Investigation of Tick-Borne Pathogens in Ixodes Ticks from Bosnia and Herzegovina.

Limited information is available regarding the presence of tick-borne pathogens and their distribution within Ixodes species in Bosnia and Herzegovina. This study aimed to identify Rickettsia spp., Babesia spp., Anaplasma phagocytophilum, and Borrelia burgdorferi sensu lato (s.l.) in Ixodes ticks collected from domestic and wild animals and vegetation in different regions across Bosnia and Herzegovina. A total of 7438 adult ticks, including 4526 Ixodes ricinus, Ixodes canisuga, and Ixodes hexagonus, were collected. Real-time PCR screening of 450 pooled I. ricinus samples revealed a 22.1% infection rate with at least one pathogen. Rickettsia spp. (6.3%) were found in ticks from dogs, cats, and goats, Babesia spp. (3.1%) in ticks from dogs and cattle, A. phagocytophilum (8.8%) in ticks from dogs, goats, and cattle, and B. burgdorferi s.l. (3.4%) in ticks from dogs and cats. Mixed infections with B. burgdorferi s.l. and A. phagocytophilum, as well as B. burgdorferi s.l. and Rickettsia spp., were found in two pools of I. ricinus from dogs and cats, respectively. Additionally, co-infection with Rickettsia spp. and A. phagocytophilum was confirmed in three tick pools from dogs and goats. Each tick from these pooled samples was individually retested to confirm the presence of pathogens. In the examined pooled samples of I. canisuga (1) and I. hexagonus (6), none of the tested pathogens were detected. Our findings represent the first detection of Rickettsia spp., Babesia spp., A. phagocytophilum, and B. burgdorferi s.l. in I. ricinus collected from domestic animals and vegetation in Bosnia and Herzegovina. Considering the established infection rates, the detection of tick-borne pathogens in adult ticks collected from domestic animals and vegetation enriches the current knowledge of the presence of tick-borne pathogens at the local, regional, national, and broader levels.

A New Endemic Locality of Dermacentor reticulatus in Central-Southern Poland and Its Potential Epidemiological Implications.

Dermacentor reticulatus (Acari: Ixodidae) is an important arthropod vector in medical and veterinary contexts. Its geographic range is divided into western and eastern populations separated by a "Dermacentor-free zone" in central Poland. Recent faunistic studies showed a new endemic locality of the species in Upper Silesia to the west of the Vistula River (central-southern Poland) and its co-occurrence with I. ricinus. The prevalence of five tick-borne pathogens (TBPs), e.g., B. burgdorferi s.l., Bartonella spp., Rickettsia spp., and Babesia spp., in the ticks was assessed with polymerase chain reaction (PCR) methods. The molecular studies revealed the presence of Rickettsia spp. in 23.8% of the D. reticulatus specimens. In turn, 94.1% of the I. ricinus adults were infected with B. burgdorferi s.l., 11.7 % with Babesia spp., and 5.8% with Rickettsia spp. Coinfections with two TBPs were noted in 17.6% of the I. ricinus. These findings highlight not only the risk of infestation by both tick species in an area previously considered Dermacentor-free, but also the high prevalence of TBPs in the study area. Increased focus on medical and veterinary services appears necessary to diagnose and prevent tick-borne diseases in this region.

Tick-Borne Pathogens in Dermacentor reticulatus Ticks from Bosnia and Herzegovina.

Dermacentor (D.) reticulatus ticks carry and transmit a wide range of pathogens to vertebrate hosts. Limited information is available about the existence of emerging tick-borne pathogens and the distribution of D. reticulatus in Bosnia and Herzegovina. The study aimed to investigate the occurrence and distribution of D. reticulatus and to detect the presence of Anaplasma spp., Borrelia (B.) burgdorferi s.l., Rickettsia spp., and Babesia spp. in samples originating from questing ticks and ticks collected from domestic animals in various regions of Bosnia and Herzegovina. A total of 402 collected D. reticulatus ticks were widely distributed throughout the country. Of the 41 pools consisting of 205 individual D. reticulatus ticks, 21 (51.2%) indicated the presence of Rickettsia spp., 17 (41.4%) of Babesia spp., 2 (4.8%) of Anaplasma spp., and 1 (2.4%) of B. burgdorferi s.l. after real-time PCR screening. Our study indicates that D. reticulatus has significantly expanded its distribution and host range in Bosnia and Herzegovina. Moreover, our results represent the first detection of Babesia spp. in D. reticulatus in Bosnia and Herzegovina. Given the demonstrated presence of emerging pathogens in questing and feeding ticks, there is an urge to establish a surveillance system for ticks and tick-borne pathogens in Bosnia and Herzegovina.

