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Ornamental fish are becoming increasingly popular, but the lack of knowledge regarding their various diseases is a major challenge. Skin diseases commonly found in freshwater fish include black spot disease (BSD), which is characterized by melanin deposits around the metacercariae of some trematode species. Since BSD remains poorly understood, this study describes an outbreak of BSD in Etroplus maculatus raised in outdoor ponds at a Brazilian fish farm. Metacercariae samples were collected, examined, and subjected to molecular phylogenetic analysis. The parasites were conspecific to an unnamed species, Crassiphiala lineage 5, recently found in Brazilian birds (Megaceryle torquata). Sequences obtained for longifurcate cercariae of the planorbid snail Biomphalaria straminea from the same region were identical to our metacercariae of Crassiphiala sp. These results suggest that Biompahalaria snails are likely an intermediate host of this parasite on farms where E. maculatus was found to be infected. We provide the first molecular evidence that Crassiphiala are the causative agents of BSD in fish from Brazil. Combatting snails and preventing access of fish-eating birds to outdoor ponds are strategies to control this disease in ornamental fish farms.

Francisella orientalis infections, known as francisellosis, are one of the most important diseases affecting the production of Nile tilapia, causing high mortality rates in the most susceptible fish stages: fingerlings and juveniles. Antibiotic therapy is the method of choice for treating the disease, as there are no commercially available vaccines. In this study, we developed an inactivated whole-cell vaccine using an isolate of F. orientalis in combination with the aqueous adjuvant Montanide IMS 1312 VG, which was administered to Nile tilapia through immersion. Two immunization trials (1 and 2) were conducted with fish at the fingerling and juvenile stages. For each trial, five different experimental groups were established: a complete vaccine (bacterin in combination with aqueous adjuvant), bacterin, aqueous adjuvant, and positive and negative controls. Thirty days after vaccination, an experimental challenge was performed through intraperitoneal injection of the same F. orientalis isolate. As a result, the vaccinated fingerlings were the only group in which mortality and progression of clinical signs of francisellosis were statistically significantly reduced, although relative percentage of survival (RPS) was low at 50%. In the juvenile group, RPS was higher at 63%, but not statistically significant. Nevertheless, an RPS of only 50% is acceptable for using vaccines in the field. The bacterin and adjuvant treatments alone were not effective, showing an RPS of 37% and 0%, respectively. Post-vaccination mortality was observed in the group exposed only to the adjuvant, which may indicate excessive immune stimulation at this stage. Interestingly, the immune response elicited by the vaccine was unable to eliminate the pathogen from the host; therefore, the surviving animals became carriers. Although the immune response elicited by the vaccine was unable to eliminate the pathogen from the host, this vaccine formulation could be a viable alternative for use in the field and serve as another means of controlling the mortality caused by the pathogen. Our study provides the first report of vaccination, using immersion, against francisellosis at the most susceptible stages of farmed Nile tilapia. Future studies should address the efficiency of immersion vaccines under field conditions.

In 2016 and 2017, we characterized outbreaks caused by Streptococcus agalactiae serotype III sequence type (ST) 283 in Nile tilapia farms in Brazil. Whole-genome multilocus sequence typing clustered the fish isolates together with the zoonotic ST283 and other STs related to cases in humans, frogs, dogs, cattle, and dolphins.

This study aimed to evaluate the influence of increasing concentrations of dietary dry oregano leaves (DOL), ranging from 0 to 0.15%, on different performance parameters of Nile tilapia. Hence, two separate experimental trials testing growth factors (Trial 1) and innate immunity and disease resistance against Streptococcus agalactiae infection (Trial 2) were conducted with a total of 464 tilapia juveniles and two to four replicates. As a result, dietary inclusion of DOL did not induce significant effects on growth performance, hematological parameters, and fillet composition. However, there were significant differences in lysozyme activity between groups of DOL at 0.075% and the control (0%). Moreover, the bactericidal index of the positive control and groups supplemented with DOL were significantly higher than baseline treatment groups. For disease resistance, there was a reduction of 15% in the relative risk of streptococcosis in fish juveniles after dietary inclusion of oregano at 0.15%. In conclusion, the inclusion of DOL in tilapia diets resulted in no significant gains in terms of growth; nevertheless, the immune system and resistance of tilapia against S. agalactiae infection were significantly improved.

