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Trypanosoma evansi, the causative agent of surra, is the most prevalent pathogenic salivarian trypanosome and affects the majority of domesticated and wild animals in endemic regions. This work aimed to analyze detergent-solubilized T. evansi proteins and identify potential diagnostic biomarkers for surra. Triton X-114-extracted membrane-enriched proteins (MEP) of T. evansi bloodstream forms were analyzed using a gel-free technique (LC-ESI-MS/MS). 247 proteins were identified following the MS analysis of three biological and technical replicates. Two of these proteins were predicted to have a GPI-anchor, 100 (40%) were predicted to have transmembrane domains, and 166 (67%) were predicted to be membrane-bound based on at least one of six features: location (WolfPSORT, DeepLoc-2.0, Protcomp-9.0), transmembrane, GPI, and gene ontology. It was predicted that 76 (30%) of proteins had membrane evidence. Typical membrane proteins for each organelle were identified, among them ISG families (64, 65, and 75 kDa), flagellar calcium-binding protein, 24 kDa calflagin, syntaxins and oligosaccharyltransferase some of which had previously been studied in other trypanosomatids. T. evansi lacks singletons and exclusive orthologous groups, whereas three distinct epitopes have been identified. Data are available via ProteomeXchange with identifier PXD040594. SIGNIFICANCE: Trypanosoma evansi is a highly prevalent parasite that induces a pathological condition known as "surra" in various species of ungulates across five continents. The infection gives rise to symptoms that are not pathognomonic, thereby posing challenges in its diagnosis and leading to substantial economic losses in the livestock industry. A significant challenge arises from the absence of a diagnostic test capable of distinguishing between Trypanosoma equiperdum and T. evansi, both of which are implicated in equine diseases. Therefore, there is a pressing need to conduct research on the biochemistry of the parasite in order to identify proteins that could potentially serve as targets for differential diagnosis or therapeutic interventions.
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Proteômica , Proteínas de Protozoários , Trypanosoma , Tripanossomíase , Trypanosoma/metabolismo , Proteínas de Protozoários/química , Proteínas de Protozoários/metabolismo , Proteínas de Protozoários/análise , Proteômica/métodos , Animais , Tripanossomíase/diagnóstico , Tripanossomíase/parasitologia , Detergentes/química , Proteínas de Membrana/química , CavalosRESUMO
Neosporosis and toxoplasmosis are important parasitic causes of abortions in small ruminants. This study verified the occurrence of these diseases in sheep fetuses from Santa Catarina State, Southern Brazil from 2015 to 2022. Sheep fetuses were necropsied with organ sampling for histopathology, and polymerase chain reaction (PCR) for Neospora caninum and Toxoplasma gondii using the Nc5 and SAG2 targets, respectively, in frozen brain tissue. Microbiological culture and RT-PCR for Pestivirus were conducted to discard other abortion causes. One positive fetus for toxoplasmosis was genotyped using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) with ten genetic markers. Fifty-five sheep fetuses were evaluated, with 10 (18.2%) cases of neosporosis and 7 (12.7%) cases of toxoplasmosis, comprising six and four flocks, respectively. Macroscopically, neosporosis abortions exhibited fetal mummification, maceration, and arthrogryposis. Toxoplasmosis abortions showed fetal mummification and maceration. The neosporosis abortions included lymphoplasmacytic myositis (70%; 7/10) and myocarditis (60%; 6/10), in addition to necrotizing encephalitis and gliosis (50%; 5/10). Toxoplasmosis abortions included lymphoplasmacytic necrotizing encephalitis (71.4%; 5/7), lymphoplasmacytic myositis (42.8%, 3/7), and myocarditis (14.3%; 1/7). Through PCR, N. caninum and T. gondii were detected in 6 (60%) and 5 (71.4%) fetuses, respectively. In one fetus, T. gondii genotyping was conducted, which was characterized as atypical genotype ToxoDB #98. All of the cases were negative for Pestivirus and bacterial agents. This study establishes the occurrence of these diseases as causes of abortions, malformations, mummification, and fetal maceration in sheep, with the characterization of an atypical T. gondii genotype in one of the fetuses.
