RESUMO
Cysteine proteases are involved in the digestion of host blood and the degradation of yolk proteins of arthropod ectoparasites. In this study, a cathepsin L-like cysteine proteinase gene (HasCPL) of Hyalomma asiaticum was cloned, and recombinant (r)HasCPL protein was generated for immunization study. Bioinformatic analysis confirmed HasCPL was a member of the papain family (clan CA) and have high sequence identities with CPLs of other Ixodid ticks. The efficacy of immunization against H. asiaticum infestations in rabbits was assessed. Rabbits (n = 3) were immunized three times with rHasCPL before challenged with 250 larvae per rabbit four weeks post-immunization. A high antibody titer was detected in immunized rabbits in comparison to control. Western blot analysis detected CPLs in midgut, salivary gland, and ovary. Increase of rejection percentage of larvae were noted in ticks fed on immunized animals in comparison to control. Overall, a 55.09% protection against larva ticks was noted.
Assuntos
Cisteína Proteases , Ixodidae , Infestações por Carrapato , Animais , Cisteína Proteases/genética , Feminino , Imunização , Coelhos , Glândulas Salivares , Infestações por Carrapato/prevenção & controle , Infestações por Carrapato/veterináriaRESUMO
Hyalomma asiaticum and H. anatolicum are tick species in Eurasia and Africa with major medical and veterinary significance. Beside their direct pathogenic effects, H. asiaticum and H. anatolicum are vectors of important diseases of livestock and in some instances of zoonoses. In search of ways to address the increasing incidence of global acaricide resistance, tick control through vaccination is regarded as a sustainable alternative approach. Cathepsin L-like cysteine protease (CPL) is a potent hemoglobinase, and plays important roles in the digestion of blood acquired from a host. CPL from H. anatolicum (HanCPL) with high similarity (> 90%) for H. asiaticum CPL (HasCPL) were aligned by in silico analysis. After further in vitro validation, the anti-HasCPL sera have cross-reactivity between the different total native protein of life stages and tissues for H. asiaticum and H. anatolicum. Furthermore, we further confirmed that recombinant HasCPL (rHasCPL) immunized rabbits were partially cross-protected (54.8%) by H. anatolicum infestation.
Assuntos
Acaricidas , Ixodidae , Infestações por Carrapato , Carrapatos , Animais , Antígenos , Catepsina L , Coelhos , Infestações por Carrapato/veterináriaRESUMO
BACKGROUND: The neutrophil-to-lymphocyte ratio (NLR) is calculated from the white cell differential blood count. Recently, NLR was identified as a potential biomarker for the prediction of acute kidney injury (AKI). We conducted this systematic review and meta-analysis to evaluate the diagnostic value of NLCR for AKI in adult patients. METHODS: Studies in the PubMed, EMBASE, Web of Science and Cochrane Library databases were systematically searched from the date of database inception to February 28, 2019. The predictive value of NLR for AKI was evaluated by the pooled sensitivity, specificity, and summary receiver operating characteristic curve (SROC) analyses. Review Manager and Stata were used for all statistical analyses. The sources of potential heterogeneity were explored by a sensitivity analysis and subgroup analysis. RESULTS: This meta-analysis returned 89 reports, of which 9 fulfilled the inclusion criteria, accounting for 9766 patients. Bivariate analysis yielded a mean sensitivity of 0.736 (95% CI 0.675-0.790) and specificity of 0.686 (95% CI 0.601-0.759). The SROC was 0.77 (95% CI 0.74-0.81). The studies had no significant heterogeneity (Q = 0.675, p = 0.357, I2 = 0). CONCLUSIONS: Our findings indicate that the NLR may be a reliable biomarker for the early detection of AKI. Our findings also provide important information and assistance for clinicians in the prediction of AKI.
