Investigation of anti-Leptospira spp. antibodies and leptospiruria in cats attended to a veterinary teaching hospital in southern Brazil.

Leptospirosis is a bacterial zoonosis that affects both humans and animals worldwide. Currently, it is known that cats may be susceptible to infection. This study aims to investigate the presence of anti-Leptospira spp. antibodies and leptospiruria in cats, using Microscopic Agglutination Test (MAT) and Real-time Polymerase Chain Reaction (PCR) techniques, respectively. A total of 76 cats, undergoing comprehensive anamnesis, general physical examination, and complementary exams were included in the investigation. Among the 76 cats tested, 9.2% (7/76) exhibited the presence of anti-Leptospira spp. antibodies, while Leptospira spp. DNA was detected in at 1.3% (1/76) of the evaluated urine samples. No significant associations were observed between the serological and molecular diagnostic results and the assessed variables, including clinical data and laboratory results of cats testing positive. This study provides insight into the occurrence of Leptospira spp. infection and leptospiruria in cats treated at a veterinary teaching hospital in southern Brazil.

Genome analysis of Pseudomonas strain 4B with broad antagonistic activity against toxigenic fungi.

Pseudomonas sp. 4B isolated from the effluent pond of a bovine abattoir was investigated as antifungal against toxigenic fungi. The complete genome of Pseudomonas 4B was sequenced using the Illumina MiSeq platform. Phylogenetic analysis and genome comparisons indicated that the strain belongs to the Pseudomonas aeruginosa group. In silico investigation revealed gene clusters associated with the biosynthesis of several antifungals, including pyocyanin, rhizomide, thanamycin, and pyochelin. This bacterium was investigated through antifungal assays, showing an inhibitory effect against all toxigenic fungi tested. Bacterial cells reduced the diameter of fungal colonies, colony growth rate, and sporulation of each indicator fungi in 10-day simultaneous growing tests. The co-incubation of bacterial suspension and fungal spores in yeast extract-sucrose broth for 48 h resulted in reduced spore germination. During simultaneous growth, decreased production of aflatoxin B1 and ochratoxin A by Aspergillus flavus and Aspergillus carbonarius, respectively, was observed. Genome analysis and in vitro studies showed the ability of P. aeruginosa 4B to reduce fungal growth parameters and mycotoxin levels, indicating the potential of this bacterium to control toxigenic fungi. The broad antifungal activity of this strain may represent a sustainable alternative for the exploration and subsequent use of its possible metabolites in order to control mycotoxin-producing fungi.

Comparative Analysis of How the Fecal Microbiota of Green-Winged Saltator (Saltator similis) Diverge among Animals Living in Captivity and in Wild Habitats.

The microbiota's alteration is an adaptive mechanism observed in wild animals facing high selection pressure, especially in captive environments. The objective of this study is to compare and predict the potential impact of habitat on the fecal bacterial community of Saltator similis, a songbird species that is a victim of illegal trafficking, living in two distinct habitats: wild and captivity. Nine wild and nine captive S. similis were sampled, and total bacterial DNA was obtained from the feces. Each DNA sample was employed to the amplification of the V4 region of the 16S rDNA following high-throughput sequencing. The most predominant phyla in all songbirds, irrespective of habitat, were Firmicutes, Bacteroidota, Proteobacteria, and Actinobacteriota. Interestingly, a microbiota profile (phylogenetic and abundance relationship) related to habitat was identified. The genera "Candidatus Arthromitus", Acinetobacter, Kocuria, and Paracoccus were exclusively identified in animals living in captivity, which can be a potential biomarker associated with birds in captive environments. This study presents the first description of the fecal bacterial community composition of S. similis living two different lifestyles. Finally, our results suggest that the lifestyle of S. similis birds significantly impacts the composition of the fecal microbiota. The animals living in captivity showed dysbiosis in the microbiota, with some bacteria genera being indicated as biological markers of environmental behavior. Thus, the present research provides a new concept of life quality measure for songbirds.

New candidate genes potentially involved in Zika virus teratogenesis.

