From Nature to Drug: Overview and CADD Approach of Anacardic Acid to Propose their Biological Potential.

Anacardic acids are natural compounds found in various plant families, such as Anacardiaceae, Geraniaceae, Ginkgoaceae, and Myristicaceae, among others. Several activities have been reported regarding these compounds, including antibacterial, antioxidant, anticancer, anti-inflammatory, and antiviral activities, showing the potential therapeutic applicability of these compounds. From a chemical point of view, they are structurally made up of salicylic acids substituted by an alkyl chain containing unsaturated bonds, which can vary in number and position, determining their bioactivity and differentiating them from the various existing forms. Our work aimed to explore the potential of anacardic acids, based on studies that address the bioactivity of these compounds, as well as to establish a greater understanding of the structure-activity relationship of these compounds through in silico methods, with a focus on the elucidation of a possible drug target through the application of computer-aided drug design, CADD.

Neutralizing antibodies to bovine and bubaline alphaherpesviruses in water buffaloes (Bubalus bubalis).

Water buffaloes (Bubalus bubalis) have been introduced in many regions of the world as a source of animal protein. In many instances, bubaline cattle are reared close to or mixed with bovine or zebuine cattle. However, little is known about infectious diseases of bubaline and the interactions that may arise involving the microbiota of those species. Alphaherpesviruses of ruminants (bovine alphaherpesviruses types 1 and 5, BoHV-1, BoHV-5; bubaline alphaherpesvirus 1, BuHV-1) are highly cross-reactive in serological assays performed with bovine or zebuine sera. However, the profile of reactivity of bubaline cattle sera to alphaherpesviruses remains unknown. As such, it is not known which virus strain (or strains) would be most appropriate to be used as the challenge virus in the laboratory in search for alphaherpesvirus-neutralizing antibodies. In this study, the profile of neutralizing antibodies to alphaherpesviruses in bubaline sera was determined against different types/subtypes of bovine and bubaline alphaherpesviruses. Sera (n=339) were screened in a 24-h serum neutralization test (SN) against 100 TCID50 of each of the challenge viruses. From those, 159 (46.9 %) neutralized at least one of the viruses assayed; 131 (38.6%) sera neutralized the three viral strains used for screening. The viral strain that was neutralized by the largest number of sera was BoHV-5b A663 (149/159; 93.7%). A few sera neutralized only one of the challenge viruses: four sera neutralized BoHV-1 LA only; another neutralized BoHV-5 A663 only and four others neutralized BuHV-1 b6 only. SN testing with two additional strains gave rise to similar results, where maximum sensitivity (defined here as the largest number of sera that neutralized the challenge viruses) was obtained by adding positive results attained with three of the challenge strains. Differences in neutralizing antibody titers were not significant to allow inferences on which would be the most likely virus that induced the antibody responses detected here.

An ELISA to Detect Antibodies to Bovine Alphaherpesviruses 1 and 5 and Bubaline Alphaherpesvirus 1 in Cattle Sera.

Bovine alphaherpesvirus 1 (subtypes 1.1, 1.2a, and 1.2b), type 5 (subtypes 5a, 5b, and 5c), and bubaline herpesvirus 1 (BuHV-1) induce highly, though not fully cross-reactive serological responses. Most types and subtypes of these viruses circulate particularly in countries of the southern hemisphere, notably Brazil and Argentina. Therefore, the detection of infected animals is important in defining prevention and control strategies, particularly when flocks are destined for international trade. Identification of infected herds is most often achieved by assays that detect antibodies, such as enzyme immunoassays (ELISAs). However, to date, no ELISA has been evaluated in its capacity to detect antibodies to these alphaherpesviruses. Here, an ELISA was developed to detect antibodies to all currently recognized BoAHV-1, BoAHV-5, and BuAHV-1 types/subtypes, and its sensitivity and specificity were determined. Six hundred bovine sera were screened in serum neutralization tests (SN) against the seven viruses. ELISAs prepared with each of the viruses were compared to SN. Subsequently, a combined assay with multiple antigens LISA was prepared by mixing five viral antigens, chosen for their highest sensitivity in the preparative assays. In comparison to SN, the mAgELISA sensitivity was 96.5% with 96.1% specificity (κ = 0.93; PPV = 95.0%; NPV = 97.3%). The findings reveal that the mAgELISA developed here is highly suitable for the detection of antibodies, comparable in sensitivity and specificity to that of SN when performed with all known types and subtypes of bovine and bubaline alphaherpesviruses.