Assuntos
Injúria Renal Aguda/diagnóstico , Linfócitos , Neutrófilos , Injúria Renal Aguda/sangue , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Reprodutibilidade dos TestesRESUMO
Identification and molecular characterization of Babesia gibsoni proteins with potential antigenic properties are crucial for the development and validation of the serodiagnostic method. In this study, we isolated a cDNA clone encoding a novel B. gibsoni 76-kDa protein by immunoscreening of the parasite cDNA library. Computer analysis revealed that the protein presents a glutamic acid-rich region in the C-terminal. Therefore, the protein was designated as B. gibsoni glutamic acid-rich protein (BgGARP). A BLASTp analysis of a translated BgGARP polypeptide demonstrated that the peptide shared a significant homology with a 200-kDa protein of Babesia bigemina and Babesia bovis. A truncated BgGARP cDNA (BgGARPt) encoding a predicted 13-kDa peptide was expressed in Escherichia coli (E. coli), and mouse antisera against the recombinant protein were used to characterize a corresponding native protein. The antiserum against recombinant BgGARPt (rBgGARPt) recognized a 140-kDa protein in the lysate of infected erythrocytes, which was detectable in the cytoplasm of the parasites by confocal microscopic observation. In addition, the specificity and sensitivity of enzyme-linked immunosorbent assay (ELISA) with rBgGARPt were evaluated using B. gibsoni-infected dog sera and specific pathogen-free (SPF) dog sera. Moreover, 107 serum samples from dogs clinically diagnosed with babesiosis were examined using ELISA with rBgGARPt. The results showed that 86 (80.4%) samples were positive by rBgGARPt-ELISA, which was comparable to IFAT and PCR as reference test. Taken together, these results demonstrate that BgGARP is a suitable serodiagnostic antigen for detecting antibodies against B. gibsoni in dogs.
Assuntos
Antígenos de Protozoários/imunologia , Babesia/química , Proteínas de Protozoários/imunologia , Sequência de Aminoácidos , Animais , Antígenos de Protozoários/genética , Babesia/genética , Babesia/imunologia , Babesiose/diagnóstico , Babesiose/parasitologia , Babesiose/veterinária , Sequência de Bases , Clonagem Molecular , Reações Cruzadas , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Cães , Ensaio de Imunoadsorção Enzimática/normas , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Expressão Gênica , Ácido Glutâmico , Soros Imunes/imunologia , Camundongos , Camundongos Endogâmicos ICR , Dados de Sequência Molecular , Proteínas de Protozoários/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade , Organismos Livres de Patógenos EspecíficosRESUMO
Theileria equi is a tick-borne intracellular apicomplexan protozoan parasite that causes equine theileriosis (ET). ET is an economically important disease with a worldwide distribution that significantly impacts international horse movement. Horses are an essential part of the economy in Xinjiang which is home to â¼10% of all the horses in China. However, there is very limited information on the prevalence and genetic complexity of T. equi in this region. Blood samples from 302 horses were collected from May to September 2021 in Ili, Xinjiang, and subjected to PCR examination for the presence of T. equi. In addition, a Bayesian latent class model was employed to estimate the true prevalence of T. equi, and a phylogenetic analysis was carried out based on the 18S rRNA gene of T. equi isolates. Seventy-two horses (23.8%) were PCR positive. After accounting for the imperfect PCR test using a Bayesian latent class model, the estimated true prevalence differed considerably between age groups, being 10.8% (95%CrI: 5.8% - 17.9%) in ≤ 3-year-old horses and 35.7% (95%CrI: 28.1% - 44.5%) in horses that were > 3 year-old. All T. equi isolates had their 18S rRNA gene (430bp) sequenced and analyzed in order to identify whether there were multiple genotypes of T. equi in the Xinjiang horse population. All of the 18S rRNA genes clustered into one phylogenetic group, clade E, which is thus probably the dominant genotype of T. equi in Xinjiang, China. To summarize, we monitored the prevalence of T. equi in horses of Xinjiang, China, with a focus on the association between age and the occurrence of T. equi by Bayesian modelling, accompanied by the genotyping of T. equi isolates. Obtaining the information on genotypes and age structure is significant in monitoring the spread of T. equi and studying the factors responsible for the distribution.