Challenges of animals shelters in caring for dogs infected with Leishmania and other pathogens.

The incidence of human Visceral Leishmaniasis (VL) has decreased in Brazil; however, the number of areas reporting human and canine cases has increased, with Leishmania infantum usually preceding human infection. This study aimed to analyze the profile of infectious diseases that are endemic for both human and canine VL, in dogs housed in a shelter located in the state of Rio Grande do Norte, Northeast Brazil. Data was obtained between November/2021 to April/2022. All dogs residing at the shelter (98 dogs) were examined and blood was collected for testing for L. infantum, Ehrlichia canis, and Babesia sp. Statistical analyses considered the clinical and laboratory findings. Of the 98 animals, approximately 43% were positive for L. infantum antibodies, 19% were positive for L. infantum kDNA, and 18% were L. infantum positive by culture. Greater levels of anti-leishmania antibodies were observed in dogs with symptoms suggestive of VL. The dogs tested positive for E. canis (19/98) and B. canis (18/98). Lutzomyia longipalpis was captured inside the shelter, representing 74.25% (n = 225) of whole sandflies in the dog shelter. Concomitant infection by L. infantum and E. canis increased the odds of death. Treatment of VL included the use of allopurinol (n = 48) and miltefosine (n = 8). Treated animals showed more signs of Leishmania infection. Tickborn parasites and Leishmania were prevalent in sheltered dogs in a VL-endemic area, which increases the odds of death and poses an additional challenge for caring for abandoned dogs and at the same time setting protocols to manage reservoirs of L. infantum.

Molecular Identification of Babesia and Theileria Infections in Livestock in the Qinghai-Tibetan Plateau Area, China.

The northwestern region of China, known as the Qinghai-Tibet Plateau Area (QTPA), is characterized by unique climate conditions that support the breeding of various highly-adapted livestock species. Tick vectors play a significant role in transmitting Babesia and Theileria species, posing serious risks to animal health as well as the economy of animal husbandry in QTPA. A total of 366 blood samples were collected from Tibetan sheep (n = 51), goats (n = 67), yaks (n = 43), cattle (n = 49), Bactrian camels (n = 50), horses (n = 65), and donkeys (n = 40). These samples were examined using conventional and nested PCR techniques to detect Theileria and Babesia species. The overall infection rates were 0.3% (1/366) for Babesia spp. and 38.2% (140/366) for Theileria spp. Notably, neither Babesia nor Theileria species were detected in donkeys and yaks. The infection rates of Babesia and Theileria species among animals in different prefectures were significantly different (p < 0.05). Furthermore, Babesia bovis, B. bigemina, B. caballi, and B. ovis were not detected in the current study. To our knowledge, this is the first documented detection of Theileria luwenshuni infection in Bactrian camels and goats, as well as T. sinesis in cattle and T. equi in horses on the Qinghai plateau. These novel findings shed light on the distribution of Babesia and Theileria species among livestock species in QTPA.

Molecular detection of Babesia vesperuginis in bats from Lithuania.

Babesia vesperuginis is an intraerythrocytic protozoan parasite that circulates among bats and ticks in many countries worldwide. However, the distribution of B. vesperuginis in the Baltic region has not been studied. A total of 86 dead bats from eight different species were collected and screened for Babesia spp. using real-time PCR. Overall, 52.3% (45/86) of the bats were found positive for Babesia spp. The prevalence of Babesia spp. in different organs varied, with the highest prevalence observed in heart tissues (37.0%) and the lowest in liver tissues (22.2%). However, the observed differences in prevalence among organs were not statistically significant. Blood samples from 125 bats of nine different species were also analyzed for Babesia spp. prevalence using real-time PCR and nested PCR. The results showed a prevalence of 35.2% and 22.4%, respectively. Moreover, 28.3% (17/60) of the examined blood samples were confirmed positive for Babesia spp. through blood smear analysis. The total of 32 partial sequences of the 18S rRNA gene derived in this study were 100% identical to B. vesperuginis sequences from GenBank. In eight species of bats, Pipistrellus nathusii, Pipistrellus pipistrellus, Pipistrellus pygmaeus, Vespertilio murinus, Eptesicus nilssonii, Eptesicus serotinus, Myotis daubentonii and Nyctalus noctula, Babesia parasites were identified. In E. nilssonii, Babesia spp. was identified for the first time.

