Artigos - Patologia Veterinária - FCAV

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  • ItemArtigo
    Molecular detection of blood-borne agents in vampire bats from Brazil, with the first molecular evidence of Neorickettsia sp. in Desmodus rotundus and Diphylla ecaudata
    (2023-08-01) de Mello, Victória Valente Califre [UNESP]; Placa, Ana Julia Vidal [UNESP]; Lee, Daniel Antonio Braga [UNESP]; Franco, Eliz Oliveira [UNESP]; Lima, Luciana; Teixeira, Marta M.G.; Hemsley, Claudia; Titball, Richard W.; Machado, Rosangela Zacarias [UNESP]; André, Marcos Rogério [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); University of Exeter
    Bats (Mammalia, Chiroptera) represent the second largest group of mammals. Due to their ability to fly and adapt and colonize different niches, bats act as reservoirs of several potentially zoonotic pathogens. In this context, the present work aimed to investigate, using molecular techniques, the occurrence of blood-borne agents (Anaplasmataceae, Coxiella burnetii, hemoplasmas, hemosporidians and piroplasmids) in 198 vampire bats sampled in different regions of Brazil and belonging to the species Desmodus rotundus (n = 159), Diphylla ecaudata (n = 31) and Diaemus youngii (n = 8). All vampire bats liver samples were negative in PCR assays for Ehrlichia spp., Anaplasma spp., piroplasmids, hemosporidians and Coxiella burnetii. However, Neorickettsia sp. was detected in liver samples of 1.51% (3/198) through nested PCR based on the 16S rRNA gene in D. rotundus and D. ecaudata. This is the first study to report Neorickettsia sp. in vampire bats. Hemoplasmas were detected in 6.06% (12/198) of the liver samples using a PCR based on the 16S rRNA gene. The two 16S rRNA sequences obtained from hemoplasmas were closely related to sequences previously identified in vampire and non-hematophagous bats from Belize, Peru and Brazil. The genotypic analysis identified a high diversity of bat-associated hemoplasma genotypes from different regions of the world, emphasizing the need for studies on this subject, in order to better understand the mechanisms of co-evolution between this group of bacteria and their vertebrate hosts. The role of neotropical bat-associated Neorickettsia sp. and bats from Brazil in the biological cycle of such agent warrant further investigation.
  • ItemArtigo
    Molecular Survey of Hemotropic Mycoplasma spp. and Bartonella spp. in Coatis (Nasua nasua) from Central-Western Brazil
    (2023-04-01) Perles, Lívia [UNESP]; Barreto, Wanessa Teixeira Gomes; Santos, Filipe Martins; Duarte, Leidiane Lima; de Macedo, Gabriel Carvalho; Barros-Battesti, Darci Moraes [UNESP]; Herrera, Heitor Miraglia; Machado, Rosangela Zacarias [UNESP]; André, Marcos Rogério [UNESP]; Universidade Estadual Paulista (UNESP); Mato Grosso do Sul Federal University; Dom Bosco Catholic University; Universidade de São Paulo (USP)
    Even though previous works showed molecular evidence of hemotropic Mycoplasma spp. (hemoplasmas) in ring-tailed coatis (Nasua nasua) from Brazil, Bartonella sp. has not been reported in these mammals so far. The present study aimed to detect the above-mentioned agents in coatis’ blood and associated ectoparasites, assessing the association between these infections and red blood parameters. Between March 2018 and January 2019, coati (n = 97) blood samples, Amblyomma sp. ticks (2242 individual ticks, resulting in 265 pools), and Neotrichodectes pallidus louse (n = 59) were collected in forested urban areas from midwestern Brazil. DNA extracted from coatis’ blood, and ectoparasite samples were submitted to quantitative PCR (qPCR) (16S rRNA) and conventional PCR (cPCR) (16S rRNA and 23S rRNA) for hemoplasmas and qPCR (nuoG gene) and culturing (only blood) for Bartonella spp. Two different hemoplasma genotypes were detected in blood samples: 71% coatis positive for myc1 and 17% positive for myc2. While 10% of ticks were positive for hemoplasmas (myc1), no louse was positive. The estimated bacterial load of hemoplasmas showed no association with anemia indicators. All coatis were negative for Bartonella sp. in qPCR assay and culturing, albeit two Amblyomma sp. larvae pools, and 2 A. dubitatum nymph pools were positive in the qPCR. The present work showed a high occurrence of hemoplasmas, with two distinct hemoplasma genotypes, in coatis from forested urban areas in midwestern Brazil.