Assuntos
Babesiose , Doenças dos Cavalos , Theileria , Theileriose , Bovinos , Cavalos , Animais , Prevalência , Filogenia , Teorema de Bayes , Theileriose/parasitologia , Variação Genética , Doenças dos Cavalos/diagnóstico , Babesiose/epidemiologiaRESUMO
BACKGROUND: Ticks in Xinjiang distribute widely and account for one third of China. Ticks can carry and transmit bacteria, virus, and parasite. However, the research of tick-borne pathogens in Xinjiang is rather little. OBJECTIVE: To understand the situation of hard tick carry Theileria equi, Babesia caballi and Rickettsia spp. of Zhaosu and Altay in Xinjiang. METHODS: In this study, 119 tick samples were obtained from horses in Xinjiang, China, Ticks were identified morphologically to determine species and PCR was used to investigate the situation of pathogens by hard ticks. RESULTS: One hundred and seven belong to Dermacentor marginatus, five belong to D. niveus, and seven belong to D. silvarum. Theileria equi and Babesia caballi were detected in one tick and 18 ticks, respectively. However, the carrying rate of Rickettsia spp. was 51.26% (61/119). Among these, the mixed carriage rate of T. equi and Rickettsia spp. was 0.8% (1/119). The mixed carriage rate of B. caballi and Rickettsia spp. was 10.1% (12/119). CONCLUSION: Our results revealed that hard tick can carry not only haeimoparasite but also many important zoonotic pathogens in Xinjiang, and this situation was worth heeding.
Assuntos
Babesia , Rickettsia , Theileria , Carrapatos , Animais , China/epidemiologia , Cavalos , Rickettsia/genética , Theileria/genética , Carrapatos/microbiologia , Carrapatos/parasitologiaRESUMO
In the present study, we screened 502 natural product compounds against the in vitro growth of Babesia (B.) bovis. Then, the novel and potent identified compounds were further evaluated for their in vitro efficacies using viability and cytotoxicity assays. The in vivo inhibitory effects of the selected compounds were evaluated using B. microti "rodent strain" in mice model. Three potent compounds, namely, Rottlerin (RL), Narasin (NR), Lasalocid acid (LA), exhibited the lowest IC50 (half-maximal inhibitory concentration) as follows: 5.45 ± 1.20 µM for RL, 1.86 ± 0.66 µM for NR, and 3.56 ± 1.41 µM for LA. The viability result revealed the ability of RL and LA to prevent the regrowth of treated parasite at 4 × IC50 and 2 × IC50, respectively, while 4 × IC50 of NR was sufficient to stop the regrowth of parasite. The hematology parameters of B. microti in vivo were different in the NR-treated groups as compared to the infected/untreated group. Interestingly, intraperitoneal administration of NR exhibiting inhibition in the growth of B. microti in mice was similar to that observed after administration of the commonly used antibabesial drug, diminazene aceturate (DA) (76.57% for DA, 74.73% for NR). Our findings indicate the richness of natural product compounds by novel potent antibabesial candidates, and the identified potent compounds, especially NR, might be used for the treatment of animal babesiosis.
RESUMO
Among bloodsucking arthropods, hard tick is a vector of transmitting the most diverse human and animal pathogens, leading to an increasing number of manifestations worldwide. The development of the anti-tick vaccine has the potential to be an environmentally friendly and cost-effective option for tick management. We have previously demonstrated the induction of both humoral and cellular response against Hyalomma asiaticum (H. asiaticum) following immunization with recombinant cathepsin L-like cysteine protease from H. asiaticum tick (rHasCPL), and could control tick infestations. Interferon-gamma (IFN-γ), is an immunomodulatory factor that plays an important role in the regulation of adaptive immunity against infection. In the present study, recombinant BALB/c mouse IFN-γ (rMus-IFN-γ) was cloned and expressed using a prokaryotic expression system, and verified by Western blotting and IFN-γ-ELISA kit analysis. Female BALB/c mice (n = 12) were used for immunization using rHasCPL (100 µg) plus IFN-γ as adjuvant (10 µg). In immunized female BALB/c mice, the levels of anti-CPL antibodies as well as cytokines were determined using ELISA analysis. Protective efficacy of immunization was evaluated by larvae H. asiaticum challenge of immunized female BALB/c mice. Using rMus-IFN-γ as an adjuvant to rHasCPL vaccine (CPL + IFN-γ) promoted specific antibody IgG (IgG1 > IgG2a) and increased production of IFN-γ and IL-4 compared to immune rHasCPL group (CPL). The protected rate of immunized mice from tick challenge was significantly higher after immunization with CPL + IFN-γ (85.11 %) than with CPL (63.28 %). Immunization using CPL + IFN-γ promoted the activation of anti-HasCPL humoral and cellular immune responses, and could provide better protection against H. asiaticum infestation. This approach may could help develop a candidate vaccine for control tick infestations.