Cardiac disease is of importance in captive chimpanzee (Pan troglodytes) health. Here we report an eosinophilic and necrotizing myocarditis in a 17-y-old chimpanzee with no previous history of cardiac disease that progressed to death within 48 h. Toxic and infectious causes were ruled out. The chimpanzee had eosinophilia at different occasions in previous years. The animal had a severe, diffuse, and acute monophasic necrotizing myocarditis, with a moderate lymphoplasmacytic infiltrate that was rich in eosinophils. Ante- and postmortem investigations are compatible with an unusual eosinophilic myocarditis with clinical evolution and morphology comparable with human eosinophilic myocarditis secondary to hypereosinophilic syndrome.

Cardiac disease is of importance in captive chimpanzee (

The objective of this work was to evaluate the effects of ultraviolet radiation (UV) and water salinity on nitrification, water quality, bacterial load, and juvenile tilapia growth in recirculating aquaculture systems (RASs). The experimental period was divided into two phases. The first one lasted 20 days and evaluated the effects of salinity (0 and 2 g L-1) and UV (with or without) on water quality during the period of substrate colonization by nitrifying bacteria. In the second phase, after the storage of juvenile tilapia, the effects of the same experimental factors were evaluated on water quality, bacterial load, and fish growth performance. The RASs employed were efficient for ammonia removal, regardless of the treatments used. During the experimental period, the nitrite concentrations increased linearly, with a more pronounced increase after fish storage until 30 days of the experiment. There were no significant effects of UV, salinity, or the interaction of both on total ammonia, nitrite, and alkalinity. The low levels of salinity (2 g L-1) and UV did not affect the nitrification process and fish performance. The use of UV is efficient to reduce the bacterial load of recirculating aquaculture systems.

Fish aquaculture is rapidly growing in Brazil. Nile tilapia is the most cultivated species, mainly through an intensive production system carried out in floating cages installed in large reservoirs. However, fish pathogens pose a major challenge to production chain sustainability, and tilapia farmers may have limited knowledge of prevailing health problems and rarely implement effective biosecurity practices to prevent introduction of economically important infectious agents. This study aimed to identify the key disease risks of tilapia farming in a tropical reservoir and characterize the dynamics of the prevalent pathogens, as a basis for development of effective control measures for tilapia health and surveillance programs. From August 2015 to October 2016, a longitudinal study was carried out at the Tres Marias reservoir, in the municipality of Morada Nova de Minas in the southeast of Brazil. Daily and monthly data were collected from six out of 32 existing fish farms, including fish samples, mortality counts, and measurements of temperature and water quality parameters. The main bacteria detected were Streptococcus agalactiae, infecting mostly adult tilapia throughout the period, with higher frequency as the average temperature increased, and Francisella noatunensis subsp. orientalis (Fno), infecting mainly younger tilapia, only during the cooler months. Coinfections with multiple pathogens were detected in 33 fish. The detection of Fno in one farm in two consecutive winters, after months of unfavorable water temperature conditions and without evidence of introduction of infected stock, strengthens the case for investigating if this pathogen can survive and remain infective causing new outbreaks. Furthermore, variation in mortality was likely associated with the dynamics of the studied pathogens.

Francisella noatunensis subsp. orientalis (FNO) is an important emerging pathogen associated with disease outbreaks in farm-raised Nile tilapia. FNO genetic diversity using PCR-based typing, no intra-species discrimination was achieved among isolates/strains from different countries, thus demonstrating a clonal behaviour pattern. In this study, we aimed to evaluate the population structure of FNO isolates by comparing whole-genome sequencing data. The analysis of recombination showed that Brazilian isolates group formed a clonal population; whereas other lineages are also supported by this analysis for isolates from foreign countries. The whole-genome multilocus sequence typing (wgMLST) analysis showed varying numbers of dissimilar alleles, suggesting that the Brazilian clonal population are in expansion. Each Brazilian isolate could be identified as a single node by high-resolution gene-by-gene approach, presenting slight genetic differences associated to mutational events. The common ancestry node suggests a single entry into the country before 2012, and the rapid dissemination of this infectious agent may be linked to market sales of infected fingerlings.

