quantification of ASFV DNA and RNA in Ornithodoros Soft Ticks.

Molecular biology methods are highly sensitive to detect the genome of pathogens and to study their biology. Polymerase chain reaction (PCR) and reverse transcription followed by a polymerase chain reaction (RT-PCR) permit the detection of the presence and the replication of African swine fever virus in soft ticks. Here, we described our techniques to detect and quantify DNA and RNA of African swine fever virus in soft ticks including a housekeeping gene of soft ticks as internal control.

First Report of the Bat Tick Carios kelleyi (Acari: Ixodida: Argasidae) From Vermont, United States.

The soft tick Carios kelleyi (Cooley and Kohls, 1941) is an ectoparasite of bats that can harbor bacteria known to cause disease in humans, such as Rickettsia spp., Bartonella spp., and relapsing fever Borrelia spp. Human-tick encounters may occur when bats occupy attics or similar dwellings with access points to human-inhabited areas. During May 2021, a partially engorged adult female C. kelleyi was collected from a Vermont home with an attic that was being used as a roost by big brown bats, Eptesicus fuscus (Chiroptera: Vespertilionidae). The source of the blood in the tick was the domestic dog, Canis lupus familiaris. Subsequently, eight C. kelleyi larvae were collected from a rescued E. fuscus adult. This is the first report of a soft tick species from Vermont and it is unknown how long C. kelleyi has been present in this state. Reports of C. kelleyi are on the rise across the northeastern United States but the implications for the health of humans, domestic animals, and bats in northern New England remain unclear. Bat management plans should consider the importance of bat exclusion in preventing tick encounters with members of the household and should include a tick monitoring component if bats are evicted.

Characterization of Keterah orthonairovirus and evaluation of therapeutic candidates against Keterah orthonairovirus infectious disease.

The species Keterah orthonairovirus is a member of the genus Orthonairovirus. Few studies have focused on this species, and there remains no treatment for Issyk-Kul fever, an infectious disease caused by a Keterah orthonairovirus. This study was performed to characterize this species using two viruses, Issyk-Kul virus (ISKV) and Soft tick bunyavirus (STBV), in cell culture and type I interferon receptor knockout (IFNAR-/-) mice and to evaluate the efficacy of serum transfusion using a mouse model of ISKV infection. The two viruses replicated in many kinds of mammal- and tick-derived cell lines but showed few different characteristics in tropism and antigenicity against anti-viral sera in cell culture. Neither virus caused clinical signs in wild-type mice, but both caused lethal infection in IFNAR-/- mice. ISKV caused more acute death than STBV in IFNAR-/- mice. In both viral infections in IFNAR-/- mice, macroscopic abnormalities were prominent in the liver. Similar levels of viral genome between ISKV- and STBV-infected IFNAR-/- mice were observed in blood, liver, lymphoid tissues and adrenal gland at moribund stages. Hematologic abnormalities in IFNAR-/- mice infected with these viruses, including leukopenia and thrombocytopenia, and biochemical abnormalities indicating liver damage were prominent. In addition, blood levels of many kinds of cytokines and chemokines such as granulocyte colony-stimulating factor, interleukin-6, tumor necrosis factor-α, interferon gamma-induced protein 10 and monocyte chemoattractant protein-1 were elevated. ISKV-immunized serum transfusion after infection delayed the time to death of IFNAR-/- mice. Thus, the present study showed that the species Keterah orthonairovirus could proliferate in most mammal-derived cell lines and cause severe liver lesions and death in IFNAR-/- mice and that serum transfusion might be effective in treatment against Issyk-Kul fever.

Record Longevity and Reproduction of an African Tick, Argas brumpti (Ixodida: Argasidae).

Argas brumpti Neumann is a large argasid (soft) tick that inhabits the drier areas of eastern and southern Africa. This species typically feeds on a wide variety of small to large mammals (including humans) and lizards, and resides in shallow caves, rocky areas, or dust-bath areas used by large mammals. Individuals of this species, collected as nymphs and adults from a semidesert area of Kenya and subsequently maintained under constant conditions in the laboratory, survived for 27 yr. Furthermore, after 8 yr of starvation and at least 4 yr after the last male died, at least one female laid eggs. The progeny developed into considerable numbers of both males and females, some of which are still living after 26 yr. The longevity of these ticks is apparently a record for any species of tick. The delay in reproduction likely represents long-term storage of viable sperm, also apparently a record for any species of tick.

Borreliae Part 2: Borrelia Relapsing Fever Group and Unclassified Borrelia.

