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Infections caused by tick-borne haemoparasites pose a significant global threat to both human and animal health. Within this category, various haemoparasites species belonging to genera like Anaplasma sp., Babesia sp., Ehrlichia sp., Hepatozoon sp., and Theileria sp., are particularly concerning due to their ability to cause diseases in a wide range of hosts, including mammals, birds, and reptiles. The present cross-sectional study involving 580 animals provides annual insights into the prevalence of major haemoparasites infections in the Bathinda region of Punjab. The observed trends indicate that haemoparasites infections were most common in cattle, followed by buffalo and canines. Risk factor analysis revealed that crossbreed cattle were more susceptible to infection, with a prevalence of 35.73% (95% CI 4.28–45.17). Amongst the cattle, adults exhibited a higher vulnerability to haemoparasites infections, with a prevalence of 35.89% (95% CI 5.50–33.64). Conversely, companion animals showed the opposite pattern, with a prevalence of 18.18% (95% CI 9.11–169.27). Furthermore, female dogs had a higher risk of haemoparasites infection, with a prevalence of 16.28% (95% CI 8.36–218.7). In light of these findings, it is imperative to emphasize early diagnosis, prompt antiprotozoals drug treatment, and effective control of tick vectors for the successful recovery of animals afflicted by haemoparasites infections. [ABSTRACT FROM AUTHOR]

Ticks (Acari: ixodid) are notorious blood sucking ecto-parasites of wide range of animals which serve as vector of different types of pathogens like viruses, bacteria, rickettsia and protozoa and cause mortality in humans and animals. This study was focused on morphological and molecular identification of ixodid ticks, using morphological keys and an internal transcribed spacer (ITS-2) Deoxyribonucleic acid (DNA). Moreover, the presence of Babesia and Theileria species was also investigated in ticks using 18S rRNA gene. Identification of ticks collected from 384 cattle and 384 buffaloes screened revealed three tick genera and six tick species: Rhipicephalus microplus, Rhipicephalus decoloratus, Rhipicephalus annulatus, Hyalomma anatolicum anatolicum, Hyalomma anatolicum excavatum and Haemaphysalis punctata. Of those four species were confirmed on morphological basis, two ticks species Rhipicephalus microplus and Hyalomma anatolicum anatolicum whose morphological feature were overlapping with other identified species, were confirmed through molecular tools amplifying ITS-2 gene. Ticks DNA were then examined by PCR employing a genetic marker that target (18S rRNA gene), for the presence of Babesia and Theileria species in identified ticks. The most common pathogen species observed in ticks was Theileria annulata. This study exposed different hard tick species are prevalent in the study area and these ticks are playing major role in transmission of protozoa (Theileria annulata). On the basis of finding of present study an area-wise control strategy for ticks and ticks borne protozoan species have been suggestive. [ABSTRACT FROM AUTHOR]

This research paper presents the epidemiological data and clinical manifestations of blood parasitic infections in cattle. A total of 130 blood samples were collected from animals exhibiting clinical signs suggestive of blood parasitic infection. Microscopic examination of blood smears revealed an overall infection rate of 46.2% among the examined cattle. Theileria sp. infections were found to be the most prevalent, with an overall prevalence of 40%. Babesia sp. infections had a lower overall prevalence of 6.2%, Microscopic examination also highlighted the intracellular localization of Theileria and Babesia parasites within red blood cells. The infected red blood cells exhibited morphological changes, appearing enlarged and distorted compared to unaffected cells. The study analyzed the risk factors for Theileria sp. and Babesia sp. infections in cattle, specifically focusing on age and gender. The prevalence of Theileria sp. infection was highest in cattle less than one year, with no significant difference observed among different age groups or genders. Similarly, there was no significant difference in Babesia sp. infection rates based on age or gender. These results provide valuable insights into the epidemiology and clinical manifestations of blood parasitic infections in cattle. The findings can contribute to the development of effective control and prevention strategies, highlighting the importance of regular surveillance and management practices in mitigating the impact of these infections on cattle health and productivity. [ABSTRACT FROM AUTHOR]

Procyonids are reservoirs of many zoonotic infectious diseases, including tick-borne pathogens. The role of coatis (Nasua nasua) in the epidemiology of piroplasmids and Rickettsia has not been fully addressed in Brazil. To molecularly study these agents in coatis and associated ticks, animals were sampled in two urban areas in Midwestern Brazil. Blood (n = 163) and tick (n = 248) DNA samples were screened by PCR assays targeting the 18S rRNA and gltA genes of piroplasmids and Rickettsia spp., respectively. Positive samples were further molecularly tested targeting cox-1, cox-3, β-tubulin, cytB, and hsp70 (piroplasmid) and ompA, ompB, and htrA 17-kDa (Rickettsia spp.) genes, sequenced and phylogenetically analyzed. All coatis' blood samples were negative for piroplasmids, whereas five pools of ticks (2%) were positive for two different sequences of Babesia spp.. The first from Amblyomma sculptum nymphs was close (i.e., ≥ 99% nucleotide identity) to a Babesia sp. previously found in capybaras (Hydrochoerus hydrochaeris); the second from Amblyomma dubitatum nymphs and Amblyomma spp. larvae was identical (100% nucleotide identity) to a Babesia sp. detected in opossums (Didelphis albiventris) and associated ticks. Four samples (0.8%) were positive by PCR to two different Rickettsia spp. sequences, being the first from Amblyomma sp. larva identical to Rickettsia belli and the second from A. dubitatum nymph identical to Rickettsia species from Spotted Fever Group (SFG). The detection of piroplasmids and SFG Rickettsia sp. highlights the importance of Amblyomma spp. in the maintenance of tick-borne agents in urban parks where humans and wild and domestic animals are living in sympatry. [ABSTRACT FROM AUTHOR]

Simple Summary: Capybaras are known hosts for various tick species, but there are limited data regarding the tick-borne pathogens they can carry. We assessed the presence of piroplasmids and Ehrlichia spp. in capybaras and their associated ticks in Goiás state, central-western Brazil. Neither capybaras nor ticks were positive for Ehrlichia spp. However, we detected an undescribed species of protozoan in both the capybaras and ticks. Further research is required for a formal delineation of this protozoan species, as well as to investigate the role of these ticks as vectors and the possible pathogenicity of this parasite to other animals, including horses. Capybaras (Hydrochoerus hydrochaeris) are the largest rodents on Earth. While capybaras are hosts for various tick species, there is limited information regarding the tick-borne pathogens they can carry. We investigated the presence of piroplasmids and Ehrlichia spp. in capybaras and their associated ticks in two peri-urban areas in Goiás state, central-western Brazil. Blood samples collected from 23 capybaras were used to investigate the presence of piroplasmids and Ehrlichia spp. in stained-blood smears and by PCR. Ticks collected from the capybaras were identified morphologically and also tested using PCR for the same pathogens. A total of 955 ticks were collected, including 822 (86.1%) Amblyomma sculptum, 132 (13.8%) Amblyomma dubitatum, and one (0.1%) unidentified larva of Amblyomma sp. Neither the capybaras nor ticks were positive for Ehrlichia spp. However, a stained-blood smear examination revealed the presence of ring-stage and pyriform-shaped merozoites in the erythrocytes of one (4.4%) capybara. In the same way, 47.8% (11/23) and 19.9% (36/181) of blood samples and ticks, respectively, were positive for piroplasmids in the PCR. We successfully sequenced a partial 18S rRNA gene fragment of four samples (two capybaras, one A. sculptum, and one A. dubitatum), and the phylogenetic reconstruction disclosed that the organism reported in the present study clusters within the genus Babesia. Further research is required for a formal delineation of this species (designated as Babesia sp. strain Capybara) and to investigate the hypothesis of A. dubitatum and A. sculptum ticks being vectors. [ABSTRACT FROM AUTHOR]

