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Objective: The goal of the article was to assess complementary feeding patterns, and associated factors, of children between 6 and 15-month old in Rio Branco-Acre, Brazil, based on the minimum acceptable diet indicator., Methods: This study was cross-sectional, including 857 children between ages 6 and 15 mo, from a 2015 birth cohort of Rio Branco, Brazil. The prevalence of complementary feeding indicators, minimum meal frequency, minimum dietary diversity, and minimum acceptable diet was estimated based on the recommendations of the World Health Organization. Sociodemographic characteristics, infant and maternal habits, prenatal information, birth characteristics, breastfeeding, and complementary feeding were evaluated. Differences between the proportions were evaluated by the χ 2 test and univariate and multiple logistic regression analyses, to determine associated factors with child minimum acceptable diet., Results: The minimum frequencies of meals and dietary diversity were observed in 81.0% and 51.8% of the children, respectively. The minimum acceptable diet prevalence was 47.1%. Also, minimum acceptable diet was inversely associated with C, D, and E social classes (adjusted odds ratio = 0.49; 95% CI, 0.33-0.72), number of living siblings (adjusted odds ratio for two or three children = 0.69; 95% CI, 0.48-0.98, and adjusted odds ratio for ≥ 4 children = 0.56; 95% CI, 0.37-0.84). Maternal regular consumption of fruits, vegetables, and legumes (adjusted odds ratio = 2.62; 95% CI, 1.69-4.05), child age from 12 to 15 mo (adjusted odds ratio = 2.05; 95% CI, 1.32-3.18), and receiving guidance regarding complementary feeding during postnatal consultations (adjusted odds ratio = 1.38; 95% CI, 1.03-1.86) were directly associated with minimum acceptable diet., Conclusions: Fewer than 50% of the children received adequate food with adequate frequency and diversity. Low socioeconomic status and having ≥ 2 living siblings reduced the chance of minimum acceptable diet, whereas maternal healthy diet, child age (12-15 mo), and complementary feeding counseling during postnatal appointments increased the chance of minimum acceptable diet., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier Inc. All rights reserved.)

The coinfections by some microorganisms have been related to severe diseases in humans and animals, where immunosuppressive agents favor opportunistic behavior of other pathogens. A 4-month-old, female mixed-breed dog with a two-week history of inappetence, prostration, emaciation, and respiratory distress was admitted at a veterinary hospital in Brazil. Tachycardia, pale mucous membranes, severe respiratory distress, and a large number of ticks (Rhipicephalus sanguineus s.l.) in different body regions were observed at clinical examination. Hematological examination of dog showed leukocytosis, neutrophilia, mild anemia, and thrombocytopenia, whereas unremarkable values in biochemical tests. Thoracic radiography revealed a pleural effusion image. Blood and the pleural fluid (purulent aspect) samples were subjected to qPCR (16S rRNA and dsb genes) and sequencing, which identified Ehrlichia canis and Anaplasma platys coinfection. An aggregate of coccoid-to-branching or long filamentous microorganisms, surrounded by pyogranulomatous inflammatory reaction was seen at the cytology of the pleural fluid. Bacteriological culture of pleural effusion showed colonies compatible with the genus Nocardia, which revealed gram-positive filamentous organisms with a tendency of fragmentation and were identified as Nocardia otitidiscaviarum in mass spectrometry (MALDI-TOF MS). Therapy of N. otitidiscaviarum isolate using levofloxacin (supported by a previous in vitro susceptibility testing) and doxycycline for E. canis and A. platys resulted in complete resolution of the clinical picture. Here, we report for the first time a triple coinfection by Nocardia otitidiscaviarum, A. platys, and E. canis in a dog with pleural effusion, where debilitating or immunosuppressive conditions induced by A. platys and E. canis coinfection probably contributed to the opportunistic behavior of N. otitidiscaviarum., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

