Anaplasma phagocytophilum in European bison (Bison bonasus) and their ticks from Lithuania and Poland.

The increasing population of European bison (Bison bonasus) can contribute to the prevalence of zoonotic pathogens. The aim of the present study was to assess the presence of A. phagocytophilum infection in European bison tissues as well as ticks removed from European bison in Lithuania and Poland. A further objective of this work was to compare the detected A. phagocytophilum strains. A total of 85 tissue samples (spleen) of European bison and 560 ticks belonging to two species, Ixodes ricinus (n = 408) and Dermacentor reticulatus (n = 152) were tested. DNA of A. phagocytophilum was detected based on RT-PCR in 40% of the European bison samples, 8.8% of the I. ricinus and 5.9% of the D. reticulatus ticks. Analysis of the obtained partial 16S rRNA gene sequences of A. phagocytophilum revealed the presence of three variants with two polymorphic sites. Furthermore, phylogenetic analysis with partial msp4 gene sequences grouped A. phagocytophilum variants into three clusters. This study revealed that the groEL gene sequences of A. phagocytophilum from European bison and their ticks grouped into ecotype I and only one sequence from Lithuanian European bison belonged to ecotype II. The results of the present study indicated that European bison may play a role as a natural reservoir of A. phagocytophilum.

First Molecular Detection of Bartonella bovis and Bartonella schoenbuchensis in European Bison (Bison bonasus).

Bartonella bacteria infect the erythrocytes and endothelial cells of mammalians. The spread of the Bartonella infection occurs mainly via bloodsucking arthropod vectors. Studies on Bartonella infection in European bison, the largest wild ruminant in Europe, are lacking. They are needed to clarify their role in the maintenance and transmission of Bartonella spp. The aim of this study was to investigate the presence of the Bartonella pathogen in European bison and their ticks in Lithuania. A total of 38 spleen samples from bison and 258 ticks belonging to the Ixodes ricinus and Dermacentor reticulatus species were examined. The bison and tick samples were subjected to ssrA, 16S-23S rRNA ITS, gltA, and rpoB partial gene fragment amplification using various variants of PCR. Bartonella DNA was detected in 7.9% of the tissue samples of European bison. All tick samples were negative for Bartonella spp. The phylogenetic analysis of 16S-23S rRNA ITS, gltA, and rpoB partial gene fragment revealed that European bison were infected by B. bovis (2.6%) and B. schoenbuchensis (5.3%). This is the first report addressing the occurrence of B. bovis and B. schoenbuchensis in European bison in Europe.

Impact of tick-borne Anaplasma phagocytophilum infections in calves of moose (Alces alces) in southern Norway.

The Gram-negative, obligate intracellular tick-transmitted pathogen Anaplasma phagocytophilum can cause acute febrile diseases in humans and domestic animals. The expansion of the tick Ixodes ricinus (Linnaeus, 1758) in northern Europe due to climate change is of serious concern for animal and human health. The aim of the present study was to investigate the impact of A. phagocytophilum infection in moose Alces alces (Linnaeus) calves by evaluating the carcass weights of infected and non-infected animals and examining animal tissues samples for co-infections with either species of Babesia Starcovici, 1893 or bacteria of the genus Bartonella. The carcasses of 68 free-ranging moose calves were weighed by hunters during the hunting seasons from 2014 to 2017 in two regions in southern Norway and spleen samples were collected. Anaplasma phagocytophilum was detected in moose sampled from locations infected with ticks with a prevalence of 82% (n = 46). The carcass weights of A. phagocytophilum-infected calves (n = 46) and non-infected (n = 22) calves were compared. Although the average weight of infected calves (45.6 kg) was lower than that of non-infected calves (46.5 kg), the difference was not statistically significant. Three different variants of the bacterium 16S rRNA gene were identified. The average weight of animals infected with variant I was 49.9 kg, whereas that of animals infected with variant III was 42.0 kg, but the difference was not statistically significant (p = 0.077). Co-infections of A. phagocytophilum with Bartonella spp. or with Babesia spp. were found in 20 and two calves, respectively. A triple infection was found in two calves. Sequence analysis of the 18S rRNA gene of Babesia-positive samples revealed the presence of Babesia cf. odocoilei (Emerson et Wright, 1970). Strains of Bartonella closely related to Bartonella bovis (Bermond, Boulouis, Heller, Laere, Monteil, Chomel, Sander, Dehio et Piemont, 2002) were identified based on phylogenetic analysis of the gltA and rpoB genes. The loss of body mass in moose calves in the tick-infected site was probably influenced by multiple factors.

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.

Bartonella washoensis infection in red squirrels (Sciurus vulgaris) and their ectoparasites in Lithuania.

This is the first study to investigate the presence of Bartonella infections in different internal organs of red squirrels and their ectoparasites in Lithuania. A total of 39 roadkill red squirrels were collected. Squirrels were infested with Ixodes ricinus ticks (191) and Ceratophyllus sciurorum fleas (36). The presence of Bartonella spp. was screened using 16 S-23 S rRNA internal transcribed spacer region and bacteria were detected in 38.5 % (15/39) samples of squirrels, 1.0 % (2/191) samples of ticks and 55.5 % (20/36) samples of fleas. The infection rate of different internal organs of squirrels varied from 11.1%-47.4%. The 16 S-23 S rRNA ITS region sequences showed that Bartonella washoensis were detected in squirrels and their ectoparasites. The results from this study support the hypothesis that S. vulgaris and their fleas, C.sciurorum, serve as a major reservoir and a vector, respectively, of zoonotic B. washoensis in Lithuania.

Prevalence and co-infection with tick-borne Anaplasma phagocytophilum and Babesia spp. in red deer (Cervus elaphus) and roe deer (Capreolus capreolus) in Southern Norway.

Anaplasma phagocytophilum and Babesia spp. are causative agents of tick-borne infections that are increasingly considered as a threat to animal and public health. To assess the role of cervids in the maintenance of zoonotic pathogens in Norway, we investigated the prevalence of A. phagocytophilum and Babesia spp. in free-ranging roe deer and red deer. Initial screening of spleen samples of 104 animals by multiplex real-time PCR targeting the major surface protein (msp2) gene and 18S rRNA revealed the presence of A. phagocytophilum infection in 81.1% red deer (Cervus elaphus) and 88.1% roe deer (Capreolus capreolus), and Babesia spp. parasites in 64.9% red deer and 83.6% roe deer, respectively. Co-infections were found in 62.2% red deer and 79.9% roe deer. Nested PCR and sequence analysis of partial msp4 and 18S rRNA genes were performed for molecular characterization of A. phagocytophilum strains and Babesia species. A total of eleven A. phagocytophilum msp4 gene sequence variants were identified: five different variants were 100% identical to corresponding A. phagocytophilum sequences deposited in the GenBank database, while other six sequence variants had unique nucleotide polymorphisms. Sequence analysis of the 18S rRNA gene demonstrated the presence of multiple Babesia species, including Babesia capreoli, Babesia divergens, Babesia venatorum and Babesia odocoilei/Babesia cf. odocoilei. This study is the first report demonstrating the prevalence and molecular characterization of A. phagocytophilum strains and Babesia species in roe deer and red deer in Norway. The high infection and co-infection rates with A. phagocytophilum and Babesia spp. in red deer and roe deer suggest that these cervids may play an important role in the transmission of single and multiple pathogens.

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.