Antimicrobial resistance profiles in Escherichia coli isolated from whole-chicken carcasses from conventional, antibiotic-free, and organic rearing systems / Perfil de resistência em Escherichia coli isoladas de carcaças de frango provenientes de sistemas de produção convencional, livre de antibióticos e orgânico

Antimicrobial resistance (AMR) is a growing concern in human and animal health. Public discussions on these issues have contributed to an increased demand for antibiotic-free food. Studies comparing the antimicrobial resistance profiles of bacteria in foodstuffs originating from farming systems with restrictions on the use of antimicrobials are scarce. This study aimed to assess the antimicrobial resistance profiles of generic Escherichia coli isolated from whole chickens originating from farming systems with and without restrictions on the use of antimicrobials. For this purpose, three groups of E. coli strains were formed: (GC) from chickens reared in conventional production systems, without restriction on the use of antimicrobials (n=72); (GL) from chickens reared in farming systems certified as free of any antibiotic use (n=72); and (GO) from chickens from an organic farming system (n=72). Whole chicken units were individually rinsed as recommended by ISO 17604:2015, and E. coli was isolated from the rinse suspension. To evaluate the resistance profile, E. coli strains were tested against 12 antimicrobials using broth microdilution or disk diffusion tests. Eighty strains (40.7%) were found to be fully susceptible to the tested antimicrobials, and 23.6% were multidrug resistant. The highest frequencies of resistance were observed to tetracycline (GC=37,5%; GL=34,7%; GO=25%) and trimethoprim (GC=27,8%; GL=34,7%; GO=22,2%). In the case of multidrug resistant strains, GC presented 32% (n=23) of strains with multidrug resistance characteristics whereas the GL and GO groups presented 22% (n=16) and 17% (n=12), respectively. As for the totally susceptible strains, a frequency of 56% of Tsus strains was observed in the organic group, whereas this frequency was 33% in the GC and GL groups. Using GC as a reference, the Poisson regression model showed a higher occurrence of fully susceptible E. coli strains, as well as lower frequencies of multidrug resistance and resistance to ampicillin and nalidixic acid in GO. The GL group exhibited the lowest frequency of ampicillin resistance. These observations suggest that the lower selection pressure for antimicrobial use in the farming system may be reflected in the resistance profile of bacteria present in foodstuffs purchased by consumers.(AU)

A resistência antimicrobiana (AMR) é uma preocupação crescente para a saúde humana e animal. A discussão pública dessas questões tem contribuído para o aumento da demanda por alimentos produzidos sem o uso de antibióticos. No entanto, estudos que comparem os perfis de resistência antimicrobiana de bactérias em alimentos oriundos de sistemas agrícolas com restrição ao uso de antimicrobianos ainda são escassos. O objetivo deste estudo foi avaliar o perfil de resistência antimicrobiana em Escherichia coli genérica isolada de carcaças de frangos inteiros oriundos de sistemas de criação com e sem restrições ao uso de antimicrobianos. Para tanto, três grupos de cepas de E. coli foram formados: (GC) isolados de carcaças de frangos criados no sistema convencional - sem restrição de uso de antimicrobianos (n=72); (GL) isolados de frangos de sistemas certificados sem uso de antimicrobianos (n=72); (GO) de frangos originados de produção orgânica (n=72). As unidades de frango inteiro foram submetidas à lavagem conforme recomendado pela ISO 17604: 2015 e E. coli foi isolada da suspensão de enxágue. Para avaliar o perfil de resistência, as cepas de E. coli foram testadas frente à 12 antimicrobianos pelos testes de microdiluição em caldo ou difusão em disco. Oitenta cepas (40,7%) foram totalmente suscetíveis aos antimicrobianos testados e 23,6% multirresistentes. As maiores frequências de resistência foram observadas frente a tetraciclina (GC=37,5%; GL=34,7%; GO=25%) e trimetoprima (GC=27,8%; GL=34,7%; GO=22,2%). No caso de cepas multirresistentes, GC apresentou 32% (n=23) das cepas com características de multirresistência enquanto os grupos GL e GO apresentaram 22% (n=16) e 17% (n=12), respectivamente. Quanto às cepas totalmente suscetíveis, foi observada uma frequência de 56% de cepas Tsus no grupo orgânico enquanto tal frequência foi de 33% nos grupos GC e GL. Utilizando GC como referência, o modelo de regressão de Poisson demonstrou maior ocorrência de cepas de E. coli totalmente suscetíveis, bem como menores frequências de multirresistência e resistência à ampicilina e ácido nalidíxico no GO. Em GL, apenas a frequência mais baixa de resistência à ampicilina pôde ser demonstrada. Essas observações sugerem que a menor pressão de seleção do uso de antimicrobianos no sistema de cultivo pode se refletir no perfil de resistência das bactérias presentes nos alimentos adquiridos pelo consumidor.(AU)