The Anaplasmataceae family includes obligate, arthropod-transmitted intracellular bacteria that can be zoonotic and potentially fatal. Studies focusing on the interaction between neotropical primates and the agents of this family are scarce. The present study aimed to identify agents of the Anaplasmataceae family in the whole blood of free-living and captive neotropical primates in the State of Mato Grosso, Central-West Brazil. Thirty-eight samples of six nonhuman primate (NHP) species were collected in seven municipalities and analysed through polymerase chain reaction (PCR), nucleotide sequencing, and phylogenetic analysis of the dsb, groEL, 16S rRNA, and gltA genes. DNA fragments similar to those of Ehrlichia canis were detected in Sapajus apella and Ehrlichia chaffeensis from Mico melanurus. The sequences generated in this study and homologous sequences retrieved from GenBank® were used for phylogenetic analyses to characterize the Ehrlichial agents detected in NHPs. The agents were then grouped into clades corresponding to different isolates from the NHP species. In addition, an Anaplasma sp. closely related to Anaplasma marginale was identified in two S. apella individuals. These findings shed light on the susceptibility of neotropical NHPs to Anaplasmataceae agents. These bacteria are known to be transmitted by ticks, which can also serve as possible sources of infection for other animals, including humans., Competing Interests: Declarations of Competing Interest None., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Background: Canine monocytic ehrlichiosis (CME) is caused by the tick-borne pathogen Ehrlichia canis, an obligate intracellular Gram-negative bacterium of the family Anaplasmataceae with tropism for canine monocytes and macrophages. The trp36 gene, which encodes for the major immunoreactive protein TRP36 in E. canis, has been successfully used to characterize the genetic diversity of this pathogen in different regions of the world. Based on trp36 sequence analysis, four E. canis genogroups, United States (US), Taiwan (TWN), Brazil (BR) and Costa Rica (CR), have been identified. The aim of this study was to characterize the genetic diversity of E. canis in Cuba based on the trp36 gene., Methods: Whole blood samples (n = 8) were collected from dogs found to be infested with the tick vector Rhipicephalus sanguineus sensu lato (s.l.) and/or presenting clinical signs and symptoms of CME. Total DNA was extracted from the blood samples and trp36 fragments were amplified by PCR. Nucleotide and protein sequences were compared using alignments and phylogenetic analysis., Results: Four of the trp36 sequences obtained (n = 8) fall within the phylogenetic cluster grouping the US genogroup E. canis strains. The other E. canis trp36 sequences formed a separate and well-supported clade (94% bootstrap value) that is phylogenetically distant from the other major groups and thus represents a new genogroup, herein designated as the 'Cuba (CUB) genogroup'. Notably, dogs infected with the CUB genogroup presented frequent hemorrhagic lesions., Conclusions: The results of this study suggest that genetic diversification of E. canis in Cuba is associated with the emergence of E. canis strains with increased virulence., (© 2022. The Author(s).)

Objectives: To determine the concentrations of total protein (TP), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and hyaluronic acid (HA) in amniotic membranes (AMs) harvested from placentas of bitches of different ages and cryopreserved for different time points. The outcomes of complicated corneal defects of dogs repaired with AMs stored for the same time points were also evaluated., Procedures: Ten cryopreserved canine AMs were stored for short term (2-50 days), middle term (92-210 days), or long term (256-357 days). TP was quantified by Bradford's test, whereas TIMP-1 and HA were quantified by ELISA. Twenty-one dogs that had an AM transplantation to restore deep or perforating corneal wounds were selected., Results: TIMP-1 levels were lower in AMs cryopreserved for middle term (p = .02) and long term (p = .0009), when compared to AMs stored for short term. TP (p = .39) and HA (p = .18) concentrations in AMs did not differ among the storage time. TIMP-1 concentration in AMs correlated with storage time (R = -.65, p < .0001), while TP (R = -.33, p = .07) and HA concentrations did not (R = -.15, p = .41). The age of donors did not correlate with the components evaluated in the AMs. Corneal defects repaired with AMs stored for short term healed sooner than the ones repaired with AMs stored for middle (p < .01) and long term (p = .02). Additionally, TIMP-1 levels in AMs correlated negatively with the epithelization time (R = -.62, p = .002). Graft opacity was severe in 55% of cases. However, the HA levels in AMs correlated negatively with the opacification score (R = -.47, p = .03). Vision was observed in more patients who presented deep ulcers and descemetoceles, than in the ones with perforations (p = .004)., Conclusions: TIMP-1 concentration in canine AMs significantly decreased over a year storage time, while TP and HA concentrations did not change during the same period. The age of donors did not correlate with the components evaluated in the AMs. Complicated corneal defects repaired with AMs cryopreserved for short term healed sooner and tended to be less opaque; however, satisfactory to optimal outcomes were achieved even in the eyes repaired with AMs stored for up to a year., (© 2021 American College of Veterinary Ophthalmologists.)

