First detection of feline sporotrichosis (Sporothrix brasiliensis) at the zoonoses control service in Campo Grande, Mato Grosso do sul, Brazil.

Sporotrichosis is a traumatic mycosis affecting the skin or subcutaneous tissues caused by Sporothrix dimorphic fungus. The fungal complex includes several pathogenic species, out of which S. brasiliensis and S. schenckii are predominant in Brazil. In Mato Grosso do Sul (MS) state, the first human and animal cases were reported in 2016 in Corumbá and Ladário cities. Accordingly, we present the first occurrences of feline sporotrichosis detected in the state capital Campo Grande, MS, by the Zoonoses Control Service (ZCS) of the Municipal Public Health Department. The study included four allochthonous cases of feline sporotrichosis originating from Corumbá, MS, attended by the ZCS. All four cats presented classical clinical signs of sporotrichosis, as ulcerative nodular cutaneous lesions. Three slides tested positive by direct microscopy and PCR, followed by Sanger sequencing confirmed Sporothrix brasiliensis in two samples. The initial suspicion and diagnosis of feline sporotrichosis at the ZCS highlights the importance of accurate surveillance of sporotrichosis in non-endemic areas to enhance the capacity to prevent, detect and respond to emerging diseases in Campo Grande.

A long-term prospecting study on giant viruses in terrestrial and marine Brazilian biomes.

The discovery of mimivirus in 2003 prompted the search for novel giant viruses worldwide. Despite increasing interest, the diversity and distribution of giant viruses is barely known. Here, we present data from a 2012-2022 study aimed at prospecting for amoebal viruses in water, soil, mud, and sewage samples across Brazilian biomes, using Acanthamoeba castellanii for isolation. A total of 881 aliquots from 187 samples covering terrestrial and marine Brazilian biomes were processed. Electron microscopy and PCR were used to identify the obtained isolates. Sixty-seven amoebal viruses were isolated, including mimiviruses, marseilleviruses, pandoraviruses, cedratviruses, and yaraviruses. Viruses were isolated from all tested sample types and almost all biomes. In comparison to other similar studies, our work isolated a substantial number of Marseillevirus and cedratvirus representatives. Taken together, our results used a combination of isolation techniques with microscopy, PCR, and sequencing and put highlight on richness of giant virus present in different terrestrial and marine Brazilian biomes.

The genomic and phylogenetic analysis of Marseillevirus cajuinensis raises questions about the evolution of Marseilleviridae lineages and their taxonomical organization.

Marseilleviruses (MsV) are a group of viruses that compose the Marseilleviridae family within the Nucleocytoviricota phylum. They have been found in different samples, mainly in freshwater. MsV are classically organized into five phylogenetic lineages (A/B/C/D/E), but the current taxonomy does not fully represent all the diversity of the MsV lineages. Here, we describe a novel strain isolated from a Brazilian saltwater sample named Marseillevirus cajuinensis. Based on genomics and phylogenetic analyses, M. cajuinensis exhibits a 380,653-bp genome that encodes 515 open reading frames. Additionally, M. cajuinensis encodes a transfer RNA, a feature that is rarely described for Marseilleviridae. Phylogeny suggests that M. cajuinensis forms a divergent branch within the MsV lineage A. Furthermore, our analysis suggests that the common ancestor for the five classical lineages of MsV diversified into three major groups. The organization of MsV into three main groups is reinforced by a comprehensive analysis of clusters of orthologous groups, sequence identities, and evolutionary distances considering several MsV isolates. Taken together, our results highlight the importance of discovering new viruses to expand the knowledge about known viruses that belong to the same lineages or families. This work proposes a new perspective on the Marseilleviridae lineages organization that could be helpful to a future update in the taxonomy of the Marseilleviridae family. IMPORTANCE: Marseilleviridae is a family of viruses whose members were mostly isolated from freshwater samples. In this work, we describe the first Marseillevirus isolated from saltwater samples, which we called Marseillevirus cajuinensis. Most of M. cajuinensis genomic features are comparable to other Marseilleviridae members, such as its high number of unknown proteins. On the other hand, M. cajuinensis encodes a transfer RNA, which is a gene category involved in protein translation that is rarely described in this viral family. Additionally, our phylogenetic analyses suggested the existence of, at least, three major Marseilleviridae groups. These observations provide a new perspective on Marseilleviridae lineages organization, which will be valuable in future updates to the taxonomy of the family since the current official classification does not capture all the Marseilleviridae known diversity.