Asymptomatic carrier of Babesia spp. among blood donors - epidemiological situation in Poland.

Introduction: Babesiosis is caused by one of several Babesia species. In Europe, B. divergens predominates in humans, while in North America it is B. microti. Babesia spp. infection in donors with a disease-free course of infection can be a major problem in blood recipients. A recipient with impaired immune system functions is at risk of full-blown development of the disease. In Poland and in most countries of the world, blood donors are not routinely tested for Babesia spp. infection. In our previous study, we detected Babesia venatorum DNA in blood donors, which was the reason for expanding the study to include more test subjects. Objective: The aim of this study was an attempt at estimating the prevalence of asymptomatic infection with Babesia spp. among blood donors from the Regional Centres for Blood Donation and Blood Treatment in Warsaw and Wroclaw. Materials and methods: The material for the study was whole blood from regular blood donors from two Regional Centre for Blood Donation and Blood Treatment in Warsaw and Wroclaw. Whole blood samples from 1,067 blood donors collected in June-July 2022 were analyzed. Blood collected directly from the donor during the blood donation procedure. All persons qualified by a doctor as a donor were selected for the study, regardless of age and sex. All subjects were informed in detail about the purpose of the study and gave their written consent. Isolation was made by using the Chelex 100 chelating resin, followed by the studying of the genetic material using the qPCR reaction. The results were analysed based on the amplification curve. Results: The protozoan Babesia spp. was not detected in the blood samples. Conclusions: The risk of blood-borne babesiosis is extremely low in Poland.

Zoonotic Microparasites in Invasive Black Rats (Rattus rattus) from Small Islands in Central Italy.

Invasive species have a detrimental impact on native populations, particularly in island ecosystems, and they pose a potential zoonotic and wildlife threat. Black rats (Rattus rattus) are invasive species that disrupt native flora and fauna on islands and serve as potential competent reservoirs for various pathogens and parasites. Microparasites screening was conducted in rat populations from small islands in central Italy (the Pontine Islands and Pianosa) with the aim of assessing the role of rats in maintaining infections, particularly in cases where key reservoir hosts were scarce or absent. We focused on microparasites of zoonotic and veterinary relevance. A total of 53 rats was kill-trapped and target tissues were analysed with molecular techniques. We observed the absence or very low prevalence of Anaplasma spp., while Babesia was found in rats from all locations, marking the first recorded instance of Babesia divergens in wild rats. Data from Pianosa strongly suggest the presence of an autochthonous Leishmania infantum cycle in the Tuscan archipelago islands. Neospora caninum was absent from all islands, even in areas where dogs, the main reservoirs, were present. Toxoplasma gondii was only recorded on the Pontine Islands, where genotyping is needed to shed light on infection dynamics. This study confirms that invasive species, such as rats, may be responsible for maintaining an increased parasitological threat to fauna and human communities in certain ecosystems.

Molecular detection of pathogens from ticks collected from dogs and cats at veterinary clinics in Finland.

BACKGROUND: Ticks carry microbes, some of which are pathogenic for humans and animals. To assess this One Health challenge, 342 ticks were collected from pet dogs and cats at 10 veterinary clinics in Finland as part of the European project "Protect Our Future Too". METHODS: The tick species were identified, and ticks were screened with quantitative PCR (qPCR) for tick-borne pathogens, including Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Ehrlichia canis, Anaplasma spp., Candidatus Neoehrlichia mikurensis, tick-borne encephalitis virus (TBEV), and Babesia spp. For comparison, a subset of tick DNA (20 qPCR-positive samples) was analysed with 16S next-generation sequencing (NGS). RESULTS: Most ticks were Ixodes ricinus (289, 84.5%), followed by Ixodes persulcatus (51, 14.9%). One hybrid tick (I. ricinus/I. persulcatus, 0.3%) and one Rhipicephalus sanguineus tick (0.3%) were identified. We found one or more of the analysed pathogens in 17% (59/342) of the ticks. The most prevalent pathogen was B. burgdorferi s.l. (36, 10.5%), followed by Anaplasma phagocytophilum (12, 3.5%), B. miyamotoi (5, 1.5%), Babesia venatorum (4, 1.2%), and TBEV (1, 0.3%). Candidatus Neoehrlichia mikurensis DNA was amplified from three (0.9%) ticks. Ehrlichia canis was not detected. In the 16S NGS, six samples produced enough reads for the analysis. In these six samples, we confirmed all the positive qPCR findings of Borrelia spp. and Ca. N. mikurensis. CONCLUSIONS: The high prevalence of pathogenic microorganisms in the ticks of this study emphasizes the importance of awareness of ticks and tick-borne diseases and prevention. Furthermore, the results show that veterinary surveillance can facilitate early detection of tick-borne pathogens and new tick species and draw attention to possible co-infections that should be considered both in symptomatic humans and animals after tick bites.