In the present study, 51 strains of Toxoplasma gondii (T. gondii) were isolated from free-range chickens in the state of Mato Grosso, Midwestern Brazil, upon conducting bioassays in mice, and genotyped them using PCR-restriction fragment length polymorphism (PCR-RFLP) and 11 markers, including SAG1, SAG2 (5'3'SAG2 and alt. SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3. Fifty isolates were completely genotyped revealing 17 genotypes of T. gondii as follows: 12 matched using ToxoDB PCR-RFLP with the previously reported genotypes, including #6 type BrI (n = 4), #8 type BrIII (n = 7), #11 type BrII (n = 3), #14 (n = 1), #19 (n = 1), #41 (n = 1), #99 (n = 1), #109 (n = 4), #116 (n = 1), #140 (n = 2), #166 (n = 9), #190 (n = 1); and five genotypes have not been described before [#313 (n = 6), #314 (n = 1), #315 (n = 1), #316 (n = 1), #317 (n = 1)]. Moreover, mixed infections were identified in five isolates (TgCkBrMT8, TgCkBrMT9, TgCkBrMT33, TgCkBrMT38, and TgCkBrMT41). Additionally, genotype #190 was reported for the first time in chickens from Brazil. Our results corroborate with previous studies on T. gondii isolates identified in chickens from Brazil, thereby confirming their diversity, a typicality, and possibility of co-infection due to different T. gondii strains present in the country., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Cats naturally exposed to Ehrlichia canis have been described in different regions of the world, but little is known about the genotypes associated with infection in these animals. To detect E. canis -specific antibodies and investigate the E. canis TRP genotypes in cats, serum samples from 76 domestic cats reactive to crude E. canis antigens by the indirect fluorescence antibody test (IFAT) were analyzed by ELISA, using E. canis -specific peptides (i.e., TRP19 and TRP36 /BR/US/CR). Of these, 25 (32.9%) cats reacted to at least one TRP peptide, confirming their specific exposure to E. canis . Eighteen (23.7%) cats reacted to TRP19, 15 (19.8%) to BRTRP36, and 11 (14.5%) to USTRP36, but none of them reacted to CRTRP36. Eight (10.5%) cats reacted to TRP19 but not to any TRP36 genotype, demonstrating the possible existence of a new E. canis genotype infecting felines. Nevertheless, this study provides the first report of anti- E. canis -specific antibodies in domestic cats.

The present study aimed to determine the seroprevalence of anti-Toxoplasma gondii antibodies using an indirect immunofluorescence assay (IFA-IgG) in 3,814 cows aged ≥ 24 months belonging to 353 beef farms in the Mato Grosso state. Additionally, we aimed to identify the possible risk factors associated with seropositivity in Mato Grosso, which encompasses three biomes (Amazon, Cerrado, and Pantanal) of Brazil. Analysis of 3,814 samples observed that 1,307 animals were positive for anti-T. gondii antibodies (IFA-IgG ≥ 64), with an animal-level seroprevalence of 34.27%. Herd-level seroprevalence was 92.07%. In the animal-level model, cows raised in the Amazon and Pantanal biomes and breed studies on European and Zebu or hybrid were found to be at risk for T. gondii seropositivity. Prevalence of anti-T. gondii antibodies in cows destined for human consumption was found to be widely distributed throughout the entire study area. Further studies are required to assess the impact of beef in the possible transmission of toxoplasmosis to humans residing in the region and establish additional control measures for this protozoan mainly on beef herds raised in the Amazon and Pantanal biomes, where the highest values of seroprevalence were observed., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

