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Background/Objective: Yellow fever (YF) is an acute infectious disease caused by the yellow fever virus which is transmitted by mosquitoes. Neotropical primates are susceptible to infection, which is often presented as epizootic outbreaks. The aim was to evaluate and characterize the immune response against YF in different species of neotropical primates from the Belo Horizonte Zoo. Methods: Vaccine 17DD was administered to 24 neotropical primates, with a single subcutaneous dose. Clinical exams, RNAemia, and detection of IgG and neutralizing antibodies against YFV were performed. In addition, an ethogram was performed to assess clinical changes and animal welfare. Results: At 4 days post-vaccination, RNAemia was detected in nine animals. There was seroconversion and persistence of immune response in Alouatta guariba clamitans, Sapajus xanthosternos, Saguinus imperator and Aotus infulatus. However, the vaccine was not immunogenic for Lagothrix cana. In Pithecia irrorata seroconversion did not persist long term, while the Ateles sp. had a transient immune response. No significant clinical manifestations were observed in any of the vaccinated animals. Conclusions: This study demonstrated a safe, immunogenic and persistent immune response induced by the attenuated 17DD vaccine strain in A. guariba clamitans, S. xanthosternos, S. imperator, and A. infulatus.

Pseudoplacentational endometrial hyperplasia (PEH) is a common uterine lesion in dogs. A high frequency of pyometra has been associated with PEH in dogs, suggesting that PEH might be related to the pathogenesis of pyometra. This study aimed to assess transcription levels and expression of Toll like receptors (TLR) 1, 2 and 4; alpha estrogen receptors (ESR1), progesterone receptors (PR) and prolactin receptors (PRLR) in uteri with PEH. Furthermore, the inflammatory response of dog endometrium with PEH against ex vivo bacterial stimulus was also investigated. Uteri were classified as controls or with PEH. Uterine explants were cultured for 6 and 12 hours after in vitro stimulus with inactivated Escherichia coli . Transcription of receptors and proinflammatory cytokines, namely interleukin-6 ( IL-6 ) and CXCL8 were evaluated. Expression of receptors was also evaluated in uterine explants and uteri from biopsy archives. CXCL8 concentration was measured in supernatants from all cultured explants. Transcription levels and expression of both PR and ESR1 were lower in uteri explants with PEH not stimulated and cultured for 6 hours. Expression of PRLR was higher in uteri with PEH from biopsy archives. Proinflammatory response by transcription levels of interleukin 6 demonstrated downregulation in uteri with PEH at 6 hours of stimulation followed by upregulation at 12 hours. However, no differences between groups were observed. Both control and uteri with PEH secreted similar concentrations of CXCL8 at 6 hours of bacterial stimulation. At 12 hours, no response to stimulation was observed in the PEH group and supernatant concentrations of CXCL8 were higher in the control group. The inflammatory response to bacterial stimulus in uteri with PEH had different pattern than the control group, with an inversion in IL-6 transcription levels between 6–12 hours of culture. Additionally, CXCL8 production ceased earlier in explants with PEH than in control.

Serological methods are valuable diagnostic tools for bovine brucellosis, a zoonotic infectious disease with a worldwide distribution. Currently employed diagnostic methods utilize crude bacterial extract or regions of LPS, a molecule that can exhibit cross-reactions with other infectious agents. The goal of this study was to develop an indirect enzyme-linked immunosorbent assay (ELISAi) using synthetic peptides or a multi-epitope protein based on in silico-predicted B cell epitopes. Peptides were synthesized on a cellulose membrane using spot synthesis, and immunoblots were performed to evaluate reactivity by densitometry against bovine sera of interest. Peptides reacting with serum from positive cattle and non-reactive to the serum from negative controls were selected, synthesized in a soluble form, and used as antigens for the development of the ELISAi. Two peptides (P1 and P2) were selected and, after standardization of the ELISAi, positive (25) and negative (175) samples were tested, resulting in a sensitivity of 84% (21/25) and specificity of 83.43% (146/175). The sequence of the two peptides in replicates with spacers were inserted into a multi-epitope protein, which was also used as antigen in an ELISAi resulting in a sensitivity of 72% (18/25) and a specificity of 61.71% (108/175). This study provided a preliminary analytical assessment of in silico-predicted epitopes for developing novel serologic diagnostic tests.