  • ItemArtigo
    Bartonella spp. in households with cats: Risk factors for infection in cats and human exposure
    (2023-06-01) Sepúlveda-García, Paulina; Alabi, Amir [UNESP]; Álvarez, Karla; Rojas, Lisbeth; Mella, Armin; Gonçalves, Luiz Ricardo [UNESP]; André, Marcos Rogerio [UNESP]; Machado, Rosangela Zacarias [UNESP]; Müller, Ananda; Monti, Gustavo; Universidad Austral de Chile; Universidade Estadual Paulista (UNESP); Ross University School of Veterinary Medicine; Wageningen University & Research
    The aim of this study was to estimate the occurrence of Bartonella spp. per household in cats and the risk factors for Bartonella spp. positivity in cats and their owners from Valdivia, Chile. A total of 464 cats (distributed within 324 households) and 326 humans (control group [n = 112] and cat owner [n = 214]) distributed in 262 households were sampled. From the cat owners (n = 214), 128 humans were in households where the cat was also sampled, totaling 84 households with dual sampling. Real-time PCR (qPCR) was used for Bartonella spp. detection in blood from cats and humans, and immunofluorescent immunoassay (IFA) anti-Bartonella henselae was performed in human serum samples. Out of the total of 324 households, 20.43% presented at least one Bartonella positive cat. From the households with dual sampling, 29.7% (25/84) presented at least one qPCR-Bartonella spp. positive cat. However, Bartonella DNA was not amplified in humans, and in 7.3% (6/82) of the households was found at least one of the cat's owners exposed to B. henselae. Cats younger than one year (Odds Ratio (OR) = 5.3), non-neutered (OR 3.46), sampled at home (OR 5.82), and with improper application of tick/flea control products (OR 3.13) showed a higher risk for Bartonella spp. presence. Humans with occupational exposure involving animal contact, were more likely to exhibit B. henselae seropositivity (OR 7.5). Bartonella spp. was present in the cats a moderate number of households, but Bartonella DNA was not detected in owners' blood, inferring that there is a low risk of recent human infection in the studied population.
  • ItemArtigo
    Contribution to the knowledge of Neotrichodectes (Nasuicola) pallidus (Piaget, 1880) (Phthiraptera: Trichodectidae)
    (2023-06-01) Perles, Lívia [UNESP]; Bassini-Silva, Ricardo [UNESP]; Jacinavicius, Fernando Castro; Barreto, Wanessa Teixeira Gomes; de Macedo, Gabriel Carvalho; Martins, Filipe Santos; Herrera, Heitor Miraglia; Machado, Rosangela Zacarias [UNESP]; Barros-Battesti, Darci Moraes [UNESP]; André, Marcos Rogério [UNESP]; Universidade Estadual Paulista (UNESP); Instituto Butantan; Mato Grosso do Sul Federal University; Dom Bosco Catholic University
    The species in the genus Neotrichodectes (Phthiraptera: Ischnocera) infest carnivores. Neotrichodectes (Nasuicola) pallidus (Piaget, 1880), which has been primarily found parasitizing Procyonidae mammals, has been recorded in ring-tailed coatis (Nasua nasua) in the Brazilian states of Minas Gerais, Pernambuco, Santa Catarina, Rio Grande do Sul and Pernambuco. We report a new record of N. pallidus in coatis in the state of Mato Grosso do Sul, central-western Brazil, using morphological (Light and Scanning Electronic Microscopy) and molecular approaches (PCR, sequencing and phylogenetic analysis). Coatis were sampled in two peri-urban areas of Campo Grande city, Mato Grosso do Sul state, Brazil, between March 2018 and March 2019, as well as in November 2021. Lice were collected and examined under light and Scanning Electron Microscopy. DNA was also extracted from nymphs and adults and submitted to PCR assays based on the 18S rRNA and cox-1 genes for molecular characterization. One hundred and one coatis were sampled from 2018 to 2019 and 20 coatis in 2021 [when the intensity of infestation (II) was not accessed]. Twenty-six coatis (26/101–25.7%) were infested with at least one louse, with a total of 59 lice collected in 2018–2019. The II ranged from one to seven lice (mean 2.2 ± SD 1.7). The louse species was confirmed based on the following morphological characteristics: female gonapophyses rounded with the setae along anterior region but not in the medial margin; the male genitalia with a parameral arch not extending beyond the endometrial plate. The same ornamentation was observed on the abdomen of the females, males, and nymphs. The nymphs and the eggs were described in detail for the first time. The obtained 18S rRNA and cox1 sequences from N. pallidus clustered in a clade with other sequences of Ischnocera species. In the present study, a new record of the louse N. pallidus in central-western Brazil was provided, along with new insights into the morphological features of this species, with the first morphology contribution of nymphal and eggs stages.
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    Bartonella spp. and Typhus Group Rickettsiae among Persons Experiencing Homelessness, São Paulo, Brazil
    (2023-02-01) Faccini-Martínez, Álvaro A.; Kmetiuk, Louise Bach; Blanton, Lucas S.; Felipetto, Laís Giuliane; Gravinatti, Mara Lúcia; Timenetsky, Jorge; Gonçalves, Luiz Ricardo [UNESP]; Machado, Rosangela Zacarias [UNESP]; André, Marcos Rogério [UNESP]; Figueiredo, Fabiano Borges; dos Santos, Andrea Pires; Labruna, Marcelo B.; Monti, Gustavo; Biondo, Alexander Welker; Walker, David H.; Hospital Militar Central; Medical Branch; Oswaldo Cruz Foundation; Federal University of Paraná; Central Paulista University Center; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); Purdue University; Wageningen University and Research
    Persons experiencing homelessness in São Paulo, Brazil, were seropositive for Bartonella spp. (79/109, 72.5%) and typhus group rickettsiae (40/109, 36.7%). Bartonella quintana DNA was detected in 17.1% (14/82) body louse pools and 0.9% (1/114) blood samples. Clinicians should consider vectorborne agents as potential causes of febrile syndromes in this population.