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Aborto Animal , Coccidiose , Neospora , Doenças dos Ovinos , Toxoplasma , Toxoplasmose Animal , Animais , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/parasitologia , Neospora/genética , Neospora/isolamento & purificação , Brasil/epidemiologia , Coccidiose/veterinária , Coccidiose/parasitologia , Coccidiose/epidemiologia , Aborto Animal/parasitologia , Aborto Animal/epidemiologia , Toxoplasma/genética , Toxoplasma/isolamento & purificação , Doenças dos Ovinos/parasitologia , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/virologia , Ovinos , Feminino , GravidezRESUMO
Bovine trypanosomosis, caused by Trypanosoma vivax, is a disease that originated in Africa and currently affects cattle in several South American countries, including almost all Brazilian states. Despite the reports on T. vivax infection in southern Brazil, data on its circulation status is currently unavailable. In this study, we aimed to detect anti-Trypanosoma spp. IgG antibodies in cattle from Rio Grande do Sul and suggest areas with T. vivax transmission risk. A total of 691 serum samples from cattle in the intermediate regions of Rio Grande do Sul were analyzed using indirect immunofluorescence assay (IFA). The overall seroprevalence of anti-Trypanosoma antibodies in cattle was 24.6% (170/691). The detection rate ranged from 0-37.3%, with a high prevalence in the intermediate regions of Ijuí (37.3%), Uruguaiana (30.7%), and Passo Fundo (28.9%). Thus, these regions were suggested as possible bovine trypanosomosis risk areas due to the high seroprevalence. This is the first serological study to determine Trypanosoma spp. infection status in cattle from Rio Grande do Sul, providing data on the epidemiology of trypanosomosis in the state.
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Doenças dos Bovinos , Trypanosoma , Tripanossomíase Bovina , Tripanossomíase , Bovinos , Animais , Brasil/epidemiologia , Estudos Soroepidemiológicos , Tripanossomíase/epidemiologia , Tripanossomíase/veterinária , Tripanossomíase Bovina/diagnóstico , Tripanossomíase Bovina/epidemiologia , Tripanossomíase Bovina/parasitologia , Trypanosoma vivax , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologiaRESUMO
Introduction: The BoLA-DRB3 gene in cattle is associated with tolerance to several infectious diseases, such as neosporosis, dermatophilosis, leukosis, and mastitis. Methods: This study used PCR-SBT and BoLA-DRB3 gene sequencing to determine the association between the presence or absence of Anaplasma marginale, Babesia bovis, and Babesia bigemina infections in 208 Crioulo Lageano cattle and alleles present in the population. The chi-square test and odds ratio analysis were employed to establish the association. Results: Of the BoLA-DRB3 gene alleles present in the population, two alleles were significantly associated with resistance to A. marginale infections: BoLA-DRB3001:01 (p < 0.001; OR = 0.224), which had a frequency of 7.93%, and BoLA-DRB3024:06 (p = 0.007; OR < 0.00001), which had a frequency of 0.72%. Regarding B. bovis infection, the BoLA-DRB3*011:01 allele (p = 0.002; OR = 0.271) had a frequency of 6% in the population and was associated with resistance to the infection. None of the alleles was associated with resistance to infection by B. bigemina. Discussion: The Crioulo Lageano breed has alleles that may confer resistance against infection by A. marginale and B. bovis.
