Molecular detection of Bartonella rochalimae and Hepatozoon canis in red foxes (Vulpes vulpes) from China.

Red foxes (Vulpes vulpes) have been recognized as natural reservoirs for multiple pathogens and a source of infection for domestic animals, wildlife and humans. To date, no reports are available on the Bartonella rochalimae and Hepatozoon canis infection in red foxes from China. In 2018-2022, a total of 16 red foxes were sampled in two counties and a city in Xinjiang Uygur Autonomous Region (XUAR) in northwest China. Subsequently analyzed by DNA extraction amplified by polymerase chain reaction (PCR). In the present study, based on nucleotide sequence and phylogenetic tree analyses, B. rochalimae and H. canis were molecularly identified in red foxes. Our findings provide the first molecular evidence of B. rochalimae and H. canis in red foxes from China.

New records of Hepatozoon and Oswaldofilaria from saltwater crocodiles (Crocodylus porosus) in Australia.

Diseases affecting wild Australian saltwater crocodiles (Crocodylus porosus) are rarely reported due to the difficulty in capturing animals and obtaining samples. In this investigation, we identified two haemoparasites (Hepatozoon and a filarial nematode) in saltwater crocodiles in Darwin, Australia. Light microscopic examination identified Hepatozoon in 7/7 (100%) wild crocodiles and in 2/20 (10%) of captive ones. When genomic DNAs from these same samples were further investigated using polymerase chain reaction (PCR)-based sequencing, we detected Hepatozoon in all 27 blood samples. Using both microscopy and PCR-based sequencing, we detected a filarial worm (proposed to be Oswaldofilaria) in one of 20 captive crocodiles. The sequence data were compared with sequence data available in public databases, and phylogenetic analyses indicated that the operational taxonomic units of Hepatozoon and Oswaldofilaria discovered here in these crocodiles are likely new species. This study is the first to use molecular tools to explore haemoparasites in Australian saltwater crocodiles and highlights the importance of health investigations in poorly studied vertebrate hosts.

Description of an intramonocytic haemoparasite, Hepatozoon lainsoni sp. nov. (Apicomplexa: Adeleorina: Hepatozoidae), infecting Ameiva ameiva lizard (Reptilia: Squamata: Teiidae) in northern Brazil.

Haemogregarine (Apicomplexa: Adeleorina) parasites are considered to be the most common and widespread haemoparasites in reptiles. The genus Hepatozoon (Apicomplexa: Adeleorina: Hepatozoidae) can be found parasitizing a broad range of species and, in reptiles, they infect mainly peripheral blood erythrocytes. The present study detected and characterized a haemogregarine isolated from the lizard species, Ameiva ameiva, collected from the municipality of Capanema, Pará state, north Brazil. Blood smears and imprints from lungs, brain, heart, kidney, liver, bone marrow and spleen were observed using light microscopy and the parasite was genetically identified by molecular analysis. Morphological, morphometric and molecular data were obtained. Parasite gamonts were found in 49.5% (55/111) of the blood smears from A. ameiva, and were characterized as oval, averaging 12.0 ± 0.8 × 5.9 ± 0.6 µm2 in size, which displaced the nuclei of parasitized monocytes laterally. Parasite forms resembling immature gamonts were observed in the spleen and bone marrow of the lizards. Furthermore, phylogenetic analyses of 18S rRNA sequences did not reveal gene similarity with other Hepatozoon spp. sequences from reptiles. Thus, morphological and molecular analyses have identified a new species of Hepatozoon parasite, Hepatozoon lainsoni sp. nov., which infects monocytes of the A. ameiva lizard.

Tick-borne pathogens Ehrlichia, Hepatozoon, and Babesia co-infection in owned dogs in Central Thailand.

Tick-borne pathogens are transmitted by a wide range of tick species and affect both human and animal health. Understanding the diversity of these pathogens and their co-infection rates in domesticated animals in urban areas is crucial for effective disease management and prevention. In this study, a total of 565 owned dogs in the central region of Thailand were investigated for the infection rate of three genera of Ehrlichia, Hepatozoon, and Babesia infection using multiplex PCR. The results revealed an overall infection rate of 19.1%, with Ehrlichia having the highest infection rate (12.2%), followed by Babesia (2.5%) and Hepatozoon (1.4%). The rate of co-infection was 3%, with mixed infections involving two or three genera. Male dogs exhibited a slightly higher infection rate compared to females, although not statistically significant. Young adult dogs (1-3 years) showed the highest infection rate of both single infections and co-infections. Monthly infection rate indicated variations throughout the year, with co-infection rate significantly associated with overall infection rate. Clinical manifestations in three genera of infected dogs included thrombocytopenia and eosinopenia. The results of this study are useful to design strategies for the management and prevention of tick-borne diseases in the study area.

