Multiple piroplasm parasites (Apicomplexa: Piroplasmida) in northeastern populations of the invasive Asian longhorned tick, Haemaphysalis longicornis Neumann (Ixodida: Ixodidae), in the United States.

Piroplasms, which include the agents of cattle fever and human and dog babesiosis, are a diverse group of blood parasites of significant veterinary and medical importance. The invasive Asian longhorned tick, Haemaphysalis longicornis, is a known vector of piroplasms in its native range in East Asia and invasive range in Australasia. In the USA, H. longicornis has been associated with Theileria orientalis Ikeda outbreaks that caused cattle mortality. To survey invasive populations of H. longicornis for a broad range of piroplasms, 667 questing H. longicornis collected in 2021 from 3 sites in New Jersey, USA, were tested with generalist piroplasm primers targeting the 18S small subunit rRNA (395­515 bp, depending on species) and the cytochrome b oxidase loci (1009 bp). Sequences matching Theileria cervi type F (1 adult, 5 nymphs), an unidentified Theileria species (in 1 nymph), an undescribed Babesia sensu stricto ('true' Babesia, 2 adults, 2 nymphs), a Babesia sp. Coco (also a 'true Babesia', 1 adult, 1 nymph), as well as Babesia microti S837 (1 adult, 4 nymphs) were recovered. Babesia microti S837 is closely related to the human pathogen B. microti US-type. Additionally, a 132 bp sequence matching the cytochrome b locus of deer, Odocoileus virginanus, was obtained from 2 partially engorged H. longicornis. The diverse assemblage of piroplasms now associated with H. longicornis in the USA spans 3 clades in the piroplasm phylogeny and raises concerns of transmission amplification of veterinary pathogens as well as spillover of pathogens from wildlife to humans.

Expanding the universe of Piroplasmids: morphological detection and phylogenetic positioning of putative novel piroplasmids in black-eared opossums (Didelphis aurita) from southeastern Brazil, with description of "South American Marsupialia Group" of Piroplasmida.

The growing proximity of wildlife to large urban niches arouses greater interest in understanding wild reservoirs in the epidemiology of diseases of importance to animal and human health. The aim of the present study was to investigate the presence of piroplasmids in opossums rescued from the metropolitan region of Rio de Janeiro state, Brazil. Blood and bone marrow samples were collected from 15 Didelphis aurita and subjected to DNA extraction and PCR using primers for the 18S rRNA, cox1, cox3, and hsp70 genes of piroplasmids. Clinical and hematological evaluation of the animals was also performed. Five (33.3%) of the 15 opossums tested positive for piroplasms in the nested PCR based on the 18S rRNA, and in two animals, it was possible to observe intra-erythrocytic structures compatible with merozoites. One of the positive animals showed clinical signs of infection such as jaundice, fever, and apathy. Anemia, low level of plasma protein, leukocytosis, and regenerative erythrocyte signs were observed in positive animals. Phylogenetic analysis based on both 18S rRNA and cox-3 genes demonstrated that the piroplasmids detected in D. aurita formed a unique sub-clade, albeit related to piroplasmids previously detected in Didelphis albiventris and associated ticks from Brazil. This study proposes the novel Piroplasmida Clade, namely "South American Marsupialia Group," and reinforces the need for new clinical-epidemiological surveys to understand the dynamics of these infections in didelphids in Brazil.

Molecular detection of piroplasmids in mammals from the Superorder Xenarthra in Brazil.

