Immunoproteomic and immunoinformatic approaches identify secreted antigens and epitopes from Staphylococcus saprophyticus.

Urinary tract infections (UTIs) are common human infections that compromise women's health around the world, even though they can affect men and women of all ages. Bacterial species are the primary causative agents of UTIs, while Staphylococcus saprophyticus, a gram-positive bacterium, is especially important for uncomplicated infections in young women. Despite the number of antigenic proteins identified in Staphylococcus aureus and other bacteria of the genus, there is no immunoproteomic study in S. saprophyticus. In this context, since pathogenic microorganisms secrete important proteins that interact with hosts during infection, the present work aims to identify the exoantigens from S. saprophyticus ATCC 15305 by immunoproteomic and immunoinformatic approaches. We identified 32 antigens on the exoproteome of S. saprophyticus ATCC 15305 by immunoinformatic tools. By using 2D-IB immunoproteomic analysis, it was possible to identify 3 antigenic proteins: transglycosylase IsaA, enolase and the secretory antigen Q49ZL8. In addition, 5 antigenic proteins were detected by immunoprecipitation (IP) approach, where the most abundant were bifunctional autolysin and transglycosylase IsaA proteins. The transglycosylase IsaA was the only protein detected by all the tools approaches used in this study. In this work it was possible to describe a total of 36 S. saprophyticus exoantigens. Immunoinformatic analysis allowed the identification of 5 exclusive linear B cell epitopes from S. saprophyticus and 5 epitopes presenting homology with other bacteria that cause UTIs. This work describes, for the first time, the profile of exoantigens secreted by S. saprophyticus and can contribute to the identification of new diagnostic targets of UTIs, as well as to develop vaccines and immunotherapies against bacterial urinary infections.

The exoproteome profiles of three Staphylococcus saprophyticus strains reveal diversity in protein secretion contents.

Staphylococcus saprophyticus is a gram-positive microorganism responsible for urinary tract infections (UTIs). Although some virulence factors are characterized, such as urease, autolysins, adhesins and hemagglutinins, large-scale proteomic studies have not been performed within this species. We performed the characterization of the exoproteome from three S. saprophyticus strains: the reference strain ATCC 15,305, a non-capsular strain 7108 and the 9325 strain containing a thick capsule which were cultured in BHI medium and culture supernatants were analysed by using mass spectrometry approach. We observed a core of 72 secreted proteins. In addition, it was possible to detect diversity in the protein profiles of the exoproteomes. Interestingly, strain 7108 presented no secretion of three antigenic proteins, including the classical SsaA antigen. In addition, the level of antigenic proteins secreted by strain 9325 was higher than in ATCC 15,305. This result was confirmed by Western blot analysis using anti-SsaA polyclonal antibodies, and no production/ secretion of SsaA was detected in strain 7108. Transcriptional data shows that 7108 strain produces transcripts encoding SsaA, suggesting post-transcriptional regulation occurs in this strain. Moreover, when compared with the other strains that were analyzed, it was possible to detect higher levels of proteases secreted by strain 7108 and higher levels of antigenic proteins and transglycosylases secreted by 9325 strain. The results reveal diversity in protein secretion among strains. This research is an important first step towards understanding the variability in S. saprophyticus exoproteome profile and could be significant in explaining differences among strains.

Early development of two commercially valuable fish from the lower Amazon River, Brazil (Characiformes: Serrasalmidae)

We described the early development of Myloplus asterias and M. lobatus, two fish species of the Serrasalmidae using morphological, meristic, and morphometric characteristics. These herbivores serrasalmids are heavily fished because of their commercial importance in the Amazon. The individuals were collected between 2016 and 2020 in the limnetic zone of open water and macrophyte stands in the Lower Amazon River floodplain. We tested different growth models to identify the development pattern of these species. During the initial ontogeny, these two species can be differentiated mainly by the myomeres total number, 37 to 39 in M. asterias vs. 40 to 41 in M. lobatus, the pigmentation pattern, and dorsal-fin ray numbers. These characteristics are effective in distinguishing these species from other serrasalmids. The morphometric relationships were also different between these two species, showing distinct patterns in growth between the analyzed features. An identification key for larvae of some sympatric Serrasalmidae species from the Amazon basin is presented. Our expectation is that this study will contribute to the knowledge of the initial ontogeny and the biology of fish species in the Neotropical region.(AU)