The Amazon catfish is one of the main farm-raised native fish in Brazil. In spite of its importance, the main health-associated issues have been poorly characterized in these fish. In this study, we describe outbreaks of S. agalactiae, S. iniae, and S. dysgalactiae in Amazon catfish farms in detail. The genetic diversity and antibiotic resistance patterns of different isolates were also evaluated. Thirty-five diseased fish were sampled from four commercial farms, and were subjected to bacteriological examinations. The isolates were genotyped by REP-PCR, and antimicrobial susceptibility of six antibiotics was evaluated using the disc diffusion assay. Fifteen Streptococcus spp. isolates were identified (S. agalactiae = 10 strains; S. dysgalactiae = 1; S. iniae = 4), which were further confirmed by MALDI-TOF. S. agalactiae and S. dysgalactiae infections were successfully achieved under experimental conditions, which confirmed Koch's postulates. Three distinct genetic patterns were verified by REP-PCR for S. agalactiae; S. iniae isolates were indistinguishable by this technique. In antimicrobial susceptibility tests, all S. agalactiae isolates were classified as wild-types to erythromycin (ERY); S. agalactiae strains, non-wild-types to amoxicillin (n = 2 isolates), florfenicol (n = 4), norfloxacin (n = 2), oxytetracycline (n = 3) (OXY), and trimethoprim-sulfamethoxazole (n = 6) (SXT); S. dysgalactiae strain, non-wild-type to OXY and ERY; and S. iniae isolates, non-wild-types to both SXT (n = 1) and OXY (n = 3). This study represents the first description of Streptococcus iniae, S. dysgalactiae, and S. agalactiae serotype II infections in farm-raised Amazon catfish. In addition, it is the first study that describes OXY resistant S. agalactiae isolates from farmed native fish in Brazil.

Streptococcus agalactiae is a highly pathogenic bacterium of farmed tilapia. Antimicrobial therapy is the principal control measure applied during outbreaks. Several reports have documented treatment failure and recurrent disease in herds treated with FLO. The present study aimed to: evaluate the profile of florfenicol resistance of Streptococcus agalactiae to calculate the provisional epidemiological cut-off value (ECV); address the therapeutic efficiency of FLO in treating streptococcosis in Nile tilapia; and determine the persister phenotype induced by FLO. Evaluation by MIC found 100 strains to possess sensitivity to FLO. The ECV was 8 μg/mL for 94% of the tested strains classified as wild-type. Therapeutic efficiency of FLO under doses of 10, 20, 40 and 60 mg/Kg for 10 days was tested. Dosages of 20 and 40 mg/Kg of LW were able to control mortality during therapy period, however, they did not cure the fish and mortality was observed post-treatment. A persistence assay was carried out and the strain SA95-10 remained viable for 10 days when exposed to 100 times its MIC value. In conclusion, the ECV for FLO, recommended dosage to control Streptococcus agalactiae mortality and the persister phenotype are described here for the first time.

Vertebral osteomyelitis (VO) is a worldwide emerging disease that affects broilers. Recently, the isolation of Enterococcus faecalis in cases of the disease has been described. This study aimed at determining the genetic diversity and antimicrobial resistance profile of 12 E. faecalis strains isolated from broilers with VO. Strains were isolated from nine flocks from six farms in a high-density poultry production area in Southeast Brazil and were evaluated using multilocus sequence typing and phylogenetic analysis. Antimicrobial susceptibility tests and PCR were performed to detect antimicrobial resistance genes. E. faecalis isolates belonged to different sequence types (ST), six of which (ST49, ST100, ST116, ST202, ST249, and ST300) have been previously described. Strains ST708 and ST709 were newly identified in this study. Strain ST49 was most frequently isolated (50% of the flocks) from the analysed VO cases. No phylogenetic or phylogeographic relationship was found among the strains. The VO isolated E. faecalis strains showed highest resistance to aminoglycosides, mainly gentamicin (40%), but were highly susceptible to vancomycin (10%). Aminoglycoside resistance genes were detected in seven E. faecalis strains, and AAC6'-APH2″ genes were most frequently detected. The results showed that E. faecalis strains isolated from recently reported VO cases were highly diverse genetically. The diversity of genotypes in circulation in the analysed flocks, without apparent relationship among them, raises questions on aetiopathogenesis of the disease in broilers and evolutionary aspects of E. faecalis.

This report describes a severe outbreak of the gill fluke Centrocestus formosanus in farm-raised platies Xiphophorus maculatus in Brazil, with mortality rate approaching 95%. Typical clinical signs of infection were observed, with microscopic examinations of fresh gills revealing multiple cysts containing a once-folded metacercaria with an X-shaped excretory bladder. The 18S subunit of the metacercariae (BR1) was amplified by PCR, sequenced and analyzed by BLAST. Subsequent phylogenetic analyses revealed that the BR1 isolate was closely related to C. formosanus from Thailand. This is the first report of C. formosanus in ornamental fish in Brazil. Our observations suggest that platies are highly sensitive to this digenetic parasite. Controlling population densities of the parasite's intermediate host, the snail Melanoides tuberculata, would help to reduce outbreaks.

