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1.
Ticks Tick Borne Dis ; 14(1): 102077, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36402047

RESUMO

Theileria orientalis is known to cause a benign infection in cattle and buffalo (Bubalus bubalis). However, the Ikeda and Chitose genotypes of the parasite cause lethal disease in beef and dairy cattle. Recently an outbreak of clinical oriental theileriosis occurred in buffalo calves in a Government Animal Husbandry and Agricultural Farm located in Uttar Pradesh, India. Examination of Giemsa stained thin blood smears revealed typical rod-shaped T. orientalis piroplasms in the erythrocytes. The clinical signs included pyrexia, nasal discharge, lacrimation, lethargy, inappetence and anaemia with varying degrees of paleness of the visible mucous membranes. Vascular congestion in internal organs, pulmonary emphysema and consolidation of lungs, focal areas of necrosis in the heart with mononuclear cell infiltration, focal mononuclear cell aggregation in the cortex and tubular degeneration of the kidney were significant necropsy findings. The T. orientalis major piroplasm surface protein (MPSP) gene was amplified by polymerase chain reaction (PCR) using specific primers. The nucleotide sequence analysis of the PCR product revealed 84.8% identity between the T. orientalis Uttar Pradesh isolate and other reference genotypes available in the public domain. Furthermore, the phylogenetic analysis of the MPSP gene sequence ratified that this is a new genotype of T. orientalis. This is the first report of a clinical outbreak of oriental theileriosis in Indian buffalo calves caused by a novel genotype of T. orientalis.


Assuntos
Theileria , Animais , Bovinos , Theileria/genética , Búfalos , Filogenia , Índia/epidemiologia
2.
Ticks Tick Borne Dis ; 14(1): 102078, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36395616

RESUMO

Central Asia, including Kazakhstan, is an endemic area of Theileria and Babesia infections in cattle. Current data on the geographic distribution, prevalence, and genetic diversity of these pathogens in vertebrate hosts are lacking in Kazakhstan. The present study aimed to fill this gap, using molecular techniques for the first time. A cross-sectional survey was performed on adult cattle from 40 villages in nine administrative districts of the provinces of Turkistan and Zhambyl, southern Kazakhstan, in summer 2020. A total of 766 blood samples were screened for Theileria annulata (enolase gene), Theileria orientalis (major piroplasm surface protein gene, MPSP) and Babesia spp. (18 S ribosomal RNA gene) using polymerase chain reaction. The genetic variability of Theileria spp. was assessed by sequencing one amplicon from each village. All Babesia spp. positive amplicons were sequenced to identify the species involved. The overall prevalence of infections with T. annulata, T. orientalis and Babesia spp. was 83.0% (40 villages positive), 33.3% (31 villages) and 13.5% (36 villages), respectively. Co-infections with two or three species were present in 48.9% of all positive cattle. Theileria annulata showing a high polymorphism of the enolase gene occurred with similar frequency in both provinces. Theileria orientalis was detected for the first time in Kazakhstan being significantly (P = 0.014) more prevalent in Zhambyl than in Turkistan. Fourteen genotypes of T. orientalis were identified; two belonged to the moderately virulent MPSP-type 1 ('Chitose') and the others to MPSP-type 3 ('Buffeli') which is considered avirulent. The prevalence of Babesia infection was significantly (P < 0.000) higher in Turkistan than in Zhambyl. An unequivocal identification of the species involved was possible in 127 sequenced samples: Babesia occultans was the most common species, followed by Babesia bigemina and Babesia major, the latter being the first record in the country. The results show that Theileria and Babesia infections in cattle are widespread and occur with remarkably high prevalence in the southern Kazakhstan. They also provide first data on the genetic diversity of the species involved.


Assuntos
Babesiose , Theileria , Bovinos , Animais , Theileria/genética , Babesiose/epidemiologia , Estudos Transversais , Cazaquistão/epidemiologia
3.
J Equine Vet Sci ; 120: 104190, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36509242

RESUMO

Among infection diseases transmitted by arthropods, the equine vector-borne diseases transmitted by ticks represent an emerging problem worldwide due to their morbidity and mortality and, in some cases, to their zoonotic relevance. Understanding the host immune/inflammatory response to the pathogens is crucial to develop effective methods of diagnosis, control and treatment. This study aimed to evaluate red blood cells (RBC), hematocrit (Hct), hemoglobin concentration (Hb), mean cell volume (MCV), mean cell hemoglobin content (MCH), mean cell hemoglobin (MCHC), platelets (PLT), white blood cells (WBC), leukocytes populations, and serum protein pattern of Sardinian donkeys scored positive to tick-borne pathogens. Thirty donkeys were dived in five groups, six subjects each: Group 1, tested negative to pathogens (control group); Group 2, donkeys infected by Theileria equi; Group 3, donkeys infected by Ehrlichia equi; Group 4, donkeys infected by Rickettsia Rickettsii; Group 5, donkeys co-infected by T. equi, E. equi and R. Rickettsii. From donkeys blood samples were collected and haematological and serum protein profiles were assessed. One-way ANOVA showed lower red blood cells, haemoglobin, platelets, lymphocytes and neutrophils values, and higher eosinophils, basophils, serum total proteins, α2-, ß- and γ- globulins values in donkeys infected by one or more pathogens compared to control group (P < .05). The study highlighted an immune and inflammatory response of donkeys positive to E. equi, T. equi, and R. Rickettsii and represents a starting point for further investigations in the field to fill the gaps currently existing on the complex interwoven relationship between the pathogen and host immune and inflammatory response in equids.