The aetiological agent of equine infectious anaemia (EIA) is the retrovirus equine infectious anemia virus (EIAV) that infects all members of the Equidae family. The EIA is widely disseminated in the Brazilian territory with a high seroprevalence in the Brazilian Pantanal and is mainly diagnosed using agar gel immunodiffusion (AGID). There are few complete EIAV genome sequences available in GenBank, which had an impact on molecular detection studies. In this study, we conducted molecular detection and sequencing of EIAV proviral DNA from Brazilian horses. We analysed the genomic region from exon 1 of tat to gag (tat-gag). Comparative serological tests, comprising AGID and two enzyme-linked immunosorbent assays (ELISAs), were also conducted. Of the 133 samples, 58 were positive in the tat-gag PCR, and 49 nucleotide sequences of 272 bp were obtained. Using this developed tat-gag PCR EIAV proviral DNA was detected in 7% of the AGID-negative samples and 26% of the AGID-negative samples were positive in at least one of the ELISA tests used. Using phylogenetic analysis, the Brazilian Pantanal EIAV sequences grouped in a different clade of EIAV sequences from other countries. Thus, the EIAV sequences can contribute to the knowledge of the tat-gag genomic region in the circulating viruses in the Brazilian Pantanal, in addition to providing new information about the genetic diversity. In addition, the serological results demonstrate the greater sensitivity of the ELISAs used in this study compared to AGID for EIA diagnosis., (© 2020 Wiley-VCH GmbH.)

Ehrlichia spp. are important tick-borne pathogens of animals in Brazil, and Ehrlichia canis is the most prevalent species infecting dogs. Moreover, Ehrlichia minasensis has also recently been identified as a novel ehrlichial agent that infects cattle in Brazil. The objective of this study was to determine whether dogs could be infected by E. minasensis. To investigate this possibility, sera (n = 429) collected from dogs in the Pantanal region were retrospectively analyzed for the presence of antibodies against E. canis and E. minasensis. Canine sera were screened by two isolates of E. canis in indirect immunofluorescence assay (IFA) and the majority (n = 298; 69.4%) had antibodies with endpoint titers ranging from 80 to 327,680. In order to further confirm E. canis-specific antibodies, IFA positive sera were analyzed by ELISA using E. canis-specific peptides (i.e. TRP19 and TRP36 US/BR/CR), which detected E. canis antibodies in 80.2% (239/298) of the dog sera. Fifty-nine (13.7%) samples had detectable antibodies to E. canis by IFA but were negative by E. canis peptide ELISA. These sera were then tested by E. minasensis IFA (Cuiaba strain) as antigen and 67.8% (40/59) were positive (titers ranging from 80 to 20,480). Eleven sera had antibody titers against E. minasensis at least two-fold higher than observed for E. canis and suggests that these dogs were previously infected with E. minasensis. The results of the present study suggest that multiple ehrlichial agents infect dogs in Brazil, which highlights the need to consider different Ehrlichia spp. in Brazilian dogs, particularly in areas where dogs are frequently exposed to multiple tick species. This investigation is the first to provide serologic evidence of E. minasensis infection in dogs from Brazil., (Copyright © 2021. Published by Elsevier B.V.)