Serologic diagnosis of canine brucellosis caused by Brucella canis remains quite challenging since the currently available methods have considerable limitations, although the relevance and awareness about this zoonotic pathogen is increasing over the past few years. Therefore, the development of novel strategies is highly desirable. In this study, in silico analyses resulted in prediction of B. canis specific B cell epitopes, which were validated by synthesizing the corresponding peptides on a membrane followed by immunobloting with sera from dogs naturally infected with B. canis or uninfected controls as well as by excluding epitopes that cross reacted with sera from cattle and sheep infected with B. abortus and B. ovis , respectively. This approach resulted in the identification of 26 epitopes that reacted exclusively with sera from dogs infected with B. canis , from which the 15 with strongest signal in the immunoblot were selected and synthesized in soluble form for further analyses. The best 10 synthetic peptides (considering the noise to signal ratio) were used in combination as antigens for development of a B. canis specific indirect enzyme-linked immunosorbent assay (iELISA) protocol, which yielded improved specificity to differentiate from other common canine pathogens ( Leishmania sp. and Babesia sp.) and an improved performance (100% specificity and 80% sensitivity) when compared to crude bacterial protein extracts used as antigen for iELISA. These selected epitopes were also incorporated in a multi- B. canis epitope protein that was expressed in E. coli and employed as antigen for detection of anti- B. canis IgG and IgM from naturally infected dogs, resulting in analytical performance similar to the iELISA protocol in which synthetic peptides were used as antigens (100% specificity and 75% sensitivity). The results clearly indicate that combination of detection of IgM and IgG may result in higher sensitivity. Therefore, this study provides novel tools for improving the accuracy of serologic diagnosis of canine brucellosis caused by B. canis .

Marsupials of the genus Didelphis are highly adapted to urban environments and are widely distributed in the Americas. Streptococcus didelphis is a bacterium that has been isolated from, and associated with disease, in the Virginia opossum ( Didelphis virginiana ) and the white-eared opossum ( Didelphis albiventris ). This study describes pathological changes associated with S. didelphis infection in white-eared opossums ( Didelphis albiventris ) and Brazilian common opossums ( Didelphis aurita ) as well as microbiological and genomic characterization of isolates. Ten opossums underwent necropsy and had ulcerative dermatitis of probable traumatic origin had S. didelphis was isolated from cutaneous lesions (n = 16) or systemic sites (n = 4). In contrast, 34 free-ranging opossums that were captured had negative culture results for S. didelphis from nasal swabs. Other lesions in opossums naturally infected with S. didelphihs included splenitis (7/10), myocarditis (6/10), interstitial nephritis and pyelonephritis (7/10), and myositis (4/10). Most isolates were susceptible to the antimicrobial drugs tested and none of them were able to form biofilm in vitro . Whole genome analysis of six isolates revealed no resistance determinants, virulence factors or plasmids, and the isolates showed high genomic similarity.

Pseudoplacentational endometrial hyperplasia (PEH) and cystic endometrial hyperplasia (CEH) are both hyperplastic conditions that may occur in the canine uterus during diestrus. CEH can impair fertility, and, although the consequences of PEH are poorly known, this condition is significantly associated with pyometra. The aim of this study was to investigate frequencies of both PEH and CEH in female dogs according to age, size, breed, and breed group. Uteri from 300 female dogs were sampled and processed for histopathologic analyses. Lesions were identified, and frequencies were statistically compared. PEH was significantly more frequent in dogs between 4 and 12 years of age and significantly less frequent in Yorkshires than Shih-tzus, as well as in the breed group that includes Brazilian Terriers and Yorkshires. CEH was significantly more frequent in dogs older than 12 years, but no breed predisposition was observed. The frequency of PEH was significantly higher than that of CEH. The frequency of pyometra in PEH cases was significantly higher than endometritis or uterus without inflammation. This study identified age, breed group, and size as predisposing factors for PEH and further demonstrated the association between this condition and pyometra in dogs.

Marsupials of the genus Didelphis are highly adapted to urban environments and are widely distributed in the Americas. Streptococcus didelphis is a bacterium that has been isolated from, and associated with disease, in the Virginia opossum (Didelphis virginiana) and the white-eared opossum (Didelphis albiventris). This study describes pathological changes associated with S. didelphis infection in white-eared opossums (Didelphis albiventris) and Brazilian common opossums (Didelphis aurita) as well as microbiological and genomic characterization of isolates. Ten opossums underwent necropsy and had ulcerative dermatitis of probable traumatic origin had S. didelphis was isolated from cutaneous lesions (n = 16) or systemic sites (n = 4). In contrast, 34 free-ranging opossums that were captured had negative culture results for S. didelphis from nasal swabs. Other lesions in opossums naturally infected with S. didelphihs included splenitis (7/10), myocarditis (6/10), interstitial nephritis and pyelonephritis (7/10), and myositis (4/10). Most isolates were susceptible to the antimicrobial drugs tested and none of them were able to form biofilm in vitro. Whole genome analysis of six isolates revealed no resistance determinants, virulence factors or plasmids, and the isolates showed high genomic similarity.