The virome of the white-winged vampire bat Diaemus youngi is rich in circular DNA viruses.

In the Neotropical region, the white-winged vampire bat (Diaemus youngi) is the rarest of the three species of vampire bats. This bat species feeds preferentially on bird blood, and there is limited information on the viruses infecting D. youngi. Hence, this study aimed to expand the knowledge about the viral diversity associated with D. youngi by sampling and pooling the lungs, liver, kidneys, heart, and intestines of all animals using high-throughput sequencing (HTS) on the Illumina MiSeq platform. A total of three complete and 10 nearly complete circular virus genomes were closely related to gemykrogvirus (Genomoviridae family), smacovirus (Smacoviridae family), and torque teno viruses (TTVs) (Anelloviridae family). In addition, three sequences of bat paramyxovirus were detected and found to be closely related to viruses reported in Pomona roundleaf bats and rodents. The present study provides a snapshot of the viral diversity associated with white-winged vampire bats and provides a baseline for comparison to viruses detected in future outbreaks.

CTX-M-15-producing Klebsiella pneumoniae ST273 associated with nasal infection in a domestic cat.

OBJECTIVES: The aim of this study was to investigate the genetic context of expanded-spectrum ß-lactam resistance in a Klebsiella pneumoniae strain causing a hard-to-treat nasal infection in a domestic cat. METHODS: A K. pneumoniae isolate was recovered from a 4-year-old male cat hospitalised in a veterinary hospital in Paraíba, Northeastern Brazil. Following phenotypic confirmation of multidrug resistance by the disk diffusion method, the genome was sequenced using an Illumina MiSeq system. Multilocus sequence typing (MLST) and structural features related to antimicrobial resistance were determined by downstream bioinformatics analyses. RESULTS: The strain was confirmed as sequence type 273 (ST273) K. pneumoniae harbouring a variety of genes conferring antimicrobial resistance to phenicols tetracyclines, aminoglycosides, ß-lactams, fosfomycin, sulfonamides and quinolones. Two plasmids were identified. Plasmid p114PB_I co-harboured a set of plasmid-borne resistance genes [blaCTX-M-15, blaTEM-1, qnrS1, tetD, tetR, sul2, aph(6)-Id, aph(3'') and cat2]. Notably, the multiresistance region was characterised as a chimeric plasmid structure sharing high sequence homology with several plasmids from Enterobacteriaceae. The second plasmid (p114PB_II) was characterised as a plasmid present in many genomes belonging to K. pneumoniae. CONCLUSION: The genetic context of the plasmid sequences harboured by a veterinary pathogenic K. pneumoniae isolate reveals the high complexity of horizontal gene transfer mechanisms in the acquisition of antimicrobial resistance genes. The emergence, dissemination and evolution of antimicrobial resistance must be investigated from a One Health perspective.

A variety of highly divergent eukaryotic ssDNA viruses in sera of pigs.

