Bacterial communities and signatures in the stomach and intestine of juvenile P enaeus (litopenaeus) vannamei shrimp affected by acute hepatopancreatic necrosis disease.

Acute hepatopancreatic necrosis (AHPND) is a severe bacterial disease affecting farmed shrimp. Although various pathogenic bacteria associated with AHPND-affected shrimp have been described, little is known about the bacterial signatures in the stomachs and intestines when the disease occurs naturally. In this study, we characterized the microbiome of P. vannamei by high-throughput sequencing (HTS). Shrimp samples were collected from a commercial farm and divided into two groups: healthy and affected by AHPND, confirmed by PCR. Stomach and intestine samples were subjected to microbiome analysis targeting the V3-V4 region of the 16S rRNA gene. PERMANOVA analysis revealed a significant disparity in the bacterial diversity between the stomach and intestine microbiomes of these two health conditions. Our results suggest that the significant abundance of Vibrio brasiliensis and V. sinaloensis in the intestines of affected shrimp plays a role in AHPND infection. This imbalance could be mitigated by the presence of Pseudoalteromonas, Gilvimarinus, and other members of the phylum Pseudomonadota such as Cellvibrionaceae, Psychromonadaceae, and Halieaceae, which showed significant abundance in healthy intestines. This study highlights the significance of the microbial community in the differentiation of specific microbial signatures in different organs of P. vannamei. These findings offer a deeper understanding of the intricate dynamics within the shrimp microbiome under these conditions, enriching our view of AHPND progression and paving the way toward future identification of probiotics tailored for more efficient management of this disease.

Draft genome sequence of Bacillus velezensis strain 3TSA-3, a potential probiotic for Pacific white shrimp Penaeus vannamei postlarvae isolated from commercial hatchery tanks.

We report the draft genome of Bacillus velezensis strain 3TSA-3, isolated from Pacific white shrimp Penaeus vannamei postlarvae collected from a hatchery tank with high survival despite the presence of pathogenic Vibrio. The strain possesses genes encoding bacteriocins and lacks virulence factor genes, characteristics for a potential aquaculture probiotic.

Positive cases of bovine genital campylobacteriosis and bovine trichomonosis in the eradication program of La Pampa, Argentina: 2008-2021.

Bovine genital campylobacteriosis, caused by the gram-negative bacteria Campylobacter fetus venerealis, and bovine trichomonosis, caused by the parasite protozoan Tritrichomonas foetus, are venereal diseases that occur with long intercalving periods and abortion. The control of both diseases relies on microbiological testing and culling infected bulls. Vaccination and antibiotic treatment may help in controlling campylobacteriosis but are not recommended for trichomonosis control. Several regions of the world have active control programs for trichomonosis, not campylobacteriosis. In Argentina, the state of La Pampa aims to eradicate trichomonosis and campylobacteriosis by imposing annual diagnostic testing of every bull and slaughtering positive animals. Prior studies indicated a declining trend in the prevalence of campylobacteriosis and trichomonosis in La Pampa. It was also proposed that the prevalence of one disease could be estimated from the prevalence of the other. The purpose of this retrospective analysis of data gathered from 2008 to 2021 was to determine the La Pampa program's efficacy. Descriptive statistics were employed to determine the reason behind the correlation between tricomonosis and campylobacteriosis diagnostic results. The outcomes refute the notion that this program of venereal eradication was a success. Furthermore, an excess of false positives in both diagnoses may have contributed to the correlation between the prevalences of campylobactriosis and trichomonosis. The practice of killing animals without verifying positive results hinders the determination of disease prevalence and results in the death of numerous healthy animals.

Design of a multi-epitope vaccine (vme-VAC/MST-1) against cholera and vibriosis based on reverse vaccinology and immunoinformatics approaches.