Enfermedades transmitidas por vectores en gatos: una mirada molecular en ambientes urbanos de Medellín, Colombia / Vector-borne diseases in cats: a molecular look at urban environments in Medellin, Colombia

RESUMEN Los microorganismos hemotrópicos en felinos son agentes infecciosos que varían desde nematodos, protozoos y bacterias. El presente estudio retrospectivo tiene como objetivo evaluar la frecuencia de agentes hemotrópicos mediante qPCR de las bases de datos de 1.418 felinos en Medellín entre julio de 2021 y marzo de 2022, periodo en el que se evidencia una frecuencia del 70%, con un número de animales infectados con uno, dos o tres agentes del 56%, 14%, y 2,3%, respectivamente. La frecuencia para cada uno de los agentes es: Rickettsia spp. 0,21%, Babesia spp. 0,35%, Ehrlichia spp. 0,49%, Dirolifaria spp. 0,64%, Anaplasma spp 0,7%, Hepatozoon spp. 5,4%, Mycoplasma spp. 24,4% y Bartonella spp. 37,9%. Las coinfecciones evidenciadas de dos agentes hemotrópicos son: Bartonella spp. y Mycoplasma spp. 7,9%, Bartonella spp. y Hepatozoon spp. 2,1%, Mycoplasma spp. y Hepatozoon spp. 2% y Ehrlichia spp. y Anaplasma spp. 0,5%. De los 15 hemogramas de felinos infectados, 11 de ellos tienen hemogramas sin alteraciones significativas. Dos de los felinos positivos evidencian anemia moderada y severa y reticu-locitos de 0,9% y 0,4%, respectivamente. Solo un individuo positivo para Mycoplasma spp. presenta trombocitopenia y tres plaquetas en limites inferiores. Se concluye que la PCR es la prueba más confiable para el diagnóstico de agentes hemotrópicos.

ABSTRACT Hemotropic microorganisms in felines are infectious agents that vary from nematodes, protozoa, and bacteria. The objective of this retrospective study is to evaluate the frequency of hemotropic agents by means of qPCR from the databases of 1,418 felines in the city of Medellín between July 2021 and March 2022, where a frequency of 70% is evidenced, with several infected animals, with one, two, or three agents of 56%, 14%, and 2.3%, respectively. With a frequency for each of the agents of: Rickettsia spp. 0.21%, Babesia spp. 0.35%, Ehrlichia spp. 0.49%, Dirolifaria spp. 0.64%, Anaplasma spp. 0.7%, Hepatozoon spp. 5.4%, Mycoplasma spp. 24.4%, and Bartonella spp. 37.9% The evidenced coinfections of two hemotropic agents is: Bartonella spp. and Mycoplasma spp. 7.9%, Bartonella spp. and Hepatozoon spp. 2.1%, Mycoplasma spp. and Hepatozoon spp. 2% and Ehrlichia spp. and Anaplasma spp. 0.5%. Of the 15 blood counts from infected cats, 11 of them had blood counts without significant changes. Two of the positive cats show moderate and severe anemia, and reticulocytes of 0.9% and 0.4%, respectively. Only one individual positive for Mycoplasma spp. presented thrombocytopenia, and three platelets in lower limits. It is concluded that PCR is the most reliable test for the diagnosis of hemotropic agents.

Prevalence of gastro-intestinal and haemoparasitic infections among domestic cats of Kerala.