Over the last decade, viral metagenomics has been established as a non-targeted approach for identifying viruses in stock animals, including pigs. This has led to the identification of a vast diversity of small circular ssDNA viruses. The present study focuses on the investigation of eukaryotic circular Rep-encoding single-stranded (CRESS) DNA viral genomes present in serum of commercially reared pigs from southern Brazil. Several CRESS DNA viral genomes were detected, including representatives of the families Smacoviridae (n=5), Genomoviridae (n=3), Redondoviridae (n=1), Nenyaviridae (n=1) and other yet unclassified genomes (n=9), plus a circular DNA molecule, which probably belongs to the phylum Cressdnaviricota. A novel genus within the family Smacoviridae, tentatively named 'Suismacovirus', comprising 21 potential new species, is proposed. Although the reported genomes were recovered from pigs with clinical signs of respiratory disease, further studies should examine their potential role as pathogens. Nonetheless, these findings highlight the diversity of circular ssDNA viruses in serum of domestic pigs, expand the knowledge on CRESS DNA viruses' genetic diversity and distribution and contribute to the global picture of the virome of commercially reared pigs.

Natural Products as Potential Agents against SARS-CoV and SARSCoV- 2.

BACKGROUND: Natural products are useful agents for the discovery of new lead- compounds and effective drugs to combat coronaviruses (CoV). OBJECTIVE: The present work provides an overview of natural substances, plant extracts, and essential oils as potential anti-SARS-CoV agents. In addition, this work evaluates their drug-like properties which are essential in the selection of compounds in order to accelerate the drug development process. METHODS: The search was carried out using PubMed, ScienceDirect and SciFinder. Articles addressing plant-based natural products as potential SARS-CoV or SARS-CoV-2 agents within the last seventeen years were analyzed and selected. The descriptors for Chemometrics analysis were obtained in alvaDesc and the principal component analysis (PCA) was carried out in SIMCA version 13.0. RESULTS: Based on in vitro assays and computational analyses, this review covers twentynine medicinal plant species and more than 300 isolated substances as potential anti-coronavirus agents. Among them, flavonoids and terpenes are the most promising compound classes. In silico analyses of drug-like properties corroborate these findings and indicate promising candidates for in vitro and in vivo studies to validate their activity. CONCLUSION: This paper highlights the role of ethnopharmacology in drug discovery and suggests the use of integrative (in silico/ in vitro) and chemocentric approaches to strengthen current studies and guide future research in the field of antiviral agents.

Complete genome characterization of porcine circovirus 3 recovered from wild boars in Southern Brazil.

In the present study, the complete nucleotide sequence of porcine circovirus 3 (PCV3) recovered from wild boars lymph nodes is described. The full genome was named PCV3-wb/Br/RS and comprises 2,000 nucleotides with two open reading frames (ORFs) with a stem-loop motif in intergenic region. The ORFs are oriented in opposite directions and encode the putative capsid (Cap) and replicase (Rep) proteins. Based on amino acid motif analysis, PCV3-wb/Br/RS as well as most of the sequences from wild boars are classified as PCV3b. Phylogenetic analysis including 97 PCV3 sequences available in databases showed that the PCV3-wb/Br/RS genome is more closely related to genomes recovered in Spain, China, Germany and Denmark. Phylogenetic inferences among PCV3-wb/Br/RS and other circoviruses confirmed that these seem to have a most recent common ancestor with bat-associated circoviruses. In addition, PCV3 infection was investigated by real-time PCR in a cohort of 80 wild boars in Southern Brazil. A total of 29 animals (36.3%) were PCV3-positive leading the conclusion that PCV3 is circulating in the wild boar population in Southern Brazil. The role played by PCV3-like infections in wild boars and the risk these could pose to commercial swine production within that region remains to be further investigated.

Detection of multiple viruses in oropharyngeal samples from Brazilian free-tailed bats (Tadarida brasiliensis) using viral metagenomics.

In this study, we analyzed the viral population in oropharyngeal samples from T. brasiliensis using a viral metagenomic approach. Genomes corresponding to members of the families Circoviridae, Genomoviridae, Herpesviridae, Paramyxoviridae, Coronaviridae, and Astroviridae were detected. This study provides the first preliminary understanding of the oropharyngeal virome of T. brasiliensis, which may guide the discovery and isolation of novel viruses in the future and highlights the need for continuing investigations in this regard.