Vibriosis and cholera are serious diseases distributed worldwide and caused by six marine bacteria of the Vibrio genus. Thousands of deaths occur each year due to these illnesses, necessitating the development of new preventive measures. Presently, the existing cholera vaccine demonstrates an effectiveness of approximately 60%. Here we describe a new multi-epitope vaccine, 'vme-VAC/MST-1' based on vaccine targets identified by reverse vaccinology and epitopes predicted by immunoinformatics, two currently effective tools for predicting new vaccines for bacterial pathogens. The vaccine was designed to combat vibriosis and cholera by incorporating epitopes predicted for CTL, HTL, and B cells. These epitopes were identified from six vaccine targets revealed through subtractive genomics, combined with reverse vaccinology, and were further filtered using immunoinformatics approaches based on their predicted immunogenicity. To construct the vaccine, 28 epitopes (24 CTL/B and 4 HTL/B) were linked to the sequence of the cholera toxin B subunit adjuvant. In silico analyses indicate that the resulting immunogen is stable, soluble, non-toxic, and non-allergenic. Furthermore, it exhibits no homology to the host and demonstrates a strong capacity to elicit innate, B-cell, and T-cell immune responses. Our analysis suggests that it is likely to elicit immune reactions mediated through the TLR5 pathway, as evidenced by the molecular docking of the vaccine with the receptor, which revealed high affinity and a favorable reaction. Thus, vme-VAC/MST-1 is predicted to be a safe and effective solution against pathogenic Vibrio spp. However, further experimental analyses are required to measure the vaccine's effects In vivo.Communicated by Ramaswamy H. Sarma.

First Report, Characterization and Pathogenicity of Vibrio chagasii Isolated from Diseased Reared Larvae of Chilean Scallop, Argopecten purpuratus (Lamarck, 1819).

Two Vibrio strains (VPAP36 and VPAP40) were isolated from moribund-settled larvae of the Chilean scallop Argopecten purpuratus during vibriosis outbreaks that occurred in two commercial scallop larvae hatcheries located in the Inglesa and Tongoy bays in Northern Chile. The strains were identified as Vibrio chagasii using phenotypic characterization and whole genome sequence analysis. Both strains exhibited the phenotypic properties associated with virulence, gelatin hydrolysis and ß-hemolysis, whereas only VPAP36 produced phospholipase and only VPAP40 produced caseinase. The whole genome analysis showed that the strains harbored genes encoding for the virulence factors, the EPS type II secretion system, and Quorum Sensing (auto-inductor 1 and auto-inductor 2), whereas genes encoding a metalloproteinase and a capsular polysaccharide were detected only in the VPAP40 genome. When challenge bioassays using healthy 11-day-old scallop larvae were performed, the V. chagasii VPAP36 and VPAP40 strains exhibited significant (p < 0.05) differences in their larval lethal activity, producing, after 48 h, larval mortalities of 65.51 ± 4.40% and 28.56 ± 5.35%, respectively. Otherwise, the cell-free extracellular products of the VPAP36 and VPAP40 strains produced larval mortalities of 20.86 ± 2.40% and 18.37 ± 2.40%, respectively, after 48 h of exposure. This study reports for the first time the isolation of V. chagasii from the massive larval mortalities of the farmed scallop (Argopecten purpuratus) in Chile, and demonstrates the pathogenic activity of V. chagasii towards the Chilean scallop, the second most important species for Chilean mariculture.

Antibacterial and anti-virulence potential of plant phenolic compounds against Vibrio parahaemolyticus.

Background: Vibrio parahaemolyticus is a pathogenic bacterium that affects shrimp aquaculture; its infection can lead to severe production losses of up to 90%. On the other hand, plant phenolic compounds have emerged as a promising alternative to combat bacterial infections. The antibacterial and anti-virulence activity of the plant phenolic compounds quercetin, morin, vanillic acid, and protocatechuic acid against two strains of V. parahaemolyticus (Vp124 and Vp320) was evaluated. Methods: The broth microdilution test was carried out to determine phenolic compounds' antibacterial activity. Moreover, the biofilm-forming ability of V. parahaemolyticus strains in the presence of phenolic compounds was determined by total biomass staining assay using the cationic dye crystal violet. The semisolid agar displacement technique was used to observe the effect of phenolic compounds on the swimming-like motility of V. parahaemolyticus. Results: Results showed that phenolic compounds inhibited both strains effectively, with minimum inhibitory concentrations (MICs) ranging from 0.8 to 35.03 mM. Furthermore, at 0.125 - 0.5 × MIC of phenolic compounds, V. parahaemolyticus biofilms biomass was reduced by 63.22 - 92.68%. Also, quercetin and morin inhibited the motility of both strains by 15.86 - 23.64% (Vp124) and 24.28 - 40.71% (Vp320). Conclusions: The results suggest that quercetin, morin, vanillic, and protocatechuic acids may be potential agents for controlling V. parahaemolyticus.