The population of domesticated cats has drastically increased during the last decades. With the recently identified rise in cat population an upsurge in the parasitic infections associated with domestic cats is evident. A total of 122 domestic cats were screened for gastro-intestinal and haemoparasites. Screening for gastro-intestinal parasites revealed an overall prevalence of 19 per cent (23/122). Ancylostoma spp. was the major gastro-intestinal parasite noticed (61 per cent) followed by Toxocara cati (13.04 per cent), Isospora spp. (8.7 per cent), Diphyllobothrium latum (4.35 per cent) and mixed infection of these (13 per cent). Blood smear examination revealed Cytauxzoon spp. in three cats (2.46 per cent) and Babesia spp. in two cats (1.6 per cent). None of the cats were positive for gamonts of Hepatozoon spp. Molecular analysis revealed Hepatozoon spp. infection in seven cats (5.7 per cent), Cytauxzoon spp. in 29 cats (23.8 per cent) and Babesia spp. in two cats (1.6 per cent). Present study highlights the inevitability of molecular techniques in the identification of haemoparasites. Establishment of proper preventive measures are required to control parasitic infection among domestic cats.

Prevalence of Borrelia, Neoehrlichia mikurensis and Babesia in ticks collected from vegetation in eastern Poland.

In Poland, tick-borne diseases constitute the majority of diseases related to exposure to biological agents with a predominance of Lyme borreliosis; therefore, research on ticks as a reservoir of various pathogens remains crucial in the epidemiology of human diseases after tick bites. This study aimed to identify the occurrence of Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Neoehrlichia mikurensis, and Babesia spp. in ticks collected from vegetation in eastern Poland. Additionally, the prevalence of co-infections in the adult Ixodes ricinus ticks was determined. Among I. ricinus ticks the predominantly detected pathogen was B. burgdorferi s.l. (23%) with B. burgdorferi sensu stricto as the most frequently identified species, followed by B. garinii. In 2013, the double or triple infections of B. burgdorferi s.s., B. afzelii, and B. garinii species did not exceed 9% in adult ticks, whereas in 2016, the prevalence of mixed infections reached 29%. The prevalence of N. mikurensis and B. miyamotoi in I. ricinus was determined at the same level of 2.8%. Four Babesia species were identified in the examined I. ricinus population: B. microti (1.5%), B. venatorum (1.2%), B. divergens (0.2%), and B. capreoli (0.1%). Co-infections were detected in 10.1% of all infected ticks with the highest prevalence of co-infections with B. burgdorferi s.l. and Babesia species. The changes in the prevalence and the distribution of particular pathogens within tick populations indicate the need for monitoring the current situation related to tick-borne pathogens from the aspect of risk to human health.

Molecular Detection and Phylogenetic Analyses of Babesia spp. and Theileria spp. in Livestock in Bangladesh.

Piroplasmosis, caused by Babesia spp. and Theileria spp., poses significant constraints for livestock production and upgradation in Bangladesh. Besides examining blood smears, few molecular reports are available from some selected areas in the country. Therefore, the actual scenario of piroplasmosis in Bangladesh is deficient. This study aimed to screen the piroplasms in different livestock species by molecular tools. A total of 276 blood samples were collected from cattle (Bos indicus), gayals (Bos frontalis) and goats (Capra hircus) in five geographies of Bangladesh. After that, screening was conducted through a polymerase chain reaction, and species were confirmed by sequencing. The prevalence of Babesia bigemina, B. bovis, B. naoakii, B. ovis, Theileria annulata and T. orientalis was 49.28%, 0.72%, 1.09%, 32.26%, 6.52% and 46.01%, respectively. The highest prevalence (79/109; 72.48%) of co-infections was observed with B. bigemina and T. orientalis. The phylogenetic analyses revealed that the sequences of B. bigemina (BbigRAP-1a), B. bovis (BboSBP-4), B. naoakii (AMA-1), B. ovis (ssu rRNA) and T. annulata (Tams-1) were included in one clade in the respective phylograms. In contrast, T. orientalis (MPSP) sequences were separated into two clades, corresponding to Types 5 and 7. To our knowledge, this is the first molecular report on piroplasms in gayals and goats in Bangladesh.

White-Toothed Shrews (Genus Crocidura): Potential Reservoirs for Zoonotic Leptospira spp. and Arthropod-Borne Pathogens?

