Comparative morphology of the deer ked Lipoptena fortisetosa first recorded from Italy.

Hippoboscidae flies parasitize various animal species. Knowledge about these insects remains sparse, although they are known to cause stress and damage to their hosts, and can also accidentally infest humans, causing different sanitary risks. Research conducted in Tuscany assessing the biology and distribution of Lipoptena cervi (Linnaeus, 1758) (Diptera: Hippoboscidae), the most common ectoparasite of ungulates in Italy, revealed the presence of Lipoptena fortisetosa Maa, 1965 in Italy for the first time. This study includes a morphological comparative description of L. cervi and L. fortisetosa, emphasizing the peculiar differences between the two species to facilitate their accurate identification. The most pertinent morphological differences between the two species are highlighted, such as the external features of the antennae, distribution of bristles, and different features in the external genitalia. In both species, scanning electron microscopy of mouthparts revealed strong adaptive convergence in the feeding apparatus. Modified palps and a very thin proboscis are described in relation to feeding behaviour.

First report of Lipoptena mazamae Rondani, 1878 (Diptera: Hippoboscidae) from Paraguay.

We report for the first time from Paraguay the louse-fly Lipoptena mazamae Rondani, 1878 (Diptera: Hippoboscidae), based on an individual retrieved from the body of a gray brocket Mazama gouazoubira (Fischer), found dead in the Tatí Yupí Biological Refuge, in Alto Paraná, Eastern Paraguay. Before this finding, in the decade of the 1940's, this species of fly was wrongly recorded from Paraguay, based on material collected in Bolivia, some 300 km from the border to Paraguay.

The New Haplotypes of Bartonella spp. and Borrelia burgdorferi Sensu Lato Identified in Lipoptena spp. (Diptera: Hippoboscidae) Collected in the Areas of North-Eastern Poland.

Deer keds are hematophagous ectoparasites (Diptera: Hippoboscidae) that mainly parasitize Cervidae. These flies are particularly important for animal health due to the occurrence of numerous pathogenic microorganisms. They may also attack humans and their bites may cause allergenic symptoms. The aim of the study was to identify the molecular characteristics of Borrelia burgdorferi sensu lato and Bartonella spp. pathogens detected in Lipoptena spp. sampled both from the hosts and from the environment. For identification of Bartonella spp and B. burgdorferi s. l., the primers specific to the rpoB and flaB gene fragments were used, respectively. The overall prevalence of B. burgdorferi s.l. DNA in Lipoptena cervi was 14.04%, including 14.8% infection in the tested group of winged specimens. The overall prevalence of Bartonella spp. was 57.02%. The presence of these bacteria was detected in 53.5% of specimens of L. cervi and 75.7% of L. fortisetosa. The phylogenetic analysis showed five new haplotypes of the rpoB gene of Bartonella sp. isolated from L. cervi/Lipoptena fortisetosa. We also identified one new haplotype of B. afzelii and three haplotypes of B. burgdorferi isolated from winged specimens of L. cervi. This is the first study to detect the genetic material of B. burgdorferi s.l. in L. cervi in Poland and the first report on the identification of these bacteria in host-seeking specimens in the environment.

Patterns of deer ked (Diptera: Hippoboscidae) and tick (Ixodida: Ixodidae) infestation on white-tailed deer (Odocoileus virginianus) in the eastern United States.