OCURRENCIA Y DISTRIBUCIÓN DE Vibrio cholerae CULTIVABLE EN LA CIÉNAGA GRANDE DE SANTA MARTA, CARIBE COLOMBIANO / Occurrence and distribution of cultivable Vibrio cholerae in Ciénaga Grande De Santa Marta, Colombian Caribbean

RESUMEN Vibrio cholerae es un potencial patógeno humano que habita ambientes acuáticos, aunque su presencia y abundancia se ha asociado al aumento de la temperatura del agua, poco se ha investigado sobre su ecología en ambientes estuarinos tropicales, donde los cambios de salinidad suelen ser más importantes. El presente estudio evaluó la distribución de V. cholerae en la Ciénaga Grande de Santa Marta, Colombia y su relación con la temperatura y la salinidad. Para ello, entre 2016 y 2018 se cuantificó bimestralmente esta especie en muestras de agua superficial, usando agar TCBS y pruebas bioquímicas. V. cholerae se detectó en 57 de 198 muestras (28,8 %), variando en densidad entre 5 y 54 800 UFC por 100 mL. Entre enero y septiembre de 2016 se presentó una alta salinidad promedio mensual (≥ 28,7) y una baja detección de la bacteria (0,01 %). La salinidad promedio se redujo drásticamente en noviembre de 2016 (9,6), coincidiendo con una proliferación de V. cholerae (promedio geométrico 36,4 UFC/100 mL). Durante 2017 y 2018 la salinidad promedio se mantuvo por debajo de 15,2 y la detección de V. cholerae fue mayor (39,4 %) que, en 2016, presentándose mayores densidades en los meses con menor salinidad. En las estaciones denominadas BVA y NVE, donde se ubican poblaciones palafíticas, se registraron las densidades promedio (geométrico) más altas, 25,3 UFC/100 mL y 15,4 UFC/ 100 mL, respectivamente. Los resultados de este estudio demuestran que la salinidad juega un papel determinante en la ocurrencia y abundancia de V. cholerae en esta laguna tropical.

ABSTRACT Vibrio cholerae is a potential human pathogen that inhabits aquatic environments, although its presence and abundance have been associated with increased water temperature, little research has been done on its ecology in tropical estuarine environments, where salinity changes tend to be more important. The present study evaluated the distribution of V. cholerae in the Ciénaga Grande de Santa Marta and its relationship with temperature and salinity; For this, between 2016 and 2018 this microorganism was quantified bimonthly in surface water samples, using TCBS agar and biochemical tests. V. cholerae was detected in 57 of 198 samples (28.8 %), varying in density between 5 CFU / 100 mL and 54,800 CFU / 100 mL. Between January and September 2016 there was a high average monthly salinity (≥ 28.7 °C) and a low detection of the bacteria (0.01 %). Average salinity dropped drastically in November 2016 (9.6), coinciding with a proliferation of V. cholerae (geometric average 36.4 CFU / 100 mL). During 2017 and 2018, the average salinity remained below 15.2 and the detection of V. cholerae was higher (39.4 %) than in 2016, with higher densities in the months with lower salinity. At the BVA and NVE stations, where palaphytic populations are located, the highest average (geometric) densities were recorded, 25.3 CFU / 100mL and 15.4 CFU / 100mL, respectively. The results of this study show that salinity plays a determining role in the occurrence and abundance of V. cholerae in this tropical lagoon.

Comparative Genomics of Clinical and Environmental Isolates of Vibrio spp. of Colombia: Implications of Traits Associated with Virulence and Resistance.

There is widespread concern about the increase in cases of human and animal infections caused by pathogenic Vibrio species due to the emergence of epidemic lineages. In Colombia, active surveillance by the National Institute of Health (INS) has confirmed the presence of Vibrio; however, in routine surveillance, these isolates are not genomically characterized. This study focused on the pangenome analysis of six Vibrio species: V. parahaemolyticus, V. vulnificus, V. alginolyticus, V. fluvialis, V. diabolicus and V. furnissii to determine the genetic architectures of potentially virulent and antimicrobial resistance traits. Isolates from environmental and clinical samples were genome sequenced, assembled and annotated. The most important species in public health were further characterized by multilocus sequence typing and phylogenomics. For V. parahaemolyticus, we found the virulent ST3 and ST120 genotypes. For V. vulnificus, we identified isolates belonging to lineages 1 and 2. Virulence gene homologues between species were found even in non-pathogenic species such as V. diabolicus. Annotations related to the mobilome, integrative mobile and conjugative elements and resistance genes were obtained from environmental and clinical isolates. This study contributes genomic information to the intensified surveillance program implemented by the INS to establish potential sources of vibriosis in Colombia.