Analysis of the Genomic Features and Evolutionary History of Pithovirus-Like Isolates Reveals Two Major Divergent Groups of Viruses.

New representatives of the phylum Nucleocytoviricota have been rapidly described in the last decade. Despite this, not all viruses of this phylum are allocated to recognized taxonomic families, as is the case for orpheovirus, pithovirus, and cedratvirus, which form the proposed family Pithoviridae. In this study, we performed comprehensive comparative genomic analyses of 8 pithovirus-like isolates, aiming to understand their common traits and evolutionary history. Structural and functional genome annotation was performed de novo for all the viruses, which served as a reference for pangenome construction. The synteny analysis showed substantial differences in genome organization between these viruses, with very few and short syntenic blocks shared between orpheovirus and its relatives. It was possible to observe an open pangenome with a significant increase in the slope when orpheovirus was added, alongside a decrease in the core genome. Network analysis placed orpheovirus as a distant and major hub with a large fraction of unique clusters of orthologs, indicating a distant relationship between this virus and its relatives, with only a few shared genes. Additionally, phylogenetic analyses of strict core genes shared with other viruses of the phylum reinforced the divergence of orpheovirus from pithoviruses and cedratviruses. Altogether, our results indicate that although pithovirus-like isolates share common features, this group of ovoid-shaped giant viruses presents substantial differences in gene contents, genomic architectures, and the phylogenetic history of several core genes. Our data indicate that orpheovirus is an evolutionarily divergent viral entity, suggesting its allocation to a different viral family, Orpheoviridae. IMPORTANCE Giant viruses that infect amoebae form a monophyletic group named the phylum Nucleocytoviricota. Despite being genomically and morphologically very diverse, the taxonomic categories of some clades that form this phylum are not yet well established. With advances in isolation techniques, the speed at which new giant viruses are described has increased, escalating the need to establish criteria to define the emerging viral taxa. In this work, we performed a comparative genomic analysis of representatives of the putative family Pithoviridae. Based on the dissimilarity of orpheovirus from the other viruses of this putative family, we propose that orpheovirus be considered a member of an independent family, Orpheoviridae, and suggest criteria to demarcate families consisting of ovoid-shaped giant viruses.

Molecular detection of feline hemoplasmas and retroviruses in free-roaming and shelter cats within a university campus.

Objectives: The aim of the present study was to assess the frequency of hemoplasma, feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) infections in cats living in an on-campus shelter and free-roaming cats within a university campus in Brazil. Methods: Blood samples were tested using quantitative PCR for hemoplasma, FIV and FeLV. Positive hemoplasma samples were sequenced. Associations between hemoplasma detection and living situation, sex, flea and/or tick parasitism, and coinfection with FIV and FeLV, were assessed using Fisher's exact test and the respective odds ratios were calculated. Results: Overall, 6/45 (13.3%) cats tested positive: four (8.9%) were infected with 'Candidatus Mycoplasma haemominutum' and two (4.4%) with Mycoplasma haemofelis. All positive samples were from free-roaming cats (6/15; 40.0%) and had statistically significantly lower packed cell volumes (P = 0.037). Although 5/23 (21.7%) males and 1/22 (4.6%) females were positive, no statistically significant association between sex and hemoplasma infection was found (P = 0.19). Viral quantitative PCR (qPCR) was performed on 43/45 samples, among which 2/43 (4.7%) were positive for FIV and none for FeLV. Only one cat (2.3%) was coinfected with hemoplasma and FIV (P = 0.26). In addition, 4/6 (66.7%) cats that tested positive for hemoplasmas were infested by fleas (P = 0.0014) and/or ticks (P = 0.25). Conclusions and relevance: These results show that even if the free-roaming cat population is clinically healthy and has adequate access to food, it may present flea infestation and hemoplasma infection with lower packed cell volume values.