Assuntos
Catepsina L/imunologia , Cisteína Proteases/imunologia , Citocinas/imunologia , Imunoglobulina G/imunologia , Memória Imunológica , Interferon gama/imunologia , Ixodidae/imunologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Catepsina L/genética , Feminino , Interferon gama/administração & dosagem , Interferon gama/genética , Ixodidae/enzimologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/imunologia , VacinaçãoRESUMO
The erythrocytic-stage surface protein equi merozoite antigen 1 (EMA-1) of Theileria equi is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In this study, BALB/c mice were immunized with purified recombinant EMA-1 to prepare monoclonal antibody (mAb) against T. equi EMA-1, and 1 mAb 5H2 was obtained that showed good reaction with infected red blood cells (RBC) in the indirect immunofluorescence assay (IFA). To develop a rapid serological detection method for T. equi infection in Xinjiang Uygur Autonomous Region, China, recombinant EMA-1 originating from the local T. equi strain and the mAb to EMA-1 were employed to develop an immunochromatographic test (ICT) to detect antibodies to T. equi in horse sera. The ICT showed high sensitivity and specificity and no cross-reaction with Babesia caballi. Ninety-two horse serum samples collected from Ili, Xinjiang, were tested by ICT and compared with the detection results of a commercial ELISA kit. The results showed that 56 of 92 (61%) serum samples were seropositive according to the ICT assay, and 50 (54%) samples were seropositive according to the ELISA kit. The ICT had a high coincidence (91.3%) but was more sensitive than the reference ELISA kit. To confirm whether the horses were infected by T. equi, 30 blood DNA samples from 92 horses were examined by PCR. The results showed that 14 of 30 (47%) horses were confirmed to be infected with T. equi by PCR, while 16 of 30 (53%) horses were seropositive by ICT. All PCR-positive horses were ICT-positive. The findings indicate that T. equi is endemic in Ili, Xinjiang, and that the ICT is reliable as a serological diagnosis method. The ICT developed in this study could be an efficient diagnostic tool to detect T. equi infection in horses in the Xinjiang area.