First description outside Europe of the emergent pathogen Vibrio europaeus in shellfish aquaculture.

Vibrio europaeus is an emergent pathogen affecting the most important bivalve species reared in Spanish and French hatcheries. Using a genomic approach, we identified V. europaeus outside Europe for the first time from massive larval mortalities of scallop (Argopecten purpuratus) in Chile and from seawater near a shellfish hatchery in the US West Coast. Results show the worldwide spreading and potential impact of V. europaeus for aquaculture; these four countries are among the 10 major producers of mollusks. Pathogenicity of V. europaeus was demonstrated for the first time towards scallop, the second most important species for Chilean mariculture.

Identification and characterization of outer membrane vesicles from the fish pathogen Vibrio ordalii.

Vibriosis outbreaks due to Vibrio ordalii occur globally, but Chilean salmon aquaculture, in particular, has suffered significant monetary losses in the last 15 years. Little is known about the virulence mechanisms employed by V. ordalii. However, most Vibrio pathogens (e.g., Vibrio anguillarum, a very close taxonomic species) present outer membrane vesicles (OMVs) that are released extracellularly and implicated in the delivery of virulence factors to host cells. This study provides the first reported evidence of the fish pathogen V. ordalii producing and releasing OMVs under normal growth conditions. Analyses were conducted with the V. ordalii strain Vo-LM-18 and the type strain ATCC 33509T . For comparative purposes, the reference strain V. anguillarum ATCC 43307 was employed. The average size for the three Vibrio strains was 0.215 ± 0.6 µm (via scanning electron microscopy) or between 0.19 and 1.8 µm (via dynamic light scattering), with each bacterium presenting a wide range. SDS-PAGE revealed similarities in OMV patterns, but neither total nor external proteins were identical. Comparing V. ordalii ATCC 33509T and Vo-LM-18, bands were most evident in the total proteins, and the greatest degree of similarity in OMV profiles was between 37 and 50 kDa. The purified OMVs demonstrated haemolytic enzyme activity, which could play a role during V. ordalii infection. These data represent an initial step towards gaining new insights into this virulence factor, of which a lot is known in other pathogenic microorganisms.

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii.

Vibrio ordalii is an extracellular, Gram-negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo-LM-18 and ATCC 33509T in the fish-cell lines SHK-1 and CHSE-214. Confocal microscopy revealed Vo-LM-18 and ATCC 33509T inside cytoplasm in both fish-cell lines at 4 hr post-inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo-LM-18 and ATCC 33509T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.

Susceptibility of Pacific white snook Centropomus viridis to Vibrio species.

To examine the pathogenicity of Vibrio strains, several doses of Vibrio harveyi (CAIM 1622 and CAIM 1508), Vibrio ponticus (CAIM 1751) and Vibrio anguillarum (CAIM 8) were used to challenge Pacific white snook Centropomus viridis Lockington, 1877 juveniles, and survival, gross signs and histological lesions were observed. Susceptibility of pathogenic vibrios CAIM 1508 and CAIM 1751 to antibiotics used in aquaculture was also evaluated. The growth ability of the tested strains was not related to their pathogenicity. One of the V. harveyi strains (CAIM 1508) was the most virulent, causing per-acute septicaemia in C. viridis even at a low dose (1.4 × 104 CFU g-1). Although the V. ponticus strain (CAIM 1751) was less virulent, this is the first report of it as a pathogen of white snook. Fish challenged with V. ponticus displayed external, generalized haemorrhaging. Necrosis of the digestive tract and intravascular haemosiderosis were the most remarkable histological lesions in fish challenged with both strains. Multifocal necrosis of the internal organs and bacterial masses was also observed. The lowest minimum inhibitory concentration of the pathogenic strains (CAIM 1508 and CAIM 1751) was calculated for enrofloxacin (20 and 10 µg ml-1, respectively), and both bacteria were resistant to amoxicillin, ampicillin and trimethoprim-sulfamethoxazole.

Isolation and Pathogenic Characterization of Vibrio bivalvicida Associated With a Massive Larval Mortality Event in a Commercial Hatchery of Scallop Argopecten purpuratus in Chile.