Assuntos
Theileria , Carrapatos , Cavalos , Animais , Equidae , Carrapatos/microbiologia , Rickettsia rickettsii
4.
Parasitol Res ; 122(2): 387-394, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36504396

RESUMO

Piroplasmosis is a disease that negatively affects equine health worldwide. Hence, 324 blood samples were collected from grazing horses in ten sites in Xinjiang and testing them for the presence of Theileria equi and Babesia caballi by PCR of the EMA-1 gene and BC48 gene, respectively. Of the 324 blood samples, 161 (49.7%) were positive for equine piroplasms. The prevalence of T. equi was 38.9% (126/324), while that of B. caballi was 30.2% (98/324). The T. equi and B. caballi co-infection rate was 19.4% (63/324). From the 126 EMA-1 gene sequences and 98 BC48 gene sequences we obtained, 21 and 27 genotypes were identified, respectively. The EMA-1 sequences together with the GenBank reference sequences grouped into four clusters, with those from the present study forming two distinct clusters. In contrast, the BC48 sequences formed eight clusters with the GenBank reference sequences, while those obtained in the present study formed five distinct clusters. Our results highlight the widespread distribution and abundant gene polymorphism of T. equi and B. caballi in grazing horses from Xinjiang.


Assuntos
Babesia , Babesiose , Doenças dos Cavalos , Theileria , Theileriose , Bovinos , Cavalos , Animais , Babesia/genética , Theileria/genética , Theileriose/epidemiologia , Prevalência , Doenças dos Cavalos/epidemiologia , Filogenia , Babesiose/epidemiologia , China/epidemiologia , Bactérias
5.
Vet Med Sci ; 9(1): 372-379, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36495157

RESUMO

BACKGROUND: Ticks are important ectoparasites of small ruminants in tropics and subtropics including Iran. They transmit serious zoonotic pathogens such as Babesia and Theileria. These parasites cause major burden on small ruminants jeopardising livelihoods of rural people in Zarrin Dasht County. OBJECTIVES: This study was carried out to investigate the diversity and distribution of hard ticks of small ruminants and their piroplasm infection in a bid to contribute to Theileria and/or Babesia detection and control in Zarrin Dasht County of Fars province, Iran. METHODS: We examined 751 sheep and goats from 10 sites of the County during four seasons for hard tick infestation. The collected hard ticks (994) were taxonomically identified before being separately confined in microtubes coded to indicate their species and host animals as well as site and date of collection. In total 50 pooled samples were analysed by PCR technique for Theileria and Babesia infection. RESULTS: The identified ticks included Hyalomma marginatum 994/362); 36.4%), Rhipicephalus turanicus 994/352); 35.51%), Hyalomma anatolicum 994/264); 26.6%), Hyalomma dromedarii 994/14); 1.41%) and Hyalomma asiaticum 994/2) 0.2%). Molecular analyses showed that 7 out of 50 pooled sample were infected with piroplasm genome in ticks shared by Theileria ovis (6:50) and Theileria lestoquardi (1:50). Babesia was absent in collected hard ticks. CONCLUSIONS: This is the first report on the presence of piroplasm infection in hard ticks of small ruminants in Zarrin Dasht County. Theileria ovis was more prevalent than Theileria lestoquardi but Babesia was absent. Piroplasm infection was detected in Hyalomma marginatum, Hyalomma anatolicum and Rhipicephalus turanicus. Hyalomma marginatum appears to be more competent to vector Theileria spp. This study may contribute to risk assessment and prevention of epizootic theileriosis in the County.


Assuntos
Babesia , Doenças dos Bovinos , Doenças das Cabras , Ixodidae , Doenças dos Ovinos , Theileria , Theileriose , Bovinos , Ovinos , Animais , Theileria/genética , Irã (Geográfico)/epidemiologia , Theileriose/epidemiologia , Theileriose/parasitologia , Ruminantes , Babesia/genética , Cabras , Doenças das Cabras/epidemiologia , Doenças das Cabras/parasitologia , Doenças dos Ovinos/diagnóstico
6.
BMC Vet Res ; 18(1): 454, 2022 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-36577977