Despite efforts to elucidate Zika virus (ZIKV) teratogenesis, still several issues remain unresolved, particularly on the molecular mechanisms behind the pathogenesis of Congenital Zika Syndrome (CZS). To answer this question, we used bioinformatics tools, animal experiments and human gene expression analysis to investigate genes related to brain development potentially involved in CZS. Searches in databases for genes related to brain development and CZS were performed, and a protein interaction network was created. The expression of these genes was analyzed in a CZS animal model and secondary gene expression analysis (DGE) was performed in human cells exposed to ZIKV. A total of 2610 genes were identified in the databases, of which 1013 were connected. By applying centrality statistics of the global network, 36 candidate genes were identified, which, after selection resulted in nine genes. Gene expression analysis revealed distinctive expression patterns for PRKDC, PCNA, ATM, SMC3 as well as for FGF8 and SHH in the CZS model. Furthermore, DGE analysis altered expression of ATM, PRKDC, PCNA. In conclusion, systems biology are helpful tools to identify candidate genes to be validated in vitro and in vivo. PRKDC, PCNA, ATM, SMC3, FGF8 and SHH have altered expression in ZIKV-induced brain malformations.

Exploring bacterial diversity and antimicrobial resistance gene on a southern Brazilian swine farm.

The bacterial composition of and the circulation of antimicrobial resistance genes (ARGs) in waste from Brazilian swine farms are still poorly understood. Considering that antimicrobial resistance (AMR) is one of the main threats to human, animal, and environmental health, the need to accurately assess the load of ARGs released into the environment is urgent. Therefore, this study aimed to characterize the microbiota in a swine farm in southern Brazil and the resistome in swine farm wastewater treated in a series of waste stabilization ponds (WSPs). Samples were collected from farm facilities and the surrounding environment, representing all levels of swine manure within the treatment system. Total metagenomic sequencing was performed on samples from WSPs, and 16S-rDNA sequencing was performed on all the collected samples. The results showed increased bacterial diversity in WSPs, characterized by the presence of Caldatribacteriota, Cloacimonadota, Desulfobacterota, Spirochaetota, Synergistota, and Verrucomicrobiota. Furthermore, resistance genes to tetracyclines, lincosamides, macrolides, rifamycin, phenicol, and genes conferring multidrug resistance were detected in WSPs samples. Interestingly, the most abundant ARG was linG, which confers resistance to the lincosamides. Notably, genes conferring macrolide (mphG and mefC) and rifamycin (rpoB_RIF) resistance appeared in greater numbers in the late WSPs. These drugs are among the high-priority antibiotic classes for human health. Moreover, certain mobile genetic elements (MGEs) were identified in the samples, notably tnpA, which was found in high abundance. These elements are of particular concern due to their potential to facilitate the dissemination of ARGs among bacteria. In summary, the results indicate that, in the studied farm, the swine manure treatment system could not eliminate ARGs and MGEs. Our results validate concerns about Brazil's swine production system. The misuse and overuse of antimicrobials during animal production must be avoided to mitigate AMR.

Genomic analysis on Brazilian strains of Anaplasma marginale / Análise genômica de cepas Brasileiras de Anaplasma marginale

Abstract Anaplasma marginale is a vector-borne pathogen that causes a disease known as anaplasmosis. No sequenced genomes of Brazilian strains are yet available. The aim of this work was to compare whole genomes of Brazilian strains of A. marginale (Palmeira and Jaboticabal) with genomes of strains from other regions (USA and Australia strains). Genome sequencing of Brazilian strains was performed by means of next-generation sequencing. Reads were mapped using the genome of the Florida strain of A. marginale as a reference sequence. Single nucleotide polymorphisms (SNPs) and insertions/deletions (INDELs) were identified. The data showed that two Brazilian strains grouped together in one particular clade, which grouped in a larger American group together with North American strains. Moreover, some important differences in surface proteins between the two Brazilian isolates can be discerned. These results shed light on the evolutionary history of A. marginale and provide the first genome information on South American isolates. Assessing the genome sequences of strains from different regions is essential for increasing knowledge of the pan-genome of this bacteria.