The VPAP30 strain was isolated as the highly predominant bacteria from an episode of massive larval mortality occurring in a commercial culture of the Chilean scallop Argopecten purpuratus. The main aims of this study were, to characterize and identify the pathogenic strain using biochemical and molecular methods, to demonstrate its pathogenic activity on scallop larvae, to characterize its pathogenic properties and to describe the chronology of the pathology. The pathogenic strain was identified as Vibrio bivalvicida based on its phenotypic properties, the multilocus sequence analysis (MLSA) of eight housekeeping genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA, and topA) and different in silico genome-to-genome comparisons. When triplicate cultures of healthy 10 days old scallop larvae were challenged with 1 × 105 colony forming units (CFU) mL-1 of the VPAP30 strain, percentages of larval survival of 78.9 ± 3.3%, 34.3 ± 4.9%, and 0% were observed at 12, 2,4 and 36 h, respectively, whereas uninfected larval cultures showed survival rates of 97.4 ± 1.2% after of 48 h. Clinical symptoms exhibited by the scallop larvae infected with the VPAP30 strain include the accumulation of bacteria around the scallop larvae, velum disruption and necrosis of digestive gland. The 50% lethal dose (LD50) of VPAP30 strain at 24 and 48 h was 1.3 × 104 and 1.2 × 103 CFU mL-1, respectively. The invasive pathogenic activity of the VPAP30 strain was investigated with staining of the bacterial pathogen with 5-DTAF and analyzing bacterial invasion using epifluorescence, and a complete bacterial dissemination inside the larvae at 24 h post-infection was observed. When scallop larvae were inoculated with cell-free extracellular products (ECPs) of VPAP30, the larval survival rate was 59.5 ± 1.7%, significantly (P < 0.001) lower than the control group (97.4 ± 1.2%) whereas larvae treated with heat-treated ECPs exhibited a survival rate of 61.6 ± 1.8% after 48 h of exposure. V. bivalvicida VPAP30 exhibits high pathogenic activity on scallop larvae, mediated both by bacterial invasion and the production of toxigenic heat-stable compounds. This report constitutes the first isolation of V. bivalvicida out of Europe and extends the host range of this species, having demonstrated its pathogenic activity on the Chilean scallop larvae (A. purpuratus). These results supporting the pathogenic potential of V. bivalvicida to kill the larvae of a broad range of bivalve species reared in hatcheries located in the Atlantic and the Pacific coasts.

Genotypic assessment of a dichotomous key to identify Vibrio coralliilyticus, a coral pathogen.

Vibrio coralliilyticus is a known pathogen to corals and larvae of bivalves. Its identification is made based on phenotypic and genotypic characters of isolated strains. To evaluate the efficiency of the phenotypic identification, 21 strains identified as V. coralliilyticus using a widely used dichotomous key were analyzed by qualitative PCR and sequencing of the 16S rDNA region. The results obtained by the behavioral test, amino acids usage, allow us to distinguish 3 A/L/O profiles: (1) A+/L-/O+; (2) A+/L+/O+; and (3) A-/L+/O+. In the genotypic tests, all strains tested positive with primers specific for the Vibrio genus. However, when primers were used for species identification, the results did not match those obtained with the dichotomous key chosen. The phenotypic characteristics taken into account to set apart V. coralliilyticus and other species were not proven to be efficient. More information about the morphological diversity of colonies and enzymatic activities should be considered in the formulation of phenotypic keys for V. coralliilyticus and related species.

First Report of Vibrio tubiashii Associated with a Massive Larval Mortality Event in a Commercial Hatchery of Scallop Argopecten purpuratus in Chile.