Completing the Puzzle: A Cluster of Hunting Dogs with Tick-Borne Illness from a Fishing Community in Tobago, West Indies.

Eight hunting dogs were visited by a state veterinarian on the island of Tobago, Trinidad and Tobago, West Indies, as owners reported anorexia and paralysis in five of their dogs. The veterinarian observed a combination of clinical signs consistent with tick-borne illness, including fever, anorexia, anaemia, lethargy and paralysis. Blood and ticks were collected from each dog and submitted to a diagnostic laboratory for analysis. Microscopic analysis revealed a mixed infection of intracytoplasmic organisms consistent with Babesia spp. (erythrocyte) and Ehrlichia spp. (monocyte), respectively, from one dog, while a complete blood count indicated a regenerative anaemia (n = 1; 12.5%), non-regenerative anaemia (n = 4; 50%), neutrophilia (n = 3; 37.5%), lymphocytosis (n = 2; 25%), thrombocytopaenia (n = 3; 37.5%) and pancytopaenia (n = 1; 12.5%). DNA isolated from the eight blood samples and 20 ticks (16 Rhipicephalus sanguineus and 4 Amblyomma ovale) were subjected to conventional PCR and next-generation sequencing of the 16S rRNA and 18S rRNA gene for Anaplasma/Ehrlichia and Babesia/Theileria/Hepatozoon, respectively. The DNA of Ehrlichia spp., closely related to Ehrlichia canis, was detected in the blood of three dogs (37.5%), Anaplasma spp., closely related to Anaplasma marginale, in two (25%), Babesia vogeli in one dog (12.5%) and seven ticks (35%) and Hepatozoon canis and Anaplasma spp., in one tick (5%), respectively. These findings highlight the need to test both the vector and host for the presence of tick-borne pathogens when undertaking diagnostic investigations. Further studies are also warranted to elucidate the susceptibility of canids to Anaplasma marginale.

CORRECTING THE NAMES OF TWO HAEMOGREGARINA SPP. FROM LIZARDS IN EGYPT.

Two haemogregarine "species" names, Haemogregarina tarentannulari and Haemogregarina rawashi, were cited by Saoud et al. (1995) as having been described by Mohammed and Ramadan (1996, in press). However, the paper by Mohammed and Ramadan (1996) was never published and, therefore, these names and their authorities must be suppressed because they violate Chapter 3 (Criteria for Publication), Article 8 (What Constitutes Published Work) of the International Code of Zoological Nomenclature. The following new names are introduced to replace them based on the Principle of Priority (Chapter 6, Article 23, 23.1, and Chapter 11, Article 51, Recommendation 51E): Hepatozoon rawashi (Mohammed and Ramadan in Saoud, Ramadan, Mohammed and Fawzi, 1995) n. comb., with gamonts in the erythrocytes and meronts in the lungs and liver of the fan-footed gecko, Ptyodactylus hasselquisiti (Donndorff, 1798) from Egypt, and Haemogregarina tarentannulari (Mohammed and Ramadan in Saoud, Ramadan, Mohammed and Fawzi, 1995), with gamonts in the erythrocytes and meronts in the lungs and liver of the white-spotted wall gecko, Tarentola annularis (St. Hilaire, 1827) also from Egypt. This latter species was determined to be a junior synonym of Haemogregarina annularis El-Naffar, Mandour, and Mohammed 1991, which was later reassigned to the genus Hepatozoon based on their phylogenetic analysis of 18S rDNA gene sequences.

Molecular survey of canine tick-borne pathogens in ticks and stray dogs in Dhaka city, Bangladesh.