This study evaluated the control of streptococcosis outbreaks in Brazil, isolated from diseased sorubim and identified as Lactococcus garvieae by genetic sequencing. This report determined the potential for lactococcosis control in sorubim Pseudoplatystoma sp. with two vaccines: an aqueous-based, whole-cell inactivated vaccine (bacterin) and an oil-adjuvanted bacterin. Their efficacy was evaluated at 30 days post-vaccination (d.p.v.) by challenge with L. garvieae, and the antibody production response at 15, 30 and 60 d.p.v. and the non-specific immune response were compared amongst treatments. High protection levels (P

Streptococcus agalactiae is a major etiologic agent of septicemia and meningoencephalitis outbreaks in Nile tilapia ( Oreochromis niloticus L.) farms and the presence of carrier fish can be a significant source of infection. Nonlethal methods to obtain samples for microbiological and molecular diagnostic tests are an option for detecting carrier fish. The aim of this study was to standardize nonlethal sampling methods for the diagnosis of S. agalactiae infection in Nile tilapia using microbiological and molecular assays. For this, 140 Nile tilapia juveniles were used in three different experimental trials. In the first trial, nonlethal sampling methods – kidney aspiration, venipuncture, nasal wash, gill mucus swabs, and fecal sample collection – were evaluated. The second trial was performed to determine the routes of S. agalactiae shedding, and the third trial was conducted to evaluate the efficiency of using nonlethal samples for the diagnosis of S. agalactiae infection in experimentally infected fish. The nonlethal sampling methods used to collect samples from Nile tilapia were successfully standardized. There was no mortality and no signs of infection observed in fish subjected to the sampling methods tested (kidney aspiration, venipuncture, nasal wash, gill mucus swabs, and fecal sample collection). S. agalactiae was successfully reisolated from all samples with the exception of nasal washes. The diagnostic methods used were able to detect the presence of S. agalactiae from carrier and diseased Nile tilapia. Our results demonstrate that nonlethal sampling methods, including kidney aspiration and venipuncture, can be used for the detection of S. agalactiae by bacteriology, PCR, and qPCR. Kidney aspiration and venipuncture in combination with qPCR were highly sensitive for the detection of S. agalactiae (93.04%). Statement of relevance The use of nonlethal sampling is an alternative method for the detection of S. agalactiae infection in diseased fish and carriers, allowing the diagnosis of this disease in broodstocks, or useful for surveillance programs and health monitoring in hatcheries without having to sacrifice fish. This diagnosis is crucial to avoid the maintenance of the pathogen in the culture system and the transmission of the disease to other farms.

Mercury, in particular its most toxic form methylmercury, poses a risk to public health. Dietary methylmercury exposure is mainly by fish, and it can vary with fish contamination and by dietary habits of the population. This study aimed to quantify total mercury levels in different fish from Brazil and to estimate Brazilian exposure to methylmercury by fish consumption.Total mercury occurrence was investigated in 18 different fish species by atomic absorption spectrometry with thermal decomposition and gold amalgamation. Dietary exposure to methylmercury was estimated by a deterministic method for different groups considering consumption by sex, different Brazilian geographical regions and habitat (rural or urban).Carnivorous fish showed higher levels of mercury (0.01 to 0.93 mg/kg) compared to non-strictly carnivorous fish (0.01 to 0.30 mg/kg). Farmed fishes showed significantly lower levels compared to wild fish. Mean Brazilian fish consumption achieves FAO/WHO health recommendation of about two portions of fish per week. However, there is a large difference between fish consumption at urban and rural homes and among Brazilian geographic regions. These differences in consumption impacted estimated methylmercury intake that was higher in the Northern (1.85 μg/kg bw week) and in the Northeastern (0.72 μg/kg bw week) regions and also by rural population (1.08 μg/kg bw week). These values were compared with the toxicological reference dose for neurotoxicity of 1.6 μg/kg bw week.Even though total levels of mercury in fish were lower than Brazilian and international legislations, in the Northern Brazilian region methylmercury intake overpassed the toxicological reference dose for neurotoxicity and in rural areas it achieved 68% of this reference dose.