  • ItemArtigo
    Multi-Locus Sequencing Reveals Putative Novel Anaplasmataceae Agents, ‘Candidatus Ehrlichia dumleri’ and Anaplasma sp., in Ring-Tailed Coatis (Carnivora: Nasua nasua) from Urban Forested Fragments at Midwestern Brazil
    (2022-12-01) Perles, Lívia [UNESP]; Herrera, Heitor M.; Barreto, Wanessa T. G.; de Macedo, Gabriel C.; Calchi, Ana C. [UNESP]; Machado, Rosangela Z. [UNESP]; André, Marcos R. [UNESP]; Universidade Estadual Paulista (UNESP); Dom Bosco Catholic University; Mato Grosso do Sul Federal University
    The Anaplasmataceae family encompasses obligate intracellular α-proteobacteria of human and veterinary medicine importance. This study performed multi-locus sequencing to characterize Ehrlichia and Anaplasma in coati’s blood samples in Midwestern Brazil. Twenty-five samples (25/165—15.1%) were positive in the screening PCR based on the dsb gene of Ehrlichia spp. and were characterized using 16S rRNA, sodB, groEL, and gltA genes and the 23S-5S intergenic space region (ITS). Phylogenetic analyses based on all six molecular markers positioned the sequences into a new clade, with a common origin of Ehrlichia ruminantium. Haplotype analyses of 16S RNA sequences revealed the presence of two distinct Ehrlichia genotypes. Six samples (6/165, 3.6%) were positive in the screening nPCR for the 16S rRNA gene of Anaplasma spp. and were submitted to an additional PCR targeting the ITS for molecular characterization. Phylogenetic analyses based on both 16S rRNA gene and ITS positioned the Anaplasma sp. detected in the present study in a large clade with other Anaplasma sp. previously detected in ticks and wild animals and in a clade with ‘Candidatus Anaplasma brasiliensis’, respectively. Based on distinct molecular markers, the present work described a putative novel Anaplasmataceae agent, namely ‘Candidatus Ehrlichia dumleri’, and Anaplasma sp. closely related to the previously described ‘Candidatus Anaplasma brasiliensis’.
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    Vivaxin genes encode highly immunogenic, non-variant antigens on the Trypanosoma vivax cell-surface
    (2022-01-01) Romero-Ramirez, Alessandra; Casas-Sánchez, Aitor; Autheman, Delphine; Duffy, Craig W.; Brandt, Cordelia; Clare, Simon; Harcourt, Katherine; André, Marcos Rogério [UNESP]; Neto, Kayo José Garcia de Almeida Castilho [UNESP]; Teixeira, Marta M. G.; Machado, Rosangela Zacharias [UNESP]; Coombes, Janine; Flynn, Robin J.; Wright, Gavin J.; Jackson, Andrew P.; University of Liverpool; Liverpool School of Tropical Medicine; Wellcome Genome Campus; University of York; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); The Robert Gordon University; Waterford Institute of Technology
    Trypanosoma vivax is a unicellular hemoparasite, and a principal cause of animal African trypanosomiasis (AAT), a vector-borne and potentially fatal livestock disease across sub-Saharan Africa. Previously, we identified diverse T. vivax-specific genes that were predicted to encode cell surface proteins. Here, we examine the immune responses of naturally and experimentally infected hosts to these unique parasite antigens, to identify immunogens that could become vaccine candidates. Immunoprofiling of host serum shows that one particular family (Fam34) elicits a consistent IgG antibody response. This gene family, which we now call Vivaxin, encodes at least 124 transmembrane glycoproteins that display quite distinct expression profiles and patterns of genetic variation. We focused on one gene (viv-β8) that encodes one particularly immunogenic vivaxin protein and which is highly expressed during infections but displays minimal polymorphism across the parasite population. Vaccination of mice with VIVβ8 adjuvanted with Quil-A elicits a strong, balanced immune response and delays parasite proliferation in some animals but, ultimately, it does not prevent disease. Although VIVβ8 is localized across the cell body and flagellar membrane, live immunostaining indicates that VIVβ8 is largely inaccessible to antibody in vivo. However, our phylogenetic analysis shows that vivaxin includes other antigens shown recently to induce immunity against T. vivax. Thus, the introduction of vivaxin represents an important advance in our understanding of the T. vivax cell surface. Besides being a source of proven and promising vaccine antigens, the gene family is clearly an important component of the parasite glycocalyx, with potential to influence host-parasite interactions.
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    Reactivity against Sarcocystis neurona and Sarcocystis falcatula-like in horses from Southeastern and Midwestern Brazil
    (2023-01-01) Silva, Thiago Merighi Vieira da [UNESP]; De Santi, Mariele [UNESP]; Gonçalves, Luiz Ricardo; Merino, Márcia Mariza Jusi; André, Marcos Rogério [UNESP]; Machado, Rosangela Zacarias [UNESP]; Universidade Estadual Paulista (UNESP)
    Equine protozoal myeloencephalitis (EPM) is a neurological disease caused by Sarcocystis neurona. Immunofluorescence antibody tests (IFATs) have been widely used to identify exposure of horses to S. neurona in Brazil. Here we used IFAT to search for IgG antibodies against Sarcocystis falcatula-like (Dal-CG23) and S. neurona (SN138) in sera from 342 horses sampled in Campo Grande, Mato Grosso do Sul state (Midwestern), and São Paulo, São Paulo state (Southeastern), Brazil. The 1:25 cutoff value was chosen to maximize sensitivity of the test. IgG antibodies against S. neurona were detected in 239 horses (69.88%), whereas IgG antibodies against S. falcatula-like were detected in 177 horses (51.75%). Sera from 132 horses (38.59%) reacted against both isolates. Absence of reactivity was evidenced in 58/342 horses (16.95%). The lower cutoff used, and the presence of opossums infected with S. falcatula-like and Sarcocystis spp. in the regions where the horses were sampled, might justify the high seroprevalence observed here. Owing to the similarity among antigens targeted in immunoassays, reports on S. neurona-seropositive horses in Brazil may also derive from the exposure of horses to other Sarcocystis species. The role of other Sarcocystis species in causing neurological diseases in horses in Brazil remains unclear.