Assuntos
Antígenos de Protozoários/imunologia , Doenças dos Cavalos/parasitologia , Proteínas de Protozoários/imunologia , Theileria/isolamento & purificação , Theileriose/parasitologia , Animais , Anticorpos Monoclonais/imunologia , Antígenos de Protozoários/genética , Antígenos de Protozoários/isolamento & purificação , Western Blotting , China , Cromatografia de Afinidade , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Eritrócitos/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Doenças dos Cavalos/sangue , Doenças dos Cavalos/diagnóstico , Cavalos , Hibridomas/citologia , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase , Proteínas de Protozoários/genética , Proteínas de Protozoários/isolamento & purificação , Sensibilidade e Especificidade , Baço/citologia , Baço/imunologia , Theileria/imunologia , Theileriose/sangue , Theileriose/diagnóstico , Células Tumorais CultivadasRESUMO
Babesia, Theileria, and Anaplasma are important causative agents of tick-borne diseases that severely affect sheep. However, there is paucity in the occurrence genetic diversity of the infections of tick-borne diseases in sheep in border regions, northwestern China. In this study, nested polymerase chain reaction (nPCR) assays and gene sequencing were used to identify tick-borne Babesia spp., Theileria spp., and Anaplasma spp. infections in border regions, northwestern China. Out of 323 samples tested in this study, 225 (69.7%) sheep were infected with Babesia spp., Theileria spp., and Anaplasma spp. Two hundred six (63.8%), 60 (18.6%), 54 (16.7%), 51 (15.8%), 32 (9.9%), 19 (5.9%), and 16 (5.0%) were positive for A. ovis, B. motasi-like, A. bovis, T. uilenbergi, A. phagocytophilum, T. luwenshuni, and B. motasi-like Xinjiang, respectively. The most common dual infection was with A. ovis and B. motasi-like while the most frequent triple coinfection was A. ovis, B. motasi-like, and T. uilenbergi with coinfection rates of 17.0% (55/323) and 5.0% (16/323), respectively. Sequencing analysis indicated that A. ovis MSP4, A. phagocytophilum epank1, A. bovis 16S rRNA, B. motasi-like rap1-b, B. motasi-like Xinjiang rap1-a, T. luwenshuni 18S rRNA, and T. uilenbergi 18S rRNA from border regions, northwestern China, showed 99-100% identity with documented isolates from other countries. To the best of the authors' knowledge, this is the first report of T. uilenbergi and T. luwenshuni infections of sheep in border regions, northwestern China. Furthermore, these findings provide important data for understanding the distribution of Babesia, Theileria, and Anaplasma in sheep between border countries and China.
RESUMO
Ticks carry and transmit a wide range of pathogens (bacteria, viruses, and protozoa) that are of importance to humans and animals globally. However, information about the tick-borne pathogens harbored by ticks in the Xinjiang Uygur Autonomous Region (XUAR), northwestern China, is scarce. This study investigated the occurrence of tick species of domestic animals and tick-borne pathogens by using morphological molecular identification and sequence analysis in Turpan, Qitai, Altay, Hejing, Nileke, and Zhaosu counties (XUAR). A total of 5822 adult ticks (females and males) from 12 tick species were identified from 5 animal species (cattle, goats, sheep, camels, and horses) in 6 counties in the XUAR. Collected tick species included Dermacentor marginatus (24.7 %), Dermacentor nuttalli (20.8 %), Hyalomma anatolicum (13.7 %), Dermacentor niveus (13.1 %), Haemaphysalis punctata (10.7 %), Dermacentor silvarum (7.1 %), Dermacentor pavlovskyi (3.9 %), Hyalomma asiaticum (2.2 %), Rhipicephalus pumilio (1.9 %), Rhipicephalus sanguineus sensu lato (0.7 %), Rhipicephalus turanicus (0.6 %), and Hyalomma asiaticum kozlovi (0.6 %). Furthermore, 750 partially engorged adult ticks (females and males), including H. anatolicum (250), D. nuttalli (250), and D. marginatus (250), were individually separated according to species and sampling site, used for DNA extraction, and then screened for tick-borne pathogens. The most common pathogen was Rickettsia raoultii (36.80 %), followed by Brucella sp. (26.2 %), Anaplasma ovis (22.4 %), Babesia caballi (14.8 %), Theileria equi (8.7 %), and Theileria ovis (8.5 %). The sequencing of 6 genes showed a 96-100 % nucleotide identity between the sequences in this study and those deposited in GenBank. This study provides a scientific reference for the prevention and control of tick-borne diseases in the XUAR.