Molecular surveillance of canine tick-borne pathogens (TBPs) in Bangladesh has constantly been undervalued. Therefore, the emergence of new pathogens often remains undetected. This study aimed to screen tick-borne pathogens in stray dogs and ticks in the Dhaka metropolitan area (DMA). Eighty-five dog blood and 53 ticks were collected in six city districts of DMA from September 2022 to January 2023. The ticks were identified by morphology. Screening of TBPs was performed by polymerase chain reaction (PCR), followed by sequencing. The PCR assays were conducted to analyze the 18S rRNA (Babesia gibsoni, B. vogeli, and Hepatozoon canis), 16S rRNA (Anaplasma phagocytophilum, A. platys, and A. bovis), gltA (Ehrlichia canis and Rickettsia spp.), flagellin B (Borrelia spp.) and 16-23S rRNA (Bartonella spp.). Three tick species, Rhipicephalus sanguineus (50/53), R. microplus (1/53), and Haemaphysalis bispinosa (2/53), were identified. Babesia gibsoni (38 out of 85) and A. platys (7 out of 85) were detected in dog blood. In contrast, four pathogens, B. gibsoni (1 out of 53), B. vogeli (1 out of 53), H. canis (22 out of 53), and A. platys (1 out of 53), were detected in the ticks. However, the detection rates of TBPs in dog blood and ticks were not correlated in this study. The phylogenetic analyses suggested that a single genotype for each of the four pathogens is circulating in DMA. This study reports the existence of B. vogeli, H. canis, and A. platys in Bangladesh for the first time.

Molecular identification of hemoparasites in ixodid ticks in Iran.

A total of 386 ticks were processed in order to investigate the occurrence of selected tick transmitted pathogens (i.e., Theileria, Babesia, Hepatozoon and Cytauxzoon) in ixodid ticks in six provinces of Iran (East Azerbaijan, Gilan, Kermanshah, Khuzestan, Sistan & Baluchestan and Tehran). Ticks identified as Dermacentor marginatus, Hyalomma aegyptium, Hy. anatolicum, Hy. asiaticum, Hy. marginatum, Ixodes ricinus, Rhipicephalus annulatus and R. sanguineus sensu lato were collected from sheep and cattle. Conventional PCR and sequencing results revealed DNA of Theileria ovis in three R. sanguineus sensu lato pools and one D. marginatus pool from sheep in Kermanshah and East Azerbaijan, T. annulata in one Hy. asiaticum pool from cattle in Kermanshah, and He. canis in an individual female Hy. aegyptium in Kermanshah. Data herein indicate the role of R. sanguineus complex and D. marginatus in the epidemiology of ovine theileriosis in western and northwestern Iran. Potential role of Hyalomma aegyptium in the transmission of He. canis is discussed. Considering non-principled movement of livestock across the country and increasing reports about the resistance of ticks to common acaricides, test-and-treatment of infected livestock, vaccination of the livestock against endemic tick-borne pathogens, and the use of non-chemical tick management strategies are recommended.

Phylogenetic analysis and haplotype networking of Hepatozoon felis infecting wild animals in Gir National Park, Gujarat, India.

The present study was aimed to determine the phylogenetic relationship, haplotype network, and demographic dynamics of H. felis infecting the endangered Asiatic lions in Gir National Park, Gujarat, India, on the basis of partial 18S rRNA gene. The phylogenetic analysis based on the partial 18S rRNA gene sequences of H. felis exhibited the presence of two distinct genotypes of H. felis (HfG1 and HfG2) infecting the Indian wild felids, viz., the Asiatic lion, Royal Bengal tiger, and Indian leopard. The HfG1 and HfG2 genotypes exhibited 97.6-100% and 99.7-100%, and 96.9-98.7% nucleotide identity within and between themselves, respectively. The HfG1 genotype exhibited a higher genetic diversity as compared to HfG2. A total of 22 molecular signatures were identified in the 18S rRNA gene between these genotypes. Further, analysis of a total 67 sequences of H. felis (13 different host species from 13 countries of Africa, South America, Asia, and Europe) that were downloaded from GenBankTM, generated 30 haplotypes. Among all the haplotypes, Hap_17 (h=12) was the most frequent followed by Hap_12 (h=09) and Hap_4 (h=05). Out of 13 location-wise populations, India (h=12) shared the highest number of haplotypes followed by Japan (h=08), and the least number of haplotypes were found in Hungary (h=02). Population dynamics study involving neutrality tests and mismatch distribution, and genetic differentiation indices, revealed the presence of phylogeographic population structure and a constant population size indicating a uniform gene flow among the populations worldwide. In conclusion, a high genetic diversity along with the presence of two distinct genotypes of H. felis were observed on the basis of 18S rRNA gene sequence analysis.

Metabarcoding using nanopore long-read sequencing for the unbiased characterization of apicomplexan haemoparasites.