Bacteria of the genus Ehrlichia are transmitted by ticks and also are an important cause of infection in wild and domestic mammals. Infection with Ehrlichia spp. has been reported in horses, especially in the USA, Nicaragua and Brazil. In this study, we report the parasitism by Amblyomma sculptum, Rhipicephalus microplus and Dermacentor nitens ticks in horses from a ranch located in south Pantanal wetland. Molecular and serological analyzes to determine infection by Ehrlichia spp. in horses and their respective ticks were carried out. A total of 12 horses were submitted to blood collection to investigate antibodies by indirect immunofluorescence assay (IFA) using Ehrlichia canis crude antigens and to be tested by polymerase chain reaction (PCR) in order to amplify fragments of the 16S rRNA, dsb, groEL and sodB gene of Ehrlichia spp. A total of 164 tick specimens were removed from horses, stored in isopropanol and later identified as D. nitens, A. sculptum and R. microplus. DNA from ticks were extracted and subjected to the same PCR assays to detect Ehrlichia spp. Anti-Ehrlichia spp. antibodies were detected in five/12 (41.7 %) horses by IFA, with antibody titers ranging from 40 to 160. All horse DNA samples were negative for the 16S rRNA, dsb, groEL and sodB of Ehrlichia spp. One A. sculptum female was positive to all target genes of Ehrlichia. This tick was parasitizing an Ehrlichia-seropositive horse with antibody titer of 80. Nucleotide sequences of 16S rRNA, dsb, groEL and sodB genes showed close relationship with different strains of Ehrlichia detected in wild mammals, Amblyomma ticks and horses from Brazil and Argentina. Detection of anti-Ehrlichia sp. antibodies suggests that horses have been exposed to an ehrlichial agent in the Pantanal. Future studies on Ehrlichia infection should be carried out to better elucidate and to bring new information about equine ehrlichiosis, since these animals are important hosts of ticks in the Brazilian Pantanal wetlands., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Tandem repeat proteins (TRPs) are major immunoreactive proteins of Ehrlichia canis , which have been used in the serological diagnosis of different genotypes of the microorganism. TRP19 is preserved among different E. canis isolates expressed on both reticulate and dense-core cells and observed in the extracellular matrix or associated with the morula membrane. TRP36 is differentially expressed only on the surface of the dense-core form of the bacterium and exhibits more divergence among isolates. The aim of this study was to evaluate the distribution of the American (USTRP36), Brazilian (BrTRP36) and Costa Rican (CRTRP36) genotypes of E. canis in Brazil, using ELISA assays. Serum samples of 814 dogs from 49 municipalities from all over Brazil were analyzed. Our results showed that 33.9% of the samples were reactive to the USTRP36 genotype and 32.6% to the BrTRP36 genotype. The two genotypes appeared to occur equally throughout Brazil, although the frequency of seropositivity was lower in the south than in the country's other regions. Dogs that reacted to at least one of the synthetic peptides (TRP19 and TRP36) were 456 (56%). A few dogs ( n = 5; 0.6%) reactive to the E. canis TRP36 genotype (CRTRP36) were also detected in the northeast and southern regions. We concluded that the American and Brazilian genotypes of E. canis are distributed evenly in Brazil, especially in the tropical region, while the temperate region in the south presented the lowest prevalence rates. This study offers the first report of dogs seropositive for the Costa Rican genotype in Brazil., Competing Interests: The authors declare no conflict of interest.

