Assessment of the Virulence of the Burkholderia mallei Strain BAC 86/19 in BALB/c Mice.

Burkholderia mallei is an aerobic, Gram-negative, non-motile bacillus. As an obligate mammalian pathogen, it primarily affects solipeds. Although rarely transmitted to humans, the disease it causes, glanders, is classified as a zoonosis. The bacterium was officially eradicated in Brazil in 1969; however, it reemerged after three decades. This study aims to assess the virulence of a specific B. mallei strain, isolated in Brazil, in BALB/c mice through intranasal infection. The strain, B. mallei BAC 86/19, was obtained from the tracheal secretion of a young mare displaying positive serology but no clinical signs of glanders. Post-mortem examinations revealed macroscopic lesions consistent with the disease, however. In mice, the LD50 was determined to be approximately 1.59 × 105 colony-forming units (CFU)/animal. Mice exposed to either 0.1 × LD50 or 1 × LD50 displayed transient weight loss, which resolved after three or five days, respectively. B. mallei persisted within the liver and lung for five days post-infection and in the spleen for seven days. These findings underscore the detectable virulence of the Brazilian B. mallei BAC 86/19 strain in mice, which are relatively resilient hosts. This research points to the importance of the continued investigation of the virulence mechanisms and potential countermeasures associated with B. mallei infections, including their Brazilian isolates.

Glanders Diagnosis in an Asymptomatic Mare from Brazil: Insights from Serology, Microbiological Culture, Mass Spectrometry, and Genome Sequencing.

This manuscript elucidates the occurrence of glanders in an asymptomatic mare from Brazil presenting positive Burkholderia mallei antibody titers. The diagnosis was established through a multi-pronged approach encompassing microbiological culture, mass spectrometry, and genome sequencing. The outbreak occurred in 2019 in Tatuí, São Paulo, Brazil, and the infected mare, despite displaying no clinical symptoms, had multiple miliary lesions in the liver, as well as intense catarrhal discharge in the trachea. Samples were collected from various organs and subjected to bacterial isolation, molecular detection, and identification. The strain was identified as B. mallei using PCR and confirmed by MALDI-TOF mass spectrometry. Whole-genome sequencing revealed a genome size of 5.51 Mb with a GC content of 65.8%, 5871 genes (including 4 rRNA and 53 tRNA genes), and 5583 coding DNA sequences (CDSs). Additionally, 227 predicted pseudogenes were detected. In silico analysis of different genomic loci that allow for differentiation with Burkholderia pseudomallei confirmed the identity of the isolate as B. mallei, in addition to the characteristic genome size. The BAC 86/19 strain was identified as lineage 3, sublineage 2, which includes other strains from Brazil, India, and Iran. The genome sequencing of this strain provides valuable information that can be used to better understand the pathogen and its epidemiology, as well as to develop diagnostic tools for glanders.

Molecular detection of Burkholderia mallei in different geographic regions of Brazil.

Glanders is a contagious disease of equids caused by the Gram-negative bacterium Burkholderia mallei. In Brazil, the disease is considered to be reemerging and has been expanding, with records of equids with positive serology in most of the federative units. However, there are few reports describing the genotypic detection of the agent. This study demonstrated the detection of B. mallei by species-specific PCR directly from tissues or from bacterial cultures, followed by amplicon sequencing in equids (equines, mules, and asinines) with positive serology for glanders in all five geographic regions of Brazil. The molecular evidence of B. mallei infection in serologically positive equids in this study expands the possibility of strain isolation and the conduction of epidemiological characterizations based on molecular information. The microbiological detection of B. mallei in cultures from nasal and palate swabs, even in equids without clinical manifestations, raises the possibility of environmental elimination of the agent.

Morphological and molecular diagnosis of Pseudoterranova decipiens (sensu stricto) (Anisakidae) in imported cod sold in Brazil.

An anisakid nematode larva found in cod sold in the state of Minas Gerais, Brazil was studied by light and scanning electron microscopy and by a molecular approach. Mitochondrial cytochrome c-oxidase subunit 2 (mtDNA cox-2), 28S rRNA and ITS1, 5.8S and ITS2 regions were amplified using the polymerase chain reaction and sequenced to evaluate the phylogenetic relationships of the larva. The genetic profile confirmed that this larva belongs to the species Pseudoterranova decipiens (sensu stricto). This is the first molecular and ultrastructural study of Pseudoterranova decipiens (sensu stricto) in imported cod sold in Brazil. The health implications of these findings are discussed.