Investigation on porcine circovirus type 3 in serum of farrowing sows with stillbirths.

Since its first identification in 2016, porcine circovirus 3 (PCV3) has been detected in healthy and/or diseased swine in many countries worldwide. In a previous study by our group, PCV3 was detected in sera of sows which had at least one stillborn piglet in the last parturition. As such, it became important to investigate if the presence of PCV3 in sows' sera could be associated to the occurrence of stillbirths. With that aim, the frequency of PCV3 infections and viral DNA loads in sows' sera was investigated through a real-time quantitative PCR in 89 serum samples of just farrowed sows with or without stillbirths. PCV3 genomes were identified in most samples, with genome loads ranging between less than 10 to 200,000 copies per mL of serum. No significant differences were observed either in the frequency of infection or PCV3 viral loads in sows with or without stillbirths. Thus, no association could be established between PCV3 infection of sows at farrowing and stillbirths' occurrence.

Viral DNA genomes in sera of farrowing sows with or without stillbirths.

A study was conducted to investigate the serum virome of sows with and without stillbirths after farrowing. Sera from sows with at least one stillbirth or with normal litters were collected immediately after farrowing. Viral DNA was extracted from serum pools and submitted to high throughput sequencing. No differences in the proportion of virus-related reads were found in both groups (p > 0.05). A variety of viral DNA genomes were identified, mostly representative of three viral families: Anelloviridae, Circoviridae and Smacoviridae. Besides, a number of novel unclassified circular Rep-encoding single stranded DNA (CRESS DNA) viruses were also identified. These findings suggest that the presence of such viral genomes in sows' sera bears no correlation with stillbirths' occurrence; it seems likely that these constitute part of the normal serum microbiome of sows at farrowing.

Phylogenetic and evolutionary analysis of HoBi-like pestivirus: Insights into origin and dispersal.

The HoBi-like pestivirus (HoBiPeV), currently classified as Pestivirus H species, is a pathogen associated with a broad spectrum of clinical manifestations in ruminants, particularly in cattle. Since HoBiPeV complete genome sequencing data is scarce, in the present study we described five nearly complete new Brazilian HoBiPeV genomes and further perform a more complete genetic and evolutionary characterization with all additional genome sequences available in the GenBank database. Entropy and selection pressure analysis showed the E2 gene, a surface glycoprotein, is the most variable gene, which also displays the greatest number of sites under positive selection. Phylogenetic and Bayesian inference based on complete genome and Npro gene, respectively, from all HoBiPeV sequences available so far, confirms the existence of three main clades (a, b, and c). The abovementioned analysis suggests that this pestivirus species probably emerged in Asia and spread to different regions including Brazil, where only strains belonging to specific genetic group 'a' have been found. The hypothesis of the HoBiPeV introduction in Brazil (between 1,890 and 1,962), formulated based on Bayesian inference, coincides with a period of intensive importation of water buffalo (Bubalus arnee) and indicine cattle (Bos taurus indicus) from Asia to Brazil, suggesting that this could be the origin of the current Brazilian HoBiPeV genetic group 'a'.

In-depth genomic characterization of a meropenem-nonsusceptible Pseudomonas otitidis strain contaminating chicken carcass

Background: The indiscriminate use of antibiotics in food-animal production has a major impact on public health, particularly in terms of contributing to the emergence and dissemination of antimicrobial resistant bacteria in the food-animal production chain. Although Pseudomonas species are recognized as important spoilage organisms in foodstuff, they are also known as opportunistic pathogens associated with hospital-acquired infections. Furthermore, Pseudomonas can play a role as potential reservoirs of antimicrobial resistance genes, which may be horizontally transferred to other bacteria. Considering that cephalosporins (3rd and higher generations) and carbapenems are critically important beta-lactam antimicrobials in human medicine, this study reports the occurrence and genomic characterization of a meropenem-nonsusceptible Pseudomonas otitidis strain recovered from a chicken carcass in Brazil. Materials, Methods & Results: During the years 2018-2019, 72 frozen chicken carcasses were purchased on the retail market from different regions in Brazil. Aliquots from individual carcass rinses were screened for Extended Spectrum Beta-lactamase (ESBL)-producing bacteria in MacConkey agar supplemented with 1mg.L-1 cefotaxime. Phenotypically resistant isolates were further tested for resistance to other antimicrobials and confirmed as ESBL-producers by means of disk-diffusion method using Müller-Hinton agar. Only one meropenen-nonsusceptible isolate was detected and submitted to whole genome sequencing (WGS) in Illumina Miseq. The strain was identified as Pseudomonas otitidis by local alignment of the 16S rRNA sequence using BLASTn and confirmed by Average Nucleotide Identity (ANI) analysis using JspeciesWS database. Genes encoding for antimicrobial resistance were detected by means...