Bovine Leptospirosis in Caatinga Biome, Brazil: New Insights into Diagnosis and Epidemiology.

Bovine leptospirosis causes economic losses and raises public health concerns. It is possible that there are peculiarities in the epidemiology of leptospirosis in regions with a semiarid climate, such as the Caatinga biome in Brazil, where the climate is hot and dry, and the etiological agent require alternative routes of transmission. This study aimed to close knowledge gaps to the diagnosis and epidemiology of Leptospira spp. infection in cows from the Caatinga biome, Brazil. Samples of the blood, urinary tract (urine, bladder and kidney) and reproductive tract (vaginal fluid, uterus, uterine tube, ovary and placenta) were collected from 42 slaughtered cows. Diagnostic tests included were the microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira spp. antibodies were found in 27 (64.3%) of the animals analyzed using MAT at a 1:50 dilution (cut-off 50), while 31 (73.8%) animals had at least one organ/fluid where the presence of Leptospira spp. DNA was identified, and 29 animals (69%) were positive at bacteriological culture. The highest sensitivity values for MAT were obtained at the cut-off point of 50. In conclusion, even under hot and dry climate conditions, it is possible that Leptospira spp. can spread through alternative routes such as venereal transmission; moreover, a cut-off of 50 is recommended for the serological diagnosis of cattle from the Caatinga biome.

Diversity of Surface Fibril Patterns in Mimivirus Isolates.

Among the most intriguing structural features in the known virosphere are mimivirus surface fibrils, proteinaceous filaments approximately 150 nm long, covering the mimivirus capsid surface. Fibrils are important to promote particle adhesion to host cells, triggering phagocytosis and cell infection. However, although mimiviruses are one of the most abundant viral entities in a plethora of biomes worldwide, there has been no comparative analysis on fibril organization and abundance among distinct mimivirus isolates. Here, we describe the isolation and characterization of Megavirus caiporensis, a novel lineage C mimivirus with surface fibrils organized as "clumps." This intriguing feature led us to expand our analyses to other mimivirus isolates. By employing a combined approach including electron microscopy, image processing, genomic sequencing, and viral prospection, we obtained evidence of at least three main patterns of surface fibrils that can be found in mimiviruses: (i) isolates containing particles with abundant fibrils, distributed homogeneously on the capsid surface; (ii) isolates with particles almost fibrilless; and (iii) isolates with particles containing fibrils in abundance, but organized as clumps, as observed in Megavirus caiporensis. A total of 15 mimivirus isolates were analyzed by microscopy, and their DNA polymerase subunit B genes were sequenced for phylogenetic analysis. We observed a unique match between evolutionarily-related viruses and their fibril profiles. Biological assays suggested that patterns of fibrils can influence viral entry in host cells. Our data contribute to the knowledge of mimivirus fibril organization and abundance, as well as raising questions on the evolution of those intriguing structures. IMPORTANCE Mimivirus fibrils are intriguing structures that have drawn attention since their discovery. Although still under investigation, the function of fibrils may be related to host cell adhesion. In this work, we isolated and characterized a new mimivirus, called Megavirus caiporensis, and we showed that mimivirus isolates can exhibit at least three different patterns related to fibril organization and abundance. In our study, evolutionarily-related viruses presented similar fibril profiles, and such fibrils may affect how those viruses trigger phagocytosis in amoebas. These data shed light on aspects of mimivirus particle morphology, virus-host interactions, and their evolution.

Clinical, histopathological, and molecular findings for Mycobacterium avium subspecies paratuberculosis (MAP) in dairy goats under semiarid conditions.