Xenarthra mammals can be found from southern North America to southern South America, including all Brazilian biomes. Although it has been shown that Xenarthra mammals can play a role as reservoirs for several zoonotic agents, few studies investigate the diversity of piroplasmids (Apicomplexa: Piroplasmida) in this group of mammals. Taking into account that piroplasmids can cause disease in animals and humans, understanding the prevalence and diversity of piroplasmids in Xenarthra mammals would contribute to conservation efforts for this group of animals as well as to infer risk areas for transmission of emergent zoonosis. The present study aimed to investigate the occurrence and molecular identity of piroplasmids in free-living mammals of the Superorder Xenarthra from four Brazilian states (Mato Grosso do Sul, São Paulo, Rondônia, and Pará). For this, DNA was extracted from blood or spleen samples from 455 animals. A nested PCR based on the 18S rRNA gene was used as screening for piroplasmids. Of the 455 samples analyzed, 25 (5.5%) were positive. Additionally, PCR assays based on 18S rRNA near-complete, cox-1, cox-3, hsp70, cytB, ß-tubulin genes and the ITS-1 intergenic region were performed. Five out of 25 positive samples also tested positive for ITS-1-based PCR. The phylogenetic analysis positioned three 18S rRNA sequences detected in Priodontes maximus into the same clade of Babesia sp. detected in marsupials (Didelphis albiventris, Didelphis marsupialis, and Monodelphis domestica) and Amblyomma dubitatum collected from opossums and coatis in Brazil. On the other hand, the 18S rRNA sequence obtained from Dasypus novemcinctus was closely related to a Theileria sp. sequence previously detected in armadillos from Mato Grosso State, grouping in a subclade within the Theileria sensu stricto clade. In the phylogenetic analysis based on the ITS-1 region, the sequences obtained from Myrmecophaga tridactyla and Tamandua tetradactyla were placed into a single clade, apart from the other piroplasmid clades. The present study demonstrated the molecular occurrence of Piroplasmida in anteaters and Babesia sp. and Theileria sp. in armadillos from Brazil.

A novel variant of Babesia sp. (Piroplasmida) as a hemoparasite in procellariiform seabirds.

Procellariiformes includes pelagic seabirds that only use land for breeding; and also, these sites mostly occur in insular habitats. These peculiar habits make the investigation of hemoparasites a challenging issue. Thus, the data on the blood parasites of Procellariiformes are still scarce. In the order Piroplasmida, 16 species of Babesia have been described in terrestrial birds and seabirds. However, there is no register for Babesia spp. in procellariiform seabirds. Hence, the objective of this survey was to investigate the occurrence of Babesia spp. in these seabirds. A total of 220 tissue samples from 18 different seabird species were analyzed; the samples comprised blood and fragments of liver and spleen. The samples were obtained from live rescued animals and carcasses found along the southern coast of Brazil. Polymerase chain reaction (PCR) was conducted, followed by phylogenetic analysis. Only one blood sample yielded a positive result, from an adult female Thalassarche chlororhynchos (Atlantic yellow-nosed albatross). The sequence obtained showed the highest identity with sequences of Babesia spp. of birds from the South Pacific, and the isolate was named Babesia sp. strain Albatross. In the phylogenetic analysis, the sequence was grouped within the Babesia sensu stricto group, and further still into a subgroup including Babesia spp. of the Kiwiensis clade (parasites from birds). The phylogenetic analysis also showed that Babesia sp. strain Albatross clustered apart from the Peircei group, a clade that includes Babesia spp. from seabirds. As far as it is known, this is the first report of Babesia sp. in procellariiform seabirds. Babesia sp. strain Albatross may constitute a novel variant of tick-borne piroplasmids associated with the Procellariiformes order.

Piroplasmid infection is not associated with clinicopathological and laboratory abnormalities in cats from Midwestern Brazil.

Feline piroplasmids include the genera Babesia spp., Cytauxzoon spp., and Theileria spp. In Brazil, there are few reports regarding these hemoprotozoans; however, clinicopathological and molecular data are scarce. This study aimed to characterize the clinical relevance of these parasites through hematological, biochemical, and molecular approaches. For this purpose, 166 cats from Brasilia, Federal District, Midwestern Brazil, were screened using a quantitative polymerase chain reaction (qPCR) for piroplasmids based on the LSU4 mitochondrial gene, which resulted in an overall prevalence of 36/166 (21.7%). Twelve of 166 samples (7.2%) were positive for C. felis, while 19/166 (11.4%) were positive for Babesia vogeli. No samples tested positive for Theileria spp. Babesia vogeli and Cytauxzoon spp. LSU4 sequences showed identities of 97-100% and 99.3%, respectively, to US isolates. The hematological and biochemical findings did not differ significantly between the cats that tested positive and negative for piroplasmids. Although the lack of abnormalities in clinical and laboratory parameters does not eliminate the possibility that these cats were sick and recovered, it may suggest that the Brazilian strain of Cytauxzoon spp. is not as pathogenic as that from the USA, despite the high molecular identity with North American isolates.

The Piroplasmida Babesia, Cytauxzoon, and Theileria in farm and companion animals: species compilation, molecular phylogeny, and evolutionary insights.