Three species of white-toothed shrews of the order Eulipotyphla are present in central Europe: the bicolored (Crocidura leucodon), greater (Crocidura russula) and lesser (Crocidura suaveolens) white-toothed shrews. Their precise distribution in Germany is ill-defined and little is known about them as reservoirs for zoonotic pathogens (Leptospira spp., Coxiella burnetii, Brucella spp., Anaplasma phagocytophilum, Babesia spp., Neoehrlichia mikurensis and Bartonella spp.). We investigated 372 Crocidura spp. from Germany (n = 341), Austria (n = 18), Luxembourg (n = 2) and Slovakia (n = 11). West European hedgehogs (Erinaceus europaeus) were added to compare the presence of pathogens in co-occurring insectivores. Crocidura russula were distributed mainly in western and C. suaveolens mainly in north-eastern Germany. Crocidura leucodon occurred in overlapping ranges with the other shrews. Leptospira spp. DNA was detected in 28/227 C. russula and 2/78 C. leucodon samples. Further characterization revealed that Leptospira kirschneri had a sequence type (ST) 100. Neoehrlichia mikurensis DNA was detected in spleen tissue from 2/213 C. russula samples. Hedgehogs carried DNA from L. kirschneri (ST 100), L. interrogans (ST 24), A. phagocytophilum and two Bartonella species. This study improves the knowledge of the current distribution of Crocidura shrews and identifies C. russula as carrier of Leptospira kirschneri. However, shrews seem to play little-to-no role in the circulation of the arthropod-borne pathogens investigated.

Molecular detection and characterization of vector-borne agents in common opossums (Didelphis marsupialis) from northeastern Brazil.

Opossums are synanthropic marsupials able to interchange among wild, peri­urban and urban environments, playing an epidemiologically important role as hosts for emerging pathogens and ectoparasites of relevance in public health. The present study aimed to detect and molecularly characterize vector-borne agents in a population of common opossums (Didelphis marsupialis) from the Island of São Luís do Maranhão, northeastern Brazil. Of the 45 animals analyzed, one (2.22%) was positive in the nested PCR assay based on the 18S rRNA gene of piroplasmids. The obtained sequence was phylogenetically positioned in a clade containing sequences of Babesia sp. previously detected in Didelphis aurita, Didelphis albiventris and associated ticks from Brazil. Eight (17.77%) samples were positive in PCR for Ehrlichia spp. based on the dsb gene; four samples were sequenced and positioned into a new clade, sister to E. minasensis and Ehrlichia sp. clade detected in Superorder Xenarthra mammals. No samples tested positive in the screening PCR assays based on the 16S rRNA gene of Anaplasma spp. Two samples were positive in the qPCR for Bartonella spp. based on the nuoG gene. Seven animals (15.56%) were positive in the nPCR based on the 16S rRNA gene of hemoplasmas. Of these, three were positive in a PCR based on the 23S rRNA gene. The phylogenies based on both 16S rRNA and 23S rRNA genes corroborated to each other and positioned the sequences in the same clade of hemoplasmas previously detected in D. aurita and D. albiventris sampled in Brazil. Finally, three (6.66%) animals were positive in the PCR for Hepatozoon spp.; the obtained 18S rRNA sequence was positioned into the H. felis clade.The present study showed, for the first time, the circulation of piroplasmids, Hepatozoon spp., Ehrlichia spp., hemoplasmas and Bartonella spp. in D. marsupialis sampled in northeastern Brazil, with description of putative novel genotypes of Ehrlichia and Hepatozoon and copositivity by different vector-borne agents. The present work consolidates the "South American Marsupialia" piroplasmid clade, adding one more genotype of Babesia sp. to this clade.

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 Babesia microti in dromedary camels in Egypt.

Babesia microti (Apicomplexa: Piroplasmida) causes a medically important tick-borne zoonotic protozoan disease. Egyptian camels are susceptible to Babesia infection; however, just a few cases have been documented. This study aimed to identify Babesia species, specifically Babesia microti, and their genetic diversity in dromedary camels in Egypt and associated hard ticks. Blood and hard tick samples were taken from 133 infested dromedary camels slaughtered in Cairo and Giza abattoirs. The study was conducted from February to November 2021. The 18S rRNA gene was amplified by polymerase chain reaction (PCR) to identify Babesia species. Nested PCR targeting the ß-tubulin gene was used to identify B. microti. The PCR results were confirmed by DNA sequencing. Phylogenetic analysis based on the ß-tubulin gene was used to detect and genotype B. microti. Three tick genera were identified in infested camels (Hyalomma, Rhipicephalus, and Amblyomma). Babesia species were detected in 3 out of 133 blood samples (2.3%), while Babesia spp. were not detected in hard ticks by using the 18S rRNA gene. B. microti was identified in 9 out of 133 blood samples (6.8%) and isolated from Rhipicephalus annulatus and Amblyomma cohaerens by the ß-tubulin gene. The phylogenetic analysis of the ß-tubulin gene revealed that USA-type B. microti was prevalent in Egyptian camels. The results of this study suggested that the Egyptian camels may be infected with Babesia spp. and the zoonotic B. microti strains, which pose a potential risk to public health.