Abstract Background Babesiosis, a tick-borne disease caused by protozoans of the genus Babesia, is widespread in subtropical and tropical countries. Mitochondria are essential organelles that are responsible for energy transduction and metabolism, calcium homeostasis and cell signaling. Mitochondrial genomes could provide new insights to help elucidate and investigate the biological features, genetic evolution and classification of the protozoans. Nevertheless, there are limited data on the mitochondrial genomes of ovine Babesia spp. in China. Methods Herein, we sequenced, assembled and annotated the mitochondrial genomes of six ovine Babesia isolates; analyzed the genome size, gene content, genome structure and cytochrome b (cytb) amino acid sequences and performed comparative mitochondrial genomics and phylogenomic analyses among apicomplexan parasites. Results The mitochondrial genomes range from 5767 to 5946 bp in length with a linear form and contain three protein-encoding genes, cytochrome c oxidase subunit 1 (cox1), cytochrome c oxidase subunit 3 (cox3) and cytb, six large subunit rRNA genes (LSU) and two terminal inverted repeats (TIR) on both ends. The cytb gene sequence analysis indicated the binding site of anti-Babesia drugs that targeted the cytochrome bc1 complex. Babesia microti and Babesia rodhaini have a dual flip-flop inversion of 184–1082 bp, whereas other Babesia spp. and Theileria spp. have one pair of TIRs, 25–1563 bp. Phylogenetic analysis indicated that the six ovine Babesia isolates were divided into two clades, Babesia sp. and Babesia motasi. Babesia motasi isolates were further separated into two small clades (B. motasi Hebei/Ningxian and B. motasi Tianzhu/Lintan). Conclusions The data provided new insights into the taxonomic relationships and drug targets of apicomplexan parasites.

Capybaras (Hydrochoerus hydrochaeris) are the largest rodents on Earth. While capybaras are hosts for various tick species, there is limited information regarding the tick-borne pathogens they can carry. We investigated the presence of piroplasmids and Ehrlichia spp. in capybaras and their associated ticks in two peri-urban areas in Goiás state, central-western Brazil. Blood samples collected from 23 capybaras were used to investigate the presence of piroplasmids and Ehrlichia spp. in stained-blood smears and by PCR. Ticks collected from the capybaras were identified morphologically and also tested using PCR for the same pathogens. A total of 955 ticks were collected, including 822 (86.1%) Amblyomma sculptum, 132 (13.8%) Amblyomma dubitatum, and one (0.1%) unidentified larva of Amblyomma sp. Neither the capybaras nor ticks were positive for Ehrlichia spp. However, a stained-blood smear examination revealed the presence of ring-stage and pyriform-shaped merozoites in the erythrocytes of one (4.4%) capybara. In the same way, 47.8% (11/23) and 19.9% (36/181) of blood samples and ticks, respectively, were positive for piroplasmids in the PCR. We successfully sequenced a partial 18S rRNA gene fragment of four samples (two capybaras, one A. sculptum, and one A. dubitatum), and the phylogenetic reconstruction disclosed that the organism reported in the present study clusters within the genus Babesia. Further research is required for a formal delineation of this species (designated as Babesia sp. strain Capybara) and to investigate the hypothesis of A. dubitatum and A. sculptum ticks being vectors.

In recent years, a new focus of the relict tick Haemaphysalis concinna was discovered in Western Poland, near Wolsztyn, Greater Poland voivodeship. This species may play an important role in the circulation of pathogens of medical and veterinary importance. In the present study we tested 880 juvenile ticks collected from rodents, including 427 H. concinna, 443 Ixodes ricinus and 10 Dermacentor reticulatus for three of the most common pathogens vectored by ticks in Poland: Rickettsia and Babesia spp. and Borrelia burgdorferi s.l. Additionally, molecular techniques were applied for accurate identification of tick host species (the voles Microtus and Alexandromys). Our study found differences in the range and prevalence of vectored pathogens between the three tick species. DNA of all three pathogens was found in I. ricinus. In juvenile H. concinna, DNA of Babesia microti, Borrelia afzelii and Rickettsia sp. was identified. Moreover, DNA of a new unnamed Babesia species related to B. crassa, was found in two H. concinna nymphs. This genotype of Babesia was previously identified in H. concinna in the Far East and then in Central Europe. DNA of Rickettsia raoulti and B. afzelii was detected in D. reticulatus nymphs. Among rodent hosts, Alexandromys oeconomus seems to be host of the highest significance for juvenile tick stages and was the only host species with B. afzelii detected in blood samples. Using phylogenetic methods, we confirmed a clear division between rodents from the genera Microtus and Alexandromys. Moreover, we found that A. oeconomus trapped in Western Poland clustered with a Central European A. oeconomus allopatric phylogroup. [ABSTRACT FROM AUTHOR]

Abstract Background Babesiosis is an economically important disease caused by tick-borne apicomplexan protists of the genus Babesia. Most apicomplexan parasites, including Babesia, have a plastid-derived organelle termed an apicoplast, which is involved in critical metabolic pathways such as fatty acid, iron-sulphur, haem and isoprenoid biosynthesis. Apicoplast genomic data can provide significant information for understanding and exploring the biological features, taxonomic and evolutionary relationships of apicomplexan parasites, and identify targets for anti-parasitic drugs. However, there are limited data on the apicoplast genomes of Babesia species infective to small ruminants. Methods PCR primers were designed based on the previously reported apicoplast genome sequences of Babesia motasi Lintan and Babesia sp. Xinjiang using Illumina technology. The overlapped apicoplast genomic fragments of six ovine Babesia isolates were amplified and sequenced using the Sanger dideoxy chain-termination method. The full-length sequences of the apicoplast genomes were assembled and annotated using bioinformatics software. The gene contents and order of apicoplast genomes obtained in this study were defined and compared with those of other apicomplexan parasites. Phylogenetic trees were constructed on the concatenated amino acid sequences of 13 gene products using MEGA v.6.06. Results The results showed that the six ovine Babesia apicoplast genomes consisted of circular DNA. The genome sizes were 29,916–30,846 bp with 78.7–81.0% A + T content, 29–31 open reading frames (ORF) and 23–24 transport RNAs. The ORFs encoded four DNA-directed RNA polymerase subunits (rpoB, rpoCl, rpoC2a and rpoC2b), 13 ribosomal proteins, one elongation factor TU (tufA), two ATP-dependent Clp proteases (ClpC) and 7–11 hypothetical proteins. Babesia sp. has three more genes than Babesia motasi (rpl5, rps8 and rpoB). Phylogenetic analysis showed that Babesia sp. is located in a separate clade. Babesia motasi Lintan/Tianzhu and B. motasi Ningxian/Hebei were divided into two subclades. Conclusions To our knowledge, this study is the first to elucidate the whole apicoplast genomic structural features of six Babesia isolates infective to small ruminants in China using Sanger sequencing. The data provide useful information confirming the taxonomic relationships of these parasites and identifying targets for anti-apicomplexan parasite drugs.