The objective of this study was to report the occurrence of Mycobacterium avium subspecies paratuberculosis (MAP) in dairy goats, via description of their clinical presentation, histopathological findings, and molecular identification of the infectious agent. Screening was performed using IS900 real-time PCR (qPCR) in milk samples from 179 properties in the semiarid of Northeast region of Brazil. Pooled milk samples from all lactating goats from processing plants were submitted to molecular diagnosis. One property had a positive result at qPCR. The production unit which had the positive sample for MAP was located, and an on-site visit to this property was performed to collect individual milk samples, seven of which tested MAP positive by IS900 qPCR. With permission from the owner, two goats (Animal 1 was positive and Animal 2 was negative on first qPCR for MAP) were acquired and euthanized. Animals 1 and 2 had milk and portions of the duodenum, ileum, colon, and mesenteric lymph nodes positive at qPCR for MAP. Animal 1 also had MAP DNA detected in part of the jejunum and cecum. In animal 2, the ileocecal valve tested positive. MAP was not detected in the blood or feces of either animal; however; it was confirmed for the association of clinical findings, histopathology, and qPCR. The gene IS900 from the positive samples were sequenced and showed a 99% similarity with MAP. The MAP was identified for the first time in the goat milk and tissues in the semiarid region of Northeast Brazil.

Molecular detection of Trypanosoma cruzi in equids in a semiarid zone of Pernambuco - Brazil.

Trypanosoma cruzi is the etiological agent of Chagas disease, a neglected and frequently occurring zoonosis in Central and South American countries. Wild mammals and domestic dogs are the main reservoirs of the parasite in the wild and domestic cycles, respectively. The vectors have a wide variety of food sources that can influence transmission cycles. The aim of this study was to determine the prevalence of T. cruzi infection in donkeys (Equus asininos) and mules (Equus mulus) living in rural areas of the Brazilian semi-arid region. Whole-blood samples from 72 equids (65 donkeys and 7 mules) were analyzed by nested polymerase chain reaction (nested PCR). A total of 51.39% of the samples (37/72) were positive. Phylogenetic analysis identified discrete typing units TcI and TcII, which suggested the possibility that donkeys and mules might be participating in domestic/peridomestic and wild transmission cycles. This was the first report of T. cruzi infection in donkeys and mules in Brazil, with high prevalence of positive animals. This places these animals as potential reservoirs for the parasite and the particular features of these hosts, the presence of vectors and the socioeconomic characteristics of the population under semiarid conditions create interactions that may favor transmission and overlapping T. cruzi infection cycles.

Detection of Leptospira spp. in genitourinary tract of female goats managed in the brazilian semiarid.

Studies related to the prevalence of leptospirosis in the semiarid region showed that even during long periods of drought, the disease has a remarkable frequency in herds in the region. It is a neglected disease and the extent of its effects in the Brazilian semiarid region is not known. The dynamics of this agent is well studied in the urinary tract, however, there are not many studies regarding the genital tract in female goats. Observing this scenario, the present work aimed to diagnose Leptospira spp. in female goats kept in the Brazilian semiarid region by means of serological, molecular and isolation techniques. Blood samples, vaginal fluid, urine and fragments of organs from the genitourinary tract were collected from 40 goats destined for slaughter. Microscopic agglutination test (MAT) was used as a serological technique, with a battery of 24 serovars. The Polymerase Chain Reaction (PCR) of the vaginal fluid, urine and organ fragments was performed, as well as the bacterial growth of these same products in a selective medium. Isolation positive samples were subjected to PCR. It was observed that two (5%) animals were serologically positive for the Pyrogenes serogroup. A total of 29 (72.5%) animals were PCR positive, with DNA present in 51/160 (31.8%) samples from the genital tract and 34/120 (28.3%) from the urinary tract, with no statistical difference. For bacterial growth, 22/40 (55%) animals were positive for growth, with morphology being observed in 19/160 (11.8%) for the genital tract and 16/120 (13.3%) for the urinary tract, with no statistical difference. Two uterus samples showed 99% similarity with L. interrogans after sequencing. Thus, female goats kept under semiarid conditions were positive for Leptospira spp, with positive samples from both the urinary and genital tracts, which possible is an alternative way of adapting and maintaining the agent for severe and adverse conditions.

Lack of serological and molecular evidences of Zika virus circulation in non-human primates in three states from Brazil.