The order Piroplasmida, including the genera Babesia, Cytauxzoon, and Theileria is often referred to as piroplasmids and comprises of dixenous hemoprotozoans transmitted by ticks to a mammalian or avian host. Although piroplasmid infections are usually asymptomatic in wild animals, in domestic animals, they cause serious or life-threatening consequences resulting in fatalities. Piroplasmids are particularly notorious for the enormous economic loss they cause worldwide in livestock production, the restrictions they pose on horse trade, and the negative health impact they have on dogs and cats. Furthermore, an increasing number of reported human babesiosis cases are of growing concern. Considerable international research and epidemiological studies are done to identify existing parasite species, reveal their phylogenetic relationships, and develop improved or new drugs and vaccines to mitigate their impact. In this review, we present a compilation of all piroplasmid species, isolates, and species complexes that infect domestic mammals and which have been well defined by molecular phylogenetic markers. Altogether, 57 taxonomic piroplasmid entities were compiled, comprising of 43 piroplasmid species, 12 well-defined isolates awaiting formal species description, and two species complexes that possibly mask additional species. The extrapolation of the finding of at least 57 piroplasmid species in only six domestic mammalian groups (cattle, sheep, goat, horse, dog, and cat) allows us to predict that a substantially higher number of piroplasmid parasites than vertebrate host species exist. Accordingly, the infection of a vertebrate host species by multiple piroplasmid species from the same and/or different phylogenetic lineages is commonly observed. Molecular phylogeny using 18S rRNA genes of piroplasmids infecting domestic mammals results in the formation of six clades, which emerge due to an anthropocentric research scope, but not due to a possibly assumed biological priority position. Scrutinizing the topology of inferred trees reveals stunning insights into some evolutionary patterns exhibited by this intriguing group of parasites. Contrary to expectations, diversification of parasite species appears to be dominated by host-parasite cospeciation (Fahrenholz's rule), and, except for piroplasmids that segregate into Clade VI, host switching is rarely observed. When only domestic mammalian hosts are taken into account, Babesia sensu lato (s.l.) parasites of Clades I and II infect only dogs and cats, respectively, Cytauxzoon spp. placed into Clade III only infect cats, Theileria placed into Clade IV exclusively infect horses, wheras Theileria sensu stricto (s.s.) of Clade V infects only cattle and small ruminants. In contrast, Babesia s.s. parasites of Clade VI infect all farm and companion animal species. We outline how the unique ability of transovarial transmission of Babesia s.s. piroplasmids of Clade VI facilitates species diversification by host switching to other host vertebrate species. Finally, a deterioration of sequence fidelity in databases is observed which will likely lead to an increased risk of artifactual research in this area. Possible measures to reverse and/or avoid this threat are discussed.

Anthemosoma garnhami in an HIV-Infected Man from Zimbabwe Living in South Africa.

An HIV-positive man from Zimbabwe living in South Africa sought treatment for multiple clinical signs, including fever, weight loss, anemia, and splenomegaly. We identified in his blood an African rodent piroplasm, Anthemosoma garnhami, related to Babesia species. This finding extends the known geographic and host range of A. garnhami.

First molecular detection of piroplasmids in non-hematophagous bats from Brazil, with evidence of putative novel species.

Piroplasmida is an order of the phylum Apicomplexa that comprises the Babesia, Cytauxzoon, and Theileria genera. These hemoparasites infect vertebrate blood cells and may cause serious diseases in animals and humans. Even though previous studies have shown that bats are infected by different species of piroplasmids, the occurrence and diversity of these hemoparasites have not been investigated in this group of mammals in Brazil. Therefore, the present work aimed to investigate the occurrence and assess the phylogenetic placement of piroplasmids infecting bats sampled in a peri-urban area from Central-Western Brazil. Seventeen (12.6%) out of 135 animals were positive by nested PCR assay for the detection of Babesia/Theileria targeting the 18S rRNA gene. Eleven sequences of the 17 positive samples could be analyzed and showed an identity of 91.8-100% with Theileria bicornis, Babesia vogeli, a Babesia sp. identified in a small rodent (Thrichomys pachyurus) from the Brazilian Pantanal and a Babesia sp. identified in a dog from Thailand as assessed by nBLAST. A phylogenetic tree was constructed from an alignment of 1399 bp length using analyzed and known piroplasmid 18S rRNA sequences. In this tree, piroplasmid 18S rRNA sequences detected in three specimens of Phyllostomus discolor (Piroplasmid n. sp., P. discolor) were placed as a sister taxon to Theileria sensu stricto (Clade V) and Babesia sensu stricto (Clade VI). An additional phylogenetic tree was generated from a shorter alignment of 524 bp length including analyzed piroplasmid 18S rRNA sequences of bat species Artibeus planirostris and A. lituratus (Piroplasmid sp., Artibeus spp.). The two 18S rRNA sequences detected in Artibeus spp. (Piroplasmid n. sp., Artibeus spp.) were placed within Babesia sensu stricto (Clade VI) into a strongly supported clade (bootstrap: 100) that included Babesia vogeli. The two 18S rRNA sequences of Piroplasmid sp., Artibeus spp. showed a single and a two-nucleotide differences, respectively, with respect to B. vogeli in a 709 pb length alignment. For the first time, the present study shows the occurrence of putative new piroplasmid species in non-hematophagous bats from Brazil.