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Enzyme-linked immunosorbent assay (ELISA) for viral antigen is commonly used for the diagnosis of progressive feline leukemia virus (FeLV) infection but is not able to determine the true prevalence of infection when used as the sole test. Additional testing to detect proviral DNA will identify regressive (antigen negative) FeLV infections as well as progressive infections. Therefore, this study aimed to determine the prevalence of progressive and regressive FeLV infection, outcome-associated factors, and hematologic changes. A cross-sectional study was performed on 384 cats selected from routine hospital care. Blood samples were subjected to complete blood count, ELISA for FeLV antigen and FIV antibody, and nested PCR amplifying the U3- LTR region and gag gene, which are conserved in most exogenous FeLV. The prevalence of FeLV infection was 45.6% (CI95% 40.6-50.6%). The prevalence of progressive infection (FeLV+P) was 34.4% (CI95% 29.6-39.1%), that of regressive infection (FeLV+R) was 10.4% (CI95% 7.4-13.4%), for discordant but positive results 0.8% (CI95% 0.75-0.84%), for FeLV+P coinfected with FIV 2.6% (CI95% 1.2-4.0%), and FeLV+R coinfected with FIV 1.5% (CI95% 0.3-2.7%). Male cats were three times more likely to be in the FeLV+P group. Cats coinfected with FIV were 4.8 times more likely to belong to the FeLV+R group. In the FeLV+P group, the main clinical changes were lymphoma (38.5%), anemia (24.4%), leukemia (17.9%), concomitant infections (15.4%), and feline chronic gingivostomatitis - FCGS (3.8%). In the FeLV+R group, the main clinical signs were anemia (45.4%), leukemia (18.2%), concomitant infections (18.2%), lymphoma (9.1%), and FCGS (9.1%). Cats in the FeLV+P and FeLV+R groups showed mainly thrombocytopenia (56.6% and 38.2%), non-regenerative anemia (32.8% and 23.5%), and lymphopenia (33.6% and 20.6%). Hemoglobin concentration, packed cell volume (PCV), platelet count, lymphocytes, and eosinophils in the FeLV+P and FeLV+R groups had lower medians than the control group (FeLV/FIV-uninfected, healthy). Erythrocyte and eosinophil counts were statistically different among the three groups, with the medians of the FeLV+P and FeLV+R groups being lower than those of the control group. In addition, the median PCV and band neutrophil counts were higher in FeLV+P than in FeLV+R. Our results show a high prevalence of FeLV, different factors associated with the course of infection, and more frequent and severe hematologic changes in progressive infections compared with regressive infections.
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Doenças do Gato , Síndrome de Imunodeficiência Adquirida Felina , Vírus da Imunodeficiência Felina , Leucemia Felina , Leucemia , Linfoma , Gatos , Animais , Masculino , Vírus da Leucemia Felina , Prevalência , Brasil/epidemiologia , Estudos Transversais , Leucemia Felina/diagnóstico , Leucemia/veterinária , Linfoma/veterinária , Fatores de Risco , Síndrome de Imunodeficiência Adquirida Felina/epidemiologia , Doenças do Gato/epidemiologiaRESUMO
Background and Aim: Flemish cattle in Brazil are on the brink of extinction and are found only in one herd in Lages, Santa Catarina State. This study aimed to uncover the reasons for the recurring abortions in the Flemish cattle herd. Materials and Methods: Seventeen Flemish fetuses underwent postmortem examinations, with samples collected for histopathology and microbiology culture tests, polymerase chain reaction (PCR) test for Neospora caninum, and reverse transcription-PCR (RT-PCR) test for bovine viral diarrhea virus (BVDV) from 2015 to 2020. Results: Of the 17 fetuses, N. caninum was the most common diagnosis and was found in 88% (15/17). One fetus (5.8%) had a coinfection with N. caninum and Citrobacter amalonaticus, leading to fibrinonecrotic pericarditis. All fetuses tested negative for BVDV by RT-PCR. Of the 107 dams tested by indirect immunofluorescence assay, 26 (25.2%) were anti-N. caninum seropositive, with 17 (65.4%) aborting and 5 (19.2%) having estrus repetition. Reverse transcription-PCR results showed that 9 (8.4%) of the serum samples collected from dams tested positive, which tested follow-up test 3 months later, indicating a BVDV transient infection. The factors that contributed to neosporosis included dogs' access to pastures and improper disposal of fetal remains, which made it easier for dogs to consume them. Conclusion: This study warns the occurrence of N. caninum as a cause of reproductive disorders that can lead to abortion in the studied Flemish cattle herd.
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Here is our proposal to improve learning in biomedical sciences for graduate and undergraduate courses with a broad vision integrating disciplines such as molecular cell biology, biochemistry, and biophysics around concepts of pathogen interaction within vertebrate and invertebrate hosts. Our paradigm is based on the possibility offered by the pandemic to have remote activities that give access to students and researchers from different places in Brazil and Latin American countries to discuss science. A multidisciplinary view of host-pathogen interaction allows us to understand better the mechanisms involved in the pathology of diseases, as well as to formulate broad strategies for the diagnosis, treatment, and control of thereof. The approach to integrating heterogeneous groups in science involves the critical analysis of national scientific resource distribution, where only some have the possibilities to conduct competitive scientific research. Solid theoretical training, contact, collaboration with groups of excellence, and training within a multidisciplinary network are our proposals for a permanent platform of scientific strengthening and dissemination for Latin America. Here we will review the concept of host-pathogen interaction, the type of institutions where it is taught and researched, new trends in active teaching methodologies, and the current political context in science.