Canine morbillivirus, also known as canine distemper virus (CDV), induces a contagious multisystemic disease caused by an enveloped RNA virus belonging to the genus Morbillivirus within the family Paramyxoviridae. CDV replicates readily in epithelial, nerve and lymphoid tissues; it is excreted in urine, feces, saliva, oral and nasal discharge; and its major route of entry for infection is through the respiratory system. Although the virus was originally believed to infect domestic dogs, new studies have shown that it can also naturally or experimentally infect non-domestic hosts. A recent blood test performed on a giant anteater (Myrmecophaga tridactyla) found Lentz inclusions in the animal's leucocytes. A rapid CDV test, an RT-PCR assay and pathology findings confirmed this report of canine morbillivirus in this species, which corresponds to the second report of CDV infection in the order Pilosa, family Myrmecophagidae in central west Brazil., (© 2020 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Caatinga is a biome exclusive to the semiarid zone of Brazil, where studies on ticks and tick-borne diseases are scarce. Herein, we investigated the occurrence of Rickettsia, Ehrlichia, and Coxiella in wild mammals, domestic dogs and their ectoparasites using molecular and serological techniques. During 2014-2016, blood samples and ectoparasites were collected from 70 small mammals (51 rodents, 18 marsupials, 1 wild canid) and 147 domestic dogs in three areas of the Caatinga. Through serological analyses of domestic dogs of the three areas, 8 to 11 % were seropositive for Rickettsia rickettsii, 9 to 37 % for Rickettsia amblyommatis, 61 to 75 % for Ehrlichia canis, and 0-5% for Coxiella burnetii. All wild mammals were seronegative for Rickettsia spp. and C. burnetii, except for one rodent (Wiedomys pyrrhorhinos) and one marsupial (Didelphis albiventris) that were seroreactive to C. burnetii, one wild canid (Cerdocyon thous) for R. amblyommatis, and two Rattus rattus for Rickettsia spp. Through PCR targeting DNA of Rickettsia, Ehrlichia or Coxiella, all blood samples were negative, except for the presence of Ehrlichia canis DNA in 8.8 % of the domestic dogs, and a recently reported novel agent, Ehrlichia sp. strain Natal, in one marsupial (Gracilinanus agilis). A total of 222 ticks, 84 fleas, and six lice were collected. Ticks were mostly Rhipicephalus sanguineus sensu lato, some Ixodes loricatus, Ornithodoros rietcorreai, Haemaphysalis sp., and Amblyomma spp.; fleas were Ctenocephalides felis felis, Pulex sp. and Polygenis (Polygenis) bohlsi jordani; and lice were Polyplax sp. and Gyropus sp. Through molecular detection of microorganisms, 9% of C. felis felis contained Rickettsia felis, 20 % of A. auricularium contained R. amblyommatis and 13 % of A. parvum contained 'Candidatus Rickettsia andeanae', whereas Ehrlichia canis DNA was detected in at least 6% of the R. sanguineus s.l. from one area. We report a variety of ectoparasites infesting small mammals and domestic dogs in the Caatinga biome, where these ectoparasites probably act as vectors of rickettsiae, ehrlichial agents (E. canis and Ehrlichia sp. strain Natal) and C. burnetii. Our results highlight to the potential risks of human infection by these tick-borne agents in the Caatinga biome., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Nocardia asiatica causing pyogranulomatous pleuropneumonia is reported for the first time in a dog coinfected with canine morbillivirus (CM), diagnosed based on epidemiological, clinical, haematological, images, microbiological, histopathological, polymerase chain reaction and hsp65 gene sequencing findings. The immunosuppression of CM probably favoured the opportunistic behaviour of N. asiatica. Despite the therapeutic measures, the animal died, mainly due to respiratory distress. The association of methods to improve early diagnosis, therapy procedures and prognosis of canine nocardiosis is discussed, as well as the close relationship between pets and their owners, which may favour the transmission of pathogens such as Nocardia from pets-to-humans, which poses an emerging public health issue., (© 2019 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Canine Monocytic Ehrlichiosis (CME) is an infectious disease caused by the rickettsia organism Ehrlichia canis which is transmitted mainly the ixodid brown dog tick Rhipicephalus sanguineus. The prevalence of E. canis infection has been increasing in recent years. The World Health Organization has been warned about antibiotics resistance and one of the way to prevent this situation is found new compound with this property. Doxycycline is the treatment of choice for this tick-borne disease. Adverse effects are noted in dogs that are sensitive to this drug. Antibiotic resistance may also occur. The present study aimed to evaluate the anti-Ehrlichia properties of the essential oil of the aerial parts of Ageratum conyzoides L. in infected DH82 cells, as well as its anti-Ehrlichia activity associated with doxycycline using the checkerboard assay. A. conyzoides is a native plant from northeast Brazil with many reports of ethnopharmacological applications. The essential oil of A. conyzoides was extracted from the aerial parts of the plant using the hydrodistillation method. E. canis-infected DH82 cells were cultured in DMEM (Dulbecco's Modified Eagle Medium), maintained at 37 °C and 5% CO 2 , and standardized at a 70% infection rate for the initiation of treatment protocols. The tests were first carried out with the aim of defining the IC 50 . The combined effect of doxycycline and A. conyzoides essential oil was then determined using the checkerboard dilution technique (checkerboard method) in which the IC 50 was 200 µg/mL. The doxycycline reduction index from the combined effect was 4.90 times resulting in a synergistic effect. To the authors' knowledge, this is the first alternative treatment (alternative therapy) based on bioactive molecules that have antibacterial activity against E. canis.