Bovine abortion by a vaccine strain of Bacillus anthracis / Aborto bovino por uma cepa vacinal de Bacillus anthracis

This paper reports the abortion of a male Aberdeen Angus bovine by a vaccine strain of Bacillus anthracis, describing the pathological and microbiological findings and the genome sequence. Necropsy findings included multifocal areas of hemorrhage in different organs. Histologically, various organs showed hemorrhage, fibrin exudation, necrosis associated with countless bacillary bacterial clumps and severe neutrophilic inflammatory infiltrate. In the microbiological examination, numerous rough, nonhemolytic, gray and dry colonies with irregular edges were isolated from liver, lung and abomasum content samples. Gram staining revealed square-ended Gram-positive rods arranged in chains. B. anthracis identification was confirmed by detection of the molecular chromosomal marker Ba813. The genomes from the isolated B. anthracis (named SPV842_15) and from the isolated vaccinal strain (Brazilian vaccinal strain), which was recovered from a commercial vaccine used in the pregnant cow, were sequenced. Genomic comparisons displayed a high level of nucleotide identity in the comparisons between B. anthracis SPV842_15 and the B. anthracis Brazilian vaccinal strain (98,2%). Furthermore, in both strains, only the plasmid pX01 sequence was detected. Although, vaccination against anthrax is characterized by an elevated protective profile and very low residual virulence, immunization with Sterne strains can cause abortion in cattle, presumably by the plasmid pX01 toxins in rare or special situations.(AU)

Este trabalho relata um aborto de um bovino, macho, Aberdeen Angus, por uma cepa vacinal de Bacillus anthracis, descreve os achados patológicos, microbiológicos e o sequenciamento do genoma. Os achados de necropsia incluíram áreas multifocais de hemorragias em diferentes órgãos. Histologicamente, órgãos afetados apresentaram hemorragia, exsudação de fibrina, necrose associada a miríades bacterianas bacilares e intenso infiltrado inflamatório neutrofílico. No exame microbiológico, foram isoladas numerosas colônias rugosas, não hemolíticas, cinzas e secas, com bordas irregulares a partir de amostras de fígado, pulmão e conteúdo do abomaso. A coloração de Gram revelou bastonetes Gram-positivos dispostos em cadeias. A identificação do B. anthracis foi confirmada pela detecção do marcador cromossômico molecular Ba813. Os genomas do isolado B. anthracis (SPV842_15) e do isolado vacinal (cepa vacinal brasileira), recuperado de uma vacina comercial utilizada na vaca prenhe, foram sequenciados. Comparações genômicas mostraram um elevado nível de identidade de nucleotídeos entre B. anthracis SPV842_15 e cepa vacinal brasileira (98,2%). Além disso, em ambas as estirpes foi detectada apenas a sequência do plasmídeo pX01. Embora a vacinação contra o antraz seja caracterizada por um perfil protetor elevado e uma virulência residual muito baixa, a imunização com estirpes de Sterne pode causar aborto em bovinos, presumivelmente pelas toxinas do plasmídeo pX01 em situações raras ou específicas.(AU)

Bovine abortion by a vaccine strain of Bacillus anthracis / Aborto bovino por uma cepa vacinal de Bacillus anthracis