Assuntos
Anaplasma ovis/isolamento & purificação , Babesia/isolamento & purificação , Brucella/isolamento & purificação , Ixodidae/microbiologia , Ixodidae/parasitologia , Rickettsia/isolamento & purificação , Theileria/isolamento & purificação , Animais , Camelus/parasitologia , Bovinos/parasitologia , China , Feminino , Cabras/parasitologia , Cavalos/parasitologia , Masculino , Carneiro Doméstico/parasitologia , Especificidade da EspécieRESUMO
Q fever, spotted fever rickettsioses and equine piroplasmosis, are some of the most serious equine tick-borne diseases caused by Coxiella burnetii, Rickettsia spp., Babesia caballi and/or Theileria equi. This study surveyed and molecularly characterized these pathogens infecting horses in ten ranches from XUAR, China using molecular technology. Among 200 horse blood samples, 163 (81.5%) were infected with at least one of the pathogens. Rickettsia spp. was the most prevalent pathogen (n = 114, 57.0%), followed by C. burnetii (n = 79, 39.5%), T. equi (n = 79, 39.5%) and B. caballi (n = 49, 24.5%). Co-infections were observed in 61.3% of positive samples in this study. Statistically significant differences were observed between the sampling regions for C. burnetii, B. caballi and T. equi, and also in different age group for C. burnetii and T. equi. The genotype analysis indicated that C. burnetii htpB, Rickettsia spp. ompA, B. caballi rap-1, B. caballi 18S rRNA, T. equi EMA-1 and T. equi 18S rRNA gene sequences from horses in XUAR were variable. To the best of our knowledge, this study is the first report of C. burnetii and Rickettsia spp. infection and co-infected with piroplasma in horses in China. Overall, this study revealed the high infection rate of the pathogens in horses in XUAR, China. The current findings are expected to provide a basis for better tick-borne disease control in the region.
Assuntos
Babesiose/epidemiologia , Coinfecção/veterinária , Febre Q/veterinária , Infecções por Rickettsia/veterinária , Animais , Babesia/genética , Babesia/patogenicidade , China/epidemiologia , Coinfecção/epidemiologia , Coxiella burnetii/genética , Coxiella burnetii/patogenicidade , DNA de Protozoário/genética , Feminino , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/microbiologia , Doenças dos Cavalos/parasitologia , Cavalos/microbiologia , Cavalos/parasitologia , Filogenia , Prevalência , Febre Q/epidemiologia , RNA Ribossômico 18S/genética , Rickettsia/genética , Rickettsia/patogenicidade , Infecções por Rickettsia/epidemiologia , Theileria/genética , Theileria/patogenicidade , Theileriose/epidemiologia , Carrapatos/microbiologiaRESUMO
Tick-borne diseases cause significant losses to livestock production in tropical and subtropical regions. However, information about the tick-borne infections in cattle in Xinjiang Uygur Autonomous Region (XUAR), northwestern China, is scarce. In this study, nested polymerase chain reaction (PCR) assays and gene sequencing were used to detect and analyze epidemiological features of Babesia bovis, B. bigemina, Coxiella burnetii and Anaplasma bovis infections in XUAR. Out of 195 samples tested, 24 (12.3%), 67 (34.4%), 40 (20.5%) and 10 (5.1%) were positive for B. bovis, B. bigemina, C. burnetii and A. bovis, respectively. Sequencing analysis indicated that B. bovis SBP-4, B. bigemina Rap1a, C. burnetii htpB and A. bovis 16S rRNA genes from XUAR showed 99%-100% identity with documented isolates from other countries. Phylogenetic analyses revealed that B. bovis SBP-4, B. bigemina Rap1a, C. burnetii htpB and A. bovis 16S rRNA gene sequences clustered in the same clade with isolates from other countries. To the best of our knowledge, this is the first report of C. burnetii infection of cattle in XUAR. Furthermore, this study provides important data for understanding the distribution of tick-borne pathogens, and is expected to improve the approach for prevention and control of tick-borne diseases in China.