BACKGROUND: White-tailed deer (Odocoileus virginianus) host numerous ectoparasitic species in the eastern USA, most notably various species of ticks and two species of deer keds. Several pathogens transmitted by ticks to humans and other animal hosts have also been found in deer keds. Little is known about the acquisition and potential for transmission of these pathogens by deer keds; however, tick-deer ked co-feeding transmission is one possible scenario. On-host localization of ticks and deer keds on white-tailed deer was evaluated across several geographical regions of the eastern US to define tick-deer ked spatial relationships on host deer, which may impact the vector-borne disease ecology of these ectoparasites. METHODS: Ticks and deer keds were collected from hunter-harvested white-tailed deer from six states in the eastern US. Each deer was divided into three body sections, and each section was checked for 4 person-minutes. Differences in ectoparasite counts across body sections and/or states were evaluated using a Bayesian generalized mixed model. RESULTS: A total of 168 white-tailed deer were inspected for ticks and deer keds across the study sites. Ticks (n = 1636) were collected from all surveyed states, with Ixodes scapularis (n = 1427) being the predominant species. Counts of I. scapularis from the head and front sections were greater than from the rear section. Neotropical deer keds (Lipoptena mazamae) from Alabama and Tennessee (n = 247) were more often found on the rear body section. European deer keds from Pennsylvania (all Lipoptena cervi, n = 314) were found on all body sections of deer. CONCLUSIONS: The distributions of ticks and deer keds on white-tailed deer were significantly different from each other, providing the first evidence of possible on-host niche partitioning of ticks and two geographically distinct deer ked species (L. cervi in the northeast and L. mazamae in the southeast). These differences in spatial distributions may have implications for acquisition and/or transmission of vector-borne pathogens and therefore warrant further study over a wider geographic range and longer time frame.

Lipoptena fortisetosa as a vector of Bartonella bacteria in Japanese sika deer (Cervus nippon).

BACKGROUND: Two species of deer ked (Lipoptena cervi and L. mazamae) have been identified as vectors of Bartonella bacteria in cervids in Europe and the USA. In an earlier study we showed that Japanese sika deer (Cervus nippon) harbor three Bartonella species, namely B. capreoli (lineage A) and two novel Bartonella species (lineages B and C); however, there is currently no information on the vector of Bartonella bacteria in sika deer. The aim of this study was to clarify potential vectors of Bartonella in Japanese sika deer. METHODS: Thirty-eight wingless deer keds (L. fortisetosa) and 36 ticks (Haemaphysalis and Ixodes species) were collected from sika deer. The prevalence of Bartonella in the arthropods was evaluated by real-time PCR targeting the 16S-23S internal transcribed spacer (ITS) and by culture of the organisms. The total number of Bartonella bacteria were quantified using real-time PCR. The distribution of Bartonella bacteria in deer ked organs was examined by immunofluorescence analysis. The relationship of Bartonella strains isolated from sika deer and arthropods were examined by a phylogenetic analysis based on concatenated sequences of the gltA, rpoB, ftsZ, and ribC genes, followed by a BLAST search for gltA and rpoB. RESULTS: Bartonella prevalence in deer keds was 87.9% by real-time PCR and 51.5% in culture and that in the ticks was 8.3% by real-time PCR and 2.8% in culture. The mean number of Bartonella bacteria per ked was calculated to be 9.2 × 105 cells. Bartonella aggregates were localized in the midgut of the keds. The phylogenetic analysis and BLAST search showed that both the host deer and the keds harbored two Bartonella species (lineages B and C), while B. capreoli (lineage A) was not detected in the keds. Two novel Bartonella species (lineages D and E) were isolated from one ked. CONCLUSIONS: Lipoptena fortisetosa likely serves as a vector of at least two Bartonella species (lineages B and C), whereas ticks do not seem to play a significant role in the transmission of Bartonella between sika deer based on the lower detection rates of Bartonella in ticks compared to keds. Bartonella species in lineages D and E appear to be L. fortisetosa-specific strains.

Molecular Characterization of Lipoptena fortisetosa from Environmental Samples Collected in North-Eastern Poland.

Recent years have witnessed an increase in the population of Lipoptenafortisetosa in Central Europe. The genetic profile of this ectoparasite has not been studied in Poland to date. The aim of the present study was to confirm the presence of L.fortisetosa in north-eastern Poland and to characterize the examined population with the use of molecular methods. Deer keds were collected between June and July 2019 in six natural, mixed forests. A fragment of the rRNA 16S gene was used as a marker to identify L.fortisetosa by polymerase chain reaction (PCR). DNA samples were sequenced in the last step. Six new locations of L. fortisetosa were confirmed. No significant differences were observed in the sex ratios of L. cervi and L. fortisetosa (L. cervi p-value = 0.74; L. fortisetosa p-value = 0.65). Significant differences were noted between the total size of L. cervi and L. fortisetosa populations (p-value < 0.001). The similarity to GenBank sequences ranged from 95.56% to 100%. The obtained nucleotide sequences were very closely related to L. fortisetosa sequences from Lithuania, the Czech Republic, and Japan. Molecular analyses revealed considerable genetic diversity, which could indicate that various ectoparasite lineages have spread throughout Europe.