Apicomplexan haemoparasites generate significant morbidity and mortality in humans and other animals, particularly in many low-to-middle income countries. Malaria caused by Plasmodium remains responsible for some of the highest numbers of annual deaths of any human pathogen, whilst piroplasmids, such as Babesia and Theileria can have immense negative economic effects through livestock loss. Diagnosing haemoparasites via traditional methods like microscopy is challenging due to low-level and transient parasitaemia. PCR-based diagnostics overcome these limitations by being both highly sensitive and specific, but they may be unable to accurately detect coinfections or identify novel species. In contrast, next-generation sequencing (NGS)-based methods can characterize all pathogens from a group of interest concurrently, although, the short-read platforms previously used have been limited in the taxonomic resolution achievable. Here, we used Oxford Nanopore Technologies' (ONT) long-read MinION™ sequencer to conduct apicomplexan haemoparasite metabarcoding via sequencing the near full-length 18S ribosomal RNA gene, demonstrating its ability to detect Babesia, Hepatozoon, Neospora, Plasmodium, Theileria and Toxoplasma species. This method was tested on blood-extracted DNA from 100 dogs and the results benchmarked against qPCR and Illumina-based metabarcoding. For two common haemoparasites, nanopore sequencing performed as well as qPCR (kappa agreement statistics > 0.98), whilst also detecting one pathogen, Hepatozoon felis, missed by the other techniques. The long-reads obtained by nanopore sequencing provide an improved species-level taxonomic resolution whilst the method's broad applicability mean it can be used to explore apicomplexan communities from diverse mammalian hosts, on a portable sequencer that easily permits adaptation to field use.

Vector-borne parasites in dogs from Ukraine translocated to Poland following Russian invasion in 2022.

INTRODUCTION: Since 24 February 2022, the day the Russian aggression against Ukraine began, millions of refugees and thousands of pets crossed the Polish-Ukrainian border. Additionally, an unknown number of shelter and stray dogs and cats were rescued and translocated to Poland by private persons and non-profit organizations. The aim of the present study was to examine rescued dogs and cats for presence of canine vector-borne parasites to determine the role of armed conflict in spreading these parasites. METHODS: In July 2022 blood samples were collected from two animal shelters in central Poland hosting dogs and cats rescued from Ukraine. Animals were imported from various regions of Ukraine, including eastern and southeastern Ukraine (military conflict area). Fifty-three dogs (51 from two shelters and two owned ones) and one shelter cat were examined by molecular methods (PCR and sequencing) for the presence of Babesia/Theileria spp., Dirofilaria spp. and Hepatozoon spp. DNA. RESULTS: We detected Dirofilaria immitis, a parasite species non-endemic in Poland, in two dogs translocated from Ukraine (2/53 = 3.8% [95% CI 0.5-13.0%]). One dog had a history of previous heartworm infection. High prevalence of Hepatozoon canis (51% [95% CI 36.8-64.9%]) was noted among translocated dogs. Prevalence of Dirofilaria repens in Ukrainian dogs (18.9% [95% CI 9.4-32.0%) was similar to prevalence in dogs from central Poland (12%). Co-infection of D. repens and D. immitis was found in two dogs and six dogs were co-infected by D. repens and H. canis. Hepatozoon canis infection was also identified in an imported cat. CONCLUSION: We confirmed that this military conflict has facilitated the spread of canine vector-borne parasites, including zoonotic species.

Molecular Survey of Hepatozoon canis Infection in Domestic Dogs from Sardinia, Italy.

Tick-borne protozoans of the genus Hepatozoon have been associated with infections of domestic and wild animals over the world. The occurrence of these apicomplexan agents in Sardinia has been poorly explored so far. In this study, the occurrence of Hepatozoon spp. has been investigated in domestic dogs from nine cities of Sardinia, Italy. Blood samples from each dog were collected and tested molecularly for the presence of Hepatozoon and Babesia/Theileria DNAs. Out of fifty-one dogs, nine were positive for Hepatozoon species based on the molecular detection of the parasite in blood samples. The phylogenetic relationships of strains detected here were also established. The PCR for amplification of the 18S rRNA fragment gene of Babesia/Theileria spp. did not give amplicons in any of the analyzed samples. Our results report the first molecular confirmation of Hepatozoon canis in Sardinian pet dogs and contribute to better understand the presence of these protozoans on the island. This study highlights the importance of recognizing and predicting the risk levels for the canine population, thus increasing the development of specific control measures. Also, since the distribution of hepatozoonosis is closely related to that of the definitive tick host, Rhipicephalus sanguineus, more accurate studies on Rhipicephalus ticks will be needed due to increasing the epidemiological knowledge of Hepatozoon species on the island.