Pathognomonic thoracic radiographic findings are lacking in cats with acute Cytauxzoonosis.

Cytauxzoon felis is a tick-borne haemoprotozoan parasite that often causes fatal disease in domestic cats. Histological studies have described substantial pulmonary pathology due to cytauxzoonosis. Published reports were not found describing the thoracic radiographic signs associated with acute cytauxzoonosis in cats. The purpose of this retrospective descriptive study was to describe thoracic radiographic findings in a group of felines with confirmed acute cytauxzoonosis. A total of 37 cats with confirmed cytauxzoonosis and with available thoracic radiographs were included. A subset of 7 cats in this sample also had histopathologic evaluation of their lung parenchyma. Thoracic radiographs were retrieved and reviewed. A bronchial pulmonary pattern was identified as the most common finding (n = 27/37; 73%). Other radiographic findings included cardiomegaly (n = 19/37; 51%), interstitial pattern (n = 17/37; 46%), pleural effusion (n = 12/37; 32%), arterial vascular distension (n = 10/37; 27%), arterial and venous distension (n = 10/37; 27%), and venous distension (n = 1/37; 3%). The primary histological features present in 7 cats with additional histopathologic evaluation, similar to previously published studies, were vascular occlusion. Our study suggests that, despite severe histologic evidence of disease, there are no pathognomonic thoracic radiographic findings in cats with acute cytauxzoonosis.

First report of Cytauxzoon sp. infection in Germany: organism description and molecular confirmation in a domestic cat.

Cytauxzoonosis is described as an emerging tick-borne disease of domestic and wild felids caused by protozoans of the genus Cytauxzoon. While in the Americas the condition is described as a fatal disease, in Europe, reports on the clinical expression of the infection are scarce. This study describes the first case of Cytauxzoon sp. infection in Germany, in a domestic cat. A 6-year-old male domestic cat living in Saarlouis (Saarland) was presented with anorexia, lethargy and weight loss. The cat had an outdoor lifestyle and had not travelled abroad. Serum clinical chemistry analysis revealed azotaemia with markedly increased symmetric dimethylarginine, hypercreatinemia, hyperphosphatemia and hypoalbuminemia. Moreover, a mild non-regenerative anaemia was present. Approximately 1 year prior to these findings, the domestic cat was diagnosed with a feline immunodeficiency virus (FIV) infection. These results pointed toward a decreased glomerular filtration rate, presumably as a result of kidney dysfunction. Round to oval signet ring-shaped intraerythrocytic organisms, morphologically suggestive for a piroplasm, were revealed during blood smear evaluation with a degree of parasitaemia of 33.0%. PCR analyses and sequencing of a region of the 18S rRNA gene confirmed the presence of a Cytauxzoon sp. infection, with 99-100% nucleotide sequence identity with previously published Cytauxzoon sp. isolates. As this is the first molecularly confirmed Cytauxzoon sp. infection in a domestic cat in Germany, these findings suggest that cytauxzoonosis should be considered as a differential diagnosis in cases of anaemia in outdoor domestic cats, particularly in areas where wild felid populations are present.

Preliminary study of Cytauxzoon felis infection in outdoor cats in Mashhad, Iran.