Two New Haplotypes of Bartonella sp. Isolated from Lipoptena fortisetosa (Diptera: Hippoboscidae) in SE Poland.

Insects of the genus Lipoptena are parasitic arthropods with a broad host range. Due to the type of parasitism (hematophagy), their potential role as vectors of pathogens, i.e., Bartonella sp., Anaplasma phagocytophilum, Rickettsia spp., and Borrelia burgdorferi is considered. As the range of their occurrence has been changing dynamically in recent years and infestations of humans have increasingly been reported, these organisms are now the subject of numerous studies. Our research aimed to present the molecular characteristics of Bartonella sp. detected in Lipoptena fortisetosa parasitizing wild cervids in south-eastern Poland. Adults of Lipoptena spp. were collected from carcasses of roe deer and red deer between spring and autumn in 2013. The PCR method was used to detect Bartonella sp. in the insects. We report two new haplotypes of the rpoB gene of Bartonella sp. isolated from L. fortisetosa feeding on wild cervids in south-eastern Poland and the presence of this invasive ectoparasitic species in the studied area since 2013. Phylogenetic analyses of newly obtained Bartonella sp. haplotypes confirmed their unique position on the constructed tree and network topology. The rpoB gene sequences found belonging to lineage B support the view that this phylogenetic lineage represents a novel Bartonella species.

Molecular characterization of Lipoptena cervi from environmental samples collected in Poland.

The activity of Lipoptena cervi has intensified in Poland in recent years. The population genetics of this ectoparasite in Poland has never been described in the literature. The objectives of this study were to investigate the population genetics of L. cervi in selected regions of Poland, to evaluate molecular differences between L. cervi populations, and to determine phylogenetic relationships with other L. cervi sequences obtained in previous studies. In 2019, louse flies were sampled in natural mixed forests in five Polish voivodeships. Seven samples of L. cervi were collected from each voivodeship, and a total of 35 insects were analyzed molecularly. In the first step, Lipoptena spp. were identified to species level under a stereoscopic microscope. A fragment of the rRNA 16S gene was used as a marker to identify L. cervi by the PCR assay. The sequences were assigned accession numbers MT337409 to MT337416. A total of eight haplotypes were identified, two of which were dominant. In the obtained sequences, intraspecific pairwise genetic distances varied between 0.000 and 0.0496 (m = 0.0135; SD = 0.0149; SE = 0.0006; V = 110.11). Mean interpopulation diversity was d = 0.0135 (SE = 0.0027). The acquired nucleotide sequences were highly similar to the sequences from the Czech Republic (MF495940, AF322437), Lithuania (MN889542-MN889544) and Poland (MF541726-MF541729). The similarity with GenBank sequences ranged from 97.24% to 100%. This study revealed two dominant haplotypes of L. cervi in Poland, MT337410 and MT337413. Fragments of the analyzed sequences were detected in only one voivodeship. These findings suggest that the two dominant sequences are the oldest sequences that gave rise to the locally identified haplotypes. The lack of significant correlations with the sequences obtained in regions situated west of the research sites suggests the presence of other genetic populations in Europe.

Molecular Detection of Selected Pathogens with Zoonotic Potential in Deer Keds (Lipoptena fortisetosa).