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    Coxiella burnetii shedding and serological status in pregnant and postpartum ewes
    (2023-08-01) Trujillo, Mayra; Conan, Anne; Calchi, Ana Cláudia [UNESP]; Mertens-Scholtz, Katja; Becker, Anne AMJ; Gallagher, Christa; Mau, Alex; Marchi, Silvia; Machado, Marcus; André, Marcos Rogério [UNESP]; Chapwanya, Aspinas; Müller, Ananda; Ross University School of Veterinary Medicine; City University of Hong Kong; Universidade Estadual Paulista (UNESP); Institute of Bacterial Infections and Zoonoses; University of California- Davis; Universidad Austral de Chile
    This study aimed to evaluate the occurence C. burnetii-DNA shedding by pregnant (vaginal mucus and feces) and postpartum (vaginal mucus, feces and milk) meat breed ewes from Saint Kitts. Additionally, antibodies anti-C. burnetii were detected in serum, and milk. Barbados Blackbelly ewes (n=187) were sampled using stratified convenience cross-sectional sampling. There were two animal groups: pregnant (n=96) and postpartum (n=91). Vaginal mucus (n=187), feces (n=177) and milk (n=83) samples were subjected to a TaqMan real time qPCR assay for C. burnetii based on the IS1111 multi copy element. IgG antibodies against C. burnetii were tested in blood serum (n=187) and milk (n=61) samples, via indirect ELISA. McNemar and Fischer exact tests were used to compare occurrence between routes and between groups, respectively. Overall, 86.6% of all the animals (162/187) were shedding C. burnetti DNA through at least one route (vaginal and/or fecal and/or milk). The DNA shedding occurrence via vaginal (73% vs 51%, p-value=0.003) and fecal routes (64% vs 47%, p-value=0.001) was higher in the pregnant compared to the postpartum animals. There was no prevalent shedding route among vaginal, fecal or milk in all ewes. Overall, 38% of the ewes were seropositive for C. burnetii IgG and a total of 19.7% of the tested postpartum ewes had IgG antibodies in milk. The vaginal and fecal DNA shedding were not associated with the blood serology, nor was milk DNA shedding related to the milk serology status, thus there was no association between C. burnetii seropositivity and bacterial DNA shedding. In short, high occurrence of C. burnetii DNA shedding was observed within ewes in St. Kitts, and represents the first detection of the Q fever agent within the Caribbean islands. Bacterial shedding was more prevalent in pregnant ewes, highlighting the importance of gestating animals as a source of C. burnetii.
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    Expanding the universe of Piroplasmids: morphological detection and phylogenetic positioning of putative novel piroplasmids in black-eared opossums (Didelphis aurita) from southeastern Brazil, with description of “South American Marsupialia Group” of Piroplasmida
    (2023-01-01) Oliveira, Ágatha Ferreira Xavier de; Calchi, Ana Cláudia [UNESP]; Stocco, Anieli Vidal; Stocco, Naiara Vidal; Costa, Alexandre Carvalho; Mureb, Elisabeth Neves; Pires, Jeferson Rocha; Guimarães, Andresa; Raimundo, Juliana Macedo; de Almeida Balthazar, Daniel; Machado, Rosangela Zacarias [UNESP]; André, Marcos Rogério [UNESP]; Baldani, Cristiane Divan; Universidade Federal Rural do Rio de Janeiro – UFRRJ; Universidade Estadual Paulista (UNESP); Universidade Estácio de Sá – UNESA
    The growing proximity of wildlife to large urban niches arouses greater interest in understanding wild reservoirs in the epidemiology of diseases of importance to animal and human health. The aim of the present study was to investigate the presence of piroplasmids in opossums rescued from the metropolitan region of Rio de Janeiro state, Brazil. Blood and bone marrow samples were collected from 15 Didelphis aurita and subjected to DNA extraction and PCR using primers for the 18S rRNA, cox1, cox3, and hsp70 genes of piroplasmids. Clinical and hematological evaluation of the animals was also performed. Five (33.3%) of the 15 opossums tested positive for piroplasms in the nested PCR based on the 18S rRNA, and in two animals, it was possible to observe intra-erythrocytic structures compatible with merozoites. One of the positive animals showed clinical signs of infection such as jaundice, fever, and apathy. Anemia, low level of plasma protein, leukocytosis, and regenerative erythrocyte signs were observed in positive animals. Phylogenetic analysis based on both 18S rRNA and cox-3 genes demonstrated that the piroplasmids detected in D. aurita formed a unique sub-clade, albeit related to piroplasmids previously detected in Didelphis albiventris and associated ticks from Brazil. This study proposes the novel Piroplasmida Clade, namely “South American Marsupialia Group,” and reinforces the need for new clinical-epidemiological surveys to understand the dynamics of these infections in didelphids in Brazil.