Reevaluation of Hemoparasites in the Black Spiny-Tailed Iguana (Ctenosaura similis) with the First Pathological and Molecular Characterizations of Lankesterella desseri n. sp. and Redescription of Hepatozoon gamezi.

Hemoprotozoa are microorganisms that parasitize the blood and possess intricate life cycles. Despite the complexity of their nature, little is known about the biology of hemoprotozoa in reptilian hosts. In this study, we conducted disease surveillance on blood samples collected from six black spiny-tailed iguanas (Ctenosaura similis) exhibiting clinical signs. We found two different types of hemoparasites in the blood films and further confirmed they belong to the genera Lakesterella and Hepatozoon through molecular methods. In the tissue section from a dead iguana infected only with Lakesterella sp., parasites were also found in melanomacrophages of the liver and kidney. Since Lakesterella sp. infection has not been reported in C. similis, we propose this hemococcidian as a new species, Lankesterella desseri n. sp. The Hepatozoon parasites discovered in this study were classified as Hepatozoon gamezi based on their morphological characteristics, particularly the notable deformation of all infected erythrocytes, and this classification was further corroborated through molecular biological and phylogenetic analyses. This is the first hemoprotozoa investigation in C. similis with pathological and molecular characterization of these pathogens. We suggest that more studies are needed to understand the epidemiology, transmission, and impact of these parasites on their hosts and ecosystems.

Hepatozoon spp. infection in wild canids in the eastern United States.

BACKGROUND: Hepatozoon spp. are apicomplexan parasites known to cause musculoskeletal disease in a variety of animals. Two species are known to infect wild and domestic canids in the US: Hepatozoon canis and H. americanum. METHODS: In this study, blood, heart, and/or spleen samples were collected from 278 wild canids (180 coyotes, 93 red foxes, and 5 gray foxes) in the eastern US and tested via PCR for Hepatozoon. Histology slides of heart and skeletal muscle were assessed for Hepatozoon cysts and associated inflammation when fresh tissue was available (n = 96). RESULTS: Hepatozoon spp. were found in 24.2% (59/278) of individuals, with Hepatozoon canis in 14.0% (34/278) and H. americanum in 10.7% (26/278). One coyote was positive for both H. canis and H. americanum. Foxes were more likely to be positive for H. canis than coyotes (23% and 7% respectively, P = 0.0008), while only coyotes were positive for H. americanum. Of the eight sampled states, H. canis was present in six (Louisiana, North Carolina, Pennsylvania, South Carolina, Tennessee, and Virginia) while H. americanum was found in two southern states (South Carolina and Louisiana). Infection status was positively correlated with myositis and myocarditis, and heart or muscle cysts were found in 83% (5/6) of H. americanum-positive coyotes. CONCLUSION: This survey showed a moderate prevalence of H. canis and H. americanum in states where the parasite was previously unrecorded including South Carolina and Pennsylvania.

Molecular Survey and Genetic Characteristics of Vector-Borne Pathogens in Domestic Dogs from Four Regions of China.

Canine vector-borne diseases are widely distributed around the world. They are transmitted by arthropods, and many seriously threaten the health of animals and humans. In China, our knowledge of Ehrlichia, Hepatozoon, and Mycoplasma species circulating in dogs is still poorly understood. Therefore, the aim of this study was to understand the prevalence and genetic characteristics of canine Ehrlichia spp., Hepatozoon spp., and Mycoplasma spp. in Chongqing (southwest), Fujian (southeast), Shandong (southeast), and Hubei (central) Provinces of China. Blood samples from healthy pet dogs were processed to detect Ehrlichia, Hepatozoon, and Mycoplasma DNA with PCR. Haplotype and phylogenetic analyses were performed on 18S rRNA sequences. Among 306 dogs, no Ehrlichia spp. or Mycoplasma spp. were detected, whereas one Hepatozoon sp. was detected in 10 (3.27%) of the animals. Only Hepatozoon canis was identified and was endemic to Chongqing (2.46%) and Hubei (8.77%). A haplotype analysis identified eight haplotypes among the H. canis isolates. A phylogenetic analysis showed that the H. canis isolates in this study clustered into four clades, together with isolates from different countries and hosts, forming a large group that was clearly separate from other Hepatozoon species. These findings provided new information on the epidemiological characteristics of canine vector-borne diseases in China and will be helpful in the development of efficient measures to safeguard the health and well-being of companion animals and their owners.