Contextualización: la piroplasmosis equina es una enfermedad transmitida por garrapatas, causada por protozoarios de la especie Babesia caballi y Theileria equi. Los animales afectados presentan diferentes signos clínicos que incluyen anemia, fiebre, ictericia y depresión. Los equinos infectados con T. equi pueden ser portadores de manera vitalicia del parasito, mientras que aquellos que contraen B. caballi pueden diseminarlo por algunos años hasta finalmente quedar libres de la infección. Vacío de investigación: en Colombia existen pocos estudios sobre el aislamiento e identificación de hemoparásitos en equinos mediante técnicas moleculares como reacción en cadena de la polimerasa. Dentro de estos parásitos se encuentran Babesia caballi y Theileria equi, agentes causantes de la Piroplasmosis una enfermedad de declaración obligatoria por la Organización Mundial de Sanidad Animal (OIE). Propósito del estudio: el objetivo de este estudio fue identificar la Piroplasmosis equina mediante PCR en un criadero localizado en el municipio de Jamundí (Valle del Cauca). Metodología: de un total de 20 ejemplares, divididos en dos grupos, (animales estabulados y animales en potrero), se colectó sangre periférica para realizar frotis teñidos con Wright. Además, se enviaron otras muestras al laboratorio para diagnóstico molecular. Resultados y conclusiones: todas las muestras con Wright fueron negativas a hemoparásitos, sin embargo, por PCR convencional fue posible identificar piroplasmosis. La PCR fue un método sensible y confiable para diagnosticar la enfermedad en un individuo asintomático. Más estudios deben ser realizados en esta región sobre Babesia sp., para identificar sus posibles factores predisponentes y causales, así como para mejorar las medidas de prevención, control y tratamiento.

Background: Babesiosis, a tick-borne disease caused by protozoans of the genus Babesia, is widespread in subtropical and tropical countries. Mitochondria are essential organelles that are responsible for energy transduction and metabolism, calcium homeostasis and cell signaling. Mitochondrial genomes could provide new insights to help elucidate and investigate the biological features, genetic evolution and classification of the protozoans. Nevertheless, there are limited data on the mitochondrial genomes of ovine Babesia spp. in China. Methods: Herein, we sequenced, assembled and annotated the mitochondrial genomes of six ovine Babesia isolates; analyzed the genome size, gene content, genome structure and cytochrome b (cytb) amino acid sequences and performed comparative mitochondrial genomics and phylogenomic analyses among apicomplexan parasites. Results: The mitochondrial genomes range from 5767 to 5946 bp in length with a linear form and contain three protein-encoding genes, cytochrome c oxidase subunit 1 (cox1), cytochrome c oxidase subunit 3 (cox3) and cytb, six large subunit rRNA genes (LSU) and two terminal inverted repeats (TIR) on both ends. The cytb gene sequence analysis indicated the binding site of anti-Babesia drugs that targeted the cytochrome bc1 complex. Babesia microti and Babesia rodhaini have a dual flip-flop inversion of 184–1082 bp, whereas other Babesia spp. and Theileria spp. have one pair of TIRs, 25–1563 bp. Phylogenetic analysis indicated that the six ovine Babesia isolates were divided into two clades, Babesia sp. and Babesia motasi. Babesia motasi isolates were further separated into two small clades (B. motasi Hebei/Ningxian and B. motasi Tianzhu/Lintan). Conclusions: The data provided new insights into the taxonomic relationships and drug targets of apicomplexan parasites. [ABSTRACT FROM AUTHOR]

This study investigated the occurrence and phylogenetic relationship of protozoan parasites and Ehrlichia infecting domestic animals from three municipalities in uMkhanyakude district of KwaZulu-Natal province, South Africa. A total of 208 blood samples collected from clinically healthy cattle, sheep, goats and dogs from uMkhanyakude district were examined by polymerase chain reaction (PCR) assays, using either genus or species-specific primers to determine the occurrence and phylogenetic relationship of various protozoan parasites and Ehrlichia of veterinary importance. A total of 5/109 (4.6%) cattle were PCR-positive for the presence of Toxoplasma gondii , 33/109 (30.3%) for Babesia bovis, 24/109 (22.02%) for Babesia bigemina and 20/109 (18.3%) for Trypanosoma sp., while 3/10 (30%) of sheep were PCR-positive for Theileria ovis and none of the goats were positive for any of the detected pathogens. The co-infection of 4/109 (3.7%) B. bovis and B. bigemina was detected in cattle. Only Ehrlichia canis was detected in dogs with infection rate of 20/48 (41.7%). Sequences of PCR-positive isolates (B. bovis, B. bigemina, E. canis, T. ovis and T. gondii) showed that they were closely related to their relevant species from various countries. These findings have expanded our knowledge about the prevalence and phylogenetic similarity between protozoan parasites and Ehrlichia isolates of South African origin. To date, this is the first study in South Africa to detect T. gondii infections from cattle blood using PCR. [ABSTRACT FROM AUTHOR]

Babesiosis is one of the most important tick-borne diseases in veterinary health, impacting mainly cattle, equidae, and canidae, and limiting the development of livestock industries worldwide. In humans, babesiosis is considered to be an emerging disease mostly due to Babesia divergens in Europe and Babesia microti in America. Despite this importance, our knowledge of Babesia sp. transmission by ticks is incomplete. The complexity of vectorial systems involving the vector, vertebrate host, and pathogen, as well as the complex feeding biology of ticks, may be part of the reason for the existing gaps in our knowledge. Indeed, this complexity renders the implementation of experimental systems that are as close as possible to natural conditions and allowing the study of tick-host-parasite interactions, quite difficult. However, it is unlikely that the development of more effective and sustainable control measures against babesiosis will emerge unless significant progress can be made in understanding this tripartite relationship. The various methods used to date to achieve tick transmission of Babesia spp. of medical and veterinary importance under experimental conditions are reviewed and discussed here.

Tick-borne pathogens cause infectious diseases that inflict much societal and financial hardship worldwide. Blacklegged ticks, Ixodes scapularis, are primary vectors of several epizootic and zoonotic pathogens. The aim was to find varius pathogens of I. scapularis and to determine their prevalence. In Ontario and Quebec, 113 I. scapularis ticks were collected from songbirds, mammals, including humans, and by flagging. PCR and DNA sequencing detected five different microorganisms: Anaplasma phagocytophilum, 1 (0.9%); Babesia odocoilei, 17 (15.3%); Babesia microti-like sp., 1 (0.9%); Borrelia burgdorferi sensu lato (Bbsl), 29 (26.1%); and Hepatozoon canis, 1 (0.9%). Five coinfections of Bbsl and Babesia odocoilei occurred. Notably, H. canis was documented for the first time in Canada and, at the same time, demonstrates the first transstadial passage of H. canis in I. scapularis. Transstadial passage of Bbsl and B. odocoilei was also witnessed. A novel undescribed piroplasm (Babesia microti-like) was detected. An established population of I. scapularis ticks was detected at Ste-Anne-de-Bellevue, Quebec. Because songbirds widely disperse I. scapularis larvae and nymphs, exposure in an endemic area is not required to contract tick-borne zoonoses. Based on the diversity of zoonotic pathogens in I. scapularis ticks, clinicians need to be aware that people who are bitten by I. scapularis ticks may require select antimicrobial regimens.