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Interações Hospedeiro-Patógeno , Pandemias , Humanos , BrasilRESUMO
Cattle trypanosomiasis negatively impacts animal husbandry due to high morbidity, productivity losses, and mortality rates. Knowledge regarding Trypanosoma evansi infections in locally adapted breeds remains limited. Some cattle breeds exhibit trypanotolerance, requiring the determination of prevalence, as well as related tolerance and resistance characteristics, for disease control programs. This study aimed to determine T. evansi prevalence in Crioula Lageana cattle and associate clinical, hematological, and biochemical aspects with the infection to further research on tolerance in this population. Blood samples from 310 Crioula Lageana cattle were tested using Polymerase Chain Reaction (PCR) and Indirect Immunofluorescence Reaction (IIFR). T. evansi prevalence was 8% (24/310) using PCR and 4% (11/310) using IIFR. Positive animals showed increased ruminal movements, elevated eosinophil counts, and reduced monocyte numbers, but both latter within the reference range for the species. Albumin concentrations were low in positive cases and remained below the reference range limit for both groups. However, triglycerides exceeded the physiological range for the species in both positive and negative groups. Increased gamma-glutamyltransferase (GGT) activity was observed in positive animals. In conclusion, Crioula Lageana cattle exhibited enzootic instability with a low T. evansi infection prevalence when assessed using PCR and IIFR techniques. Furthermore, the animals did not display clinical, hematological, or biochemical alterations attributable to the presence of hemoparasites.
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Doenças dos Bovinos , Trypanosoma , Tripanossomíase , Animais , Bovinos , Tripanossomíase/epidemiologia , Tripanossomíase/veterinária , Tripanossomíase/parasitologia , Prevalência , Trypanosoma/genética , Reação em Cadeia da Polimerase/métodos , DNA de Protozoário/genética , Doenças dos Bovinos/epidemiologiaRESUMO
To date, only a few studies have examined the impacts of feline leukemia virus (FeLV) subgroups on disease development in spontaneously infected cats. The present study identified FeLV-A and FeLV-B subgroups in cats with lymphoma and leukemia and explored the phylogenetic relationships of env sequences. Twenty-six cats with lymphoma (n=16) or leukemia (n=10) were selected. FeLV p27 antigen positivity was determined using ELISA, and proviral DNA in blood samples was detected using nested PCR. Positive animals in both tests were classified as cases of FeLV progressive infection and subjected to a second nested PCR for env amplification and subgroup determination. Six samples of FeLV-A and five samples of FeLV-B were sequenced using the Sanger method, and the results were used to build a phylogenetic tree and estimate evolutionary divergence. Among cats with lymphoma, 68.8% carried FeLV-AB and 31.2% FeLV-A. Among cats with leukemia, 70% carried FeLV-AB and 30% FeLV-A. Regarding cat characteristics, 50% were young, 30.8% young adults, and 19.2% adults; 88.5% were mixed-breed and 11.5% pure breed; and 42.3% were males and 57.7% were females. Among lymphomas, 62.5% were mediastinal, 31.3% multicentric, and 6.3% extranodal. Regarding histological classification, lymphoblastic and small non-cleaved-cell lymphomas were the most frequently detected. Among leukemia cases, 30% were acute lymphoid, 30% chronic myeloid, and 40% acute myeloid. Phylogenetic analysis showed that FeLV-A SC sequences were closely related to the Arena, Glasgow-1, and FeLV-FAIDS variants. Meanwhile, FeLV-B SC sequences were divergent from one another but similar to the endogenous FELV env gene (enFeLV). In conclusion, FeLV-AB is prevalent in cats with lymphoma and leukemia, highlighting the genetic diversity involved in the pathogenesis of these neoplasms in Brazil.