First report on detection of Hepatozoon ayorgbor in Rhipicephalus haemaphysaloides and Hepatozoon colubri in Haemaphysalis sulcata and Hyalomma anatolicum: risks of spillover of Hepatozoon spp. from wildlife to domestic animals.

This study aimed to detect Hepatozoon spp. in ticks infesting asymptomatic domestic animals and to provide insight into their potential spillover from wild to domestic animals. In total, 537 tick specimens were collected in Khyber Pakhtunkhwa, Pakistan, and morphologically identified. The most prevalent tick species was Haemaphysalis cornupunctata (69; 12.8%), followed by Haemaphysalis kashmirensis (62; 11.5%), Rhipicephalus microplus (58; 10.8%), Haemaphysalis montgomeryi (51; 9.5%), Rhipicephalus sanguineus (49; 9.1%), each Haemaphysalis bispinosa and Haemaphysalis sulcata (43; 8.0%), each Hyalomma anatolicum and Rhipicephalus turanicus (37; 6.9%), Rhipicephalus haemaphysaloides (33; 6.1%) Hyalomma scupense (30; 5.6%), and Hyalomma isaaci (25; 4.7%). The extracted DNA from a subset of each tick species was subjected to PCR to amplify 18S rRNA fragments of Hepatozoon spp. By BLAST analysis, the Hepatozoon sp. detected in Hy. anatolicum infesting cows and in Ha. sulcata infesting sheep showed 99.7% maximum identity with Hepatozoon colubri. Similarly, the Hepatozoon sp. detected in R. haemaphysaloides infesting goats shared 99.49% maximum identity with Hepatozoon ayorgbor, and the Hepatozoon sp. detected in R. sanguineus infesting dogs exhibited 99.7% identity with Hepatozoon canis. Having an overall infection rate (9.3%; 16/172), the highest infection rate was recorded for each H. canis, and H. colubri (3.5%; 6/172), followed by H. ayorgbor (2.3%; 4/172). In the phylogenetic tree, H. colubri clustered with corresponding species from Iran, H. ayorgbor clustered with the same species from Croatia, Ghana, and Portugal, and H. canis clustered with the conspecifics from Iran, Israel, Romania, and Zambia. Regarding the potential spillover of Hepatozoon spp. from wildlife through ticks, free ranging animals was at higher risk compared to confined animals (RR = 3.05), animals consuming food from wildlife habitats were at higher risk compared to those consuming domestic food (RR = 3.06), and animals residing in farm buildings located in wildlife habitats were at higher risk compared to those residing in farm buildings located in villages (RR = 3.28). In addition to the first report on H. canis in R. sanguineus in Pakistan, this is the earliest data showing H. ayorgbor in R. haemaphysaloides and H. colubri in Ha. sulcata and Hy. anatolicum. These preliminary findings suggest a potential spillover of Hepatozoon spp. from wild to domestic animals via ticks under certain risk factors.

Epidemiological survey of tick-borne pathogens in dogs in Anhui Province, China.

Tick-borne diseases have continued to increase worldwide in both developing and many developed countries due to the widespread of different tick species and tick's adaptability to different climatic weather. In order to investigate the prevalence of the tick-borne pathogens, EDTA-anticoagulated whole blood samples were aseptically collected from 765 pet dogs in twenty veterinary clinics located in sixteen prefecture-level cities in Anhui Province, China, and the samples were examined and analyzed for tick-borne pathogens using both microscopy and PCR. Our result analysis revealed 17(2.22%) positive samples to Babesia spp and 4(0.52%) positive samples to Hepatozoon spp, of which case of co-infection was recorded in Lu'An and Chuzhou. The BLAST analysis results of the 18S rRNA gene revealed that the dogs were infected with Babesia gibsoni and Hepatozoon canis. All samples were negative for Anaplasma spp., Ehrlichia spp., and Rickettsia spp. This is the first molecular report of B. gibsoni and H. canis in dogs in Anhui, China.

The first report on Hepatozoon canis in dogs and wolves in Poland: clinical and epidemiological features.