The objectives of the current study were to assess the preliminary status of Cytauxzoon felis (C. felis) infection among outdoor cats in Mashhad, Iran and also to compare clinicopathological findings between C. felis infected and non-infected cats. Blood samples were collected from 100 outdoor domestic cats between April and September in 2019. Infection with C. felis was determined using microscopic observation of giemsa-stained blood smears and molecular analysis. The piroplasms was microscopically detected in 5 (5%) of the blood smears with low parasitemia. The presence of C. felis was confirmed in one positive microscopy sample by PCR. The molecular assay revealed that 19 cats (19%) were infected with C. felis. Hematological and some serum biochemical factors were evaluated in both of the infected and non-infected cats. There was no association between C. felis infection and age, gender, and laboratory findings except for hematocrit (Hct) and concentration of total protein and globulin. Clinical signs such as fever, dehydration, lethargy, and icterus were observed only in 15.78% (3/19) of the infected cats, while 84.22% (16/19) were asymptomatic. Laboratory findings such as non-regenerative anemia, thrombocytopenia, neutrophilic leukocytosis hyperproteinemia, hypoalbuminemia, and hyperbilirubinemia were detected in the clinically infected cats. This study revealed the relatively high frequency of C. felis infection in outdoor domestic cats in Mashahd, Iran. The predominance of asymptomatic infection likely indicates that these cats may be infected with low-virulent strains of C. felis.

Cytauxzoon felis Infection in Domestic Cats, Yunnan Province, China, 2016.

We performed a molecular survey for Cytauxzoon felis infection in 311 domestic cats in Yunnan Province, China, in 2016 and found a prevalence of 21.5%. C. felis infection in domestic and wild cats in other provinces should be investigated to determine parasite prevalence and genetic diversity among cats throughout China.

Detection of Rangelia vitalii (Piroplasmida: Babesiidae) in asymptomatic free-ranging wild canids from the Pampa biome, Brazil.

Canine rangeliosis is a tick-borne disease caused by the protozoan Rangelia vitalii, which has only been reported in South America. With this knowledge, we hypothesized that neotropical foxes could act as asymptomatic natural carriers of R. vitalii. To test this, we captured 44 free-ranging foxes and investigated the presence of R. vitalii DNA, and whether the infected animals presented any clinical findings or hematological changes. Eight foxes (18%), seven Cerdocyon thous (7/27-25%), and one Lycalopex gymnocercus (1/17-5%) were positive for R. vitalii. All foxes were clinically healthy and showed no hematological abnormalities. Thus, we propose that neotropical canids, particularly C. thous, could be the natural carriers of R. vitalii.

Ticks and accompanying pathogens of domestic and wild animals of Kerala, South India.

The objective of the present study was to detect the chosen nucleotide DNA or RNA sequences of the pathogens in ticks of domestic and wild animals of Kerala, South India based on molecular techniques. Among 602 ticks collected, 413 were from bovines (cattle and buffalo), 26 from goats, 101 from dogs and 62 from wild animals. Amblyomma integrum, Am. gervaisi, Dermacentor auratus, Haemaphysalis bispinosa, Ha. intermedia, Ha. shimoga, Ha. spinigera, Rhipicephalus annulatus, Rh. microplus, Rh. haemaphysaloides and Rh. sanguineus s.l. were identified from various domestic and wild animals of Kerala. The cDNA synthesized from the RNA isolated from fully or partially engorged adult female/nymphal ticks was used as template for the specific polymerase chain reactions (PCR). Out of 602 ticks examined, nucleotide sequences of pathogens were detected in 28 ticks (4.65%). The nucleotide sequences of tick-borne pathogens like Theileria orientalis, Babesia vogeli, Hepatozoon canis, Anaplasma marginale, An. bovis, Rickettsia sp. closely related to Ri. raoultii, Ri. massiliae, Ri. africae and Ri. slovaca were detected. The identification of the previously unreported nucleotide sequences of rickettsial pathogens from India is of particular interest due to their zoonotic significance. The phylogenetic analysis of the major piroplasm surface protein (MPSP) gene of T. orientalis amplified from Rh. annulatus ticks revealed that they were genetically close to type 7, which belong to the highly pathogenic Ikeda group.

Two Tales of Cytauxzoon felis Infections in Domestic Cats.

Cytauxzoonosis is an emerging infectious disease that affects wild felids as well as the domestic cat; it is caused by the apicomplexan protozoan parasites belonging to the genus Cytauxzoon. Cytauxzoonfelis is the species of major concern, whose transmission occurs via the bite of an infected tick. Cytauxzoonosis of the domestic cat has historically been considered uniformly fatal, with a short course of illness, and most domestic cats die within 9 to 15 days postinfection. However, increasing evidence of domestic cats surviving C. felis infection suggests the existence of different strains with various levels of pathogenicity. Although wild felids are considered natural reservoirs for this parasite, a number of studies suggest that domestic cats that have survived nonlethal infections may serve as an additional reservoir. The current article comprehensively reviews the parasite and its life cycle, geographic distribution, genetic variability, and pathogenesis, as well as host immunology and the diagnosis, treatment, and prevention of infection in the domestic cat. This information should provide a basis for better understanding the parasite as well as the pathogenesis of the disease.