Borreliae of the relapsing fever group (RFG) are heterogenous and can be divided mainly into three groups according to vectors, namely the soft-tick-borne relapsing fever (STBRF) Borreliae, the hard-tick-borne relapsing fever (HTBRF) Borreliae, the louse-borne relapsing fever (LBRF) Borreliae, and the avian relapsing fever ones. With respect to the geographical distribution, the STBRF Borreliae are further subdivided into Old World and New World strains. Except for the Avian relapsing fever group Borreliae, which cause avian spirochetosis, all the others share infectivity in humans. They are indeed the etiological agent of both endemic and epidemic forms of relapsing fever, causing high spirochaetemia and fever. Vectors are primarily soft ticks of Ornithodoros spp. in the STBRF group; hard ticks, notably Ixodes sp., Amblyomma sp., Dermacentor sp., and Rhipicephalus sp., in the HTBRF group; and the louse pediculus humanus humanus in the TBRF one. A recent hypothesis was supported for a common ancestor of RFG Borreliae, transmitted at the beginning by hard-body ticks. Accordingly, STBRF Borreliae switched to use soft-bodied ticks as a vector, which was followed by the use of lice by Borrelia recurrentis. There are also new candidate species of Borreliae, at present unclassified, which are also described in this review.

Carios kelleyi (Acari: Ixodida: Argasidae) Infected With Rickettsial Agents Documented Infesting Housing in Kansas, United States.

During September-December 2018, 25 live ticks were collected on-post at Fort Leavenworth, Kansas, in a home with a history of bat occupancy. Nine ticks were sent to the Army Public Health Center Tick-Borne Disease Laboratory and were identified as Carios kelleyi (Cooley and Kohls, 1941), a species that seldom bites humans but that may search for other sources of blood meals, including humans, when bats are removed from human dwellings. The ticks were tested for numerous agents of human disease. Rickettsia lusitaniae was identified by multilocus sequence typing to be present in two ticks, marking the first detection of this Rickettsia agent in the United States and in this species of tick. Two other Rickettsia spp. were also detected, including an endosymbiont previously associated with C. kelleyi and a possible novel Rickettsia species. The potential roles of C. kelleyi and bats in peridomestic Rickettsia transmission cycles warrant further investigation.

A new species of soft tick from dry tropical forests of Brazilian Caatinga.

Ornithodoros tabajara n. sp. is described from laboratory-reared larvae and adult specimens collected in the Brazilian Caatinga. This new species shares the ecological niche with Ornithodoros rietcorreai and is likely associated with colonial rodents of genus Kerodon. However, O. tabajara n. sp. is morphologically easy to distinguish from O. rietcorreai and other Neotropical Ornithodoros by a unique combination of characters: larva with 17 pairs of dorsal setae (seven anterolateral, three central and seven posterolateral), sub-oval dorsal plate, hypostome blunt apically with dentition formula 2/2 along its extension, only one pair of posthypostomal setae, six pairs of sternal setae, posteromedian setae absent, and leave-shaped anal valves; alive adults with whitish islands of mammillae symmetrically distributed on dorsum (not visible in ethanol-preserved specimens), and median disk merging with posteromedian file. A phylogenetic analysis performed with mitochondrial 16S rDNA sequences points O. tabajara n. sp. as O. rietcorreai's sister taxon, which rises the hypothesis of sympatric speciation.

Identification of Host Bloodmeal Source in Ornithodoros turicata Dugès (Ixodida: Argasidae) Using DNA-Based and Stable Isotope-Based Techniques.

The ecology and host feeding patterns of many soft ticks (Ixodida: Argasidae) remain poorly understood. To address soft tick-host feeding associations, we fed Ornithodoros turicata Dugès on multiple host species and evaluated quantitative PCR (qPCR) and stable isotope analyses to identify the vertebrate species used for the bloodmeal. The results showed that a qPCR with host-specific probes for the cytochrome b gene successfully identified bloodmeals from chicken (Gallus gallus L.), goat (Capra aegagrus hircus L), and swine (Sus scrofa domesticus) beyond 330 days post-feeding and through multiple molting. Also, qPCR-based bloodmeal analyses could detect multiple host species within individual ticks that fed upon more than one species. The stable isotope bloodmeal analyses were based on variation in the natural abundance of carbon (13C/12C) and nitrogen (15N/14N) isotopes in ticks fed on different hosts. When compared to reference isotope signatures, this method discerned unique δ13C and δ15N signatures in the ticks fed on each host taxa yet could not discern multiple host species from O. turicata that fed on more than one host species. Given the significance of soft tick-borne zoonoses and animal diseases, elucidating host feeding patterns from field-collected ticks using these methods may provide insight for an ecological basis to disease management.

Detection of Rickettsia lusitaniae Among Ornithodoros sawaii Soft Ticks Collected From Japanese Murrelet Seabird Nest Material From Gugul Island, Republic of Korea.