RESUMO

BACKGROUND: There had been isolated reports of the presence of novel Theileria annulata genotypes based on the 18S rRNA gene sequence data from India, Pakistan and Saudi Arabia; but, these studies were restricted to limited field samples. Additionally, no comparative study has been conducted on all the isolates of this parasite from different countries whose sequences are available in the nucleotide databases. Therefore, we aimed to study the genetic diversity of T. annulata based on all available nearly complete 18S rRNA gene sequences in the GenBank™. Out of a total of 312 gene sequences of T. annulata available in the NCBI database, only 70 nearly complete sequences (> 1527 bp) were used for multiple sequence alignment. RESULTS: The maximum likelihood tree obtained using TN93 + G + I model manifested two major clades. All the valid host-cell transforming Theileria species clustered in one clade. The T. annulata designated sequences occupying this clade clustered together, excluding two isolates (DQ287944 and EU083799), and represented the true T. annulata sequences (n = 54). DQ287944 and EU083799 exhibited close association with Theileria lestoquardi. In addition, 14 Indian sequences formed a large monophyletic group with published Theileria orientalis sequences. The broad range of sequence identity (95.8-100%) of T. annulata designated sequences indicated the presence of different Theileria spp. A closer analysis revealed the presence of three Theileria spp., namely, T. annulata, T. orientalis, and two isolates (DQ287944 and EU083799) closely related to T. lestoquardi. The true T. annulata sequences manifested 98.8-100% nucleotide identity within them. EU083799 and 14 misidentified Indian T. annulata sequences exhibited the highest similarity with T. lestoquardi (98.6-98.8%) and T. orientalis (98.0-99.9%) in comparison with the other Theileria spp. of domestic and wild ruminants. CONCLUSION: In the course of analyzing the genetic diversity of T. annulata, we identified the nearly complete 18S rRNA gene sequences of other Theileria spp. that have not only been misidentified as T. annulata in the GenBank™, but are also published as T. annulata. Moreover, a high level of sequence conservation was noticed in the 18S rRNA gene of true T. annulata and T. orientalis sequences.


Assuntos
Doenças dos Bovinos , Theileria annulata , Theileria , Theileriose , Bovinos , Animais , Theileria/genética , Theileria annulata/genética , RNA Ribossômico 18S/genética , Theileriose/epidemiologia , Theileriose/parasitologia , Filogenia , Nucleotídeos , Doenças dos Bovinos/parasitologia
7.
Ann Parasitol ; 68(3): 531-536, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36586724

RESUMO

Ovine theileriosis as a critical agent in small ruminant production, can cause lethal infections. Different species of Theileria have been reported in various parts of the world, and each species causes different diseases in the host. This is the first molecular study to investigate the prevalence of ovine theileriosis and identify the dominant Theileria species in northern Iran. A number of 220 small ruminants, including sheep and goats, were randomly sampled from 22 flocks. Peripheral blood smears were stained by the Giemsa staining method. As well as for species identification, all samples were examined by PCR. From 220 samples, 160 and 60 were sheep and goat, respectively. By the Giemsa staining method, Theileria parasite was observed in 20 (9%) samples. But by polymerase chain reaction (PCR) method, 30 (13.6%) samples were positive for Theileria species. Theileria lestoquardi was the most common species found in these animals. The high prevalence of theileriosis in small ruminants demonstrates the emergence of ovine theileriosis in Mazandaran and Golestan provinces in northern Iran.


Assuntos
Doenças dos Ovinos , Theileria , Theileriose , Bovinos , Ovinos , Animais , Theileria/genética , Theileriose/epidemiologia , Theileriose/parasitologia , Irã (Geográfico)/epidemiologia , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/parasitologia , Cabras
8.
Trop Anim Health Prod ; 54(6): 383, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36380247

RESUMO

Theileriosis is one of the top ten economically important diseases in cattle in India. Cytokines are considered important mediators and regulators of the immune response to an infection. In the present study, the gene expression profiles of fourteen cytokines (IL1A, IL1B, IL2, IL4, IL6, IL8, IL10, IL12A, IL12B, IL16, TGFB1, TNFA, IFNA and IFNB) were compared in Theileria annulata-infected and healthy crossbred cattle. Blood samples were obtained from the District Disease Diagnostic Laboratory, Karnal. The presence/absence of T. annulata infection in the animals was determined on the basis of blood smear examination and molecular detection through PCR using the genus-specific primers. Total RNA was extracted from peripheral blood mononuclear cells, which was further reverse transcribed to cDNA. Primer3 software was employed to design the primers for Real-Time qPCR. The results were examined using 2-∆∆Ct method with RPS15 and GAPDH as the reference genes. The expression of IL1B, IL6, IL8, IL10, IL12A, IL12B, TNFA, IFNA and IFNB was significantly higher, whereas the expression of IL2 was lower in the infected animals. The transcript levels of IL1A and TGFB1 were also higher in the diseased animals, but the results were non-significant. This study profiles the expression kinetics of various pro-inflammatory and anti-inflammatory cytokine genes in response to bovine theileriosis.


Assuntos
Doenças dos Bovinos , Theileria annulata , Theileria , Theileriose , Bovinos , Animais , Theileria annulata/genética , Leucócitos Mononucleares/metabolismo , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Interleucina-2/metabolismo , Interleucina-8 , Citocinas/genética , Citocinas/metabolismo , Primers do DNA
9.
Trop Anim Health Prod ; 54(6): 368, 2022 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-36322222