Surface immunoglobulins of erythrocytes and platelets in dogs naturally infected by Rangelia vitalii.

Rangelia vitalii is a protozoan of the Babesiidae family that parasitizes domestic and wild dogs in South American countries. The main laboratory findings in blood samples from animals infected by R. vitalii are anemia and thrombocytopenia. The aim of this study was to detect IgM and IgG immunoglobulins on the surface of red blood cells and platelets, as well as to determine the percentage of reticulated platelets and reticulocytes in dogs naturally infected by R. vitalii. Blood samples from twenty dogs seen at the Veterinary Hospital of the Federal University of Santa Maria (UFSM) were divided into two groups: the diseased group consisted of blood samples from 10 animals with the diagnosis of rangeliosis, and the healthy group (control) consisted of samples from 10 healthy animals. All diseased dogs showed normocytic normochromic anemia but showed no differences (p > 0.05) in reticulocyte counts compared to healthy dogs. Moreover, IgM and IgG immunoglobulins were detected on the surface of the plasma membrane of red blood cells from both groups, but the amounts did not differ between groups (p > 0.05). Thrombocytopenia in infected animals was classified as severe. The percentage of reticulated platelets was higher (p < 0.001) in diseased dogs than in healthy animals. Diseased animals showed more IgM immunoglobulins bound to the surface of platelets than did the healthy group (p < 0.001). However, the amount of IgG bound to the surface of platelets was not different between groups. In conclusion, we showed that R. vitalii caused immune-mediated thrombocytopenia since IgM immunoglobulins were found on the surface of platelets of diseased dogs. We suggest that the binding of immunoglobulins on platelet surfaces contributes to early destruction of these cells and, consequently, alterations in hemostasis. An increase in reticulated platelets was noted in response to thrombocytopenia, indicating active thrombopoiesis.

Half a century after its discovery, new insights on Anthemosoma garnhami (Sporozoa, Piroplasmida): morphology, molecular characterisation and phylogenetic position.

Here, we report new insights on the erythrocytic murine parasite Anthemosoma garnhami, which was first described from Ethiopia in 1969. Its classification has been debated for years, as this parasite presents some intermediate characters between the Haemosporidia and the Piroplasmida. Based on electron-microscopy, immunological, biochemical and drug sensitivity studies, it was finally assigned to the piroplasms, in the family Anthemosomatidae. In 1985, Anthemosoma sp. was reported from Namibia, and since then, no investigation has involved this parasite. We re-examined the original material, illustrate the blood stages with a set of coloured microphotographs and performed a morphometric analysis. As no type material was designated at the time of the original description, we designate syntypes. This study provides also the first molecular data on A. garnhami with the amplification and sequencing of two genes: the nuclear 18S small subunit ribosomal RNA and the mitochondrial cytochrome c oxydase subunit I. The phylogenetic analyses of both genes confirm that A. garnhami belongs to the Piroplasmida and appears on its own new branch distinct from both Babesids and Theilerids. This result supports the placement of the genus Anthemosoma in its own family but also invalidates the order Anthemosomida. Being paraphyletic with Babesia, the conundrum about the systematics of the piroplasms is discussed as well as the records, the hosts and the possible vectors of Anthemosoma spp.

Tick infestation and occurrence of Anaplasma phagocytophilum and piroplasms in cattle in the Republic of Serbia.

