Five centuries of genome evolution and multi-host adaptation of Campylobacter jejuni in Brazil.

Consumption of raw, undercooked or contaminated animal food products is a frequent cause of Campylobacter jejuni infection. Brazil is the world's third largest producer and a major exporter of chicken meat, yet population-level genomic investigations of C. jejuni in the country remain scarce. Analysis of 221 C. jejuni genomes from Brazil shows that the overall core and accessory genomic features of C. jejuni are influenced by the identity of the human or animal source. Of the 60 sequence types detected, ST353 is the most prevalent and consists of samples from chicken and human sources. Notably, we identified the presence of diverse bla genes from the OXA-61 and OXA-184 families that confer beta-lactam resistance as well as the operon cmeABCR related to multidrug efflux pump, which contributes to resistance against tetracyclines, macrolides and quinolones. Based on limited data, we estimated the most recent common ancestor of ST353 to the late 1500s, coinciding with the time the Portuguese first arrived in Brazil and introduced domesticated chickens into the country. We identified at least two instances of ancestral chicken-to-human infections in ST353. The evolution of C. jejuni in Brazil was driven by the confluence of clinically relevant genetic elements, multi-host adaptation and clonal population growth that coincided with major socio-economic changes in poultry farming.

Multi-virulence of Campylobacter jejuni carried by chicken meat in Brazil.

Campylobacter jejuni is the most frequent cause of bacterial gastroenteritis; therefore, the characteristics of its epidemiology must be continuously investigated to support possible mitigating measures. This is particularly important when evaluating representative strains from the world's leading chicken meat exporter, Brazil. We evaluated a panel of 14 virulence genes in 359 strains of C. jejuni isolated from chilled broiler carcasses in Brazil. The genes were classified into five virulence categories (B: biofilm/motility; SS: secretion/cytotoxicity system; CI: invasion/colonization; GB: Guillain-Barré; and AE: adaptation to stress). The percentage of strains with stress adaptation genes (86.07%) indicates the ability to survive in unfavorable environments; in addition, the strains showed a risk of causing infections in humans due to the frequency of the hcp gene (97.77%). Genes related to Guillain-Barre syndrome (GBS) in 77.44% of strains are an additional concern, which must be monitored. The gene panel showed the presence of 124 virulence profiles. Individual analyses by carcass, slaughter establishment, and municipalities in which they were located showed high index variabilities (I.Var.) of 0.82, 0.87, and 0.78, respectively. Georeferencing indicated the state of Paraná as a hotspot for virulent strains. Higher levels of isolation and multi-virulence were identified in the summer, which is hot and humid in Brazil. Together, our results showed that the studied strains are a potential danger to public health and that there is an urgent need for their surveillance and the adoption of control measures, especially in the state of Paraná.

Survey for Leptospira and Brucella in Amazonian manatees, Amazon river dolphins, and a tucuxi in the Brazilian Amazon.

Leptospirosis and brucellosis are zoonotic diseases with global distributions that represent severe hazards to humans and animals. We investigated exposure to Leptospira spp. and Brucella spp. in samples from Amazonian manatees Trichechus inunguis, Amazon river dolphins Inia geoffrensis, and a tucuxi Sotalia fluviatilis. The animals were free-ranging or undergoing in situ rehabilitation in the mid-Solimões River region, Brazilian Amazon. Serum samples from 19 Amazonian manatees were tested by microscopic agglutination test, Rose Bengal test, and 2-mercaptoethanol Brucella agglutination test. Antibodies against Leptospira spp. were detected in 63% of the manatees tested and serovar Patoc was considered the infecting serovar in all positive samples. Titers were generally low, indicating chronic exposure, but higher titers indicative of an active infection were detected in 3 animals. Anti-Brucella spp. antibodies were not detected. Tissue and/or body fluid samples from 12 Amazon river dolphins, a tucuxi, and 2 Amazonian manatees were investigated by multiplex PCR and bacteriology for Leptospira spp. and Brucella spp. All samples were negative. However, Enterococcus faecalis was isolated from uterine fluid, lymph node, and lung of 3 Amazon river dolphins. Bacillus spp. were isolated from milk and synovial fluid from 2 Amazon river dolphins and from a milk sample from 1 Amazonian manatee. Knowledge of the pathogens present in Amazonian manatees, Amazon river dolphins, and tucuxis is of great relevance to species conservation and environmental health. Although no clinical signs were noted, further research is needed to elucidate the clinical relevance of infection by Leptospira sp. serovar Patoc in Amazonian aquatic mammals.

Evaluation of response to anti-leptospira bacterin vaccination in pregnant ewes and the passive transfer of antibodies to their offspring / Avaliação da resposta à vacinação com bacterina anti-leptospira em ovelhas gestantes e a transferência passiva de anticorpos a suas crias