El objetivo de este trabajo es comparar los resultados relevados y obtenidos en el 2016, con los obtenidos en el 2017, evaluando la evolución de las enfermedades por hemoparásitos en los burros de la región. Los hemoparásitos son el grupo de enfermedades más importante en burros de la región norte de Nicaragua (Departamentos de Madriz y Nueva Segovia), con una prevalencia cercana al 50% de los burros existentes. Sin embargo, las causas de muerte se deben mayormente a la falta de control de garrapatas en burros por parte de sus dueños, sumado a la falta de control y tratamiento de la hemoparasitosis. Con el fallecimiento de los burros afectados por Babesia sp., la Anaplasmosis pasa a ser la enfermedad más relevante dentro de los hematozoarios, con más de 30% de prevalencia. La presencia de Plasmodium sp., no descrito hasta el momento en la literatura internacional es relevante, no solo porque afecta al 16.88% de los burros de las diferentes comarcas de la cuenca del Río Tapacale en Somoto, sino por los riesgos inherentes a la salud pública. Se precisa en forma urgente capacitar a los dueños de los burros sobre control y tratamiento de garrapatas y hemoparásitos, así como conseguir los fondos para costear los tratamientos.

Background: Babesiosis is an economically important disease caused by tick-borne apicomplexan protists of the genus Babesia. Most apicomplexan parasites, including Babesia, have a plastid-derived organelle termed an apicoplast, which is involved in critical metabolic pathways such as fatty acid, iron-sulphur, haem and isoprenoid biosynthesis. Apicoplast genomic data can provide significant information for understanding and exploring the biological features, taxonomic and evolutionary relationships of apicomplexan parasites, and identify targets for anti-parasitic drugs. However, there are limited data on the apicoplast genomes of Babesia species infective to small ruminants. Methods: PCR primers were designed based on the previously reported apicoplast genome sequences of Babesia motasi Lintan and Babesia sp. Xinjiang using Illumina technology. The overlapped apicoplast genomic fragments of six ovine Babesia isolates were amplified and sequenced using the Sanger dideoxy chain-termination method. The full-length sequences of the apicoplast genomes were assembled and annotated using bioinformatics software. The gene contents and order of apicoplast genomes obtained in this study were defined and compared with those of other apicomplexan parasites. Phylogenetic trees were constructed on the concatenated amino acid sequences of 13 gene products using MEGA v.6.06. Results: The results showed that the six ovine Babesia apicoplast genomes consisted of circular DNA. The genome sizes were 29,916–30,846 bp with 78.7–81.0% A + T content, 29–31 open reading frames (ORF) and 23–24 transport RNAs. The ORFs encoded four DNA-directed RNA polymerase subunits (rpoB, rpoCl, rpoC2a and rpoC2b), 13 ribosomal proteins, one elongation factor TU (tufA), two ATP-dependent Clp proteases (ClpC) and 7–11 hypothetical proteins. Babesia sp. has three more genes than Babesia motasi (rpl5, rps8 and rpoB). Phylogenetic analysis showed that Babesia sp. is located in a separate clade. Babesia motasi Lintan/Tianzhu and B. motasi Ningxian/Hebei were divided into two subclades. Conclusions: To our knowledge, this study is the first to elucidate the whole apicoplast genomic structural features of six Babesia isolates infective to small ruminants in China using Sanger sequencing. The data provide useful information confirming the taxonomic relationships of these parasites and identifying targets for anti-apicomplexan parasite drugs. [ABSTRACT FROM AUTHOR]

Arthropod-borne diseases (ABD) are of increasing interest in veterinary and public health. Eurasian badgers (Meles meles) are known to harbor a wide range of pathogens, but information on their role as ABD reservoirs and their potential epidemiological relevance is limited. This study aimed to investigate the occurrence of arthropod-borne pathogens, specifically piroplasmids and the bacteria Anaplasma phagocytophilum, Ehrlichia canis, Coxiella burnetii, Francisella tularensis and Bartonella spp., in badgers from Great Britain (GB). Blood and heart samples from 18 badgers were examined using PCR and sequencing. A neighbour-joining (NJ) phylogram was also produced. Nine animals tested positive for Babesia sp., while none of the samples was positive for the investigated bacteria. The sequences obtained clustered with other sequences of Babesia sp. from badgers from GB and elsewhere, including China, Hungary, Spain and Italy, showing a widespread distribution of this parasite in badgers. Badger-associated Babesia DNA was also found recently in a wild cat in Bosnia Herzegovina, in a wolf in Italy and in dogs in Hungary. Further investigations are needed to understand the epidemiology of this putative pathogen and its impact on the health of wild and domestic carnivores.

Atranorin (ATR), is a compound with multidirectional biological activity under different in vitro and in vivo conditions and it is effective as an antibacterial, antiviral, antiprotozoal and anti-inflammatory agent. In the current study, the in vitro as well as in vivo chemotherapeutic effect of ATR as well as its combined efficacy with the existing antibabesial drugs (diminazene aceturate (DA), atovaquone (AV) and clofazimine (CF)) were investigated on six species of piroplasm parasites. ATR suppressed B. bovis, B. bigemina, B. divergens, B. caballi and T. equi multiplication in vitro with IC50 values of 98.4 ± 4.2, 64.5 ± 3.9, 45.2 ± 5.9, 46.6 ± 2.5, and 71.3 ± 2.7 µM, respectively. The CCK test was used to examine ATR’s cytotoxicity and adverse effects on different animal and human cell lines, the main hosts of piroplasm parasites and it showed that ATR affected human foreskin fibroblasts (HFF), mouse embryonic fibroblast (NIH/3T3) and Madin-Darby Bovine Kidney (MDBK) cell viability in a dose-related effect with a moderate selective index. The combined efficacy of ATR with DA, CF, and AV exhibited a synergistic and additive efficacy toward all tested species. In the in vivo experiment, ATR prohibited B. microti multiplication in mice by 68.17%. The ATR-DA and ATR-AV combination chemotherapies were more potent than ATR monotherapy. These results indicate the prospects of ATR as a drug candidate for piroplasmosis treatment.

Hemoparasites can cause serious morbidity in humans and animals and often involve wildlife reservoirs. Understanding patterns of hemoparasite infections in natural populations can therefore inform about emerging disease risks, especially in the light of climate change and human disruption of natural ecosystems. We investigated the effects of host age, sex, host group size and season on infection patterns of Plasmodium sp., Babesia sp. and filarial nematodes in a population of wild Malagasy primates, Verreaux's sifakas (Propithecus verreauxi), as well as the effects of these infections on hematological variables. We tested 45 blood samples from 36 individuals and identified two species of Plasmodium, one species of Babesia and two species of filarial nematodes. Plasmodium spp. and Babesia sp. infections showed opposite patterns of age-dependency, with babesiosis being prevalent among young animals, while older animals were infected with Plasmodium sp. In addition, Babesia sp. infection was a statistically significant negative predictor of Plasmodium sp. infection. These results suggest that Plasmodium and Babesia parasites may interact within the host, either through cross-immunity or via resource competition, so that Plasmodium infections can only establish after babesiosis has resolved. We found no effects of host sex, host group size and season on hemoparasite infections. Infections showed high prevalences and did not influence hematological variables. This preliminary evidence supports the impression that the hosts and parasites considered in this study appear to be well-adapted to each other, resulting in persistent infections with low pathogenic and probably low zoonotic potential. Our results illustrate the crucial role of biodiversity in host-parasite relationships, specifically how within-host pathogen diversity may regulate the abundance of parasites.