ABSTRACT: This paper reports the abortion of a male Aberdeen Angus bovine by a vaccine strain of Bacillus anthracis, describing the pathological and microbiological findings and the genome sequence. Necropsy findings included multifocal areas of hemorrhage in different organs. Histologically, various organs showed hemorrhage, fibrin exudation, necrosis associated with countless bacillary bacterial clumps and severe neutrophilic inflammatory infiltrate. In the microbiological examination, numerous rough, nonhemolytic, gray and dry colonies with irregular edges were isolated from liver, lung and abomasum content samples. Gram staining revealed square-ended Gram-positive rods arranged in chains. B. anthracis identification was confirmed by detection of the molecular chromosomal marker Ba813. The genomes from the isolated B. anthracis (named SPV842_15) and from the isolated vaccinal strain (Brazilian vaccinal strain), which was recovered from a commercial vaccine used in the pregnant cow, were sequenced. Genomic comparisons displayed a high level of nucleotide identity in the comparisons between B. anthracis SPV842_15 and the B. anthracis Brazilian vaccinal strain (98,2%). Furthermore, in both strains, only the plasmid pX01 sequence was detected. Although, vaccination against anthrax is characterized by an elevated protective profile and very low residual virulence, immunization with Sterne strains can cause abortion in cattle, presumably by the plasmid pX01 toxins in rare or special situations.

RESUMO: Este trabalho relata um aborto de um bovino, macho, Aberdeen Angus, por uma cepa vacinal de Bacillus anthracis, descreve os achados patológicos, microbiológicos e o sequenciamento do genoma. Os achados de necropsia incluíram áreas multifocais de hemorragias em diferentes órgãos. Histologicamente, órgãos afetados apresentaram hemorragia, exsudação de fibrina, necrose associada a miríades bacterianas bacilares e intenso infiltrado inflamatório neutrofílico. No exame microbiológico, foram isoladas numerosas colônias rugosas, não hemolíticas, cinzas e secas, com bordas irregulares a partir de amostras de fígado, pulmão e conteúdo do abomaso. A coloração de Gram revelou bastonetes Gram-positivos dispostos em cadeias. A identificação do B. anthracis foi confirmada pela detecção do marcador cromossômico molecular Ba813. Os genomas do isolado B. anthracis (SPV842_15) e do isolado vacinal (cepa vacinal brasileira), recuperado de uma vacina comercial utilizada na vaca prenhe, foram sequenciados. Comparações genômicas mostraram um elevado nível de identidade de nucleotídeos entre B. anthracis SPV842_15 e cepa vacinal brasileira (98,2%). Além disso, em ambas as estirpes foi detectada apenas a sequência do plasmídeo pX01. Embora a vacinação contra o antraz seja caracterizada por um perfil protetor elevado e uma virulência residual muito baixa, a imunização com estirpes de Sterne pode causar aborto em bovinos, presumivelmente pelas toxinas do plasmídeo pX01 em situações raras ou específicas.

The virome of an endangered stingless bee suffering from annual mortality in southern Brazil.

Meliponiculture - the management of stingless bee colonies - is an expanding activity in Brazil with economic, social and environmental potential. However, unlike in apiculture, the pathogens that impact on meliponiculture remain largely unknown. In southern Brazil, every year at the end of the summer, managed colonies of the stingless bee Melipona quadrifasciata manifest a syndrome that eventually leads to collapse. Here we characterize the M. quadrifasciata virome using high-throughput sequencing, with the aim of identifying potentially pathogenic viruses, and test whether they are related to the syndrome outbreaks. Two paired viromes are explored, one from healthy bees and another from unhealthy ones. Each virome is built from metagenomes assembled from sequencing reads derived either from RNA or DNA. A total of 40 621 reads map to viral contigs of the unhealthy bees' metagenomes, whereas only 11 reads map to contigs identified as viruses of healthy bees. The viruses showing the largest copy numbers in the virome of unhealthy bees belong to the family Dicistroviridae - common pathogenic honeybee viruses - as well as Parvoviridae and Circoviridae, which have never been reported as being pathogenic in insects. Our analyses indicate that they represent seven novel viruses associated with stingless bees. PCR-based detection of these viruses in individual bees (healthy or unhealthy) from three different localities revealed a statistically significant association between viral infection and symptom manifestation in one meliponary. We conclude that although viral infections may contribute to colony collapses in the annual syndrome in some meliponaries, viruses spread opportunistically during the outbreak, perhaps due to colony weakness.