Coronaviruses are single-stranded positive-sense RNA viruses associated with important avian diseases. Their relatively high rates of mutation and recombination frequencies allow them to adapt to new hosts and ecological niches. Although Brazil has 18% of global avian species diversity, studies regarding the presence of avian viral diseases in wild birds in South America are scarce. In this study, we performed a retrospective analysis of the presence of CoVs in 746 wild birds. Oropharyngeal and cloacal swabs were obtained and placed together in vials containing VTM transport medium collected in different regions of Brazil between 2006 and 2013. Screening for viral nucleic acid was performed using conventional RT-PCR and pancoronavirus nested PCR. Positive samples were characterized by partial sequencing of the RNA-dependent RNA polymerase (RdRp) gene, and ensuing phylogenetic analysis was performed to investigate the association between virus epidemiology and bird migration routes. Coronavirus RNA were detected and sequenced from six samples, in which three were related to gammacoronaviruses group and the other three to deltacoronavirus group. Our study documents the presence of CoVs related to avian gamma- and deltacoronaviruses circulating in both urban- and poultry-farm regions of Brazil, implicating wild birds as potential carriers of CoVs which may represent a risk to poultry farms and public health in Brazil.

This study focused on the isolation and characterization of parvovirus in an infected dog in midwestern Brazil. Non-enveloped icosahedral parvovirus-like particles were isolated in CRFK cells and were allocated to a clade comprised of strains of CPV-2c, based on genome analysis. This is the first isolate of CPV-2c genomically characterized in Brazil.

Equine infectious anemia (EIA) has a worldwide distribution, and is widespread in Brazil. The Brazilian Pantanal presents with high prevalence comprising equine performance and indirectly the livestock industry, since the horses are used for cattle management. Although EIA is routinely diagnosed by the agar gel immunodiffusion test (AGID), this serological assay has some limitations, so PCR-based detection methods have the potential to overcome these limitations and act as complementary tests to those currently used. Considering the limited number of equine infectious anemia virus (EIAV) sequences which are available in public databases and the great genome variability, studies of EIAV detection and characterization molecular remain important. In this study we detected EIAV proviral DNA from 23 peripheral blood mononuclear cell (PBMCs) samples of naturally infected horses from Brazilian Pantanal using a semi-nested-PCR (sn-PCR). The serological profile of the animals was also evaluated by AGID and ELISA for gp90 and p26. Furthermore, the EIAV PCR amplified DNA was sequenced and phylogenetically analyzed. Here we describe the first EIAV sequences of the 5' LTR of the tat gene in naturally infected horses from Brazil, which presented with 91% similarity to EIAV reference sequences. The Brazilian EIAV sequences also presented variable nucleotide similarities among themselves, ranging from 93,5% to 100%. Phylogenetic analysis showed that Brazilian EIAV sequences grouped in a separate clade relative to other reference sequences. Thus this molecular detection and characterization may provide information about EIAV circulation in Brazilian territories and improve phylogenetic inferences.

Given the fact that numerous microbial species can be detected in pregnant female dogs, the objective of this study was to assess the transplacental transmission of Brucella canis, Ehrlichia canis, Neospora caninum and Toxoplasma gondii in stillborn puppies. This study involved 41 stillborn puppies, 78.6% of which were positive for T. gondii, 52.4% for N. caninum and 59.5% for B. canis. E. canis was not detected in any of the analyzed puppies. Pregnancy is an important physiological condition for the transmission of infectious agents to puppies and transplacental transmission may be epidemiologically relevant in the spread of these opportunistic agents., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Neosporosis is considered a major cause of abortion among cattle worldwide. The aim of this study was to investigate the prevalence of anti-Neospora caninum antibodies in dairy cattle and correlate them with possible risk factors on 63 small farms (family farms) in the municipality of Ji-Paraná, the main milk-producing region of the state of Rondônia, northern Brazil. For this purpose, 621 serum samples were collected from cows and were evaluated by means of the indirect fluorescence antibody test (IFAT). The overall herd prevalence of N. caninum antibodies among the farms (38/63; 60.31%) showed that N. caninum are widespread among the dairy herds in this region, despite only infecting a small proportion of animals (66/621, 10.62%). Occurrences of abortion and birth of weak calves were the only variables that showed as risk factors for the presence of N. caninum. The result from the spatial lag model strongly indicated that birth of weak calves and presence of N. caninum are occurring on farms that are located close to each other, indicating aggregation of disease occurrence.