RESUMO

During February 2020 to October 2020, four outbreaks of theileriosis in small ruminants were recorded with overall morbidity, mortality and case fatality rates of 27.95%, 17.46% and 62.5%, respectively. The disease was characterized by high fever (up to 106°F), superficial lymphadenopathy, anaemia, anorexia, lethargy, respiratory distress and death. The presence of pleomorphic intra-erythrocytic piroplasms of Theileria species in Giemsa's stained blood smears was a common finding in all the episodes. Significant haematological alterations including high total leucocyte count and low haemoglobin and packed cell volume were characteristic. Necropsy findings of the icteric liver, enlarged spleen, pulmonary oedema and abomasal ulcerations were observed in three flocks. Smear-positive blood samples from all the episodes were screened by PCR using 18S rRNA gene-specific primer sets for T. lestoquardi, T. luwenshuni, T. uilenbergi and T. ovis. T. lestoquardi which was detected in all four flocks, while there was co-infection of T. ovis in two flocks. Phylogenetic analysis revealed that T. ovis and T. lestoquardi identified in this study had 100% and ~ 99.86% homology, respectively, with the published sequences used for comparison. This is the first confirmed report of outbreaks of malignant ovine theileriosis in the Haryana state of India which caused high morbidity, mortality and case fatality among sheep and goats. Further studies on theilerioses in small ruminants are required to understand epidemiology better.


Assuntos
Doenças dos Bovinos , Doenças das Cabras , Doenças dos Ovinos , Theileria , Theileriose , Bovinos , Ovinos , Animais , Theileriose/epidemiologia , Cabras , Filogenia , Doenças dos Ovinos/epidemiologia , Theileria/genética , Surtos de Doenças/veterinária , Doenças dos Bovinos/epidemiologia , Doenças das Cabras/epidemiologia
10.
Parasit Vectors ; 15(1): 401, 2022 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-36316753

RESUMO

BACKGROUND: The apicomplexan haemoparasite Theileria equi, a causative agent of equine piroplasmosis, is an established pathogen of significant welfare and economic concern within the Croatian equine population. A previous large surveillance study of T. equi has identified two distinct parasite populations, one in the north and one in the south, geographically separated by the Dinaric Alps, which traverse the country. This study aimed to further investigate the genetic diversity within these two populations, focussing on allelic variability of the equi merozoite antigen gene, ema-1. METHODS: Following nested PCR of DNA isolates, the generated ema-1 amplicons were subsequently sequenced and compared by phylogenetic analysis to available sequences representing previously described ema-1 genotypes (groups A-C). RESULTS: Isolates from the southern T. equi population clustered with the existing ema-1 groups A and B. Strikingly, isolates from the northern population clustered into two novel ema-1 genotypes, named groups D and E. CONCLUSIONS: This detection of hitherto unreported genotypes suggests that historic geographical isolation has led to a degree of divergent evolution in this northern T. equi population. Additionally, current global regulatory testing of equine piroplasmosis relies heavily on EMA-1 based immunodiagnostics, and the discovery of unique ema-1 genotypes may question the efficacy of current diagnostics in international equine movement, with ramifications for the global equine community.


Assuntos
Babesiose , Doenças dos Cavalos , Theileria , Theileriose , Cavalos , Animais , Bovinos , Merozoítos , Theileriose/parasitologia , Croácia/epidemiologia , Babesiose/parasitologia , Filogenia , Antígenos de Protozoários , Doenças dos Cavalos/diagnóstico
11.
Vet Parasitol Reg Stud Reports ; 36: 100792, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36436901

RESUMO

In all equids worldwide, Theileria equi and Babesia caballi are believed to be two important erythrocytic protozoa that cause equine piroplasmosis. In addition, it was recently discovered that Theileria haneyi is another potential equine piroplasmosis (EP) agent. Ixodid ticks are the major vectors of these parasites. Equine piroplasmosis is of international importance and affects enormously the equine industry. In this study, for the first time, molecular prevalence and genetic diversity of piroplasma parasites (T. equi and B. caballi) in horses from Fars province (south of Iran) were determined. Also, hematological alterations of naturally infected horses were analyzed. PCR positive horses showed anemia, thrombocytopenia, leukocytosis with a left shift of neutrophilia, and monocytosis. PCR results revealed that, from 133 blood samples of horses, 40 samples were positive (30.07%). The occurrence of T. equi in this area (30.07%) was more than the national average prevalence of T. equi (24.11%), but B. caballi prevalence in study area (0%) was less than the average of previous studies in Iran (5.47%). Our findings revealed that the T. equi was widespread in Fars province of Iran. PCR products of 18S rDNA and EMA-1 genes of T. equi strains were sequenced successfully. All 18S rDNA sequences collected in this experiment revealed 100% similarity together. According to the phylogenetic tree constructed using the 18S rDNA gene, Iranian T. equi is clustered with strains from Cuba (KY111762, KY111761) and USA (CP001669, JX177672). So, this could be concluded that T. equi studied in this research, and those strains are initiated from a common T. equi ancestor at an unknown time ago. Also, the phylogenetic tree based on EMA-1 gene demonstrated a genetically diverse population of Iranian T. equi strains (10 different genotypes). As EMA-1 is one of the most immunogenic antigens in this parasite, such variability could be a concern about the efficacy of T. equi vaccines. Finally, more studies on equine piroplasmosis in the provinces of the southern region of Iran are recommended to create a better vision of disease in this region.