Ticks transmit important pathogens affecting cattle such as intracellular bacteria of the genus Anaplasma or protozoa of the order Piroplasmida. This study aimed at assessing tick species present on pastures and cattle and determining occurrence of the tick-borne pathogens Anaplasma phagocytophilum and Babesia spp. in cattle. During May and June 2013, ticks and EDTA blood were collected from 185 dairy cows at six locations in Serbia. Ticks were also collected directly from the pastures at four of these locations. The occurrence of A. phagocytophilum was investigated by serological (immunofluorescence antibody test (IFAT)) and molecular methods (real-time PCR) and of piroplasms by traditional PCR followed by sequencing. The most prevalent tick species on animals was Ixodes ricinus, (n = 167), followed by Haemaphysalis punctata (n = 146) and Dermacentor marginatus (n = 122). On the pasture, however, the most common species was H. punctata (n = 41), I. ricinus (n = 37), and D. marginatus (n = 2). Altogether, 4 out of 163 (2.45%) serum samples of cows were positive for A. phagocytophilum-specific antibodies by IFAT. However, the 135 blood samples tested for A. phagocytophilum DNA were all negative. Altogether, 5 out of these 135 samples were positive for piroplasm DNA. PCR products were sequenced and identified as a benign Theileria spp. with 100% identity with GenBank entries from Italy (Theileria sergenti), China (Theileria spp.), and Korea (Theileria buffeli isolate HS252). The results provide evidence for the presence of several hard tick species infesting cattle in Serbia which can carry pathogens potentially influencing animal health, as well as evidence of contact with tick-borne pathogens Theileria spp. and A. phagocytophilum.

Prevalence of Tick-Borne Pathogens from Ticks Collected from Cattle and Wild Animals in Tanzania in 2012.

This study was aimed to disclose the prevalence rate of tick-borne pathogens from ticks collected from cattle and wild animals in Tanzania in 2012. Ticks were collected from slaughtered cattle and dead wild animals from November 5 to December 23, 2012 and identified. PCR for detecting Anaplasmataceae, Piroplamidae, Rickettsiaceae, Borrelia spp., and Coxiella spp. were done. Among those tested, Rickettsiaceae, Piroplasmidae, and Anaplasmataceae, were detected in ticks from the 2 regions. Rickettsiaceae represented the major tick-borne pathogens of the 2 regions. Ticks from animals in Maswa were associated with a higher pathogen detection rate compared to that in ticks from Iringa. In addition, a higher pathogen detection rate was observed in ticks infesting cattle than in ticks infesting wild animals. All examined ticks of the genus Amblyomma were infected with diverse pathogens. Ticks of the genera Rhipicephalus and Hyalomma were infected with 1 or 2 pathogens. Collectively, this study provides important information regarding differences in pathogen status among various regions, hosts, and tick species in Tanzania. Results in this study will affect the programs to prevent tick-borne diseases (TBD) of humans and livestock in Tanzania.

First clinical case report of Cytauxzoon sp. infection in a domestic cat in France.

BACKGROUND: Feline cytauxzoonosis is an emerging infection caused by tick-transmitted apicomplexan parasites of the genus Cytauxzoon. The association of clinical disease with Cytauxzoon infection appears to be limited to C. felis infections in the Americas. Sporadic infections of wild and domestic felids with Cytauxzoon sp. were recently described in European countries but clinical reports of the infection are rare and incomplete. This case report brings new interesting information on cytauxzoonosis expression in Europe. CASE PRESENTATION: A 9-years-old castrated European shorthair cat living in rural area of north-eastern France (Saint Sauveur, Bourgogne-Franche-Comté region), without any travel history was presented for consultation due to hyperthermia, anorexia, depression and prolonged fever that didn't respond to antibiotic therapy. The cat had outdoor access with a history of vagrancy and was adequately vaccinated (core vaccines and FeLV vaccine). During biological investigations, intraerythrocytic inclusions were observed on blood smear and were further investigated by PCR analysis and sequencing. Molecular analyses confirmed Cytauxzoon sp. infection. The cat was treated with a subcutaneous injection of imidocarb dipropionate (3.5 mg/kg). One week after treatment, the cat improved clinically, although parasitic inclusions within erythrocytes persisted, and only a mild lymphocytosis was found. Two weeks after treatment, the cat appeared in excellent health, appetite was normal and parasitemia was negative. However, one month after treatment the cat relapsed with hyperthermia, anorexia, and depression. Blood smears and PCR were once again positive. Subsequently, the cat received an additional dose of imidocarb dipropionate (3.5 mg/kg SC) and recovered rapidly without other clinical signs. Two weeks after the second imidocarb injection, the cat was hit by a car and died. CONCLUSION: This case provides the first clinical description of infection by Cytauxzoon sp. in a domestic cat in France. These findings support the fact that cytauxzoonosis should be considered in the differential diagnosis of acute febrile illness which does not respond to antibiotic in cats with outdoor access especially in areas where populations of wild felids are present.