BACKGROUND: Zika virus (ZIKV) was discovered in 1947 with the virus isolation from Rhesus monkey (Macaca mulatta) in Uganda forest, Africa. Old World Primates are involved in a sylvatic cycle of maintenance of this arbovirus, however a limited knowledge about the role of New World primates in ZIKV transmission cycles has been established. OBJECTIVE: This work aimed to investigate the presence of enzootic circulation of ZIKV in New World Primates from three Brazilian states: São Paulo, Paraíba, and Paraná. METHODS: We analyzed 100 non-human primate samples collected in 2018 and 2020 from free-ranging and captive environments from São Paulo (six municipalities belonging to Sorocaba region), Paraíba (João Pessoa municipality), and Paraná (Foz do Iguaçu municipality) using reverse transcriptase quantitative polymerase reaction (RT-qPCR) assays, indirect enzyme-linked immunosorbent assay (ELISA), and plaque reduction neutralization test (PRNT). FINDINGS: All samples (n = 141) tested negative for the presence of ZIKV genome from tissue and blood samples. In addition, all sera (n = 58) from Foz do Iguaçu' non-human primates (NHPs) were negative in serological assays. MAIN CONCLUSION: No evidence of ZIKV circulation (molecular and serological) was found in neotropical primates. In addition, the absence of antibodies against ZIKV suggests the absence of previous viral exposure of NHPs from Foz do Iguaçu-PR.

New insights on Leptospira sp. infection in ewes maintained in field semiarid conditions.

Leptospirosis is poorly studied in small ruminants raised in field semiarid conditions. In this study we compared serological, bacteriological and molecular diagnostic methods in ewes maintained in field Brazilian semiarid conditions. Blood, vaginal fluid and urine samples were collected from 60 Morada Nova ewes raised in a semi-intensive system in the Brazilian semiarid. Diagnostic tests performed were microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira sp. antibodies were found in eight (13.33%) animals analyzed by MAT at reciprocal titer 25 (cut-off 25), while Leptospira sp. DNA was detected in urine or vaginal fluid of 56 animals (93.33%). There was growth of leptospires in 10 urine cultures and in 11 vaginal fluid cultures, however, two of urine (2/60-3.33%) and eight cultures of vaginal fluid (8/60-13.33%) were confirmed by PCR. Two samples of vaginal fluid (one of each animal) were submitted to sequencing demonstrating 99% similarity with L. santarosai and L. interrogans. The highest MAT sensitivities were obtained with reciprocal titer 25 (cut-off 25) compared to 50 and 100. The performance of different diagnostic techniques for leptospirosis in ewes raised in field semiarid conditions allowed a better evaluation of the herd, as well as made it possible to identify carrier animals. Genital route may be important for efficient transmission and without dependence on environmental factors in ewes from semiarid, as well as it's highlighted that titer 1:25 in serology was more efficient, indicating its use in ewes in field semiarid conditions.

Impact of SARS-CoV-2 Gamma lineage introduction and COVID-19 vaccination on the epidemiological landscape of a Brazilian city.

Background: The emergence of the Brazilian variant of concern, Gamma lineage (P.1), impacted the epidemiological profile of COVID-19 cases due to its higher transmissibility rate and immune evasion ability. Methods: We sequenced 305 SARS-CoV-2 whole-genomes and performed phylogenetic analyses to identify introduction events and the circulating lineages. Additionally, we use epidemiological data of COVID-19 cases, severe cases, and deaths to measure the impact of vaccination coverage and mortality risk. Results: Here we show that Gamma introduction in São José do Rio Preto, São Paulo, Brazil, was followed by the displacement of seven circulating SARS-CoV-2 variants and a rapid increase in prevalence two months after its first detection in January 2021. Moreover, Gamma variant is associated with increased mortality risk and severity of COVID-19 cases in younger age groups, which corresponds to the unvaccinated population at the time. Conclusions: Our findings highlight the beneficial effects of vaccination indicated by a pronounced reduction of severe cases and deaths in immunized individuals, reinforcing the need for rapid and massive vaccination.

