Prvi dokaz invazije pasa lutalica na Filipinima protozoonom Babesia gibsoni pomoću imunokromatografskog testa.

A total of 46 stray dogs in an impounding facility comprising 17 males and 29 females were diagnosed using the Babesia gibsoni P50 truncated antigen immunochromatographic test (P50t-ICT). Thirteen dogs (28.0%) were serologically positive. There was no cross-reactivity with serum samples from Babesia canis (= B. vogeli)-infected dogs and none of the ICT strips showed invalid results, which reinforce the sensitivity and specificity of the P50t antigen and the reliability and accuracy of the P50t-ICT. Thirty-seven (80.4%) dogs had mixed tick infestations principally of the genus Rhipicephalus and Boophilus. From 11 seropositive and 20 seronegative dogs a total of 80 Rhipicephalus ticks were pooled. Among the 33 seronegative dogs, 48.5% had infestation with Rhipicephalus sp. only, 12.1% with Boophilus sp. only, 12.1% with mixed Rhipicephalus sp. and Boophilus sp., and 19.6% were un-infested. This paper documents the first account of serological detection of B. gibsoni in stray dogs and their infestation mainly with Rhipicephalus sp. suggestive of their role as putative key vectors of B. gibsoni in Philippine stray dogs.Istraživanje je provedeno kako bi se imunokromatografskim testom (P50t-ICT) dokazala prisutnost protozoona Babesia gibsoni u pasa lutalica na Filipinima. Ukupno je bilo pretraženo 46 pasa, 17 mužjaka i 29 ženki, na prisutnost antigena p50 protozoona Babesia gibsoni. Trinaest pasa (28%) bilo je serološki pozitivno. Nije ustanovljen nijedan slučaj križne reaktivnosti s uzorcima seruma pasa prirodno invadiranima protozoonom Babesia canis (Babesia vogeli). Test se pokazao prikladnim, osjetljivim, specifičnim, pouzdanim i točnim. U 37 pasa (80,4%) dokazane su mješovite infestacije krpeljima rodova Rhipicephalus i Boophilus. Na 11 serološki pozitivnih i 20 serološki negativnih pasa pronađeno je ukupno 80 krpelja iz roda Rhipicephalus. Od 33 serološki negativna psa krpelji roda Rhipicephalus dokazani su u 48,5%, roda Boophilus u 12,1%, a jedna mješovita infestacija vrstama tih rodova dokazana je također u 12,1% pasa. U 27,3% pasa krpelji nisu dokazani. Ovaj rad prvi put prikazuje mogućnost serološkoga dokaza infekcije vrstom B. gibsoni u pasa lutalica i njihovu infestaciju pretežito krpeljima roda Rhipicephalus kao ključnih prijenosnika B. gibsoni u pasa lutalica na Filipinim

Prvi dokaz invazije pasa lutalica na Filipinima protozoonom Babesia gibsoni pomoću imunokromatografskog testa.

A total of 46 stray dogs in an impounding facility comprising 17 males and 29 females were diagnosed using the Babesia gibsoni P50 truncated antigen immunochromatographic test (P50t-ICT). Thirteen dogs (28.0%) were serologically positive. There was no cross-reactivity with serum samples from Babesia canis (= B. vogeli)-infected dogs and none of the ICT strips showed invalid results, which reinforce the sensitivity and specificity of the P50t antigen and the reliability and accuracy of the P50t-ICT. Thirty-seven (80.4%) dogs had mixed tick infestations principally of the genus Rhipicephalus and Boophilus. From 11 seropositive and 20 seronegative dogs a total of 80 Rhipicephalus ticks were pooled. Among the 33 seronegative dogs, 48.5% had infestation with Rhipicephalus sp. only, 12.1% with Boophilus sp. only, 12.1% with mixed Rhipicephalus sp. and Boophilus sp., and 19.6% were un-infested. This paper documents the first account of serological detection of B. gibsoni in stray dogs and their infestation mainly with Rhipicephalus sp. suggestive of their role as putative key vectors of B. gibsoni in Philippine stray dogs.Istraživanje je provedeno kako bi se imunokromatografskim testom (P50t-ICT) dokazala prisutnost protozoona Babesia gibsoni u pasa lutalica na Filipinima. Ukupno je bilo pretraženo 46 pasa, 17 mužjaka i 29 ženki, na prisutnost antigena p50 protozoona Babesia gibsoni. Trinaest pasa (28%) bilo je serološki pozitivno. Nije ustanovljen nijedan slučaj križne reaktivnosti s uzorcima seruma pasa prirodno invadiranima protozoonom Babesia canis (Babesia vogeli). Test se pokazao prikladnim, osjetljivim, specifičnim, pouzdanim i točnim. U 37 pasa (80,4%) dokazane su mješovite infestacije krpeljima rodova Rhipicephalus i Boophilus. Na 11 serološki pozitivnih i 20 serološki negativnih pasa pronađeno je ukupno 80 krpelja iz roda Rhipicephalus. Od 33 serološki negativna psa krpelji roda Rhipicephalus dokazani su u 48,5%, roda Boophilus u 12,1%, a jedna mješovita infestacija vrstama tih rodova dokazana je također u 12,1% pasa. U 27,3% pasa krpelji nisu dokazani. Ovaj rad prvi put prikazuje mogućnost serološkoga dokaza infekcije vrstom B. gibsoni u pasa lutalica i njihovu infestaciju pretežito krpeljima roda Rhipicephalus kao ključnih prijenosnika B. gibsoni u pasa lutalica na Filipinim