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    Concomitant occurrence of multicentric hemangiosarcoma and histiocytic sarcoma in a dog
    (2023-03-01) Gouveia, Bethânia Almeida [UNESP]; Perossi, Isabela Fernanda Spinelli [UNESP]; da Silva, Marina Carla Bezerra [UNESP]; Martinelli, Paulo Edson Baptista [UNESP]; Ramos, Fernanda Ramalho [UNESP]; De Nardi, Andrigo Barboza [UNESP]; Vasconcelos, Rosemeri de Oliveira [UNESP]; Universidade Estadual Paulista (UNESP)
    Histiocytic sarcoma (HS) is uncommon malignant neoplasia of round cells with marked predilection in Rottweiler and Bernese Mountain. The disseminated form, which mainly affects the spleen, lungs, lymph nodes, bone marrow, skin, and subcutis, presents a quick and aggressive clinical behavior. Hemangiosarcoma (HAS) is a malignant neoplasm of endothelial vessel cells commonly reported to affect the right atrium of dogs’ hearts. A male Rottweiler, five years old, presented flaccid paraplegia and progressive muscular atrophy in the temporal, masseter, and limbs muscles; Due to the clinical stage of the animal, euthanasia was conducted. During the necroscopic examination, it was noticed that several masses presented different sizes; some were whitish, and others were reddish and spread in multiple organs (lungs, heart, spleen, stomach, kidneys, brain, medulla, skeletal muscle, and pre-scapular lymph node). Microscopically, in some organs such as the stomach, right ventricle, lungs, and medulla, it was noticed a proliferation of myeloid cells, highly cellular, with poor demarcation, no encapsulation, and with the infiltrative growth pattern of cells with high pleomorphism. Numerous tumoral emboli were observed in the spleen, brain, skeletal muscle, and lymph node. These cells were submitted for immunohistochemistry and were positive for CD18 (HS antibody). In the right atrium, liver, and kidney it was observed malignant and infiltrative endothelial proliferation (HSA) and emboli in the medulla. Therefore, we conclude that both neoplasms (HS and HSA) cause the animal’s paraplegia due to their embolism and metastasis to the spinal cord and skeletal muscle.
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    Canine piroplasmids: Molecular detection and laboratory characterization in dogs from Brasilia, Brazil, with the first molecular evidence of dog exposure to a novel opossum-associated Babesia sp.
    (2023-07-01) de Oliveira, Camila Manoel; Yang, Tzushan Sharon; Duarte, Matheus Almeida; Marr, Henry; McManus, Concepta Margaret; André, Marcos Rogério [UNESP]; Birkenheuer, Adam Joseph; Paludo, Giane Regina; Universidade de Brasília (UnB); North Carolina State College of Veterinary Medicine; Universidade Estadual Paulista (UNESP)
    Canine piroplasmid infections can be caused by Babesia spp., Theileria spp. and Rangelia vitalii. In Brazil, canine babesiosis caused by Babesia vogeli is endemic and reported throughout the country. On the other hand, Rangeliosis caused by R. vitalii has only been described so far in the South and Southeast regions. Despite that, studies analyzing the laboratory and molecular characterization of these hemoprotozoa are still scarce. To investigate the occurrence, the laboratory features, the molecular characterization, and the diversity of piroplasmids from Midwestern Brazil, a survey was performed using blood samples obtained from 276 domestic dogs from Brasília, Federal District, Midwestern Brazil. A broad-range quantitative PCR (qPCR) targeting the mitochondrial large subunit ribosomal DNA (LSU4) was used to detect piroplasmid DNA. The overall molecular occurrence of piroplasmids was 11.2% (31/276), with 9.7% (27/276) of the sequences identified as Babesia vogeli (98–100% identity to B. vogeli isolate from the USA). Based on a partial 18S rRNA sequence pairwise alignment (-250 bp), 1.4% (4/276) of the sequences showed only 76.8% identity with B. vogeli but 100% identity with opossum-associated Babesia sp. (MW290046–53). These findings suggest the exposure of dogs from Brazil to a recently described Babesia sp. isolated from white-eared opossum. None of the analyzed dogs was positive for Theileria spp. or R. vitalii. Subsequently, all positive sequences were submitted to three additional PCR assays based on the 18S rRNA, cox-1, and cytb genes, aiming at performing a haplotype network analysis. Haplotype network using cox-1 sequences showed the presence of six different haplotypes of B. vogeli; one of them was shared with isolates from Brazil, the USA, and India. When including animals co-infected with other vector-borne diseases, piroplasmid-positive dogs had 2.3 times higher chance of having thrombocytopenia than the negative ones. The molecular results demonstrated that the compared Babesia vogeli sequences showed a low variability as well as evidence of exposure to a putative novel opossum-associated Babesia sp. in dogs from Midwestern Brazil.