Fish farming represents an important and healthy source of protein for human nutrition. Currently, Nile tilapia (Oreochromis niloticus) is the second most raised fish species worldwide, and its fillet plays an important role in the international trade of fish meat. In Brazil, fish slaughterhouses submitted to the Federal Meat Inspection Service detected lesions on tilapia fillets, which cause condemnation and economic losses to farmers and companies. The main etiologic agents associated with these fillet lesions have never been addressed. This study aimed to characterize the anatomopathology and etiologic agents involved in Nile tilapia fillet lesions. From May to September 2017, 70 fillets of Nile tilapia discarded as improper for human consumption at the inspection line were sampled, macroscopically characterized, and submitted to histopathology. In addition, Streptococcus agalactiae and Francisella noatunensis subsp. orientalis (Fno) were detected by PCR and qPCR, respectively. Out of all analyzed samples, 67% were positive either for Streptococcus agalactiae, Fno, or both. Three macroscopic patterns were identified, being two associated with bacterial pathogens. 85% of the Fno positive fillets had a black dot pattern and 90% of the S. agalactiae positive fillets had a cavity pattern on the surface of the fillet. Irregular white areas were not positive for any agent and were trauma related. The microscopic findings were pyogranulomatous and necrotizing myositis of varying degrees and distribution. The rate of discard in the lots affected by bacteria ranged from 0.2% to 2% of fillets. This is the first report of Fno as an important cause of fillet discharge in Nile tilapia slaughterhouses. The macroscopic patterns identified may be helpful in suggesting the etiology of these lesions and preventing economic loss.

Considering the increasing popularity of reptiles as pets and their possible role as reservoirs of pathogenic microorganisms, the aim of this study was to isolate Escherichia coli, Salmonella spp., Clostridium perfringens, and C. difficile strains from reptiles in Brazil and to characterize the isolated strains. The characterization was based on phylogenetic typing of E. coli, identification of virulence genes of E. coli, C. perfringens, and C. difficile, serotyping of Salmonella spp., ribotyping and MLST of C. difficile and antimicrobial susceptibility test of pathogenic strains. Cloacal swabs were collected from 76 reptiles, of which 15 were lizards, 16 chelonians, and 45 snakes, either living in captivity, in the wild, or as companion animals. E. coli was isolated from 52 (68.4%) reptiles, of which 46 (88.4%) were characterized as phylogroup B1. The virulence factor CNF1 of E. coli was found in seven (9.2%) sampled animals, whereas the gene of EAST1 was found in isolates from two (2.6%) reptiles. Three isolates positive for CNF1 were resistant to cephalothin, one of which was also resistant to ciprofloxacin, trimethoprim/sulfamethoxazole, and chloramphenicol, being then classified as multidrug resistant strain (MDR). Salmonella enterica was identified in 26 (34.2%) reptiles, of which 13 belonged to the subspecies enterica. Serotypes such as S. Mbandaka, S. Panama, S. Infantis, S. Heidelberg, and S. Anatum were identified. One isolate of S. enterica subsp. houtenae was resistant to cephalothin and ciprofloxacin. C. perfringens type A was isolated from six (7.8%) animals. C. difficile was isolated from three (3.9%) reptiles. Two of these isolates were toxigenic and classified into ribotypes/MLST 081/ST9 and 106/ST42, which have been previously reported to infect humans. In conclusion, reptiles in Brazil can harbor toxigenic C. difficile and potentially pathogenic E. coli and Salmonella enterica subsp. enterica, thus representing a risk to human and animal health.

One of the major challenges in Nile tilapia (Oreochromis niloticus L.) farming is the occurrence of bacterial infections, and the Francisella noatunensis subsp. orientalis (FNO) is an important pathogen that has emerged in last decades. Francisellosis outbreaks have been reported in the literature as occurring seasonally when water temperature is below 24 °C. The aim of this study was to quantify the median lethal doses (LD50) of FNO in experimental challenges at 28 °C and 22 °C, and to investigate the impact of temperature changes in whole genome expression using microarray technology. The LD50 for Nile tilapia at 28 °C was ∼105.7, whereas at 22 °C, the LD50 was ∼102.2, showing that the decrease in temperature enhanced disease outcome. Out of 1917 genes screened, a total of 31 and 19 genes were down- and up-regulated at 22 °C, respectively. These genes were grouped by orthology into functional categories of: amino acid, inorganic ion, and carbohydrate transport and metabolism; transcription; and posttranslational modification, protein turnover, and chaperones. Expression of genes related to metabolism, oxidative stress, and thermal shock were regulated by temperature changes, reflecting an ability of FNO to adapt to the environment. Expression of virulence genes usually required for the Francisella genus was not changed between tested temperatures, including that of genes located on the Francisella Pathogenicity Island.