Data about the spreading of arthropod-borne pathogens among hare populations are very scant, so the aim of the present preliminary study was to investigate, through molecular analysis, the occurrence of Anaplasma phagocytophilum , Bartonella sp., Borrelia burgdorferi sensu lato, Coxiella burnetii , Francisella tularensis , Leishmania spp. and piroplasms DNA in blood of 51 wild hares ( Lepus europaeus ) living in protected areas in Tuscany. All hares resulted negative for A . phagocytophilum , Bartonella sp., B . burgdorferi s.l., C . burnetii and F . tularensis . Five animals (9.8%) were positive for Leishmania and one hare (1.9%) tested positive for piroplasms. Sequencing of this sample showed a piroplasm similar to one Babesia isolate from the same animal species in Turkey. Therefore, to the best of our knowledge, this is the first molecular report of piroplasms occurring in wild hares from Italy, and the second worldwide. The examined hares appeared to be in good health status, corroborating the hypothesis of a chronic carrier state of some vector borne agents for this animal species. [ABSTRACT FROM AUTHOR]

Babesia sp. is a protozoan hemoparasite that affects livestock worldwide. The Colombian Middle Magdalena is an enzootic region for babesiosis, but there is no previous research providing detail on its transmission cycle. This study aims to assess some Babesia sp. infection indicators in cattle and ticks from the area, by using direct microscopic and molecular techniques to detect the infection. In the cattle, 59.9% and 3.4 % positivity values for B. bigemina and mixed infection (B. bovis + B. bigemina) were found respectively. In ticks, the positivity of B. bigemina reached 79.2% and 9.4% for the mixed infection. The degree of infestation in the region was 3.2 ticks per bovine. There was positive correlation between tick control acaricide frequencies and infestation in bovines. This leads us to infer that control periodicity greater than 90 days, in stable zones, is an abiotic factor that benefits the acquisition of protective immunity in calves, the natural control of the infection and eventual disease absence. It is necessary to monitor the disease by applying new entomological and parasitological indicators showing the complexity of this phenomenon.

Tick-borne apicomplexan haemoparasites infect wild and domestic animals, but studies on their distribution among freeliving animals are comparatively fewer. Tissues from 241 wild carnivores of eight Mustelidae, two Canidae, one Viverridae and one Felidae species were collected in Northern Spain, and analysed by real-time PCR targeting the 18S rRNA gene and sequencing. Babesia vulpes (formerly known as Theileria annae) was the only piroplasm detected in red foxes (Vulpes vulpes). Badgers (Meles meles) were shown to harbour two novel Babesia sp. sequence types (A and B) that only shared 96.7% homology between them and were closely related (ca. 97-98%) to, but distinct from B. vulpes and other babesia from carnivores. Analysis of PCR-derived sequences also revealed the presence of Cystoisospora sp. and Hepatozoon sp. in badgers and showed that wild cats (Felis silvestris catus) were infected with Cytauxzoon sp. Forty-two per cent of the animals subjected to a detailed external examination were parasitized by ixodid ticks, being Ixodes hexagonus and Ixodes ricinus the most abundant species. This study provided novel data on the different haemoparasites that can infect European wild carnivores and showed that they can be hosts for a range of haemoparasites and pose a potential risk for transmission to domestic animals. [ABSTRACT FROM AUTHOR]

This is the first reported case of Babesia sp. in Antarctic penguins, specifically a population of Chinstrap penguins ( Pygoscelis antarctica ) in the Vapour Col penguin rookery in Deception Island, South Shetlands, Antarctica. We collected peripheral blood from 50 adult and 30 chick Chinstrap penguins. Examination of the samples by microscopy showed intraerythrocytic forms morphologically similar to other avian Babesia species in 12 Chinstrap penguin adults and seven chicks. The estimated parasitaemias ranged from 0.25 × 10 −2 % to 0.75 × 10 −2 %. Despite the low number of parasites found in blood smears, semi-nested PCR assays yielded a 274 bp fragment in 12 of the 19 positive blood samples found by microscopy. Sequencing revealed that the fragment was 97% similar to Babesia sp. 18S rRNA from Australian Little Penguins ( Eudyptula minor ) confirming presence of the parasite. Parasite prevalence estimated by microscopy in adults and chicks was higher (24% vs. 23.3%, respectively) than found by semi-nested PCR (16% vs. 13.3% respectively). Although sampled penguins were apparently healthy, the effect of Babesia infection in these penguins is unknown. The identification of Babesia sp. in Antarctic penguins is an important finding. Ixodes uriae , as the only tick species present in the Antarctic Peninsula, is the key to understanding the natural history of this parasite. Future work should address the transmission dynamics and pathogenicity of Babesia sp. in Chinstrap penguin as well as in other penguin species, such as Gentoo penguin ( Pygoscelis papua ) and Adélie penguin ( Pygoscelis adeliae ), present within the tick distribution range in the Antarctic Peninsula. [ABSTRACT FROM AUTHOR]

Background: Babesiosis is an uncommon but emerging tick-borne disease caused by the genus Babesia. In this case study, we report a case of human infection with a novel Babesia sp. in China. Findings: The patient in question had been suffering from repetitive occurrences of mild fever of unknown origin and fatigue for 10 years. Ring forms, tetrads, and one or two dots of chromatin or trophozoite-like organisms were observed in the patient's thin blood smears and bone marrow smears. Using a confocal laser-scanning microscope, it was observed that the patient's serum had reactivity with the surface proteins of the B. microti strain. Electron microscopy revealed oval red blood cells with 1 ~ 2 µm of knob protrusions in the cellular membrane. The results of the Babesia-specific nested PCR assay for 18S rRNA confirmed the presence of Babesia infection. The construction of a phylogenetic relationship showed clustering with B. microti and B. duncani, which was identified as a novel Babesia species and named as Babesia sp. XXB/HangZhou. Azithromycin, doxycycline, and moxifloxacin hydrochloride were shown to relieve symptoms but were not as effective after continuous usage. After atovaquone (Mepron®) administration, the patient recovered from fever and tested negative for detection of Babesia-specific genes. Conclusion: Babesia sp. XXB/HangZhou is a novel Babesia species, which causes mild babesiosis in an immunocompetent patient. [ABSTRACT FROM AUTHOR]

Foi analisada a ocorrência de babesiose em pequenos roedores nos municípios de Silva Jardim e Nova lguaçu, Estado do Rio de Janeiro. Foram capturados 44 roedores de seis espécies diferentes e entre eles a prevalência da infecção foi de 27,3%. Rattus norvegicus foi considerado o principal reservatório (50,0%) e Oligoryzomys nigripes como novo hospedeiro para Babesia sp. Este foi o primeiro relato de Babesia sp. em roedores no Brasil. A freqüência de roedores positivos e o risco de infecção dos roedores não diferiram entre as áreas estudadas.The occurrence of babesiosis was studied in 44 small rodents of six species captured in Silva Jardim and Nova lguaçu counties, State of Rio de Janeiro, Brazil. The prevalence of injection was 27.3%. Rattus norvegicus was considered as the main reservoir and Oligoryzomys nigripes as a new host to Babesia sp. The frequency and the risk of rodent infection were considered equal among the studied areas. This is the first report of Babesia sp in small rodents in Brazil.