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    Epidemiological, anatomopathological, and immunophenotypical aspects of cutaneous lymphomas in dogs
    (2023-01-01) Mazaro, Renata D.; Lorenzetti, Douglas M.; Leite, Suzana M.G.; Masuda, Eduardo K.; Da Costa, Leonardo D.; Vasconcelos, Rosemeri O. [UNESP]; Dantas, Antonio Flávio M.; Lacerda, Luciana C.; Da Silva, Tereza Cristina; Motta, Marco Aurélio A.; Cogliati, Bruno; Fighera, Rafael A.; Universidade Federal de Sergipe (UFS); Veterinarian; Axys Análises – Diagnóstico Veterinário e Consultoria Ltda.; Universidade Estadual Paulista (UNESP); Universidade Federal de Campina Grande (UFCG); Célula – Laboratório Veterinário; Universidade de São Paulo (USP); Centro Clínico e Cirúrgico Veterinário (CCCV)
    Cutaneous lymphomas are uncommon tumors in dogs that can occur as epitheliotropic and non-epitheliotropic types. The epitheliotropic type comprises three, already well established, distinct clinicopathological presentations. However, the non-epitheliotropic lymphoma, despite its poor characterization, represents a heterogeneous group of not yet correlated presentations that can mimic different skin tumors, configuring a diagnostic challenge. Therefore, this study’s main aim was to establish whether there is a correlation between the macroscopic presentation and the histological subtypes of cutaneous non-epitheliotropic lymphoma in the population of dogs involved in this study. Additionally, we aimed to determine the prevalence of each type and histological subtype of canine cutaneous lymphoma and describe the epidemiological and anatomopathological characteristics of the presented cases. From a total of 38 cases of cutaneous lymphoma diagnosed in dogs, 17 (44.7%) were considered as epitheliotropic and 21 (55.3%) as non-epitheliotropic. From the 17 cases of cutaneous epitheliotropic lymphoma, 13 (34.2%) and four (10.5%) were subclassified as mycosis fungoides and pagetoid reticulosis, respectively. The cases of cutaneous non-epitheliotropic lymphoma included were: anaplastic large T-cell lymphoma (ALTCL – 9/21, 23.9%), peripheral T-cell lymphoma, unspecified (PTCL-NOS – 4/21, 10.5%), subcutaneous panniculitis-like T-cell lymphoma (SPTCL – 4/21, 10.5%), diffuse large B-cell lymphoma – immunoblastic type (DLBCL – 2/21, 5.2%), lymphomatoid granulomatosis (LYG – 1/21, 2.6%), and marginal zone lymphoma (MZL) of mucosa-associated lymphoid tissue (MALT) - lymphoplasmacytic variant (1/21, 2.6%). Based on the anatomopathological findings, it was possible to infer that when faced with multiple, nodular or placoid skin lesions, predominantly on the trunk and limbs, the diagnosis is more likely to be consistent with ALTCL. Whereas, with solitary skin nodules or plaques, PTCL-NOS will be the most frequently observed histological type. When these lesions are exclusively located in the subcutaneous tissue, one should first think about SPTCL and, more rarely, DLBCL. Regarding to epitheliotropic cutaneous lymphomas, the most commonly observed type in dogs is the cutaneous form of mycosis fungoides, especially in the pre-mycotic and mycotic phases. We hope that this information can assist veterinary clinicians and pathologists in their diagnostic routines and contribute to the characterization of non-epitheliotropic cutaneous lymphomas in the canine species.
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    Feeding on a Bartonella henselae Infected Host Triggers Temporary Changes in the Ctenocephalides felis Microbiome
    (2023-03-01) Moore, Charlotte; Lashnits, Erin; Neupane, Pradeep; Herrin, Brian H.; Lappin, Michael; André, Marcos Rogério [UNESP]; Breitschwerdt, Edward B.; North Carolina State University; University of Wisconsin-Madison; The University of North Carolina at Chapel Hill; Kansas State University; Colorado State University; Universidade Estadual Paulista (UNESP)
    The effect of Bartonella henselae on the microbiome of its vector, Ctenocephalides felis (the cat flea) is largely unknown, as the majority of C. felis microbiome studies have utilized wild-caught pooled fleas. We surveyed the microbiome of laboratory-origin C. felis fed on B. henselae-infected cats for 24 h or 9 days to identify changes to microbiome diversity and microbe prevalence compared to unfed fleas, and fleas fed on uninfected cats. Utilizing Next Generation Sequencing (NGS) on the Illumina platform, we documented an increase in microbial diversity in C. felis fed on Bartonella-infected cats for 24 h. These changes returned to baseline (unfed fleas or fleas fed on uninfected cats) after 9 days on the host. Increased diversity in the C. felis microbiome when fed on B. henselae-infected cats may be related to the mammalian, flea, or endosymbiont response. Poor B. henselae acquisition was documented with only one of four infected flea pools having B. henselae detected by NGS. We hypothesize this is due to the use of adult fleas, flea genetic variation, or lack of co-feeding with B. henselae-infected fleas. Future studies are necessary to fully characterize the effect of endosymbionts and C. felis diversity on B. henselae acquisition.