Whole genome sequence and diversity in multigene families of Babesia ovis

Ovine babesiosis, caused by Babesia ovis, is an acute, lethal, and endemic disease worldwide and causes a huge economic loss to animal industry. Pathogen genome sequences can be utilized for selecting diagnostic markers, drug targets, and antigens for vaccine development; however, those for B. ovis have not been available so far. In this study, we obtained a draft genome sequence for B. ovis isolated from an infected sheep in Turkey. The genome size was 7.81 Mbp with 3,419 protein-coding genes. It consisted of 41 contigs, and the N50 was 526 Kbp. There were 259 orthologs identified among eight Babesia spp., Plasmodium falciparum, and Toxoplasma gondii. A phylogeny was estimated on the basis of the orthologs, which showed B. ovis to be closest to B. bovis. There were 43 ves genes identified using hmm model as well. They formed a discriminating cluster to other ves multigene family of Babesia spp. but showed certain similarities to those of B. bovis, B. caballi, and Babesia sp. Xinjiang, which is consistent with the phylogeny. Comparative genomics among B. ovis and B. bovis elucidated uniquely evolved genes in these species, which may account for the adaptation

Imunokromatografski test za dokaz invazije vrstama Babesia caballi i Babesia equi Laveran 1901 (Theileria equi Mehlhorn i Schein, 1998) (Phylum Apicomplexa) u fi lipinskih konja u usporedbi s dokazom parazita u krvnim razmascima

Sera collected from 71 slaughtered and 33 racing horses were assayed for Babesia spp. infection using immunochromatographic (ICT) assay. The ICT strips which were developed at the National Research Center for Protozoan Diseases (NRCPD), Obihiro University, Hokkaido, Japan contained a recombinant B. caballi 48-kDa rhoptry protein (rBc48) and a recombinant truncated B. equi merozoite antigen 2 (rEMA-2t) for the detection of anti-horse Babesia spp. antibodies. The 63 sero-positive blood samples consisted of 41(57.7%) and 22 (66.7%) cases in slaughtered and racing horses, respectively. Twelve sera (19.0%) reacted with both B. caballi and B. equi antigens, 45 sera (71.0%) reacted with rBc48 antigen only, and six sera (10.0%) were positive for B. equi antibodies only. Babesia caballi infection accounted for 90.5% cases. Infection with B. caballi and/or B. equi confirmed in Giemsa-stained blood smears prepared from racing horse samples only revealed 22 (66.7%) seropositive cases. Paired pear or crescent-shaped merozoites (0.5-1.25 μm), characteristic of B. caballi were observed in 20 blood smears, while only two seropositive cases revealed the presence of both B. caballi and the Maltese cross or tetrad-shaped merozoites (0.62-0.95 μm) generally associated with Theileria sp. (B. equi) parasite. To our knowledge, this is the first immunochromatographic assay of equine babesiosis in the Philippines validated by the detection of specific etiologic agent(s) in blood smears.Uzorci seruma 71 zaklanog konja i 33 športska konja bili su pretraženi na prisutnost protutijela za babezije imunokromatografskim testom (ICT). Testovi razvijeni u Nacionalnom istraživačkom centru za protozojske bolesti u sklopu Sveučilišta Obihiro u Hokaidu u Japanu sadržavali su rekombinantni protein od 48-kDa (rBc48) vrste B. caballi i rekombinantni krnji merozoitski antigen 2 (rEMA-2t) vrste B. equi za određivanje protutijela za vrste roda Babesia. Od ukupno 63 serološki pozitivna konja, 41 (57,7%) pripadao je skupini kojoj je krv bila uzeta pri klanju, a 22 (66,7%) bila su iz skupine športskih konja. Dvadeset uzoraka seruma (19,0%) bilo je pozitivno na oba antigena (B. caballi i B. equi), 45 uzoraka (71,0%) samo na antigen rBc48, dok je svega šest uzoraka (10%) bilo pozitivno na protutijela za vrstu B. equi. Protutijela za vrstu B. caballi bila su dokazana u 90,5% pretraženih uzoraka. Pretragom krvnih razmazaka obojenih po Giemzi babezije su bile dokazane u svega 22 (66,7%) športska konja. Kruškaste tvorevine (0,5-1,25 μm), karakteristične za merozoite protozoona B. caballi bile su dokazane u 20 razmazaka krvi. Samo u dva serološki pozitivna uzorka dokazana je vrsta B. caballi i merozoiti razmješteni u obliku malteškoga križa (0,62-0,95 μm) što je i karakteristika protozoa iz roda Theileria (B. equi). Ovim istraživanjem prvi put je dokazana prikladnost imunokromatografskoga testa za određivanje protutijela za babezije konja na Filipinima, a rezultati su uspoređeni s nalazom uzročnika u krvnim razmascima