Descrevemos o desenvolvimento inicial de Myloplus asterias e M. lobatus, duas espécies de peixes da família Serrasalmidae usando características morfológicas, merísticas e morfométricas. Esses serrasalmídeos herbívoros são fortemente pescados devido à sua importância comercial na Amazônia. Os indivíduos foram coletados entre 2016 e 2020 na zona limnética de águas abertas e em bancos de macrófitas na planície de inundação do baixo rio Amazonas. Testamos diferentes modelos de crescimento para identificar o padrão de desenvolvimento dessas espécies. Durante a ontogenia inicial, essas duas espécies podem ser diferenciadas pelo número total de miômeros; 37 a 39 em M. asterias vs. 40 a 41 em M. lobatus; pelo padrão de pigmentação e número de raios da nadadeira dorsal. Essas características são eficazes para distinguir essas espécies de outros serrasalmídeos. As relações morfométricas também foram diferentes entre as duas espécies, com distintos padrões de crescimento entre as características analisadas. Uma chave de identificação para larvas de algumas espécies simpátricas de Serrasalmidae da bacia amazônica é apresentada. Nossa expectativa é que este estudo contribua para o conhecimento da ontogenia inicial e da biologia das espécies de peixes da região Neotropical.(AU)

Larvae of migratory Characiformes species in an archipelago in the Lower Amazon River

In the Amazon basin, most of the migratory Characiformes species represent an important fishing resource for local people. However, the lack of information about the main areas and periods of reproduction and the importance of certain environments for the maintenance of species may jeopardize the renewal of fish stocks. Thus, the aim was to examine the spatial and seasonal variation in the density of larvae of migratory Characiformes species in an archipelago in the Lower Amazon River and to assess the importance of this environment for the biological recruitment of the studied species. The capture of larvae was carried out in places close to restinga swamps, steep ruts,and inlet areas with a plankton net (300 μm) in monthly sampling from January to December 2013. An analysis of variance was used to verify differences in larval density between seasons and sampling months. A total of 30,997 larvae were captured and showed no significant differences in their distributions between habitats. However, a variationbetween the phases of the hydrological cycle was evident, in which the highest larval concentrations were observed during the flooding (17.72 larvae.10m-3) and the drought (1.33 larvae.10m-3). The study area has a great capacity to assist in the maintenance and renewal of regional fish stocks, as it is an important retention and nursery site for larvae of Characiformes that drift along the main channel of the Amazon River.(AU)

Larvae of migratory Characiformes species in an archipelago in the Lower Amazon River

In the Amazon basin, most of the migratory Characiformes species represent an important fishing resource for local people. However, the lack of information about the main areas and periods of reproduction and the importance of certain environments for the maintenance of species may jeopardize the renewal of fish stocks. Thus, the aim was to examine the spatial and seasonal variation in the density of larvae of migratory Characiformes species in an archipelago in the Lower Amazon River and to assess the importance of this environment for the biological recruitment of the studied species. The capture of larvae was carried out in places close to restinga swamps, steep ruts,and inlet areas with a plankton net (300 μm) in monthly sampling from January to December 2013. An analysis of variance was used to verify differences in larval density between seasons and sampling months. A total of 30,997 larvae were captured and showed no significant differences in their distributions between habitats. However, a variationbetween the phases of the hydrological cycle was evident, in which the highest larval concentrations were observed during the flooding (17.72 larvae.10m-3) and the drought (1.33 larvae.10m-3). The study area has a great capacity to assist in the maintenance and renewal of regional fish stocks, as it is an important retention and nursery site for larvae of Characiformes that drift along the main channel of the Amazon River.