Molecular Detection of Babesia gibsoni in Cats in China.

As there are few studies of Babesia spp. infection in cats in China, or anywhere in the world, the aim of this study was to explore the epidemic features of babesiosis in pet cats in China. In total, 429 blood samples were randomly collected in four different geographical regions. The 18S rRNA gene fragment of Babesia spp. was amplified by nest polymerase chain reaction (PCR), and haplotype and phylogenetic analysis of Babesia were performed to analyze the relationship of this protozoa. The total positive rate of infection was 2.8%. BLAST analysis indicated that Babesia gibsoni was detected in 12 cats. Among these, 4.3%, 3.1%, 0.8% and 2.0% were from Chongqing, Fujian, Hubei and Shandong, respectively. Haplotype and phylogenetic analysis showed that there were nine haplotypes and no obvious genetic variation among B. gibsoni populations. These findings will be helpful for understanding the epidemiology of Babesia spp. in China, and provide a foundation for developing effective preventative strategies.

Detection of Ticks and Tick-Borne Pathogens of Urban Stray Dogs in South Africa.

This study aimed to identify ticks infesting dogs admitted to the Potchefstroom Animal Welfare Society (PAWS) and to detect tick-borne pathogens they are harbouring. A total of 592 ticks were collected from 61 stray dogs admitted to PAWS originating from several suburbs in and near Potchefstroom, South Africa. The dog ticks were identified as Haemaphysalis elliptica (39%) and Rhipicephalus sanguineus (61%) by both morphological and DNA analyses. Of these ticks, H. elliptica consisted of 67.5% (156/231) and 32.5% (75/231) female and male ticks, respectively, whilst R. sanguineus consisted of 48.5% (175/361) and 51.5% (186/361) female and male ticks, respectively. Microscopic examination of blood smears from engorged female ticks indicated overall occurrences of 0.5% (1/204) for Babesia spp. from R. sanguineus, 1% (2/204) of Anaplasma spp. from H. elliptica, and 22% (45/204) of Rickettsia spp. from both H. elliptica and R. sanguineus. Using pooled samples molecular detection of tick-borne pathogens indicated overall occurrences of 1% (1/104) for A. phagocytophilum in H. elliptica, 9.6% (10/104) of Rickettsia spp. in H. elliptica and R. sanguineus, 5.8% (6/104) of Ehrlichia canis in H. elliptica and R. sanguineus, and 13.5% (14/104) of Coxiella spp. in both H. elliptica and R. sanguineus. Additionally, PCR detected 6.5% (2/31) of Coxiella spp. DNA from H. elliptica eggs. Our data indicate that urban stray dogs admitted at PAWS are infested by H. elliptica and R. sanguineus ticks which are harbouring several pathogenic organisms known to cause tick-borne diseases.

Molecular detection of pathogens in the ova and unfed larvae of Rhipicephalus annulatus and Haemaphysalis bispinosa ticks infesting domestic cattle of south India.

This study aimed to investigate the presence of pathogens in the engorged ticks infesting domestic cattle, their ova, and unfed larvae. The engorged female ticks infesting domestic cattle of Wayanad district of Kerala, south India were collected and kept for oviposition. The dead females after the complete oviposition, their egg masses, and unfed larvae were screened for the presence of various pathogens by specific PCRs. The presence of Babesia bigemina, Anaplasma marginale, A. phagocytophilum, and Rickettsia spp. similar to R. raoultii was confirmed in Rhipicephalus annulatus ticks, their egg masses, and unfed larvae. Theileria orientalis was detected in Rh. annulatus females, but not in their egg masses or progenies. The presence of A. phagocytophilum and Rickettsia spp. similar to R. raoultii was confirmed in Haemaphysalis bispinosa ticks, their egg masses, and unfed larvae too. The presence of coinfections of B. bigemina with A. phagocytophilum and A. marginale were detected in Rh. annulatus ticks and their progenies.