A 5 years old crossbred cow was brought to the Veterinary Clinical Complex of Lala Lajpat Rai University of Veterinary and Animal Sciences Hisar with history of progressive weakness, pale mucous membrane, anorexia, high fever (105 °F), tachycardia, laboured breathing and coffee coloured urine. Analysis of haematological parameters revealed severe anaemia, leucocytopenia, marked poikilocytosis of erythrocytes. Blood smear examination showed presence of signet ring shaped Theileria organisms, Babesia piroplasms and condensed dot forms of Anaplasma marginale in the stained erythrocytes. Further animal was treated with buparvaquone @ 2.5 mg/kg b.wt deep I/M in neck region and long acting oxytetracycline at 25 mg/kg b. W. slow I/V daily in normal saline solution for 5 days. Berenil (Diminazene aceturate 5%) injection was also administered @ 1 ml/20 kg b.wt. I/M along with supportive therapy. Clinical signs started to subside 3 days post treatment. Complete recovery was achieved by 4 weeks post treatment however animal succumbed to death due to immunosuppression. [ABSTRACT FROM AUTHOR]

The infection rates of Babesia sporokinetes in engorged Boophilus microplus were evaluated during a 2-year period in a dairy farm located in an area of enzootic stability. Every 14 days engorged females were collected from calves and from adult animals. Ticks were incubated at 27 ± 0.5ºC and 80-90% relative humidity and Babesia infection rates were determined by microscopic examination of Giemsa-stained hemolymph smears. After 52 collections, 2105 ticks were obtained, from which 982 were collected from calves and 1123 from cows. The total Babesia infection rate was 10%, however the incidence was higher (p < 0.05) in ticks collected from calves (17.5%) than in those collected from cows (3.6%). Females collected from cows showed the highest infection rates in January, March, and August, and absence of infection in April and May. Ticks feeding on calves were infected throughout the experimental period. The infection rates of engorged females collected from naturally infected calves that were artificially infested with Babesia-free-larvae of B. microplus gradually decreased until the calves were four months old. No differences were observed among infection rates of ticks collected from calves maintained under natural conditions.

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Hemoparasites can cause serious morbidity in humans and animals and often involve wildlife reservoirs. Understanding patterns of hemoparasite infections in natural populations can therefore inform about emerging disease risks, especially in the light of climate change and human disruption of natural ecosystems. We investigated the effects of host age, sex, host group size and season on infection patterns of Plasmodium sp., Babesia sp. and filarial nematodes in a population of wild Malagasy primates, Verreaux's sifakas ( Propithecus verreauxi ), as well as the effects of these infections on hematological variables. We tested 45 blood samples from 36 individuals and identified two species of Plasmodium , one species of Babesia and two species of filarial nematodes. Plasmodium spp. and Babesia sp. infections showed opposite patterns of age-dependency, with babesiosis being prevalent among young animals, while older animals were infected with Plasmodium sp. In addition, Babesia sp. infection was a statistically significant negative predictor of Plasmodium sp. infection. These results suggest that Plasmodium and Babesia parasites may interact within the host, either through cross-immunity or via resource competition, so that Plasmodium infections can only establish after babesiosis has resolved. We found no effects of host sex, host group size and season on hemoparasite infections. Infections showed high prevalences and did not influence hematological variables. This preliminary evidence supports the impression that the hosts and parasites considered in this study appear to be well-adapted to each other, resulting in persistent infections with low pathogenic and probably low zoonotic potential. Our results illustrate the crucial role of biodiversity in host-parasite relationships, specifically how within-host pathogen diversity may regulate the abundance of parasites. [ABSTRACT FROM AUTHOR]

The article offers information on a study which aims to determine several Babesia sp. infection indicators in cattle and ticks from the middle Magdalena region in Colombia. It discusses the use of direct microscopic as well as molecular techniques in detecting the infection. Information regarding the positive correlation between tick control acaricide frequencies and infestation in bovines is provided.

Capybaras ( Hydrochoerus hydrochaeris ) are the largest rodents on Earth. While capybaras are hosts for various tick species, there is limited information regarding the tick-borne pathogens they can carry. We investigated the presence of piroplasmids and Ehrlichia spp. in capybaras and their associated ticks in two peri-urban areas in Goiás state, central-western Brazil. Blood samples collected from 23 capybaras were used to investigate the presence of piroplasmids and Ehrlichia spp. in stained-blood smears and by PCR. Ticks collected from the capybaras were identified morphologically and also tested using PCR for the same pathogens. A total of 955 ticks were collected, including 822 (86.1%) Amblyomma sculptum , 132 (13.8%) Amblyomma dubitatum , and one (0.1%) unidentified larva of Amblyomma sp. Neither the capybaras nor ticks were positive for Ehrlichia spp. However, a stained-blood smear examination revealed the presence of ring-stage and pyriform-shaped merozoites in the erythrocytes of one (4.4%) capybara. In the same way, 47.8% (11/23) and 19.9% (36/181) of blood samples and ticks, respectively, were positive for piroplasmids in the PCR. We successfully sequenced a partial 18S rRNA gene fragment of four samples (two capybaras, one A. sculptum , and one A. dubitatum ), and the phylogenetic reconstruction disclosed that the organism reported in the present study clusters within the genus Babesia . Further research is required for a formal delineation of this species (designated as Babesia sp. strain Capybara) and to investigate the hypothesis of A. dubitatum and A. sculptum ticks being vectors.

Babesia spp. were detected from 4 asymptomatic pukus captured on a game ranch in central Zambia in October 2008. Blood smears were examined in 4 species of aymptomatic free-ranging antelopes, namely the puku (Kobus vordanii), reedbuck (Redunca arundinum), bushbuck (Tragelaphus sylvaticus), and kudu (Tragelaphus strepsiceros), and showed the presence of Babesia parasites only in the puku. In the puku, the prevalence of babesiosis was estimated at 33.3% (n=12), while the overall prevalence in all examined animals was 8.5% (n=47). The parasites showed morphological characteristics of paired ring-like stages with the length varying between 1.61 µm and 3.02 µm (mean=2.12 µm, n= 27; SD=0.76 µm). Both the infected and non-infected pukus showed good body condition scores (BCS), while the dominant tick species detected from all animals were Rhipicephalus appendiculatus, Rhipicephalus spp., and Boophilus spp. To our knowledge this is the first report of Babesia spp. infection in pukus in Zambia. These findings suggest that wildlife could play an important role in the epidemiology of babesiosis in Zambia. [ABSTRACT FROM AUTHOR]

The article presents a study which examines the presence of parasites Babesia spp. in exotic and neotropical carnivores in Brazilian zoos using molecular and serologic techniques. The study collects blood and sera samples from Brazilian wild felids, exotic felids and European wolves (Canis lupus) in Brazilian zoos in São Paulo and Mato Grosso to detect Babesia spp. parasites. Results show that antibodies to Babesia spp. have been detected in the sera of wild canids and felids and the parasites have no impact on the health of wild animals. However, it may trigger a benign babesionis into a clinical disease when the animals are subjected to stressful situations such as capture and captivity.

Abstract: Babesiosis and Theileriosis are tick-borne diseases widespread in tropical and sub-tropical regions with high economic impact worldwide. In Portugal there are at least 4 tick vectors known to be competent for the transmission of Babesia and Theileria sp. identified: Rhipicephalus bursa, Rhipicephalus (Boophilus) annulatus, Ixodes ricinus and Haemaphysalis punctata. All these potential Babesia and Theileria tick vectors are widely distributed in Portugal, although they are predominant in the Southern region. In this study, 1104 cattle blood samples were randomly collected from Central and Southern regions of Portugal and analyzed by PCR–reverse line blotting (RLB) for the detection of Babesia and Theileria sp. Testing indicated that 74.7% of the bovines tested were positive for either Babesia and/or Theileria sp. In addition, five different apicomplexan species, namely, Theileria buffeli, Theileria annulata, Babesia divergens, Babesia bovis, and Babesia bigemina were detected by RLB among the bovines tested. T. buffeli was the most frequently found species, being present in 69.9% of the positive samples either as single infections (52.4%), or as mixed infections (17.5%). The Babesia specie most frequently found was B. divergens, detected in 4.2% of the infected bovines. Overall, infected bovines were found in all regions tested; however the highest number of infected bovines was observed in Évora district (96.2%) and in cattle from Limousin breeds (81.7%). The results indicate widespread Babesia and Theileria infections in Portuguese bovines, suggesting the need for improved control of ticks and tick-borne diseases. [ABSTRACT FROM AUTHOR]