Assuntos
Babesia , Babesiose , Doenças dos Bovinos , Doenças dos Cavalos , Theileria , Theileriose , Bovinos , Cavalos , Animais , Irã (Geográfico)/epidemiologia , Babesiose/parasitologia , Theileriose/parasitologia , Filogenia , Doenças dos Cavalos/parasitologia , Babesia/genética , Variação Genética , DNA Ribossômico
12.
Prev Vet Med ; 209: 105777, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36272258

RESUMO

Tick-borne diseases (TBD) are a major constraint to livestock health and productivity in sub-Saharan Africa. Nonetheless, there are relatively few robust epidemiologic studies documenting TBD and its management in different endemic settings in Kenya. Therefore, a cross-sectional study using multi-stage cluster sampling was undertaken to characterize the epidemiology of TBD and management factors among zebu cattle reared under an extensive system in coastal Kenya. Blood samples from 1486 cattle from 160 herds in 14 villages were screened for the presence of tick-borne bacterial and protozoan pathogens using PCR with high-resolution melting analysis and sequencing. Standardized questionnaires were used to collect data on herd structure and herd management practices, and a mixed-effect logistic regression model to identify risk factors for tick-borne pathogens (TBPs). The application of chemical acaricide was the primary method for tick control (96.3%, 154/160), with the amidine group (mainly Triatix®, amitraz) being the most frequently used acaricides. Respondents identified East Coast fever as the most important disease and Butalex® (buparvaquone) was the most commonly administered drug in response to perceived TBD in cattle. The overall animal- and herd-level prevalence for TBPs were 24.2% (95% confidence interval (CI): 22.0-26.4%) and 75.6% (95% CI: 68.2-82.1%), respectively. Cattle were infected with Anaplasma marginale (10.9%, 95% CI: 9.4-12.6), Theileria parva (9.0%, 95% CI: 7.5-10.5), Anaplasma platys (2.6%, 95% CI: 1.9-3.6), Theileria velifera (1.1%, 95% CI: 0.7-1.8), Babesia bigemina (0.5%, 95% CI: 0.2-1.0), and Anaplasma sp. (0.1%, 95% CI: 0.0-0.4). Moreover, 21 cattle (1.4%) were co-infected with two TBPs. None of the assessed potential risk factors for the occurrence of either A. marginale or T. parva in cattle were statistically significant. The intra-herd correlation coefficients (lCCs) computed in this study were 0.29 (A. marginale) and 0.14 (T. parva). This study provides updated molecular-based information on the epidemiological status of TBPs of cattle and herd management practices in coastal Kenya. This information can be used in designing cost-effective control strategies for combating these TBD in the region.


Assuntos
Anaplasmose , Doenças dos Bovinos , Theileria , Theileriose , Doenças Transmitidas por Carrapatos , Carrapatos , Bovinos , Animais , Carrapatos/microbiologia , Quênia/epidemiologia , Controle de Ácaros e Carrapatos/métodos , Estudos Transversais , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Theileriose/epidemiologia , Theileriose/prevenção & controle , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/prevenção & controle , Doenças Transmitidas por Carrapatos/veterinária , Anaplasmose/epidemiologia , Anaplasmose/microbiologia
13.
Parasit Vectors ; 15(1): 356, 2022 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-36199104

RESUMO

BACKGROUND: When Theileria annulata infects host cells, it undertakes unlimited proliferation as tumor cells. Although the transformed cells will recover their limited reproductive characteristics and enter the apoptosis process after treatment with buparvaquone (BW720c), the metabolites and metabolic pathways involved are not clear. METHODS: The transformed cells of T. annulata were used as experimental materials, and the buparvaquone treatment group and DMSO control group were used. Qualitative and quantitative analysis was undertaken of 36 cell samples based on the LC-QTOF platform in positive and negative ion modes. The metabolites of the cell samples after 72 h of drug treatment were analyzed, as were the different metabolites and metabolic pathways involved in the BW720c treatment. Finally, the differential metabolites and metabolic pathways in the transformed cells were found. RESULTS: A total of 1425 metabolites were detected in the negative ion mode and 1298 metabolites were detected in the positive ion mode. After drug treatment for 24 h, 48 h, and 72 h, there were 56, 162, and 243 differential metabolites in negative ion mode, and 35, 121, and 177 differential metabolites in positive ion mode, respectively. These differential metabolites are mainly concentrated on various essential amino acids. CONCLUSION: BW720c treatment induces metabolic disturbances in T. annulata-infected cells by regulating the metabolism of leucine, arginine, and L-carnitine, and induces host cell apoptosis.