The Discovery of a New Mimivirus Isolate in Association with Virophage-Transpoviron Elements in Brazil Highlights the Main Genomic and Evolutionary Features of This Tripartite System.

Mimiviruses are giant viruses of amoeba that can be found in association with virophages. These satellite-like viruses are dependent on the mimivirus viral factory to replicate. Mimiviruses can also be associated with linear DNA molecules called transpovirons. Transpovirons and virophages are important drivers of giant virus evolution although they are still poorly studied elements. Here, we describe the isolation and genomic characterization of a mimivirus/virophage/transpoviron tripartite system from Brazil. We analyzed transmission electron microscopy images and performed genome sequencing and assembly, gene annotation, and phylogenetic analysis. Our data confirm the isolation of a lineage A mimivirus (1.2 Mb/1012 ORFs), called mimivirus argentum, and a sputnik virophage (18,880 bp/20 ORFs). We also detected a third sequence corresponding to a transpoviron from clade A (6365 bp/6 ORFs) that presents small terminal inverted repeats (77 nt). The main genomic features of mimivirus argentum and of its virophage/transpoviron elements corroborates with what is described for other known elements. This highlights that this triple genomic and biological interaction may be ancient and well-conserved. The results expand the basic knowledge about unique and little-known elements and pave the way to future studies that might contribute to a better understanding of this tripartite relationship.

Molecular detection of equid herpesvirus in bronchoalveolar lavage fluid from asymptomatic horses in Southern Brazil.

Background and Aim: Molecular approaches to diagnose respiratory viruses have provided an opportunity for early and subclinical pathogen detection, particularly in samples from the upper respiratory tract. This study aimed to investigate the presence of herpesviruses, particularly equid herpesvirus (EHV)-2 and EHV-5, in samples from the lower respiratory tract of healthy racehorses from Southern Brazil. Materials and Methods: Samples from the lower respiratory tract (i.e., bronchoalveolar lavage fluid [BALF]) were assessed by video endoscopy, cytological evaluation of BALF, and tracheal aspirates (TA), along with quantitative polymerase chain reaction (qPCR), to detect equine herpesvirus infection in the lower respiratory tract samples and compare corresponding cytological and endoscopic findings. Results: At least one abnormality per horse during endoscopy examination was observed, including, but not limited to, mucous secretion in the airways and pharyngeal lymphoid hyperplasia. The presence of EHV-2 and/or EHV-5 was detected by qPCR in 3/10 animals. One horse was positive for EHV-2 alone, one for EHV-5 alone, and one for both. Conclusion: To the authors' knowledge, this is the first molecular detection of EHV-2 and EHV-5 in Brazilian thoroughbred horses. These findings may provide new insights into the epidemiology of EHV-2 and EHV-5 in Brazilian horses, evidencing the importance of the molecular investigation, early detection, and prevention of respiratory diseases.

Hemotropic mycoplasmas (hemoplasmas) in wild boars, hunting dogs, and hunters from two Brazilian regions.

Haemotropic mycoplasmas (haemoplasmas) are small pleomorphic bacteria infecting erythrocytes of several mammalian species, including human beings. No study to date has focused on the risk of bacteria exposure in hunting activities, particularly in natural environments of highly tick-infested areas. Accordingly, the present study aimed to assess haemoplasma occurrence in the complex encompassing wild boars, hunting dogs and hunters of Brazil. A total of 38/65 (58.5%) wild boars and 94/159 (59.1%) dogs were positive by qPCR for at least one haemoplasma. All 25 hunters were negative. Dogs with high hunting frequency were 2.4 more likely to be infected. Sequencing revealed a probable novel haemoplasma species in wild boars. Although exposure to haemoplasma species was present, the study herein found no evidence of cross-species transmission.

