This work evaluated the effects of dietary supplementation of A-Live (phytogenic) either individually or in combination with Aquaform (potassium diformate, acidifier) on juvenile Nile tilapia (Oreochromis niloticus) growth performance, innate immune parameters, gut microbiome, and resistance against Francisella noatunensis subsp. orientalis challenge. Each experimental group contained 140 fishes (34.3 ± 0.33) in two 150L tanks. The experimental design consisted of five groups: a negative control; treated groups (G1, G2, G3) supplemented with different concentrations of A-Live and Aquaform in the feed; and a positive control (PC) for pathogen infection. Groups G1, G2, G3, and PC were challenged with Francisella spp. after 15 days. After infection, the mortality was significantly lower in groups G1, G2, and G3 (p < 0.01). Furthermore, these groups showed significant increase (p < 0.05) in daily weight gain, feed conversion rate, and specific growth rate. The PC group presented increase (p < 0.05) in the leukocytes and neutrophils number. Innate immunity parameters showed no difference between treatments after infection. Microbiome analysis revealed an increased number of bacteria belonging to the Vibrionaceae family after pathogen infection suggesting a secondary pathogen function of these bacteria. These results validate the beneficial effects of these products in tilapia farming.

Research background. Despite the great properties of bacterial cellulose, its manufacture is still limited due to difficulties in large-scale production. These problems are mainly related to low production yields and high overall costs of the conventional culture media normally used. To surpass these problems, it is necessary to identify new cheap and sustainable carbon sources. Thus, this work aims to isolate and select a high cellulose-producing Komagataeibacter strain from vinegar industry, and study its potential for bacterial cellulose synthesis in an industrial soybean co-product, known as soybean molasses, used as fermentation medium. Experimental approach. One isolated strain was able to produce high amount of cellulose in the standard Hestrin-Schramm medium, so we tested its ability to produce this biopolymer in a soybean molasses medium. The characteristics and properties of the produced bacterial cellulose membranes were analyzed by thermogravimetric analysis, X-ray diffraction, infrared spectroscopy, water-holding capacity and rehydration ratio. Genetic analysis of the selected strain served to determine its genus and species. Results and conclusions. An isolated strain that produced the highest amount of cellulose in Hestrin-Schramm medium (3.7 g/L) was genetically identified as Komagataeibacter intermedius V-05. This strain produced 10.0 g/L of cellulose in soybean molasses medium. Membranes from both substrates had similar chemical structure, crystallinity and thermal degradation. Soybean molasses proved to be a suitable alternative medium for biosynthesis of cellulose in comparison with the standard medium. In addition to providing higher production yield, the membranes showed great structural characteristics, similar to those obtained from standard medium. Novelty and scientific contribution. In this research, we have isolated and identified a Komagataeibacter strain which exhibits a high capacity for cellulose production in soybean molasses. The isolation and selection of strains with high capacity for microbial metabolite production is important for decreasing bioprocess costs. Furthermore, as there is a necessity today to find cheaper carbon sources to obtain microbial products at a lower cost, soybean molasses represents an interesting alternative medium to produce bacterial cellulose for its industrial application.

The serovar Hardjo belongs to the serogroup sejroe and this serovar is the most prevalent in bovine herds worldwide. The sejroe serogroup is the most frequently detected by serology in Brazilian cattle herds suggesting that it is due serovar Hardjo. In the molecular classification, this serovar has two genotypes: Hardjobovis and Hardjoprajitno. This serovar is as considered as fastidious pathogens, and their isolatin is one of the bottlenecks in leptospirosis laboratories. In addition, its molecular characterization using genomic approaches is oftentimes not simple and time-consuming. This study describes a method for isolating the two genotypes of serovar Hardjo using culture medium formulations and suggests a get-at-able molecular characterization. Ten cows naturally infected which were seropositive were selected from small dairy farms, and their urine was collected for bacterial isolation. We evaluated three modifications of liquid Leptospira medium culture supplemented with sodium pyruvate, superoxide dismutase enzyme and fetal bovine serum, and the isolates were characterized by molecular techniques. After isolation and adaptation in standard culture medium, the strains were subcultured for one week in the three modified culture media for morphologic evaluation using electronic microscopy. Strains were molecularly identified by multilocus variable-number tandem-repeat analysis (MLVA), partial sequencing and phylogenic analyses of gene sec Y. Combining the liquid culture medium formulations allowed growth of the Leptospira serovar Hardjo in three tubes. Two isolates were identified as genotype Hardjobovis, and the other as genotype Hardjoprajitno. Morphologically, compared with control media, cells in the medium supplemented with the superoxide dismutase enzyme were more elongated and showed many cells in division. The cells in the medium supplemented with fetal bovine serum were fewer and lost their spirochete morphology. This indicated that the additional supplementation with fetal bovine serum assisted in the initial growth and maintenance of the viable leptospires and the superoxide dismutase enzyme allowed them to adapt to the medium. These culture strategies allowed for the isolation and convenient molecular characterizationof two genotypes of serovar Hardjo, creating new insight into the seroepidemiology of leptospirosis and its specific genotypes. It also provides new information for the immunoprophylaxis of bovine leptospirosis.

Streptococcus agalactiae is an important pathogen to world aquaculture due to its high mortality rates in fish farms and consequent economic losses. Our study presents the complete genome sequence of strain S13, isolated from a tilapia farm outbreak in southern Brazil.

Most epidemiologic studies on bovine leptospirosis are based on serological tests that use antibodies against several serotypes, including the serovar Hardjo, which is widespread and considered to be the most adapted to bovine hosts. However, using only serological studies is not sufficient to identify and distinguish species of leptospires. The aim of this study was report the first isolation in Brazil of two strains serovar Hardjo obtained in urine samples from naturally infected cows in a small Brazilian dairy herd and find the genetic species and consequently the type strain Hardjobovis by molecular characterization. Fifteen dairy cows with a history of reproductive failure, such as abortion and infertility, were selected. Urine samples obtained from each animal were immediately seeded in tubes containing Ellinghausen-McCullough-Johnson-Harris culture medium. The identification of the isolates was performed by Multilocus variable-number tandem-repeat analysis (MLVA) technique and phylogenetic analysis of partial sequence of gene sec Y. From the 15 urine samples evaluated, two Leptospira were found and identified as the Londrina 49 and Londrina 54 strains. The MLVA profiles and sequencing of gene sec Y characterized the isolates as L. borgpetersenii serovar Hardjo strain Hadjobovis because it has different genetic pattern of Leptospira interrogans serovar Hardjo strain Hardjoprajitno. Therefore, more studies are needed including isolation and molecular characterization from regional strains to obtain a better knowledge about epidemiology of serovar Hardjo in bovine which may assist in future strategies of prevention and control of bovine leptospirosis.