Assuntos
Theileria annulata , Theileria , Theileriose , Animais , Arginina/uso terapêutico , Carnitina/uso terapêutico , Bovinos , Dimetil Sulfóxido/uso terapêutico , Leucina/uso terapêutico , Naftoquinonas , Theileriose/tratamento farmacológico
14.
Parasitol Res ; 121(11): 3321-3326, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36114837

RESUMO

Tick-borne pathogens are causing severe diseases in livestock, wild animals, and humans. Wild animals play a crucial role in tick-borne pathogens' transmission life cycle by serving as reservoir hosts or intermediate hosts, posing a continuous risk for domestic animals and humans. The presence of tick-borne pathogens is often ignored in wild animals kept in zoos, which is a public health concern. In the present study, we investigated these pathogens in tick-infested captive wild animals at the Lohi Bher zoo, Pakistan. Blood samples were collected from 22 animals, which include urials (4) (Ovis aries vignei), blackbucks (3) (Antilope cervicapra), fallow deer (1) (Dama dama), hog deer (6) (Axis porcinus), chinkaras (4) (Gazella bennettii), white tiger (2) (Panthera tigris tigris), a giraffe (Giraffa camelopardalis), and African lions (2) (Panthera leo). The samples were screened for Piroplasm and Anaplasma spp. by polymerase chain reaction targeting different gene loci. We detected three Theileria spp. and one Anaplasma sp. from the investigated captive wild animals. The Theileria sp. dama gazelle was detected from chinkara, Theileria sp. NG-2012b from chinkara and giraffe and T. parva from African lion, and Anaplasma bovis was identified in a giraffe. Moreover, Theileria sp. and Anaplasma sp. coinfection was detected in one giraffe. Overall, this study shows that Theileria spp. and Anaplasma spp. are circulating in captive wild animals, which can play an important role in their spread. Further studies are required to monitor tick-borne pathogens in zoo animals and their potential to spread from exotic wild captive animals to local wild and domestic.


Assuntos
Antílopes , Cervos , Girafas , Theileria , Doenças Transmitidas por Carrapatos , Carrapatos , Anaplasma/genética , Animais , Animais Selvagens , Humanos , Paquistão/epidemiologia , Filogenia , Ovinos , Theileria/genética , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/veterinária
15.
Vet Pathol ; 59(6): 1031-1046, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36052867

RESUMO

Cases of Theileria-associated mortality are rarely reported in African wild artiodactyls. Descriptions of lesions are limited, particularly in endangered hippotraginids. Here, we analyzed retrospectively the gross and histologic findings in 55 roan antelope (Hippotragus equinus) with fatal natural theileriosis. The most frequently recorded gross findings in 40 cases were widespread petechiae and ecchymoses (72.5%), probable anemia (67.5%), icterus (60%), splenomegaly (60%), hepatomegaly (52.5%), and pulmonary edema (50%). Histologic lesions in 34 cases were characterized by multi-organ infiltrates of parasitized and nonparasitized mononuclear leukocytes (MLs), and fewer multinucleate giant cells (MNGCs). Liver, lung, kidney, adrenal gland, and heart were most consistently infiltrated, followed by spleen and lymph nodes. Leukocytes were phenotyped in lung, liver, kidney, and heart specimens from 16 cases, using immunohistochemistry to detect CD20, CD3, myeloid/histiocyte antigen (MAC387), IBA-1, and CD204 surface receptors. A roan polyclonal anti-Theileria sp. (sable) antibody was applied to the same tissues to identify intraleukocytic parasite antigens. Similar proportions of intravascular and extravascular IBA-1-, CD204-, and MAC387-reactive putative monocyte-macrophages and fewer CD3-positive putative T-lymphocytes were identified in all organs, especially the lungs in infected roan. CD20-positive putative B-lymphocytes were significantly scarcer than in uninfected controls. Intraleukocytic Theileria parasites labeled consistently in affected tissues. Some parasitized and nonparasitized MLs and the MNGCs failed to label with selected leukocyte markers. Fatal theileriosis in roans may largely be the result of multi-organ monocyte-macrophage activation with associated tissue injury and overwhelming systemic inflammation. The identity of the parasitized leukocytes and characteristics of the lymphohistiocytic response require further clarification in roans.


Assuntos
Antílopes , Artiodáctilos , Doenças dos Bovinos , Theileria , Theileriose , Animais , Bovinos , Estudos Retrospectivos , Theileriose/parasitologia
16.
Acta Parasitol ; 67(4): 1697-1707, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36178614

RESUMO

PURPOSE: The parasites of genera such as Babesia and Theileria are called piroplasmids due to the pear-shaped morphology of the multiplying parasite stages in the blood of the vertebrate host. Because of the enormous number of parasite species and the challenges of multiplex PCR, initial screening of samples using piroplasmid-specific PCR may be a more cost-effective and efficient technique to identify parasite species, especially during epidemiological studies. Accordingly, 18S rRNA PCR was standardized and optimized on common piroplasmids of different animals like cattle, buffaloes, sheep, goats, dogs, horses, and leopards. METHODS: Bloods samples from 1250 animals were collected from different animals in Junagadh district of Gujarat, India. 18S rRNA PCR was standardized and optimized as a primary method for molecular screening of piroplasms in domestic and wild animals. The method was checked for its analytical sensitivity and specificity. Parasite species-specific PCR and sequencing was used to validate the test. Moreover, in-silico restriction enzyme (RE) analysis was also done to assess its applicability in PCR-RFLP. RESULTS: Piroplasm infections were recorded in 63.3% of animals in Junagadh. The 18S rRNA PCR detected the piroplasmid DNA in as low as 39 picograms (pg) of whole blood genomic DNA isolated from microscopically Theileria positive blood samples and no reactivity was recorded from common but unrelated haemoparasites viz., Trypanosoma evansi, Hepatozoon spp., Anaplasma spp., and Ehrlichia canis was observed. The 18S rRNA PCR assay findings were confirmed by species-specific PCR and sequencing. Analysis of different sequences generated using 18S rRNA PCR revealed that the amplicon size of Babesia spp. is nearly 400 bp (393-408 bp) whereas Theileria spp. were more than 400 bp (418-424 bp). The percentage of sequence divergence among Babesia and Theileria spp. was 7.3-12.2% and 0.7-12.2%, respectively. In-silico restriction enzyme (RE) analysis reveals the presence of at least one site for a commercially available RE in 18S rRNA fragments of every parasite, which can differentiate it from its congeners. CONCLUSIONS: The presented universal oligonucleotide-based PCR assay provides a highly sensitive, specific, cost-effective, and rapid diagnostic tool for the initial screening of piroplasmids infecting domestic and wild animals and is potentially helpful for large-scale epidemiological studies.