Leptospira spp. of the urinary tract of female carrier goats in semi-arid conditions

Background: Leptospirosis is an important infectious disease in goat farming, with a worldwide distribution. It is usually transmitted by rodents and the genital route, may cause reproductive losses, negatively impacting goat farming. The diagnosis lies on serological, molecular and isolation techniques. Considering the importance of this disease for small ruminants, this work aimed to evaluate the serological, molecular findings and isolation of pathogenic leptospires in the urinary tract (kidney and bladder tissues) of goats. Materials, Methods & Results: Thirty-four adult goats were used for slaughter. Renal samples (n = 34), bladder (n = 34), were collected for isolation of the agent and molecular detection of Leptospira sp. and blood samples (n = 34) for serological testing. The polymerase chain reaction (PCR) was used as a molecular test and the microscopic serum agglutination test (MAT) was used as a serological test. Samples with DNA amplification were subjected to genetic sequencing. The presence of Leptospira DNA was found in the tissues of 8 (23.4%) goats, and of these, only 2 were positive in PCR and MAT. There was a slight agreement between the PCR and MAT techniques (k = 0.150; P = 0.436). In 6 (17.6%) samples of renal tissue and 2 (5.8%) bladder samples, Leptospira DNA was detected. The genes in a kidney tissue sample were sequenced and demonstrated 99% similarity to Leptospira interrogans. Anti-Leptospira sp. were detected in 6 (17.6%) of the animals tested. Discussion: Serology identified 3 predominant serogroups: Icterohaemorrhagiae, Tarassovi and Autumnalis, serogroups that are related to the presence of rodents that coexist in rural environments. Autumnalis has been reported in small ruminants, raising the hypothesis that goats are adapted, becoming chronic carriers and possible maintenance hosts. The frequency obtained (17.6%) may be the result of the mixed breed pattern and rustic characteristics inherent to the goat species. Given the characteristics of the semi-arid region, such as low rainfall and high solar incidence, it is essential to use an adapted methodology, with a lower cut-off point (1:50), as the serological titer is an established relationship between the animal species, the level of exposure throughout its evolution and the region studied. Molecular findings and bacterial isolation reveal the agent's ability to colonize the urinary tract of goats. These data show the importance that urine has in the epidemiological chain, being able to transmit the agent through direct contact with this product or through contamination of soil and water. There was no statistical agreement between the diagnostic techniques used in this study, in this case, an association between PCR and MAT is recommended to obtain data with high sensitivity and specificity. A bladder sample was sequenced and showed 99% similarity to Leptospira interrogans. In the semiarid region, the most common form of leptospirosis spread is through the sale of animals in business fairs for breeding, rearing or slaughter, as well as sharing the same property with several breeders. The introduction of chronic and asymptomatic carriers on the properties represents a serious risk for the spread of the disease. The results show the presence of Leptospira spp. in semi-arid goat herds, having as risk factors the presence of rodents and intercropping. The association of MAT and PCR is necessary for a better diagnosis of the disease.

Mycobacterium avium subsp. paratuberculosis (MAP) in goat milk in the semiarid region of the Brazilian Northeast - molecular characterization