Resumo Anaplasma marginale é um patógeno transmitido por vetores que causam uma doença conhecida como anaplasmose. Até a presente data, não há genomas sequenciados de cepas brasileiras. O objetivo deste estudo foi comparar o genoma completo das cepas brasileiras de A. marginale (Palmeira e Jaboticabal) com os genomas de cepas de outras regiões (cepas dos EUA e Austrália). As sequências dos genomas das cepas brasileiras foram obtidas mediante sequenciamento de nova geração. As "reads" foram mapeadas usando-se como referência o genoma de A. marginale da cepa Florida. Foram identificados polimorfismos de nucleotídeo único (SNPs) e analisadas inserções/deleções (INDELs). As duas linhagens brasileiras se agruparam em um clado particular que, por sua vez, agrupou-se em um grupo maior junto com as linhagens norte-americanas. Além disso, foram identificadas diferenças significativas nas proteínas de superfície entre os dois isolados brasileiros. Esses resultados lançam luz sobre a história evolutiva de A. marginale e fornecem as primeiras informações de genomas de isolados sul-americanos. Avaliar as sequências de genomas de cepas de diferentes regiões é essencial para aumentar o conhecimento do pan-genoma dessa bactéria.

Tuberculosis outbreak in intensive swine farming from southern Brazil / Surto de tuberculose em suínos de criação intensiva no Sul do Brasil

ABSTRACT: Mycobacterium tuberculosis var. bovis is the etiologic agent of animal tuberculosis (aTB), a neglected zoonotic disease. Animal tuberculosis can affect many species, including swine. aTB-consistent granulomas in these animals lead to carcass disposal, generating economic losses and posing risks to human health. In the present study, an aTB outbreak was identified at an intensive swine farming operation in Southern Brazil. Inspection during swine slaughter revealed aTB-suspected lesions, which were collected for diagnosis by histology, PCR, and bacterial isolation. The animals had no clinical signs of tuberculosis. Granulomatous lesions were identified in 0.73% (59/8,071) of the slaughtered swine, and were confirmed by histology. Nine samples were further examined by PCR and bacterial isolation, with 44.4% and 55.5% positive results, respectively. Data from abattoirs subjected to federal surveillance show an aTB prevalence in Brazil of <0.001%. The present data thus indicate a swine aTB outbreak in intensive breeding. Swine infection can be related to exposure to infected animals or to contaminated food or environment. Biosecurity measures must be taken to avoid aTB transmission. Although certified swine breeding farms adopt such measures, this report indicates that constant monitoring is crucial, and greater control in swine breeding and finishing units is required to prevent outbreaks and spread of tuberculosis.

RESUMO: Mycobacterium bovis é o agente etiológico da tuberculose animal (aTB), uma doença zoonótica negligenciada. A tuberculose animal pode afetar muitas espécies, incluindo suínos. Os granulomas compatíveis com a aTB nesses animais levam ao descarte de carcaças, gerando perdas econômicas e trazendo riscos à saúde humana. No presente estudo, um surto de aTB foi identificado em um sistema de criação intensiva de suínos na região Sul do Brasil. A inspeção durante um abate de suínos revelou lesões suspeitas de tuberculose, as quais foram coletadas para diagnóstico por histologia, PCR e isolamento bacteriano. Os animais não apresentavam sinais clínicos de tuberculose. Lesões granulomatosas foram identificadas em 0,73% (59/8.071) dos suínos abatidos, e foram confirmadas pela histologia. Nove amostras foram posteriormente examinadas por PCR e isolamento bacteriano, com 44,4% e 55,5% de resultados positivos, respectivamente. Dados de frigoríficos submetidos à vigilância federal demostram prevalência da aTB de < 0,001% no Brasil. Os dados presentes indicam, portanto, um surto de aTB em suínos de criação intensiva. A infecção em suínos pode estar relacionada à exposição a animais infectados, alimentos ou ambientes contaminados. Medidas de biossegurança devem ser tomadas para evitar a transmissão da aTB. Embora as granjas suinícolas certificadas adotem tais medidas, este relato indica que o monitoramento constante é crucial, e maior controle nas unidades de criação e terminação de suínos é necessário para prevenir surtos e disseminação da tuberculose.