Assuntos
Babesia , Babesiose , Theileria , Theileriose , Ovinos , Cavalos , Cães , Bovinos , Animais , RNA Ribossômico 18S/genética , Genes de RNAr , Filogenia , Reação em Cadeia da Polimerase/veterinária , Cabras , Búfalos , Babesiose/diagnóstico , Babesiose/epidemiologia , Theileriose/diagnóstico , Theileriose/epidemiologia
17.
Parasit Vectors ; 15(1): 329, 2022 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-36123705

RESUMO

BACKGROUND: An innovative approach has been introduced for identifying and developing novel potent and safe anti-Babesia and anti-Theileria agents for the control of animal piroplasmosis. In the present study, we evaluated the inhibitory effects of Malaria Box (MBox) compounds (n = 8) against the growth of Babesia microti in mice and conducted bioinformatics analysis between the selected hits and the currently used antibabesial drugs, with far-reaching implications for potent combinations. METHODS: A fluorescence assay was used to evaluate the in vivo inhibitory effects of the selected compounds. Bioinformatics analysis was conducted using hierarchical clustering, distance matrix and molecular weight correlation, and PubChem fingerprint. The compounds with in vivo potential efficacy were selected to search for their target in the piroplasm parasites using quantitative PCR (qPCR). RESULTS: Screening the MBox against the in vivo growth of the B. microti parasite enabled the discovery of potent new antipiroplasm drugs, including MMV396693 and MMV665875. Interestingly, statistically significant (P < 0.05) downregulation of cysteine protease mRNA levels was observed in MMV665875-treated Theileria equi in vitro culture in comparison with untreated cultures. MMV396693/clofazimine and MMV665875/atovaquone (AV) showed maximum structural similarity (MSS) with each other. The distance matrix results indicate promising antibabesial efficacy of combination therapies consisting of either MMV665875 and AV or MMV396693 and imidocarb dipropionate (ID). CONCLUSIONS: Inhibitory and hematology assay results suggest that MMV396693 and MMV665875 are potent antipiroplasm monotherapies. The structural similarity results indicate that MMV665875 and MMV396693 have a similar mode of action as AV and ID, respectively. Our findings demonstrated that MBox compounds provide a promising lead for the development of new antibabesial therapeutic alternatives.


Assuntos
Babesia microti , Babesiose , Cisteína Proteases , Malária , Theileria , Animais , Atovaquona/farmacologia , Atovaquona/uso terapêutico , Babesiose/tratamento farmacológico , Babesiose/parasitologia , Clofazimina/farmacologia , Clofazimina/uso terapêutico , Cisteína Proteases/farmacologia , Reposicionamento de Medicamentos , Imidocarbo/análogos & derivados , Camundongos , Theileria/fisiologia
18.
Acta Trop ; 235: 106656, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35988819

RESUMO

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


Assuntos
Babesia , Ixodidae , Rhipicephalus , Rickettsia , Theileria , Doenças Transmitidas por Carrapatos , Animais , Babesia/genética , Bovinos , Feminino , Ixodidae/microbiologia , Larva , Theileria/genética , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/microbiologia , Doenças Transmitidas por Carrapatos/veterinária
19.
Parasit Vectors ; 15(1): 308, 2022 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-36042502

RESUMO

BACKGROUND: Theileria annulata, a transforming parasite, invades bovine B cells, dendritic cells and macrophages, promoting the uncontrolled proliferation of these cells. This protozoan evolved intricate strategies to subvert host cell signaling pathways related to antiapoptotic signaling to enable survival and proliferation within the host cells. However, the molecular mechanisms of the cell transformation induced by T. annulata remain largely unclear. Although some studies have predicted that the subtelomere-encoded variable secreted protein (SVSP) family plays roles in host-parasite interactions, the evidence for this is limited. METHODS: In the present study, the SVSP455 (TA05545) gene, a member of the SVSP gene family, was used as the target molecule. The expression pattern of SVSP455 in different life-cycle stages of T. annulata infection was explored using a quantitative real-time PCR assay, and the subcellular distribution of SVSP455 was observed using confocal microscopy. The host cell proteins interacting with SVSP455 were screened using the Y2H system, and their interactions were verified in vivo and in vitro using both bimolecular fluorescence complementation and confocal microscopy, and co-immunoprecipitation assays. The role played by SVSP455 in cell transformation was further explored by using overexpression, RNA interference and drug treatment experiments. RESULTS: The highest level of the SVSP455 transcript was detected in the schizont stage of T. annulata, and the protein was located both on the surface of schizonts and in the host cell cytoplasm. In addition, the interaction between SVSP455 and heat shock protein 60 was shown in vitro, and their link may regulate host cell apoptosis in T. annulata-infected cells. CONCLUSION: Our findings are the first to reveal that T. annulata-secreted SVSP455 molecule directly interacts with both exogenous and endogenous bovine HSP60 protein, and that the interaction of SVSP455-HSP60 may manipulate the host cell apoptosis signaling pathway. These results provide insights into cancer-like phenotypes underlying Theilera transformation and therapeutics for protection against other pathogens.