Corynebacterium pseudotuberculosis has been widely studied in an effort to understand its biological evolution. Transcriptomics has revealed possible candidates for virulence and pathogenicity factors of strain 1002 (biovar Ovis). Because C. pseudotuberculosis is classified into two biovars, Ovis and Equi, it was interesting to assess the transcriptional profile of biovar Equi strain 258, the causative agent of ulcerative lymphangitis. The genome of this strain was re-sequenced; the reassembly was completed using optical mapping technology, and the sequence was subsequently re-annotated. Two growth conditions that occur during the host infection process were simulated for the transcriptome: the osmotic and acid medium. Genes that may be associated with the microorganism's resilience under unfavorable conditions were identified through RNAseq, including genes present in pathogenicity islands. The RT-qPCR was performed to confirm the results in biological triplicate for each condition for some genes. The results extend our knowledge of the factors associated with the spread and persistence of C. pseudotuberculosis during the infection process and suggest possible avenues for studies related to the development of vaccines, diagnosis, and therapies that might help minimize damage to agribusinesses. Fil: Gomide, Anne Cybelle Pinto. Universidade Federal de Minas Gerais; Brasil Fil: Ibraim, Izabela Coimbra. Universidade Federal de Minas Gerais; Brasil Fil: Alves, Jorianne T. C.. Universidade Federal do Pará; Brasil Fil: de Sá, Pablo Gomes. Federal Rural University Of Amazonia; Brasil Fil: de Oliveira Silva, Yuri Rafael. Universidade Federal do Pará; Brasil Fil: Santana, Mariana Passos. Universidade Federal de Minas Gerais; Brasil Fil: Marques Da Silva, Wanderson. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Folador, Edson Luiz. Universidade Federal Da Paraiba; Brasil Fil: Mariano, Diego C. B.. Universidade Federal de Minas Gerais; Brasil Fil: de Paula Castro, Thiago Luiz. Universidade Federal da Bahia; Brasil Fil: Barbosa, Silvanira. Universidade Federal do Pará; Brasil Fil: Dorella, Fernanda Alves. Universidade Federal de Minas Gerais; Brasil Fil: Carvalho, Alex F.. Universidade Federal de Minas Gerais; Brasil Fil: Pereira, Felipe L.. Universidade Federal de Minas Gerais; Brasil Fil: Leal, Carlos A. G.. Universidade Federal de Minas Gerais; Brasil Fil: Figueiredo, Henrique C. P.. Universidade Federal de Minas Gerais; Brasil Fil: Azevedo, Vasco. Universidade Federal de Minas Gerais; Brasil Fil: Silva, Artur. Universidade Federal do Pará; Brasil Fil: Folador, Adriana Ribeiro Carneiro. Universidade Federal do Pará; Brasil

Infection with Streptococcus agalactiae causes mortality and major economic losses in Nile tilapia ( Oreochromis niloticus ) farming worldwide. In Brazil, serotype strains Ia, Ib and III have been isolated in streptococcosis outbreaks, but serotype Ib is the most prevalent. Vaccination is considered an effective method to prevent economically-important diseases in aquaculture and has been associated with decreased use of antibiotics and improvements in fish survival. We developed a flexible partial-budget model to undertake an economic appraisal of vaccination against Streptococcus agalactiae in Nile tilapia farmed in net cages in large reservoirs. The model considers the benefits and costs that are likely to be associated with vaccination at the farm-level, in one production cycle. We built three epidemiological scenarios of cumulative mortality attributable to S. agalactiae (5%, 10%, and 20%, per production cycle) in a non-vaccinated farm. For each scenario, we applied a stochastic model to simulate the net return of vaccination, given a combination of values of “vaccine efficacy”, “gain in feed conversion ratio”, “feed price”, “fish market price “, and “cost of vaccine dose”. In the 20% cumulative mortality scenario, the net return would break-even (benefits ≥ costs) in at least 97.9% of interactions. Should cumulative mortality be lower than 10%, the profitability of vaccination would be more dependent on better feed conversion ratio . The inputs “feed price” and “cost of vaccine” had minor effects on the output, in all pre-vaccination mortality scenarios. Although our simulations are based on conservative values and consider uncertainty about the modeled parameters, we conclude that vaccination against S. agalactiae is likely to be profitable in Nile tilapia farms, under similar production conditions.