Valvular endocarditis associated with Helcococcus ovis in a cow in Southern Brazil / Endocardite vegetativa associada à Helcococcus ovis em uma vaca no Sul do Brasil

ABSTRACT: A 4-year-old cow with a history of breathing difficulty, progressive weight loss, and muffled heart sound was treated for a period of two weeks, but died and underwent necropsy examination. Macroscopic examination revealed heart with vegetative proliferative lesion firmly adhered to pulmonary valve, lungs with marked consolidation, and the presence of thrombus in vessels. There were grade II ulcers in abomasal mucosa associated to blood clots. Microscopic examination revealed marked thickening of the pulmonary valve due to the proliferation of fibrous connective tissue, inflammatory neutrophil infiltration, fibrin deposition, and a significant number of coccoid basophilic bacteria. Septic thromboemboli were present in the large and small pulmonary blood vessels suggestive of embolic pneumonia. The bacterial culture of the valve showed growth of small, nonhemolytic colonies that demonstrated satellitism to coagulase-negative staphylococci contaminating colonies, which were subjected to 16S gene sequencing and were compatible with Helcococcus ovis in GenBank. This was the first report of H. ovis endocarditis in cattle in South America.

RESUMO: Um bovino, fêmea, 4 anos com histórico de dificuldade respiratória, perda de peso progressiva e som cardíaco abafado, foi tratado por um período de duas semanas, porém veio a óbito e foi submetido a exame de necropsia. No exame macroscópico, notou-se coração com lesão proliferativa vegetativa firmemente aderida em valva pulmonar, pulmões com consolidação acentuada, e presença de trombos no interior de vasos. Havia ainda úlcera abomasal grau II em mucosa associada a coágulo sanguíneo. No exame microscópico notou-se acentuado espessamento da valva pulmonar por proliferação de tecido conjuntivo fibroso, infiltrado inflamatório de neutrófilos, deposição de fibrina e acentuado número de miríades bacterianas basofílicas cocoides. Tromboembolia séptica foi vista no interior de vasos pulmonares de pequeno e médio calibre, sugestivo de pneumonia embólica. Em cultivo bacteriano da valva notou-se crescimento de colônias pequenas, não hemolíticas que demonstravam satelitismo a colônias contaminantes de estafilococos coagulase negativa, essas foram submetidas ao sequenciamento do gene 16S e foram compatíveis com Helcococcus ovis no GenBank. Este foi o primeiro relato de endocardite por H. ovis em bovino na América do Sul.

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.

Genome sequencing of two Bacillus anthracis strains: a virulent strain and a vaccinal strain

ABSTRACT Bacillus anthracis strain SPV842_15 was isolated from bovine fetus, while B. anthracis strain Brazilian vaccinal was recovered from a commercial vaccine. We report here the genome sequences of both strains. The SPV842_15 genome is composed of a single circular chromosome with a length of 5,228,664 base pairs, and comprises 5911 coding sequences. In turn, the Brazilian vaccinal genome remains in 201 contigs with 5733 coding sequences. Both genomes have an overall C + G content of 35.4%, and 11 genes encoding the ribosomal RNAs (rRNAs) 5S, 16S and 23S. Only the plasmid pX01 sequence, which carries genes for toxins synthesis, was detected and completely assembled for both strains. These plasmids have a length of 181,684 base pairs and a C + G content of 32.5%. These genomic data generate insights about vaccinal B. anthracis virulence.

Long-term restoration of alpha-L-iduronidase activity in fibroblasts from patients with mucopolysaccharidosis type I after non-viral gene transfer

Mucopolysaccharidosis type I (MPS I) is a lysosomal storage disorder caused by deficiency of alpha-L-iduronidase (IDUA). Limitations such as the need for weekly injections, high morbidity and mortality, and high cost of current treatments show that new approaches to treat this disease are required. In this study, we aimed to correct fibroblasts from a patient with MPS I using non-viral gene therapy. Using a plasmid encoding the human IDUA cDNA, we achieved stable high IDUA levels in transfected fibroblasts up to 6 months of treatment. These results serve as proof of concept that a non-viral approach can correct the enzyme deficiency in cells of patients with lysosomal storage disorders, which can be used as a research tool for a series of disease aspects. Future studies should focus on showing if this approach can be useful in small animals and clinical trials (AU)