Babesia ovis secreted antigen-1 is a diagnostic marker during the active Babesia ovis infections in sheep

Ovine babesiosis caused by Babesia ovis is an economically significant disease. Recently, a few B. ovis-specific proteins, including recombinant B. ovis secreted antigen-1 (rBoSA1), have been identified. Immunological analyses revealed that rBoSA1 resides within the cytoplasm of infected erythrocytes and exhibits robust antigenic properties for detecting anti-B. ovis antibodies. This protein is released into the bloodstream during the parasite’s development. It would be possible to diagnose active infections by detecting this secretory protein. For this purpose, a rBoSA1-specific polyclonal antibody-based sandwich ELISA was optimized in this study. Blood samples taken from the naturally (n: 100) and experimentally (n: 15) infected sheep were analyzed for the presence of native BoSA1. The results showed that native BoSA1 was detectable in 98% of naturally infected animals. There was a positive correlation between parasitemia level in microscopy and protein density in sandwich ELISA. Experimentally infected animals showed positive reactions from the first or second day of inoculations. However, experimental infections carried out by Rhipicephalus bursa ticks revealed the native BoSA1 was detectable from the 7th day of tick attachment when the parasite began to be seen microscopically. Sandwich ELISA was sensitive enough to detect rBoSA1 protein at a 1.52 ng/ml concentration. Additionally, no serological cross-reactivity was observed between animals infected with various piroplasm species, including Babesia bovis, B. bigemina, B. caballi, B. canis, B. gibsoni, Theileria equi, and T. annulata. Taken collectively, the findings show that the rBoSA1-specific polyclonal antibody-based sandwich ELISA can be successfully used to diagnose clinical B. ovis infections in sheep at the early stage

Scavenger Receptor Mediates Systemic RNA Interference in Ticks

RNA interference is an efficient method to silence gene and protein expressions. Here, the class B scavenger receptor CD36 (SRB) mediated the uptake of exogenous dsRNAs in the induction of the RNAi responses in ticks. Unfed female Haemaphysalis longicornis ticks were injected with a single or a combination of H. longicornis SRB (HlSRB) dsRNA, vitellogenin-1 (HlVg-1) dsRNA, and vitellogenin receptor (HlVgR) dsRNA. We found that specific and systemic silencing of the HlSRB, HlVg-1, and HlVgR genes was achieved in ticks injected with a single dsRNA of HlSRB, HlVg-1, and HlVgR. In ticks injected first with HlVg-1 or HlVgR dsRNA followed 96 hours later with HlSRB dsRNA (HlVg-1/HlSRB or HlVgR/HlSRB), gene silencing of HlSRB was achieved in addition to first knockdown in HlVg-1 or HlVgR, and prominent phenotypic changes were observed in engorgement, mortality, and hatchability, indicating that a systemic and specific double knockdown of target genes had been simultaneously attained in these ticks. However, in ticks injected with HlSRB dsRNA followed 96 hours later with HlVg-1 or HlVgR dsRNAs, silencing of HlSRB was achieved, but no subsequent knockdown in HlVgR or HlVg-1 was observed. The Westernblot and immunohistochemical examinations revealed that the endogenous HlSRB protein was fully abolished in midguts of ticks injected with HlSRB/HlVg-1 dsRNAs but HlVg-1 was normally expressed in midguts, suggesting that HlVg-1 dsRNA-mediated RNAi was fully inhibited by the first knockdown of HlSRB. Similarly, the abolished localization of HlSRB protein was recognized in ovaries of ticks injected with HlSRB/HlVgR, while normal localization of HlVgR was observed in ovaries, suggesting that the failure to knock-down HlVgR could be attributed to the first knockdown of HlSRB. In summary, we demonstrated for the first time that SRB may not only mediate the effective knock-down of gene expression by RNAi but also play essential roles for systemic RNAi of ticks