Molecular detection of apicomplexan parasites in splenic samples of red foxes collected from northeastern Poland was conducted by PCR amplification of a fragment of the 18S rRNA spanning the V4 gene region of Apicomplexa. Positive PCR products were further analysed by restriction fragment length polymorphism (RFLP) and sequencing to identify species. One hundred and eleven red foxes ( Vulpes vulpes) were acquired from 15 localities in the Mazovian Province and 27 foxes were acquired from the Mazurian Lakeland. Apicomplexan 18S rDNA was detected in 15.9% of 138 fox spleens examined. Three apicomplexan species were identified: Hepatozoon canis was detected in 11.6% of the spleen samples, Toxoplasma gondii was detected in 3.6% of the spleen samples and a Babesia sp. was sequenced from 1 sample (0.7%). This data represent the first record of H. canis, T. gondii and a B. sp. from naturally infected red foxes in Poland. Infected foxes may act as sylvatic reservoirs of these apicomplexan parasites as well as serving as a source of infection for arthropod definitive hosts and vectors. [ABSTRACT FROM AUTHOR]

This study was conducted between January 2006 and September 2008 to identify Babesia and Theileria species in sheep and goats in Central Anatolia via reverse line blotting (RLB). In total, 421 sheep and 152 goats were randomly selected from herds located in Kayseri, Yozgat, and Sivas provinces (all in central Turkey). The amplified hypervariable V4 region of the 18S rRNA gene of ovine piroplasm species, including Theileria lestoquardi, T. ovis, Babesia ovis, B. motasi, and B. crassa, was hybridized against species-specific probes. Theileria ovis and B. ovis were observed in the study animals. Among the 573 small ruminants included in the study, 182 (43.2%) of 421 sheep and 18 (11.8%) of 152 goats, (in total 200 [34.9%]) were catch-all positive based on RLB. Among the sheep, T. ovis was observed in 170 (40.4%), B. ovis in 5 (1.2%), and B. ovis + T. ovis in 7 (1.7%). Theileria ovis, B. ovis, and B. ovis + T. ovis were observed in 15 (9.9%), in 1 (0.7%), and in 2 (1.3%) of the examined goats, respectively. As a result, the molecular prevalence of B. ovis was as follows: 2.9% in sheep (12/421), 2% in goats (3/152), and 2.6% in the total study population (15/573), versus 42% in sheep (177/421), 11.2% in goats (17/152), and 33.9% in the total population (194/573) for T. ovis. [ABSTRACT FROM AUTHOR]

Abstract: We recently reported that feral raccoons (Procyon lotor) with splenomegaly native to Japan were carriers of a Babesia microti-like parasite identical to that found in the United States, which was likely introduced to Japan from North America via raccoons imported as pets. Thus, we attempted extensive molecular survey for piroplasma infections of feral raccoon with normal spleen in Hokkaido, Japan using nested PCR that target broadly to 18S ribosomal RNA gene (SSU-rDNA) of all the parasites in the genus Babesia, Theileria, Cytauxzoon and B. microti group. Of the 348 raccoon samples analyzed, 9 gave positive signals. Cloning and phylogenetic analysis on SSU-rDNA sequences revealed that six of nine positives were found to be infected with Babesia and the remaining three with previously unreported Sarcocystis. Babesia sequences were further separated into two distantly related groups, those that reside in a novel phylogenetic group were consisted solely of four parasites found in this study, while those which included one identical sequence found in the three of our specimens were assembled together with both Babesia parasites of tick''s in Japan and of raccoon''s in U.S. These results may indicate that not only a B. microti-like parasite but also at least two yet undescribed Babesia species are being established in their new life cycles in the feral raccoon populations in Japan. [Copyright &y& Elsevier]

Abstract: The first and second internal transcribed spacers (ITS1, ITS2) as well as the intervening 5.8S coding region of the rRNA gene for six Babesia spp. isolated from different geographic origins were characterized. Varying degrees of ITS1 and ITS2 intra- and inter-species sequence polymorphism were found among these isolates. Phylogenetic analysis of the ITS1-5.8S gene-ITS2 region clearly separated the isolates into two clusters. One held an unidentified Babesia sp. transmitted by Hyalomma anatolicum anatolicum. The second held five other isolates, which were considered to be Babesia motasi. Each Babesia species cluster possessed ITS1 and ITS2 of unique size(s) and species specific nucleotide sequences. The results showed that ITS1, ITS2 and the complete ITS1-5.8S-ITS2 region could be used to discriminate these ovine Babesia spp. effectively. [Copyright &y& Elsevier]

Abstract: The loop-mediated isothermal amplification (LAMP) reaction is a method that amplifies with high sensitivity, efficiency, and rapidity, deoxyribonucleic acid (DNA) under isothermal condition in simple incubators. Two primer sets for the LAMP method were designed using the nucleotide sequences of 18S rRNA gene of Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005 isolated in China. The primers were used to detect parasite DNA extracted from infected blood and purified parasites by LAMP. The specific ladder bands were amplified from the autologous genomic DNA of two Babesia species, respectively, and did not cross-react with the genomic DNA of Theileria sp. China 1, Theileria sp. China 2, B. bovis, Theileria sp. (Japan) and sheep. The LAMP was sensitive enough to detect 0.02pg and 0.2pg genomic DNA of Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005, respectively, from 10-fold serially diluted samples corresponding to the amount of DNA present in 50μl of 0.000002% and 0.00002% parasitemic erythrocytes. Furthermore, DNA extracted from blood of intact (non-splenectomized) sheep experimentally infected with Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005 was amplified by the LAMP from week 1 to 9 and week 2 and 3 post-infection, respectively, demonstrating the high sensitivity of these primers. Of 365 samples collected from Gansu province, 14.3% (52/365) were positively detected by the LAMP. Of 145 samples collected on filter papers (Whatman) from the grazing sheep in Xinjiang province, 3.5% (5/145) were positive. These results show that the LAMP could be an alternative diagnostic tool for the detection of babesial infection in sheep and goats. [Copyright &y& Elsevier]

Abstract: Seven healthy sheep and 10 sheep diagnosed with piroplasmosis based on clinical signs were tested for the presence of babesiae and theileriae. Using the molecular techniques, two species of theileriae were detected and characterized. Theileria ovis was present mostly in healthy sheep and in Rhipicephalus ticks collected from infected sheep. Theileria sp. OT3 parasite was detected mostly in ill animals which represent additional evidence to the possible pathogenic nature of Theileria sp. OT3. The presence of babesiae in sheep or in ticks was not determined. The results of this study showed that ovine piroplasmosis due to Theileria is present in Southern Croatia. It was concluded that clinical diagnosis of ovine piroplasmosis should be confirmed by molecular analysis in order to identify the species of piroplasm, to select the appropriate treatment and to exclude the threat for public health. [Copyright &y& Elsevier]