Streptococcus agalactiae is one of the most important pathogens associated with streptococcosis outbreaks in Nile tilapia farms worldwide. High water temperature (above 27°C) has been described as a predisposing factor for the disease in fish. At low temperatures (below 25°C), fish mortalities are not usually observed in farms. Temperature variation can modulate the expression of genes and proteins involved in metabolism, adaptation, and bacterial pathogenicity, thus increasing or decreasing the ability to infect the host. This study aimed to evaluate the transcriptome and proteome of a fish-pathogenic S. agalactiae strain SA53 subjected to in vitro growth at different temperatures using a microarray and label-free shotgun LC-HDMSE approach. Biological triplicates of isolates were cultured in BHIT broth at 22 or 32°C for RNA and protein isolation and submitted for transcriptomic and proteomic analyses. In total, 1,730 transcripts were identified in SA53, with 107 genes being differentially expressed between the temperatures evaluated. A higher number of genes related to metabolism, mainly from the phosphotransferase system (PTS) and ATP-binding cassette (ABC) transport system, were upregulated at 32°C. In the proteome analysis, 1,046 proteins were identified in SA53, of which 81 were differentially regulated between 22 and 32°C. Proteins involved in defense mechanisms, lipid transport and metabolism, and nucleotide transport and metabolism were upregulated at 32°C. A higher number of interactions were observed in proteins involved in nucleotide transport and metabolism. We observed a low correlation between the transcriptome and proteome datasets. Our study indicates that the transcriptome and proteome of a fish-adapted S. agalactiae strain are modulated by temperature, particularly showing differential expression of genes/proteins involved in metabolism, virulence factors, and adaptation.

The study aimed to describe the Bubalus bubalis seminal plasma proteome using a label-free shotgun UDMSE approach. A total of 859 nonredundant proteins were identified across five biological replicates with stringent identification. Proteins specifically related to sperm maturation and protection, capacitation, fertilization and metabolic activity were detected in the buffalo seminal fluid. In conclusion, we provide a comprehensive proteomic profile of buffalo seminal plasma, which establishes a foundation for further studies designed to understand regulation of sperm function and discovery of novel biomarkers for fertility. MS data are available in the ProteomeXchange with identifier PXD003728.

Flavobacterium columnare is responsible for disease outbreaks in freshwater fish farms. Several Brazilian native fish have been commercially exploited or studied for aquaculture purposes, including Amazon catfish Leiarius marmoratus × Pseudoplatystoma fasciatum and pacamã Lophiosilurus alexandri. This study aimed to identify the aetiology of disease outbreaks in Amazon catfish and pacamã hatcheries and to address the genetic diversity of F. columnare isolates obtained from diseased fish. Two outbreaks in Amazon catfish and pacamã hatcheries took place in 2010 and 2011. Four F. columnare strains were isolated from these fish and identified by PCR. The disease was successfully reproduced under experimental conditions for both fish species, fulfilling Koch's postulates. The genomovar of these 4 isolates and of an additional 11 isolates from Nile tilapia Oreochromis niloticus was determined by 16S rRNA restriction fragment length polymorphism PCR. The genetic diversity was evaluated by phylogenetic analysis of the 16S rRNA gene and repetitive extragenic palindromic PCR (REP-PCR). Most isolates (n = 13) belonged to genomovar II; the remaining 2 isolates (both from Nile tilapia) were assigned to genomovar I. Phylogenetic analysis and REP-PCR were able to demonstrate intragenomovar diversity. This is the first report of columnaris in Brazilian native Amazon catfish and pacamã. The Brazilian F. columnare isolates showed moderate diversity, and REP-PCR was demonstrated to be a feasible method to evaluate genetic variability in this bacterium.

The effect of different dietary oil sources on the innate immunity and resistance of Nile tilapia, Oreochromis niloticus, to Streptococcus agalactiae infection were evaluated. Fish were fed with diets containing different lipid sources (soybean oil [SO], corn oil, linseed oil [LO], fish oil [FO], and olive oil [OO]). Fish fed SO presented the highest (P 0.05) was found between diets in alternative complement activity. Fish fed the SO diet had the highest (P < 0.05) survival rate against S. agalactiae challenge. In conclusion, diets with LO oil and FO, rich in ω-3 fatty acids, and OO, rich in ω-9 fatty acids, have an immunomodulatory effect in Nile tilapia juveniles. The use of SO in the Nile tilapia diet improved immune function and resistance against S. agalactiae.

The species Corynebacterium renale, Corynebacterium pilosum, and Corynebacterium cystitidis were initially thought to be the same species C. renale, but with different immunological types. These bacteria are the causative agent of cystitis, urethritis and pyelonephritis and are found usually as constituents of the normal flora in the lower urogenital tract of cattle. Therefore, we present the draft genome sequences of two pathogenic Corynebacterium species: C. renale CIP 52.96 and C. pilosum CIP 103422. The genome sequences of these species have 2,322,762 bp with 2,218 protein encoding genes and 2,548,014 bp with 2,428 protein encoding genes, respectively. These genomes can help clarify the virulence mechanisms of these unknown bacteria and enable the development of more effective methods for control.