Background: Goat farming has been on the rise in Brazil in recent years. Overall, 93% of the national herd is concentrated in the Northeast, with the state of Paraíba being the largest goat milk producer in the country. Considering Mycobacterium avium subsp. paratuberculosis (MAP) as a sanitary issue for the development of animal farming with risks for human health and that is a notifiable disease, this research was structured with the objective of confirming the presence and performing a molecular characterization of MAP in goat milk destined for processing plants in the semiarid region of the Brazilian Northeast. Materials, Methods & Results: Samples from 179 production units and 5 collective bulk tanks and 4 samples of pasteurized goat milk were analyzed through Real-time polymerase chain reaction (qPCR). Genetic material (DNA) for MAP was found in the goat milk sample from 1 production unit (1/179). From this positive sample, 9 lactating goats were identified in the original property, 7 of which showed MAP DNA in milk samples (77.77%). The characterization of the nucleotide sequence detected in the positive sample has 99% identity with KJ173784. Discussion: One sample (1/179), from the production units, had MAP genetic material (DNA) detected using the molecular test. Samples from these production units represent the milk from all lactating goats from each producer. Therefore, it was possible to identify from which farm the samples originated, allowing individual animals to then be tested, with milk samples collected from 9 goats and MAP DNA detected in 7 of them (77.77%) via PCR. Control and/or prevention programs need this type of surveillance in reason that it allows the tracking of possible foci from milk samples collected from dairy products or cooling stations. The use of PCR to detect MAP foci via goat milk is thus advantageous because samples are obtained in a non-invasive manner, with faster results when compared to the culture technique. The low detection via PCR in goat milk may be related to factors such as the small amount of MAP eliminated and the intermittent excretion in asymptomatic animals, as also false-positive samples. Samples from the collective bulk tanks was negative. It is possible that the combination of milk from all the properties diluted the amount of MAP. This suggests that the sensitivity of the PCR can be improved if the samples are obtained from the pooled milk from the same property. In some regions of Brazil, for example, showed the frequency of Zona da Mata region of the state of Minas Gerais, Brazil, found 1.94% of positive samples (9/464) and 9.76% (4/41) of properties with at least 1 positive sample for MAP. Different results to what were found in the semiarid region of Paraíba, where climate and production characteristics are different. Goats are susceptible to 3 strains: type "S" (Sheep), "Bison type" and type "C" (Cattle). Previous contact with this species may explain the similarity between the strain found in goat milk and those detected from bovine samples. This must also be taken into consideration during diagnosis and upon implementation of control measures for paratuberculosis in goats. Mycobacterium avium subsp. paratuberculosis was recorded for the first time in goat milk in the semiarid region, which may reveal a potential biological risk to humans and suggests the need for active surveillance of the agent.

Lack of serological and molecular evidences of Zika virus circulation in non-human primates in three states from Brazil

BACKGROUND Zika virus (ZIKV) was discovered in 1947 with the virus isolation from Rhesus monkey (Macaca mulatta) in Uganda forest, Africa. Old World Primates are involved in a sylvatic cycle of maintenance of this arbovirus, however a limited knowledge about the role of New World primates in ZIKV transmission cycles has been established. OBJECTIVE This work aimed to investigate the presence of enzootic circulation of ZIKV in New World Primates from three Brazilian states: São Paulo, Paraíba, and Paraná. METHODS We analyzed 100 non-human primate samples collected in 2018 and 2020 from free-ranging and captive environments from São Paulo (six municipalities belonging to Sorocaba region), Paraíba (João Pessoa municipality), and Paraná (Foz do Iguaçu municipality) using reverse transcriptase quantitative polymerase reaction (RT-qPCR) assays, indirect enzyme-linked immunosorbent assay (ELISA), and plaque reduction neutralization test (PRNT). FINDINGS All samples (n = 141) tested negative for the presence of ZIKV genome from tissue and blood samples. In addition, all sera (n = 58) from Foz do Iguaçu' non-human primates (NHPs) were negative in serological assays. MAIN CONCLUSION No evidence of ZIKV circulation (molecular and serological) was found in neotropical primates. In addition, the absence of antibodies against ZIKV suggests the absence of previous viral exposure of NHPs from Foz do Iguaçu-PR.

Molecular detection of Leishmania infantum in donkeys and mules under semiarid conditions in Brazil.

Visceral leishmaniasis is a parasitic zoonosis that mainly affects poorest and most vulnerable populations, and domestic dogs are considered to be the main source of infection to the vector and therefore humans. However, several studies have investigated the role of other vertebrate hosts in the disease cycle. In this context, the aim of the present study was to conduct a survey of Leishmania infantum infection in donkeys and mules living in a semiarid region of Brazil. Whole blood sampled from 72 equids (65 donkeys and 7 mules) was used to perform molecular diagnosis using the real-time polymerase chain reaction (qPCR) technique. A total of 25% of the samples (18/72) were positive through qPCR, but there were no significant differences between the species (donkeys or mules), sex (male or female) and abandonment situation of the animals (yes or no). Donkeys and mules living under semiarid conditions have high frequency of L. infantum infection. It is therefore worth assigning importance to these species in the epidemiological cycle of visceral leishmaniasis, either as potential reservoirs or just as an abundant food source for vectors.