High frequency and extensive genetic heterogeneity of TTSuV1 and TTSuVk2a in PCV2- infected and non-infected domestic pigs and wild boars from Uruguay.

Torque teno sus virus (TTSuV) infection is common worldwide in both healthy and diseased swine and a relationship between this virus and a particular disease in pigs has not been established. This work aimed to investigate the presence of TTSuV1 and TTSuVk2a in Porcine Circovirus type 2 (PCV2)-infected and non-infected domestic pigs and free-living wild boars from Uruguay. Our data evidenced a high frequency of detection and a wide circulation of TTSuV among pig herds and wild boar populations. Furthermore, TTSuV1+TTSuVk2a co-infection was more frequent than single infections in domestic pigs. In addition, we thoroughly characterized at the molecular level TTSuV strains by extensive sequence data analysis. Our findings revealed an extremely high genetic heterogeneity among Uruguayan isolates. On the basis of detailed analyses, we proposed a more comprehensive criterion of TTSuV classification which would contribute to shedding light over the genetic diversity of these viruses worldwide. On the other hand, data obtained suggested that neither TTSuV1 nor TTSuVk2a frequency of infection or viral loads have any correlation with PCV2 infection, health status or age. The role of TTSuV during co-infection with other pathogens and the age-related dynamics of TTSuV infection are currently under debate. Therefore, taking into account the controversial epidemiological data regarding these viruses and their ubiquitous infection, a likely role as components of the host microbiota should be brought into discussion.

Quillaja brasiliensis saponin-based nanoparticulate adjuvants are capable of triggering early immune responses.

Commercially available saponins are extracted from Quillaja saponaria barks, being Quil A® the most widely used. Nanoparticulate immunostimulating complexes (ISCOMs or ISCOMATRIX) formulated with these, are able to stimulate strong humoral and cellular immune responses. Recently, we formulated novel ISCOMs replacing QuilA® by QB-90 (IQB-90), a Quillaja brasiliensis leaf-extracted saponin fraction, and reported that IQB-90 improved antigen uptake, and induced systemic and mucosal antibody production, and T-cell responses. However, its mechanism of action remains unclear. In this study we provide a deeper insight into the immune stimulatory properties of QB-90 and ISCOMATRIX-like based on this fraction (IMXQB-90). We show herein that, when used as a viral vaccine adjuvant, QB-90 promotes an "immunocompetent environment". In addition, QB-90 and IMXQB-90 induce immune-cells recruitment at draining-lymph nodes and spleen. Subsequently, we prove that QB-90 or IMXQB-90 stimulated dendritic cells secret IL-1ß by mechanisms involving Caspase-1/11 and MyD88 pathways, implying canonical inflammasome activation. Finally, both formulations induce a change in the expression of cytokines and chemokines coding genes, many of which are up-regulated. Findings reported here provide important insights into the molecular and cellular mechanisms underlying the adjuvant activity of Q. brasiliensis leaf-saponins and its respective nanoparticles.

Mamastrovirus 5 detected in a crab-eating fox (Cerdocyon thous): Expanding wildlife host range of astroviruses.

Astroviruses are a common cause of gastroenteritis in children worldwide and can also cause infection in a range of domestic and wild animal species. Canine astrovirus (formally named as Mamastrovirus 5, MAstV5) has been reported worldwide, and its role as an enteric pathogen is still controversial. Herein, we describe the genomic characterization of a MAstV5 (strain crab-eating fox/2016/BRA) identified in a wild canid (Cerdocyon thous) diagnosed with canine distemper virus (CDV) as causa mortis. The nearly complete genome comprised 6579 nt in length and displayed the archetypal organization of astroviruses. The present report is the first evidence of MAstV5 infection in an animal species other than the dog and highlights a possible natural astrovirus spillover between domestic and wild canids. Moreover, these results show the first evidence of extra-intestinal MAstV5, suggesting a virus systemic spread. This work is expected to contribute to a better understanding of the astroviruses biology and their interactions with the wildlife health.