BACKGROUND: Canine hepatozoonosis caused by Hepatozoon canis is a common infection in dogs, with frequent case reports from the Mediterranean region and more recently from several Central European countries, such as Hungary and Germany. Despite the high prevalence of H. canis in red foxes, no infections have been reported to date in dogs in Poland. We describe here the first autochthonous cases of H. canis infection in dogs, including their clinical features, and report the prevalence of H. canis in grey wolves from different regions of Poland. METHODS: Thin smears prepared from blood samples collected from dogs were evaluated by microscopic examination. A total of 60 wolves and 47 dogs were tested. Infections were confirmed by PCR and sequencing. RESULTS: Gamonts of H. canis were found in > 50% of the neutrophils of two dogs and in < 10% of the neutrophils in another five dogs. Molecular typing by PCR sequencing of the 18S ribosomal RNA gene fragment confirmed infections in 11 dogs from different regions of Poland, in 2.7% of dogs attending veterinary practices in central Poland and in 35% of wolves from various geographical regions of Poland. Clinical features manifested mostly in older dogs, and the most common signs were anaemia and apathy. Young dogs usually remained asymptomatic. CONCLUSIONS: This is the first report of H. canis infection in dogs and wolves in Poland. Although the exact vector of the parasite is not known, veterinary practitioners should be aware of this new parasitosis and should consider appropriate diagnostics to confirm/exclude this infection. Further studies are needed to understand the transmission routes of H. canis in domestic and wild canids in Poland.

Molecular prevalence and genetic diversity of Hepatozoon spp. in stray cats of Izmir, Türkiye.

Hepatozoon spp. are an apicomplexan protozoan parasites that infect vertebrates including mammals, marsupials, birds, reptiles, and amphibians. Among Hepatozoon species, H. canis and H. felis are causative agents of hepatozoonosis in dogs and cats, respectively and have veterinary importance. This study aimed to determine the prevalence of Hepatozoon spp. in stray cats living in Izmir and investigate genetic diversity among positive samples. To achieve this aim, the prevalence of Hepatozoon spp. 18S rRNA gene was screened by PCR in DNA samples extracted from blood samples of stray cats (n = 1012). Then, Hepatozoon-positive samples were sequenced and the generated data were used for species identification, phylogenetic and haplotype analyses. According to the results, among the samples screened, 2.37 % (24/1012) of them were found to be Hepatozoon-positive, and of these positive samples, 18 (18/24; 75 %) were successfully sequenced. BLAST and phylogenetic analyses revealed that all of these samples were H. felis. Also, phylogenetic analysis showed that H. felis samples were genotype I. Within H. felis samples isolated from cats living in different countries/regions, 9 haplotypes were detected and among these haplotypes, H-1 was found to be prevalent (n = 20 H. felis isolates). In conclusion, this study showed that the prevalence of Hepatozoon spp. was low in stray cats analyzed. Also, H. felis genotype I was predominant in comparison to other Hepatozoon species.

Molecular Detection of Tick-Borne Pathogens in Dogs from Indigenous Communities, Amazon, Brazil.

Background: There are few reports of tick-borne pathogens infecting dogs living in indigenous communities of Brazil. Herein, we aimed to molecularly detect vector-borne pathogens in dogs from two indigenous communities in the Brazilian Amazon. Materials and Methods: We surveyed 327 dogs raised in Amazon region at 2 distinct indigenous ethnicities for the molecular detection of tick-borne pathogens (114 from Tapirapé and 213 from Karajá indigenous ethnicity). Whole blood samples were subjected to PCR and sequencing for Ehrlichia, Babesia, and Hepatozoon. Univariate and multivariate analyses were used to investigate the factors affecting the pathogen infection patterns in dogs. Results: Among the 327 blood samples, 40 were positive for Ehrlichia canis (12.2%), 2 for Anaplasma platys (0.61%), and 204 were positive for Hepatozoon canis (66.5%). Binary Logistic Regression showed association between E. canis infection and ethnicity (p = 0.010) and tick attachment (p = 0.041). Karajá dogs were 3.4 times (95% CI 1.3-8.5) more likely to be positive for E. canis than Tapirapé dogs. Dogs with ticks were 2.5 times more likely (95% CI 1.0-7.6) to be positive for E. canis than dogs without ticks. Conclusions: Our survey expands the knowledge regarding the presence of vector-borne pathogens in dogs from indigenous communities in the Amazon region.