In a follow-up to the investigations of soft ticks identified from seabird nest soil and litter collected from coastal islands of the Republic of Korea (ROK), Ornithodoros sawaii and Ornithodoros capensis were assessed for the presence and identification of rickettsiae. Ticks collected from samples of 50-100 g of nest litter and soil from seabird nests were identified individually by morphological techniques, and species confirmed by sequencing of the mt-rrs gene. Subsequently, tick DNA preparations were screened for the presence of rickettsiae using a genus-specific nested PCR (nPCR) assay targeting the 17 kDa antigen gene. The amplicons from the 17 kDa assay and two additional nPCR assays targeting the gltA and ompB gene fragments were sequenced and used to identify the rickettsiae. A total of 134 soft ticks belonging to two species, O. sawaii Kitaoka & Suzuki 1973 (n = 125) and O. capensis Neumann 1901 (n = 9), were collected. Rickettsia lusitaniae DNA was detected and identified among O. sawaii ticks (n = 11, 8.8%) collected from nest litter and soil of the Japanese murrelet (Synthliboramphus wumizusume Temminck 1836) at Gugul Island along the western coastal area of the ROK. This study confirmed for the first time the presence of R. lusitaniae associated with O. sawaii collected from migratory seabird nests in the ROK.

Differential vector competence of Ornithodoros soft ticks for African swine fever virus: What if it involves more than just crossing organic barriers in ticks?

BACKGROUND: Several species of soft ticks in genus Ornithodoros are known vectors and reservoirs of African swine fever virus (ASFV). However, the underlying mechanisms of vector competence for ASFV across Ornithodoros species remain to be fully understood. To that end, this study compared ASFV replication and dissemination as well as virus vertical transmission to descendants between Ornithodoros moubata, O. erraticus, and O. verrucosus in relation to what is known about the ability of these soft tick species to transmit ASFV to pigs. To mimic the natural situation, a more realistic model was used where soft ticks were exposed to ASFV by allowing them to engorge on viremic pigs. METHODS: Ornithodoros moubata ticks were infected with the ASFV strains Liv13/33 (genotype I) or Georgia2007/1 (genotype II), O. erraticus with OurT88/1 (genotype I) or Georgia2007/1 (genotype II), and O. verrucosus with Ukr12/Zapo (genotype II), resulting in five different tick-virus pairs. Quantitative PCR (qPCR) assays targeting the VP72 ASFV gene was carried out over several months on crushed ticks to study viral replication kinetics. Viral titration assays were also carried out on crushed ticks 2 months post infection to confirm virus survival in soft ticks. Ticks were dissected. and DNA was individually extracted from the following organs to study ASFV dissemination: intestine, salivary glands, and reproductive organs. DNA extracts from each organ were tested by qPCR. Lastly, larval or first nymph-stage progeny emerging from hatching eggs were tested by qPCR to assess ASFV vertical transmission. RESULTS: Comparative analyses revealed higher rates of ASFV replication and dissemination in O. moubata infected with Liv13/33, while the opposite was observed for O. erraticus infected with Georgia2007/1 and for O. verrucosus with Ukr12/Zapo. Intermediate profiles were found for O. moubata infected with Georgia2007/1 and for O. erraticus with OurT88/1. Vertical transmission occurred efficiently in O. moubata infected with Liv13/33, and at very low rates in O. erraticus infected with OurT88/1. CONCLUSIONS: This study provides molecular data indicating that viral replication and dissemination in Ornithodoros ticks are major mechanisms underlying ASFV horizontal and vertical transmission. However, our results indicate that other determinants beyond viral replication also influence ASFV vector competence. Further research is required to fully understand this process in soft ticks.

Confirmación de Argas neghmei (Ixodida: Argasidae) en Perú y reporte del carpintero andino (Colaptes rupicola) como nuevo hospedero / Confirmation of Argas neghmei (Ixodida: Argasidae) in Peru and report of the Andean flicker (Colaptes rupicola) as a new host

Resumen Este informe describe el hallazgo de garrapatas blandas (Ixodida, Argasidae) parasitando a dos carpinteros andinos (Colaptes rupicola) provenientes del distrito de Nuñoa, provincia de Melgar en Puno, Perú (14°31'11.77"S; 70°32'15.95"W; 3967 m de altitud). Un total de 29 larvas de garrapatas fueron colectadas directamente de las aves y posteriormente identificadas morfológicamente como Argas neghmei. El diagnóstico fue basado en la morfología de las larvas y confirmado mediante el análisis molecular del gen 16S ARNr mitocondrial de la garrapata. Un fragmento de 363 pares de bases de este gen mostró una identidad del 100% con secuencias registradas para A. neghmei de Argentina y Chile (GenBank: FJ853598 y DQ295781). Este hallazgo confirma la presencia de A. neghmei en Perú y agrega al carpintero andino como nuevo hospedero para esta garrapata.

Abstract This report describes the finding of soft ticks (Ixodida, Argasidae) parasitizing two Andean flickers (Colaptes rupicola) from the Nuñoa district, Melgar province in Puno, Peru (14°31'11.77"S; 70°32'15.95''W; elevation 3967 m). A total of 29 larval ticks were collected directly from the birds and morphologically identified as Argas neghmei. Diagnosis was based on the larval morphology and confirmed by molecular analysis of the tick mitochondrial 16S rRNA gene. A sequence of 363 base pair of this gene showed to be 100% identical with sequences of A. neghmei from Argentina and Chile (GenBank: FJ853598 and DQ295781). This finding confirms the presence of A. neghmei in Peru and adds the Andean flicker as a new host for this species.