Gram-positive cocci, such as Streptococcus agalactiae, Lactococcus garvieae, Streptococcus iniae, and Streptococcus dysgalactiae subsp. dysgalactiae, are found throughout the world, particularly in outbreaks in farmed fish, and are thus associated with high economic losses, especially in the cultivation of Nile Tilapia. The aim of this study was to evaluate the efficacy of matrix-assisted laser desorption ionization (MALDI)-time of flight (TOF) mass spectrometry (MS) as an alternative for the diagnosis of these pathogens. One hundred and thirty-one isolates from Brazilian outbreaks assisted by the national authority were identified using a MALDI Biotyper from Bruker Daltonics. The results showed an agreement with respect to identification (Kappa = 1) between this technique and 16S ribosomal RNA gene sequencing for S. agalactiae and L. garvieae. However, for S. iniae and S. dysgalactiae subsp. dysgalactiae, perfect agreement was only achieved after the creation of a custom main spectra profile, as well as further comparisons with 16S ribosomal RNA and multilocus sequence analysis. MALDI-TOF MS was shown to be an efficient technology for the identification of these Gram-positive pathogens, yielding a quick and precise diagnosis.

Weissella ceti is an emerging bacterial pathogen that affects rainbow trout, Oncorhynchus mykiss (Walbaum), farms. The aims of this study were to genotype W. ceti strains isolated from distinct geographical origins and to determine the efficacy of an oil-adjuvanted vaccine against the disease. Between 2010 and 2012, outbreaks were recorded in five Brazilian farms, and 34 W. ceti isolates were genetically characterized by repetitive extragenic palindromic PCR, enterobacterial repetitive intergenic consensus sequences PCR and pulsed-field gel electrophoresis. Two different W. ceti vaccines were tested: an aqueous-based whole-cell inactivated vaccine (bacterin) and oil-adjuvanted vaccine. Their efficacy was evaluated in rainbow trout at 30 and 60 days post-vaccination (d.p.v.). W. ceti was found to be a highly homogeneous population in Brazil, with clonally related genotypes. Oil-adjuvanted vaccine exhibited the best (P

Francisella noatunensis subsp orientalis (FNO) is an emerging pathogen of warm water tilapia in a number of different countries. The disease caused by this bacterium in fish is characterized by a systemic granulomatous infection that causes high mortality rates during outbreaks. FNO has been previously described in Asia, Europe, and Central and North America. Its occurrence in South America has never been described. Since 2012, outbreaks of a granulomatous disease have been recorded in cage farms of Nile tilapia (Oreochromis niloticus L.) in Brazil. The current study aimed to identify the etiologic agent of recent francisellosis outbreaks at Brazilian tilapia farms, and to characterize the genetic diversity of the pathogen from farms with distinct geographic origins and without epidemiological connections. Bacteriological analysis of 44 diseased Nile tilapia collected from five cage farms in Brazil was performed during 2012 and 2013. The farms were in different locations and had no recent history of animal or biological material transport between each other. Sixty-two FNO isolates were identified on the basis of FNO-specific qPCR. The main predisposing factors for the occurrence of outbreaks on Brazilian farms were lower water temperature (

Streptococcus agalactiae (Lancefield group B; group B streptococci) is a major pathogen that causes meningoencephalitis in fish, mastitis in cows, and neonatal sepsis and meningitis in humans. The available prophylactic measures for conserving human and animal health are not totally effective and have limitations. Effective vaccines against the different serotypes or genotypes of pathogenic strains from the various hosts would be useful. We used an in silico strategy to identify conserved vaccine candidates in 15 genomes of group B streptococci strains isolated from human, bovine, and fish samples. The degree of conservation, subcellular localization, and immunogenic potential of S. agalactiae proteins were investigated. We identified 36 antigenic proteins that were conserved in all 15 genomes. Among these proteins, 5 and 23 were shared only by human or fish strains, respectively. These potential vaccine targets may help develop effective vaccines that will help prevent S. agalactiae infection.

We present here the genome sequence of the attenuated Corynebacterium pseudotuberculosis strain T1. The sequencing was performed with an Ion Torrent Personal Genome Machine platform. The genome is a circular chromosome of 2,337,201 bp, with a G+C content of 52.85% and a total of 2,125 coding sequences (CDSs), 12 rRNAs, 49 tRNAs, and 24 pseudogenes.