Marsupials of the genus Didelphis are highly adapted to urban environments and are widely distributed in the Americas. Streptococcus didelphis is a bacterium that has been isolated from, and associated with disease, in the Virginia opossum (Didelphis virginiana) and the white-eared opossum (Didelphis albiventris). This study describes pathological changes associated with S. didelphis infection in white-eared opossums (Didelphis albiventris) and Brazilian common opossums (Didelphis aurita) as well as microbiological and genomic characterization of isolates. Ten opossums underwent necropsy and had ulcerative dermatitis of probable traumatic origin had S. didelphis was isolated from cutaneous lesions (n = 16) or systemic sites (n = 4). In contrast, 34 free-ranging opossums that were captured had negative culture results for S. didelphis from nasal swabs. Other lesions in opossums naturally infected with S. didelphihs included splenitis (7/10), myocarditis (6/10), interstitial nephritis and pyelonephritis (7/10), and myositis (4/10). Most isolates were susceptible to the antimicrobial drugs tested and none of them were able to form biofilm in vitro. Whole genome analysis of six isolates revealed no resistance determinants, virulence factors or plasmids, and the isolates showed high genomic similarity.

Pseudoplacentational endometrial hyperplasia (PEH) is a common uterine lesion in dogs. A high frequency of pyometra has been associated with PEH in dogs, suggesting that PEH might be related to the pathogenesis of pyometra. This study aimed to assess transcription levels and expression of Toll like receptors (TLR) 1, 2 and 4; alpha estrogen receptors (ESR1), progesterone receptors (PR) and prolactin receptors (PRLR) in uteri with PEH. Furthermore, the inflammatory response of dog endometrium with PEH against ex vivo bacterial stimulus was also investigated. Uteri were classified as controls or with PEH. Uterine explants were cultured for 6 and 12 hours after in vitro stimulus with inactivated Escherichia coli. Transcription of receptors and proinflammatory cytokines, namely interleukin-6 (IL-6) and CXCL8 were evaluated. Expression of receptors was also evaluated in uterine explants and uteri from biopsy archives. CXCL8 concentration was measured in supernatants from all cultured explants. Transcription levels and expression of both PR and ESR1 were lower in uteri explants with PEH not stimulated and cultured for 6 hours. Expression of PRLR was higher in uteri with PEH from biopsy archives. Proinflammatory response by transcription levels of interleukin 6 demonstrated downregulation in uteri with PEH at 6 hours of stimulation followed by upregulation at 12 hours. However, no differences between groups were observed. Both control and uteri with PEH secreted similar concentrations of CXCL8 at 6 hours of bacterial stimulation. At 12 hours, no response to stimulation was observed in the PEH group and supernatant concentrations of CXCL8 were higher in the control group. The inflammatory response to bacterial stimulus in uteri with PEH had different pattern than the control group, with an inversion in IL-6 transcription levels between 6-12 hours of culture. Additionally, CXCL8 production ceased earlier in explants with PEH than in control.

Canine distemper virus (CDV; Morbilivirus canis ) is a morbillivirus with worldwide distribution. The virus belongs to the family Paramyxoviridae, which includes highly pathogenic viruses such as measles virus (MeV) ( Measles virus ) and rinderpest virus ( Rinderpest morbillivirus ). Canine distemper is primarily a disease in domestic dogs, but it has been described in terrestrial and aquatic wild mammals, demonstrating the ability of CDV to cross species barriers. Here, we describe a lethal CDV outbreak in free‐ranging marmosets ( Callithrix penicillata ) in Brazil. The marmosets were captured during a targeted epidemiological surveillance program aimed at monitoring viral agents in wildlife in urban parks of Belo Horizonte, Minas Gerais, Brazil. Affected marmosets presented with neurological signs, and others were found dead in the same area. In this report, we detail the occurrence of the outbreak and describe clinical manifestations, gross and histopathological lesions, tissue distribution of the virus by immunohistochemistry (IHC) and molecular methods, and phylogenetic analyses of the virus. This study demonstrated that CDV can infect free‐ranging black‐tufted marmosets and lead to neurologic signs, cutaneous lesions, and death. Viral genomic sequences and antigens were detected in several organs, indicating a pantropic distribution of CDV in this neotropical primate species. Additionally, the marmosets were also tested for MeV and Lyssavirus , which yielded negative results. Coinfections with CDV and Callitrichinae gammaherpesvirus 3 were detected in three marmosets. The CDV sequences identified here were closely related to both South American sequences and Vero cell‐adapted lineages. These findings have significant implications from a One Health perspective.