Assuntos
Anaplasma/isolamento & purificação , Babesia/isolamento & purificação , Doenças dos Bovinos/epidemiologia , Coxiella burnetii/isolamento & purificação , Febre Q/veterinária , Doenças Transmitidas por Carrapatos/veterinária , Anaplasma/genética , Anaplasmose/epidemiologia , Animais , Babesia/genética , Babesiose/epidemiologia , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/parasitologia , China/epidemiologia , Coxiella burnetii/genética , DNA Bacteriano/genética , DNA de Protozoário/genética , Filogenia , Febre Q/epidemiologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Doenças Transmitidas por Carrapatos/epidemiologia , Carrapatos/microbiologiaRESUMO
Few studies have been conducted on the distribution of Cryptosporidium species and subtypes in equine animals. In this study, 878 stool specimens were collected during 2015-2019 from 551 donkeys and 327 horses in Shandong, Xinjiang, and Inner Mongolia, China and screened for Cryptosporidium spp. by PCR analysis of the small subunit rRNA gene. The Cryptosporidium species presented were identified by sequence analysis of the PCR products and subtyped by sequence analysis of the 60â¯kDa glycoprotein gene. The infection rates of Cryptosporidium spp. in horses and donkeys were 3.1% (10/327) and 14.5% (80/551), respectively. Four Cryptosporidium species/genotypes were identified, including C. parvum (in 5 horses), C. hominis (in 75 donkeys), Cryptosporidium horse genotype (in 5 horses and 4 donkeys) and a new genotype that is genetically related to Cryptosporidium mink genotype (in 1 donkey). All C. parvum isolates were subtyped as IIdA19G1, C. hominis as IkA16G1, and horse genotype as VIaA15G4. Data from this study indicate that four Cryptosporidium species are circulating in horses and donkeys in the study areas, with C. hominis as a dominant Cryptosporidium species in only donkeys. Attention should be paid to reduce the transmission of these zoonotic Cryptosporidium spp.
Assuntos
Cryptosporidium/isolamento & purificação , Equidae/parasitologia , Cavalos/parasitologia , Animais , Cryptosporidium/classificação , Cryptosporidium/genética , Genes de ProtozoáriosRESUMO
Babesia caballi (Nuttal, 1910) is one of the causative agents of equine piroplasmosis which causes economic losses to horse industry in China. There is an urgent need for rapid detection method for B. caballi infection in Xinjiang Province, China. To prepare monoclonal antibodies (mAbs) against Bc48 gene of B. caballi (Xinjiang local strains) and establish colloidal gold-immunochromatographic (ICT) assay for diagnosis of the disease, recombinant Bc48 was expressed and purified from Escherichia coli. With purified Bc48 as immunogen in mice, three hybridoma cells named 11F4, 1H2 and 7F4 secreting mAbs against Bc48 of B. caballi were obtained, which showed strong reaction with recombinant Bc48 and Bc48 gene transfected cells. Furthermore, colloidal gold labelled ICT assay based on purified Bc48 recombinant antigen and its mAb was developed. The ICT assay showed high sensitivity and specificity and no cross-reaction with Theileria equi (Laveran, 1901). Total of 56 horse serum samples collected from Xinjiang were tested by ICT and compared with the detection by commercial ELISA kit. The results showed that 32 out of 56 serum samples were positive by ICT and 33 were positive by ELISA. ICT assay had high coincidence (98%) to the reference ELISA kit. mAbs and ICT developed in this study could be provided as an efficient diagnosis tool for infection with B. caballi in horse in Xinjiang area.
Assuntos
Anticorpos Antiprotozoários/sangue , Babesia/imunologia , Babesiose/diagnóstico , Doenças dos Cavalos/diagnóstico , Imunoensaio/veterinária , Proteínas de Protozoários/imunologia , Animais , Anticorpos Monoclonais/sangue , Babesiose/parasitologia , China , Feminino , Doenças dos Cavalos/parasitologia , Cavalos , Imunoensaio/métodos , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Protozoários/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Sensibilidade e EspecificidadeRESUMO
Tick DNA samples from cattle in Xinjiang Uygur Autonomous Region Area, China, were examined for Rickettsia infection by citrate synthase gene-based PCR and sequencing. Four positive samples were detected from Haemaphysalis danieli and high levels of similarity were found with recently detected 'Candidatus Rickettsia principis.'