Deer keds are obligatory hematophagous ectoparasites of birds and mammals. Cervids serve as specific hosts for these insects. However, ked infestations have been observed in non-specific hosts, including humans, companion animals, and livestock. Lipoptena fortisetosa is a weakly studied ectoparasite, but there is evidence to indicate that it continues to spread across Europe. The existing knowledge on deer keds' impact on wildlife is superficial, and their veterinary importance is enigmatic. Lipoptena fortisetosa is a species with vectorial capacity, but potential pathogen transmission has not been assessed. The objective of this study was to evaluate the prevalence of selected pathogens in L. fortisetosa collected from cervids and host-seeking individuals in the environment. Out of 500 acquired samples, 307 (61.4%) had genetic material from at least one tested pathogen. Our research suggests that L. fortisetosa may be a potential vector of several pathogens, including A. phagocytophilum, Babesia spp., Bartonella spp., Borellia spp., Coxiella-like endosymbionts, Francisiella tularensis, Mycoplasma spp., Rickettsia spp., and Theileria spp.; however, further, more extensive investigations are required to confirm this. The results of the study indicate that keds can be used as biological markers for investigating the prevalence of vector-borne diseases in the population of free-ranging cervids.

The prevalence, abundance, and molecular characterization of Lipoptena deer keds from cervids.

The objectives of this study were to investigate the prevalence and abundance of deer keds on various cervids in Lithuania, to molecularly characterize the deer ked species based on mitochondrial COI and 16S rRNA genes, and to compare them with Lipoptena species found in other countries. A total of 11,939 deer keds (Lipoptena cervi and Lipoptena fortisetosa) was collected from the fur of 30 cervids in Lithuania between 2015 and 2019. The values of infestation with deer keds differed among the species of the hosts. Moose and red deer were more frequently infested with L. cervi than with L. fortisetosa, while L. fortisetosa was found more often on roe deer. Phylogenetic analysis of the COI and 16S rRNA genes of five Lipoptena species revealed 110 and 55 variable nucleotide positions, respectively. Among Lithuanian samples, three COI haplotypes of L. cervi and three haplotypes of L. fortisetosa were detected, while there was no variation observed in the 16S rRNA sequences analyzed with one haplotype of L. cervi and one haplotype of L. fortisetosa. This is the first study on L. cervi and L. fortisetosa parasitizing cervids and the first molecular characterization of these deer ked species in Lithuania.

The northernmost record of a blood-sucking ectoparasite, Lipoptena fortisetosa Maa (Diptera: Hippoboscidae), in Estonia.

BACKGROUND: Deer keds are obligatory haematophagous parasites of large homeothermic animals, particularly cervids. Two of the five known species occurring in Europe-Lipoptena cervi (Linnaeus) and L. fortisetosa Maa-are known to have a relatively wide distribution. Lipoptena fortisetosa is considered to have been introduced into Europe with sika deer from the Eastern Palaearctic and is continuously expanding its range. Little is known about the medical importance of deer keds, but they can cause hair loss in cervids and are suspected to be vectors of several diseases. NEW INFORMATION: Details of the distribution of Lipoptena fortisetosa in Europe, including its northernmost record, are provided. This species has been shown to have a viable population in Southern Estonia. Furthermore, the differences from allied L. cervi are discussed, based on morphological and molecular characters.

Infections with Bartonella spp. in free-ranging cervids and deer keds (Lipoptena cervi) in Norway.

Bartonella bacteria are arthropod-borne and can cause long-term bacteremia in humans and animals. The predominant arthropod vectors and the mode of transmission for many novel Bartonella species remain elusive or essentially unstudied. The aim of this study was to investigate the prevalence of Bartonella spp. in Norwegian cervids and deer keds (Lipoptena cervi) and to characterise the bacteria by sequencing of the partial gltA gene and 16 S-23 S rRNA intergenic spacer region (ITS) in order to evaluate a possible transmission route. A total of 260 spleen samples and 118 deer keds were collected from cervids by hunters in the Southern part of Norway. Bartonella DNA was detected in 10.5% of spleen samples of roe deer (n = 67), in 35.1% red deer (n = 37), in 35.9% moose (n = 156), and in 85% pools of adult wingless deer ked (n = 59). Two Bartonella lineages were identified based on phylogenetic analysis of the gltA gene and ITS region sequences.