Abstract: A novel large Babesia sp. from an infected dog was cultivated in vitro by microaerophilous stationary phase culture methodology. A primary culture initiated in enriched RPMI-1640 medium supplemented with 40% canine serum and incubated in a 2% oxygen environment supported parasite growth in vitro. Subsequent subcultures into enriched HL-1 medium with 20% fetal bovine serum also supported parasite propagation. Cultures were successfully introduced to 5% carbon dioxide in air atmosphere at passage 4. To date, the parasites have been continuously cultured through 35 passages, although the parasitemias are low, ranging from 0.2 to 0.3%. Parasites cultured in RPMI with canine serum were cryopreserved and successfully recovered from liquid nitrogen storage. The small subunit ribosomal rRNA gene sequence was identical in blood-derived and culture-derived parasites, differing in a single base position from the previously reported sequence for this Babesia sp. The ultrastructure of the parasite was consistent with that of other large Babesia spp., except that the spherical body contained numerous round particles unlike the inclusions previously described in Babesia spp. [Copyright &y& Elsevier]

Abstract: A fatal disease of sheep and goats in the northern part of China has been reported to be due to Babesia ovis. However, some characteristics of the causative agent in recent reports are not in accordance with the original attributes ascribed to this parasite. Therefore, the 18S small subunit ribosomal RNA (18S rRNA) genes of a number of Babesia isolates in China were sequenced and compared with that of other Babesia and Theileria species in an attempt to clarify their taxonomic position. In the present study, seven Babesia isolates were collected from distinct areas of northern China, and the 18S rRNA genes were amplified and sequenced. The phylogenetic trees were inferred based on 18S rRNA gene sequences of the Chinese ovine Babesia isolates and some of ovine Babesia and Theileria species available in GenBank. In the phylogenetic tree, Babesia sp. isolates from Madang, Tianzhu, Lintan, Ningxian, Hebei and Liaoning all grouped with B. motasi with 88.2–99.9% identity, while Babesia sp. Xinjiang grouped in a separate clade between B. ovis and B. crassa with 79.7–81.2% identity. The results indicated that there are at least two distinct Babesia species groups—B. motasi and Babesia sp. Xinjiang, the latter was dinstinctly different from other ovine Babesia isolates from China with less than 86.6% identity. [Copyright &y& Elsevier]

A cat was presented with anorexia, vomiting and diarrhoea for past three days. Clinical examination revealed pale mucous membrane and swollen popliteal lymph nodes. Hemotology revealed severe anemia, thrombocytopenia and neutropenia. Blood smear examination revealed presence of Babesia sp. organisms. Mild elevation of AST and ALT were noticed. The cat was treated initially with Doxycycline. Later, Primaquine phosphate was administered once orally as a total dose of 1mg/ cat along with Inj. Ranitidine 0.5 ml. i.m. The cat recovered uneventfully. [ABSTRACT FROM AUTHOR]

The present paper is the continuation of our previous studies dealing with the genetic characterization of piroplasmid species found in southern Europe. We report in this work new data concerning sequences of the 18s rRNA gene in Spanish piroplasms not studied (or not totally sequenced) in our former surveys.Molecular data analysis indicated that

Experiments were undertaken to determine the mode of transmission to cattle of an unnamed Babesia sp. by Hyalomma anatolicum anatolicum, Hyalomma detritum, Hy. rufipes koch, Haemaphysalis longicornis and Boophilus microplus. The unnamed Babesia species designated Babesia U sp. was isolated by infesting cattle with nymphs from female Hy. a. anatolicum ticks collected from Xinjiang province. Adults of laboratory reared Hy. a. anatolicum, Hy. detritum and Hy. rufipes koch were infected with Babesia U sp. by feeding on infected cattle, isolated with nymphal ticks of Hy. a. anatolicum derived from females collected from field. The experiments revealed that Hy. a. anatolicum was capable of transmitting Babesia U sp. transovarially in larval (2 of 4 calves), nymphal (6 of 6 calves) and adult (3 of 8 calves) stages, with prepatent periods of 16, 12, and 8 days, respectively, and that this Babesia was also transovarially transmitted by both the nymphal and adult stages of Hy. detritum and Hy. rufipes. Attempts to transmit this Babesia U sp. transovarially with Hae. longicornis and B. microplus, and transstadially with Hyalomma spp., were carried out, and the results proved to be negative. [Copyright &y& Elsevier]

Following a study on molecular epizootiology of Hepatozoon canis and piroplasmids (Babesia spp. and Theileria spp.) in southern Europe, newly obtained sequences of 18s rRNA gene were used for phylogenetic analysis. Partial sequences were analysed in isolates showing high degree of homology (>99%) with previous GenBank™ entries: H. canis, B. canis vogeli, B. equi (two isolates, Spain1 and Spain2), T. annulata and Theileria sp. The complete gene sequences were used for B. ovis and B. bovis, that showed lower homology (<95%) with rapport to previously reported species or isolates.A first set of phylogenetic trees constructed with partial 18s rRNA sequences showed that most European isolates clustered unambiguously with previously described species, so that minor sequence dissimilarities found are due probably to strain variations.The second set of phylogenetic trees was made using the complete 18s rRNA sequences of 44 species from GenBank™ and the newly sequenced B. ovis and B. bovis. The analysis revealed for the first time a division of piroplasmids in five clades:(1)B. microti group, with B. rodhaini, B. felis, B. leo, B. microti and T. annae (proposed name for the group, without taxonomic value: Archaeopiroplasmids),(2)Western USA Theilerid-like group (proposed name: Prototheilerids),(3)Theileria group, containing all Theileria species from Bovinae (proposed name: Theilerids),(4)A first group of Babesia species including B. canis and B. gibsoni from canids together with B. divergens and B. odocoilei (proposed name: Babesids),(5)A second group composed mainly by Babesia species from ungulates: B. caballi, B. bigemina, B. ovis, B. bovis and Babesia sp. from cow (proposed name: Ungulibabesids).The bootstrap support obtained with several analytical procedures for this new dicotomy of Babesiidae was always very high. Taking into account the present phylogenetic analysis and additional paleogeographic, parasitological and zoological evidences, two hypothesis on the origin and evolution of piroplasmids groups are presented. [Copyright &y& Elsevier]

Molecular epizootiology of piroplasmids (Babesia spp., Theileria spp.) and Hepatozoon canis was studied in mammals from southern Europe (mainly from Spain, but also from Portugal and France). Partial amplification and sequencing of the 18s rRNA gene was used for molecular diagnosis. In some particular cases (B. ovis and B. bovis) the complete 18s rRNA gene was sequenced.Blood samples were taken from domestic animals showing clinical symptoms: 10 dogs, 10 horses, 10 cows, 9 sheep and 1 goat. In addition, DNA samples were isolated from blood of 12 healthy dogs and from spleen of 10 wild red foxes (Vulpes vulpes). The results of the survey were the following:

Babesiosis is one of the most important tick-borne diseases in veterinary health, impacting mainly cattle, equidae, and canidae, and limiting the development of livestock industries worldwide. In humans, babesiosis is considered to be an emerging disease mostly due to Babesia divergens in Europe and Babesia microti in America. Despite this importance, our knowledge of Babesia sp. transmission by ticks is incomplete. The complexity of vectorial systems involving the vector, vertebrate host, and pathogen, as well as the complex feeding biology of ticks, may be part of the reason for the existing gaps in our knowledge. Indeed, this complexity renders the implementation of experimental systems that are as close as possible to natural conditions and allowing the study of tick-host-parasite interactions, quite difficult. However, it is unlikely that the development of more effective and sustainable control measures against babesiosis will emerge unless significant progress can be made in understanding this tripartite relationship. The various methods used to date to achieve tick transmission of Babesia spp. of medical and veterinary importance under experimental conditions are reviewed and discussed here. [ABSTRACT FROM AUTHOR]