Assuntos
Doenças dos Bovinos/parasitologia , Filogenia , Rickettsia , Infestações por Carrapato/veterinária , Carrapatos/microbiologia , Animais , Bovinos , China , Citrato (si)-Sintase/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Feminino , Amplificação de Genes , Masculino , Reação em Cadeia da Polimerase/veterinária , Rickettsia/classificação , Rickettsia/genética , Alinhamento de Sequência , Homologia de Sequência , Infestações por Carrapato/parasitologiaRESUMO
In order to found the epidemiological situation of T. equi in the horse herds in Ili Prefecture of Xinjiang Province, 723 blood samples collected from 4 counties and districts were test for T. equi through microscopic detection and Polymerase chain Reaction (PCR). In the result, we found that the 295 of 723 blood samples (40.8%) were positive for T. equi infection. The results showed that the choosed counties have a varying degrees infection. To our knowledge, this is the first time that we detected T. equi infection using the molecular techniques from Ili in Xinjiang Uygur Autonomous region.
Assuntos
Doenças dos Cavalos/epidemiologia , Theileria/isolamento & purificação , Theileriose/epidemiologia , Animais , China/epidemiologia , Estudos Epidemiológicos , Doenças dos Cavalos/parasitologia , Cavalos , Reação em Cadeia da Polimerase/veterinária , Theileria/genética , Theileriose/parasitologiaRESUMO
The prevalence of Babesia (Theileria) equi and B. caballi infections in donkeys in western Xinjiang China was investigated. In total, 93 serum samples were randomly taken from donkeys in the Kashi and Ili areas, and examined for B. equi and B. caballi infections by enzyme-linked immunosorbent assays using recombinant antigens. Of the 93 samples, 9 (9.6%) and 36 (38.7%) samples were positive for B. equi infection and B. caballi infection, respectively. In addition, 2 (2.2%) samples were positive for both B. equi and B. caballi infections. These results indicate that equine babesiosis might be extensively prevalent in donkeys in western Xinjiang.
Assuntos
Babesiose/veterinária , Equidae/sangue , Equidae/parasitologia , Animais , Babesiose/sangue , Babesiose/epidemiologia , Babesiose/parasitologia , China/epidemiologia , Prevalência , Estudos SoroepidemiológicosRESUMO
Serological methods were utilized to detect Anaplasma and Ehrlichia infection in domestic animals in Xinjiang Uygur Autonomous Region, China. By using an indirect immunofluorescence assay (IFA), antibodies that reacted with Anaplasmaphagocytophilum and Ehrlichiachaffeensis were detected mainly in ruminants kept on pastureland in Altai, Ili and Kashgar area. Antibody titers up to 1:320 were recorded. These results indicate that ruminants kept in these areas may be infected with some species of Anaplasma and Ehrlichia.
Assuntos
Anaplasma phagocytophilum/isolamento & purificação , Anaplasmose/epidemiologia , Animais Domésticos/microbiologia , Ehrlichia chaffeensis/isolamento & purificação , Ehrlichiose/veterinária , Anaplasmose/microbiologia , Animais , Anticorpos Antibacterianos/sangue , China/epidemiologia , Ehrlichiose/epidemiologia , Ehrlichiose/microbiologia , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Estudos SoroepidemiológicosRESUMO
The surface antigen 1-related sequence 2 of Neospora caninum (NcSRS2) is considered as an immunodominant antigen. In this study, the gene encoding truncated NcSRS2 (NcSRS2t) lacking an N-terminal signal peptide and C-terminal hydrophobic regions was expressed in Escherichia coli, and its diagnostic potential in an enzyme-linked immunosorbent assay (ELISA) was evaluated. ELISA could discriminate clearly between known N. caninum-positive and -negative sera from cattle. Field serum samples collected from cattle in Brazil were examined for the diagnosis of N. caninum infection using ELISA. Of the 197 samples analyzed, 64 (32.5%) samples were positive for antibodies to N. caninum. Of the 64 ELISA-positive samples, 58 (90.6%) were confirmed as positive by Western blot analysis with whole-parasite antigens. These results suggest that ELISA with recombinant NcSRS2t is an effective method for diagnosis of N. caninum infection in cattle.