Assuntos
Theileria annulata , Theileria , Theileriose , Animais , Bovinos , Chaperonina 60 , Interações Hospedeiro-Parasita , Imunoprecipitação , Esquizontes , Theileria annulata/genética , Theileria annulata/metabolismo , Theileriose/prevenção & controle
20.
Parasite ; 29: 38, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35861542

RESUMO

Ticks and tick-borne diseases pose a significant threat to public health. In this study, we aimed to determine the tick species distribution and pathogens carried by ticks in Yanbian, China. A total of 2673 questing ticks were collected from eight counties and cities in Yanbian and were morphologically identified. The presence of Candidatus Rickettsia tarasevichiae (CRT), spotted fever group Rickettsia (SFGR), severe fever thrombocytopenia syndrome virus (SFTSV), Theileria, and other pathogens was confirmed using polymerase chain reaction (PCR) and real-time quantitative PCR assays, followed by phylogenetic and genotypic analyses. According to the morphological identification, the tick species in Yanbian consisted of Haemaphysalis longicornis, Ixodes persulcatus, Dermacentor silvarum, H. japonica, and H. concinna. In H. longicornis, CRT, SFGR, SFTSV and Theileria orientalis were detected, while CRT, SFGR, and SFTSV were detected in I. persulcatus, H. japonica, and D. silvarum. Only SFTSV was detected in H. concinna. Mixed infection with CRT and SFTSV was observed in I. persulcatus and H. japonica. The gene sequences of all tested pathogens exhibited 95.7%-100% identity with the corresponding sequences deposited in GenBank. Phylogenetic analysis showed that different SFGR and SFTSV genotypes were closely related to the Korean strains. This study is the first to describe the genetic diversity of SFGR Candidatus Rickettsia longicornii in H. longicornis in Yanbian, China, using the ompA, ompB, sca4, and rrs genes. These results provide epidemiological data to support the prevention and control of ticks and tick-borne diseases in the border areas of China, North Korea, and Russia.


Title: Enquête sur les espèces de tiques et détection moléculaire de certains agents pathogènes transmis par les tiques à Yanbian, en Chine. Abstract: Les tiques et les maladies transmises par les tiques constituent une menace importante pour la santé publique. Dans cette étude, nous avons cherché à déterminer la distribution des espèces et les agents pathogènes portés par les tiques à Yanbian, en Chine. Un total de 2 673 tiques errantes ont été collectées dans huit comtés et villes de Yanbian et identifiées morphologiquement. La présence de Candidatus Rickettsia tarasevichiae (CRT), de Rickettsia du groupe de la fièvre boutonneuse (SFGR), du virus du syndrome de la fièvre thrombocytopénique sévère (SFTSV), de Theileria et d'autres agents pathogènes a été confirmée à l'aide d'une réaction en chaîne par polymérase (PCR) et de PCR quantitative en temps réel, suivies par des analyses phylogénétiques et génotypiques. Selon leur identification morphologique, les espèces de tiques à Yanbian se composaient de Haemaphysalis longicornis, Ixodes persulcatus, Dermacentor silvarum, H. japonica et H. concinna. Chez H. longicornis, CRT, SFGR, SFTSV et Theileria orientalis ont été détectés, tandis que CRT, SFGR et SFTSV ont été détectés chez I. persulcatus, H. japonica et D. silvarum. Seul le SFTSV a été détecté chez H. concinna. Une infection mixte par CRT et SFTSV a été observée chez I. persulcatus et H. japonica. Les séquences des gènes de tous les agents pathogènes testés présentaient une identité de 95,7 % à 100 % avec les séquences correspondantes déposées dans GenBank. L'analyse phylogénétique a montré que différents génotypes SFGR et SFTSV étaient étroitement liés aux souches coréennes. Cette étude est la première à décrire la diversité génétique de SFGR Candidatus Rickettsia longicornii chez H. longicornis à Yanbian, en Chine, en utilisant les gènes ompA, ompB, sca4 et rrs. Ces résultats fournissent des données épidémiologiques pour soutenir la prévention et le contrôle des tiques et des maladies transmises par les tiques dans les zones frontalières de la Chine, de la Corée du Nord et de la Russie.


Assuntos
Ixodes , Ixodidae , Rickettsia , Theileria , Doenças Transmitidas por Carrapatos , Animais , China/epidemiologia , Filogenia , Rickettsia/genética , Theileria/genética , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/microbiologia
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