Morphological and molecular diagnosis of Pseudoterranova decipiens (sensu stricto) (Anisakidae) in imported cod sold in Brazil / Diagnose morfológica e molecular de Pseudoterranova decipiens (sensu stricto) (Anisakidae) em bacalhau importado vendido no Brasil

An anisakid nematode larva found in cod sold in the state of Minas Gerais, Brazil was studied by light and scanning electron microscopy and by a molecular approach. Mitochondrial cytochrome c-oxidase subunit 2 (mtDNA cox-2), 28S rRNA and ITS1, 5.8S and ITS2 regions were amplified using the polymerase chain reaction and sequenced to evaluate the phylogenetic relationships of the larva. The genetic profile confirmed that this larva belongs to the species Pseudoterranova decipiens (sensu stricto). This is the first molecular and ultrastructural study of Pseudoterranova decipiens (sensu stricto) in imported cod sold in Brazil. The health implications of these findings are discussed.

Uma larva de Nematoda anisakídeo encontrada em bacalhau comercializado no Estado de Minas Gerais, Brasil foi estudada por microscopias de luz e eletrônica de varredura e por uma abordagem molecular. As regiões da subunidade 2 da citocromo c-oxidase mitocondrial (mtDNA cox-2), 28S rRNA e ITS1, 5.8S e ITS2 foram amplificadas usando a reação em cadeia da polimerase e sequenciadas para avaliar as relações filogenéticas da larva. O perfil genético confirmou que esta larva pertence à espécie Pseudoterranova decipiens (sensu stricto). Esse é o primeiro estudo molecular e ultraestrutural de Pseudoterranova decipiens (sensu stricto) de bacalhau importado vendido no Brasil. As implicações destes resultados para a saúde são discutidas.

Occurrence of Ehrlichia canis in free-living primates of the genus Callithrix.

Bacteria of the genus Ehrlichia are Gram-negative and coccoid-shaped microorganisms that cause ehrlichiosis - a serious infectious disease that often leads to death. These bacteria present a strong zoonotic potential and primates may act as reservoir hosts. This study involved a molecular analysis to detect these microorganisms in blood samples collected from nineteen primates of the genus Callithrix living free in an Atlantic Forest fragment in the municipality of Viçosa, state of Minas Gerais, Brazil. One of the 19 primates was found to be infected with Ehrlichia canis. This finding points to a new wild host of E. canis with a strong potential for transmission to humans because of its increasing contact with people. This is the first report of Ehrlichia spp. in primate of the genus Callithrix.

Occurrence of Ehrlichia canis in free-living primates of the genus Callithrix / Ocorrência de Ehrlichia canis em primatas de vida livre pertencentes ao gênero Callithrix

Bacteria of the genus Ehrlichia are Gram-negative and coccoid-shaped microorganisms that cause ehrlichiosis a serious infectious disease that often leads to death. These bacteria present a strong zoonotic potential and primates may act as reservoir hosts. This study involved a molecular analysis to detect these microorganisms in blood samples collected from nineteen primates of the genus Callithrix living free in an Atlantic Forest fragment in the municipality of Viçosa, state of Minas Gerais, Brazil. One of the 19 primates was found to be infected with Ehrlichia canis. This finding points to a new wild host of E. canis with a strong potential for transmission to humans because of its increasing contact with people. This is the first report of Ehrlichia spp. in primate of the genus Callithrix.

Bactérias do gênero Ehrlichia são gram-negativas em forma de cocos. Provocam uma doença infecciosa grave denominada erliquiose que, muitas vezes, causa morte. Essas bactérias apresentam grande potencial zoonótico, e os primatas podem agir como reservatórios. Este estudo objetivou a detecção molecular desse microorganismo em amostras de sangue coletadas de dezenove primatas de vida livre num fragmento de Mata Atlântica pertencentes ao gênero Callithrix, no município de Viçosa, Minas Gerais, Brasil. Entre os 19 espécimes de sagui, um estava infectado com Ehrlichia canis. Esse achado aponta para um novo hospedeiro selvagem para E. canis com grande potencial de transmissão ao homem, devido ao seu crescente contato com pessoas. Este é o primeiro relato de primata do gênero Callithrix infectado com Ehrlichia spp.