Leptospirosis in sheep is often underestimated, and leads to great economic losses for the sheep farming industry. The aim of this study was to evaluate the humoral immune response in pregnant ewes, after the injection of a commercial polyvalent vaccine for leptospirosis, and to observe the transmission of anti-Leptospira antibodies through the colostrum to the offspring. For this, 24 pregnant ewes were vaccinated for leptospirosis. Blood samples were collected prior to vaccination and then 7, 14, 21, 28, 35, 42 and 49 days after vaccination. In order to evaluate passive immunity transfer, blood samples of 32 lambs were collected during the first 48 hours after birth, and another collection was performed 10 to 21 days after birth. The lambs were placed into 2 groups: Group A (n=16): singleton lambs; and group B (n=16): twins. The sera samples were submitted to the Microscopic Agglutination Test (MAT), in which 21 Leptospira serovars were tested. The results were analyzed in a descriptive form. The number of sheep reactive to MAT gradually increased until 21 days after vaccination, and decreased right after. Of all the serovars contained in the vaccine, the largest proportion of animals were seroconverted to Hardjoprajtino serovar, Serjoe serogroup. Anti-Leptospira antibodies transferred through colostrum to lambs were detected by MAT in the serum collected 24-48 hours after birth. It was observed that 65.6% (21 out of 32) of the lambs were reactive. In the subsequent collections that occurred from 10 to 21 days after birth, a decrease in the number of animals reactive to the MAT was detected. There was no significant statistical difference for the passive transfer of antibodies between single or twin lambs.

A leptospirose é uma doença frequentemente subestimada em rebanhos ovinos e leva a grandes prejuízos à ovinocultura. O objetivo deste estudo foi avaliar a resposta imune humoral em ovelhas gestantes, após imunização com uma vacina polivalente comercial contra leptospirose, bem como verificar a transmissão dos anticorpos adquiridos às crias, pelo colostro. Para isto, 24 ovelhas gestantes foram vacinadas. Amostras de sangue foram coletadas pré-imunização, bem como 7, 14, 21, 28, 35, 42 e 49 dias após. Para avaliar a transferência passiva de anticorpos, os 32 cordeiros que nasceram dessas ovelhas foram amostrados nas primeiras 48 horas após o nascimento, bem como com 10 e 21 dias de vida. Os cordeiros foram divididos em dois grupos: partos simples (Grupo A; n=16) e partos gemelares (Grupo B, n= 16). O soro sanguíneo foi submetido à prova de Soroaglutinação Microscópica (MAT), na qual 21 sorovares de Leptospira foram testados. O número de ovelhas reativas à MAT aumentou gradualmente até os 21 dias após imunização, com posterior decréscimo. De todos os sorovares presentes na vacina, a maior parte dos animais soroconverteu para o sorovar Hardjoprajtino, do sorogrupo Serjoe. Anticorpos anti-Leptospira passaram pelo colostro e puderam ser detectados no soro dos cordeiros entre 24 e 48 horas após o nascimento. Foi observado que 65,5% (21 de 32) cordeiros foram reativos. Em coletas subsequentes, realizadas 10 e 21 dias após o nascimento, houve um decréscimo no número de animais reativos à MAT. Não houve diferença significativa na transferência de anticorpos entre cordeiros de partos simples e gemelares.

Consumption of minas frescal cheese may be a source of human infection by Campylobacter jejuni / Consumo de queijo minas frescal pode ser fonte de infecção humana por Campylobacter jejuni

Campylobacter spp. is an emerging pathogen that causes gastroenteritis in humans and the consumption of dairy food can characterize sources of infection. We aimed to verify the viability and a presence of transcripts associated with characteristics of virulence and adaptation of C. jejuni isolated from Minas Frescal cheeses, produced with contaminated milk and stored under refrigeration for up to ten days. The samples were analyzed for bioindicators, Campylobacter spp., pH, acidity, moisture and sodium chloride. Campylobacter spp. recovered were evaluated for the production of transcripts of: ciaB, dnaJ, p19 and sodB. The results were correlated with the viability of C. jejuni and changes in their transcriptome. Storage at lowtemperatures reduced C. jejuni from the first to the fourth day. The variations in humidity, pH and acidity influenced the decreasing of C. jejuni. There was a reduction in transcripts' production of the four genes, more pronounced on the fourth day, indicating the inability of the microorganism to perform its metabolic activities, due to the conditions of injury. Despite the presence of mechanisms of virulence and adaptation, C. jejuni could not remain viable four days after production. However, consumption of fresh cheese contaminated with Campylobacter jejuni can be a source of infection when consumed up to four days after production.

Campylobacter spp. é um patógeno emergente que causa gastroenterite em seres humanos e o consumo de produtos lácteos pode caracterizar fontes de infecção. O objetivo deste estudo foi verificar a viabilidade e a presença de transcritos associadas a características de virulência e adaptação de C. jejuniisoladas de queijos frescos, produzidos com leite contaminado e mantidos refrigeradas por dez dias. Foram analisados bioindicadores, Campylobacter spp., pH, acidez, umidade e cloreto de sódio. Campylobacter spp. recuperados foram avaliados quanto à produção dos transcritos: ciaB, dnaJ, p19 e sodB. Os resultados foram correlacionados com a viabilidade de C. jejuni e alterações no transcriptoma. O armazenamento em baixas temperaturas reduziu C. jejuni do primeiro ao quarto dia. As variações na umidade, pH e acidez influenciaram a queda de C. jejuni. Houve uma redução na produção de transcritos dos quatro genes, mais pronunciada no quarto dia, indicando a incapacidade do micro-organismo em realizar suas atividades metabólicas, devido às condições de injúria. Apesar da presença de mecanismos de virulência e adaptação, C. jejuni não permaneceu viável quatro dias após a produção. Porém, o consumo de queijo fresco contaminado com Campylobacter jejunipode ser uma fonte de infecção quando consumido até quatro dias após a produção.