Ehrlichiosis is a worldwide distributed disease caused by different bacteria of the Ehrlichia genus that are transmitted by arthropod vectors. Its occurrence in dogs is considered endemic in several regions of Brazil. Regarding cats, however, few studies have been done and, consequently, there is not enough data available. In order to detect Ehrlichia spp. in cats from the central-western region of Brazil, blood and serum samples were collected from a regional population of 212 individuals originated from the cities of Cuiabá and Várzea Grande. These animals were tested by the Immunofluorescence Assay (IFA) and the Polymerase Chain Reaction (PCR) designed to amplify a 409 bp fragment of the dsb gene. The results obtained show that 88 (41.5%) cats were seropositive by IFA and 20 (9.4%) cats were positive by PCR. The partial DNA sequence obtained from PCR products yielded twenty samples that were found to match perfectly the Ehrlichia canis sequences deposited on GenBank. The natural transmission of Ehrlichia in cats has not been fully established. Furthermore, tick infestation was not observed in the evaluated cats and was not observed any association between age, gender and positivity of cats in both tests. The present study reports the first serological and molecular detection of E. canis in domestic cats located in the endemic area previously mentioned.

Background: West Nile virus (WNV) is an emergent pathogen that is widely distributed in North and Central America. The recent introduction in South America has focused attention on the spread of WNV across Southern American countries. The transmission network involves mosquitoes, birds, horses and humans., Methods: The serological evaluation of sera from 678 equids and 478 birds was performed using a WNV-specific blocking ELISA, and only the positive results were confirmed by plaque reduction neutralisation tests (PRNTs). Molecular analysis was performed on sera from 992 healthy equids and on 63 macerates of brains from equids that died of encephalitis and had previously tested negative for other pathogens. We also tested swabs from 928 birds. The samples analysed were collected in different biomes of Brazil., Results: We identified WNV antibodies by ELISA in thirteen equids and five birds, and PRNT90 confirmed WNV positivity in four equid samples collected in 2009 in an area between the Amazon and the Pantanal. None of the ELISA positive bird samples were confirmed by PRNT90, and all samples tested by RT-PCR were negative., Conclusion: WNV circulation is confirmed by this large scale survey even in the absence of detection of clinical cases.

The literature contains several studies on feline ehrlichiosis. However, information about the characteristics of Ehrlichia infection in cats is still scanty. This study evaluated the association between Ehrlichia spp. infection and the hematologic data of 93 cats treated at the Federal University of Mato Grosso Veterinary Hospital in Cuiabá, state of Mato Grosso, Brazil. The presence of or exposure to Ehrlichia spp. infection was evaluated by Polymerase Chain Reaction (PCR) targeting the dsb and 16S rRNA gene of Ehrlichia, and by detection of anti-Ehrlichia canis IgG antibodies in Indirect Fluorescence Assay (IFA), respectively. Eight (8.6%) cats tested positive by PCR and the partial DNA sequence obtained from PCR products was a 100% match to E. canis. Forty-two (45.1%) cats showed antibody reactivity against Ehrlichia spp. Hematological alterations such as low erythrocyte count, thrombocytopenia, lymphopenia and monocytosis were observed in PCR positive cats. Among them, low erythrocyte counts were associated with IgG antibody titers of 40 to 640 and five cats also tested positive by PCR. Furthermore, PCR-positive cats showed a tendency to be lymphopenic. No correlation was found between age and sex, and no ticks were observed in any of the examined cats.