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    Three chigger species (Trombidiformes: Trombiculidae) co-parasitizing a domestic cat in Brazil, including pathogen monitoring
    (2023-01-01) Jacinavicius, Fernando de; Pereira Pesenato, Isabella; Takatsu, Julia Carvalho; Cousandier, Gabriela; Ochoa, Ronald; Welbourn, Cal; Barros-Battesti, Darci Moraes [UNESP]; Instituto Butantan; Universidade de São Paulo (USP); Clínica Veterinária Saúde Animal; Agricultural Research Service; National Insect and Mite Collection; Universidade Estadual Paulista (UNESP)
    The ecology of chigger mites has many unknown aspects, even with several recent studies addressing the taxonomy and systematics of this group. In Brazil, the chigger species recorded coparasitizing vertebrates include opossums in the Pernambuco State, lizards and rodents in the Piauí State, and rodents in the São Paulo State. Chiggers collected on a domestic cat, Felis catus Linnaeus, 1758 (Mammalia: Felidae), from Caxias do Sul, Rio Grande do Sul State, were sent to the Acarological Collection of Instituto Butantan (IBSP) to be identified. The species were identified as eight larvae of Eutrombicula tinami, two larvae of Parasecia valida Brennan, 1969, and two specimens of the genus Eutrombicula Ewing, 1938, which have been described here as Eutrombicula bassiniin. sp. Additionally, we amplified partial 18S rRNA gene sequences for E. tinami and E. bassiniin. sp. However, the attempts to amplify fragments of the gltA gene of Rickettsia were unsuccessful. The present study reports the coparasitism in a domesticated feline with these three species, the second record of E. tinami, the first record P. valida to the Rio Grande do Sul State, and the description of E. bassiniin. sp.
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    Molecular detection of hemoplasmas in rescued black-eared opossums (Didelphis aurita Wied-Neuwied, 1826) from southeastern Brazil, with evidence of a novel genotype infecting marsupials
    (2023-04-01) de Oliveira, Ágatha Ferreira Xavier; Calchi, Ana Cláudia [UNESP]; Mongruel, Anna Claudia Baumel [UNESP]; Stocco, Anieli Vidal; Stocco, Naiara Vidal; Costa, Alexandre Carvalho; Mureb, Elizabeth Neves; Pires, Jeferson Rocha; Guimarães, Andresa; Balthazar, Daniel de Almeida; Machado, Rosangela Zacarias [UNESP]; André, Marcos Rogério [UNESP]; Baldani, Cristiane Divan; Universidade Federal Rural do Rio de Janeiro – UFRRJ; Universidade Estadual Paulista (UNESP); Universidade Estácio de Sá – UNESA; Instituto Nacional da Mata Atlântica – INMA
    There is a growing concern about the participation of wild hosts and reservoirs in the epidemiology of several pathogens, particularly within the context of environmental changes and the expansion of the One Health concept. The aim of this study was to investigate the presence of hemoplasmas in opossums rescued from the metropolitan region of Rio de Janeiro state, Brazil. Blood samples were collected from 15 Didelphis aurita and subjected to DNA extraction and PCR using primers for the 16S rRNA and 23S rRNA genes. Physical examination and hematological analysis were also performed. Three out of 15 opossums tested positive for hemotropic Mycoplasma spp. by PCR and showed hematological alterations such as anemia and leukocytosis. Clinical signs were non-specific and associated to traumatic lesions. The phylogenetic analysis indicated that the hemoplasma detected was positioned between ‘Ca. Mycoplasma haemodidelphis’ detected in D. virginiana from North American and hemoplasmas recently detected in D. aurita from the state of Minas Gerais, Brazil. This study indicates the existence of hemoplasma infections in D. aurita from the metropolitan region of Rio de Janeiro, and reinforce the need for new epidemiological inquiries to clarify the participation of these in the dynamics of circulation of tick-borne pathogens.
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    Eutrombicula cochinocaensis n. sp.: a chigger (Trombidiformes: trombiculidae) parasite of camelids (Cetartiodactyla: camelidae) in Argentina
    (2023-01-01) Bassini-Silva, Ricardo [UNESP]; Acuña, Francisco; Welbourn, Cal; Ochoa, Ron; Barros-Battesti, Darci Moraes; Jacinavicius, Fernando de Castro [UNESP]; Universidade Estadual Paulista (UNESP); Instituto Butantan; Instituto Nacional de Tecnología Agropecuaria; National Insect and Mite Collection; Agricultural Research Service
    The known chigger fauna of Argentina comprises 17 species organized in the families Trombiculidae (10) and Leeuwenhoekiidae (7). The present study describes a new species of Eutrombicula with drawings, phase contrast microscopy images, and SEM images. This chigger was collected parasitizing a vicuna in Abra Pampa City, Jujuy Province. It is the first time a chigger from Argentina has been collected from the family Camelidae.
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    Epidemiology and Integrative Taxonomy of Helminths of Invasive Wild Boars, Brazil
    (2023-02-01) Perin, Patricia Parreira [UNESP]; Lapera, Ivan Moura [UNESP]; Arias-Pacheco, Carmen Andrea [UNESP]; Mendonça, Talita Oliveira [UNESP]; Oliveira, Wilson Junior [UNESP]; de Souza Pollo, Andressa [UNESP]; dos Santos Silva, Carolina [UNESP]; Tebaldi, José Hairton [UNESP]; da Silva, Bruna [UNESP]; Lux-Hoppe, Estevam Guilherme [UNESP]; Universidade Estadual Paulista (UNESP)
    Wild boars (Sus scrofa) are a significant invasive species in Brazil. We evaluated the helminth diversity of 96 wild boars in São Paulo state. Helminth infection descriptors were calculated, the species were identified and their 18S, 28S rDNA and internal transcribed spacer (ITS) regions were amplified for phylogenetic analyses. Ascarops strongylina, Strongyloides ransomi, Globocephalus urosubulatus, Oesophagostomum dentatum, Trichuris suis, Metastrongylus salmi, Metastrongylus pudendotecus, Ascaris suum and Stephanurus dentatus and Macracanthorhynchus hirudinaceus were identified. Globocephalus urosubulatus had the highest prevalence and mean abundance, and most animals had mixed infections with three parasite species. There was no association between parasite intensity and prevalence and host sex and body condition index (p > 0.05). Novel DNA sequences were obtained from G. urosubulatus, A. strongylina, and S. dentatus. This is the first study on the helmint diversity of non-captive wild boars in Brazil, and the first report of the occurrence of M. hirudinaceus, G. urosubulatus and S. dentatus in Brazilian wild boars. Non-captive wild boars of São Paulo State did not act as capture hosts for native helminth species but maintained their typical parasites, common to domestic pigs. They may act as parasite dispersers for low-tech subsistence pig farming and for native Tayassuidae.
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    Helminths of South American fur seals (Arctocephalus australis) from the Subtropical Convergence Zone of the Southwestern Atlantic
    (2023-01-01) Vivian, Itatiele Farias [UNESP]; Perin, Patricia Parreira [UNESP]; de Amorim, Derek Blaese; Benatti, Danise [UNESP]; Tebaldi, José Hairton [UNESP]; Hoppe, Estevam Guilherme Lux [UNESP]; Universidade Estadual Paulista (UNESP); Universidade Federal do Rio Grande do Sul – UFRGS
    Parasites are important components of ecosystems and may contribute to the ecological aspects of their hosts and indicate the integrity of their environment. To identify the gastrointestinal helminths of the South American fur seal, Arctocephalus australis, 52 animals found dead on the Rio Grande do Sul coast, Southern Brazil, were necropsied. All studied animals were parasitized, and 104,670 specimens of helminths from three phyla and 14 taxa were collected. Adult specimens represented five of the identified species: Contracaecum ogmorhini, Adenocephalus pacificus, Stephanoprora uruguayense, Ascocotyle (Phagicola) longa, and Corynosoma australe; and one of the identified genera: Strongyloides sp. Immature forms represented the other eight taxa: Anisakidae gen. sp., Anisakis sp., Pseudoterranova sp., Contracaecum sp., Tetrabothriidae gen. sp., Cestoda gen. sp., Corynosoma cetaceum, and Bolbosoma turbinella. The acanthocephalan C. australe was the most prevalent and abundant parasite, whereas Strongyloides sp. had the highest intensity. This is the first record of the nematode Anisakis sp., digenean S. uruguayense, and acanthocephalan B. turbinella in this host. Trophic generalist species such as A. australis can be good indicators of the composition of the helminth fauna of their ecosystems, indicating the presence of zoonotic parasites transmitted by the consumption of fish.
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    Molecular diversity of Sarcocystis spp. in opossums (Didelphis spp.) from Southeastern and Midwestern Brazil
    (2023-01-01) De Santi, Mariele [UNESP]; André, Marcos Rogério [UNESP]; Werther, Karin [UNESP]; Gonçalves, Luiz Ricardo; Soares, Rodrigo Martins; Herrera, Heitor Miraglia; Santos, Filipe Martins; Sano, Nayara Yoshie; de Assis, William Oliveira; Rucco, Andreza Castro; Machado, Rosangela Zacarias [UNESP]; Universidade Estadual Paulista (UNESP); Imunodot Diagnósticos; Universidade de São Paulo (USP); Universidade Católica Dom Bosco – UCDB; Universidade Federal de Mato Grosso do Sul (UFMS)
    South American opossums (Didelphis spp.) are definitive hosts of Sarcocystis neurona, Sarcocystis speeri, Sarcocystis lindsayi and Sarcocystis falcatula. In Brazil, diverse studies have demonstrated a high frequency of Sarcocystis falcatula-like in sporocysts derived from opossums, and high genetic diversity has been observed in surface antigen-encoding genes (SAGs). In this study, genetic diversity of Sarcocystis spp. derived from Didelphis albiventris and Didelphis aurita from the cities of Campo Grande and São Paulo, was accessed by sequencing SAG2, SAG3, SAG4, the first internal transcribed spacer (ITS-1) and cytochrome c oxidase subunit I (cox1). Molecular identification was performed for 16 DNA samples obtained from sporocyst or culture-derived merozoites. The ITS-1, cox1, and SAG3 fragments were cloned, whereas SAG2 and SAG4 were sequenced directly from PCR products. Four alleles variants were found for SAG2, 13 for SAG3 and seven for SAG4, from which four, 13 and four, respectively, were novel. Twentyseven allele variants were found for ITS-1, all phylogenetically related to S. falcatula-like previously described in Brazil. Sarcocystis sp. phylogenetically related to Sarcocystis rileyi was evidenced by cox1 in three opossums. Further studies are needed to clarify the role of Didelphis spp. as definitive hosts of Sarcocystis spp. other than that previous described.