The VPAP30 strain was isolated as the highly predominant bacteria from an episode of massive larval mortality occurring in a commercial culture of the Chilean scallop Argopecten purpuratus. The main aims of this study were, to characterize and identify the pathogenic strain using biochemical and molecular methods to demonstrate its pathogenic activity on scallop larvae, to characterize its pathogenic properties and to describe the chronology of this pathology. The pathogenic strain was identified as Vibrio tubiashii based on its phenotypic properties and the sequence analysis of its 16S rRNA and housekeeping genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA and topA). When triplicate cultures of healthy 10-day-old scallop larvae were challenged with 1 × 105 colony forming units (CFU) mL-1 of the VPAP30 strain, percentages of larval survival of 78.87 ± 3.33%, 34.32 ± 4.94%, and 0% were observed at 12, 24, and 36 h, respectively; whereas uninfected larval cultures showed survival rates of 97.4 ± 1.24% after of 48 h. Clinical symptoms exhibited by the scallop larvae infected with the VPAP30 strain include the accumulation of bacteria around the scallop larvae, velum disruption and necrosis of digestive gland. The 50% lethal dose (LD50) of VPAP30 strain at 24 and 48 h was 1.3 × 104 and 1.2 × 103 CFU mL-1, respectively. The invasive pathogenic activity of the VPAP30 strain was investigated with staining of the bacterial pathogen with 5-DTAF and analyzing bacterial invasion using epifluorescence, and a complete bacterial dissemination inside the larvae at 24 h post-infection was observed. When scallop larvae were inoculated with cell-free extracellular products (ECPs) of VPAP30, the larval survival rate was 59.5 ± 1.66%, significantly (P < 0.001) lower than the control group (97.4 ± 1.20%) whereas larvae treated with heat-treated ECPs exhibited a survival rate of 61.6 ± 1.84% after 48 h of exposure. This is the first report of the isolation of V. tubiashii from the diseased larvae of the scallop A. purpuratus, occurring in a commercial culture in Chile, and it was demonstrated that the VPAP30 strain exhibits high pathogenic activity on scallop larvae, mediated both by bacterial invasion and the production of toxigenic heat-stable compounds.

Dietary supplementation with probiotic and butyrate for Litopenaeus vannamei in pre-nursery / Suplementação dietética com probiótico e butirato de sódio no pré-berçário de Litopenaeus vannamei

This study aimed to evaluate the diet supplementation with sodium butyrate, probiotic (Lactobacillus plantarum) and their interaction, in the diet for post-larvae of the Pacific white shrimp (Litopenaeus vannamei) in pre-nursery. We used the following treatments during the entire experimental period: (1) Probiotic 1x108 UFC/g, sodium butyrate 2%, (3) probiotic 1x108 UFC/g + sodium butyrate 2%, (4) control (Base diet). In all treatment we added culture medium in the same doses as in the probiotic treatments. We used 16 experimental units of 60L each, with the bottom Ushaped, stocked with 2,880 post-larvae 5. Fifteen days later, regardless the presence or absence of the probiotic, shrimps from treatment with butyrate had higher survival (p = 0.0039) and lowest individual dry weight (p = 0.0043). No morphological changes were observed in the gut of postlarvae in any treatment. Therefore, the diet supplementation with sodium butyrate increases the survival of shrimp post-larvae of in the pre-nursery phase, without causing morphological changes in its gut.

Este estudo teve por objetivo avaliar o uso suplementar de probiótico (Lactobacillus plantarum) e butirato de sódio, e a interação deles na dieta de póslarvas de camarão-branco-do-pacífico (Litopenaeus vannamei) em fase de pré-berçário. Durante o experimento foram aplicados nas dietas os tratamentos: (1) probiótico 1x108 UFC/g, (2) butirato de sódio 2%, (3) probiótico 1x108 UFC/g + butirato de sódio 2%, (4) controle (dieta base). Todos os tratamentos tiveram a adição de meio de cultura na mesma dosagem utilizada nos tratamentos com probiótico. Foram utilizadas 16 unidades experimentais de 60 L cada uma, com fundo em formato U, povoadas com 2.880 póslarvas 5. Após quinze dias, independentemente da presença ou não do probiótico, os camarões dos tratamentos com butirato apresentaram maior sobrevivência (p = 0,0039) e menor peso seco individual (p = 0,0043). Não foram observadas alterações morfológicas no intestino das pós-larvas em nenhum dos tratamentos. Portanto, a suplementação com butirato de sódio na ração aumenta a sobrevivência das pós-larvas de camarões na fase de pré-berçário sem causar alteração morfológica em seu intestino.

Dietary supplementation with probiotic and butyrate for Litopenaeus vannamei in pre-nursery / Suplementação dietética com probiótico e butirato de sódio no pré-berçário de Litopenaeus vannamei

This study aimed to evaluate the diet supplementation with sodium butyrate, probiotic (Lactobacillus plantarum) and their interaction, in the diet for post-larvae of the Pacific white shrimp (Litopenaeus vannamei) in pre-nursery. We used the following treatments during the entire experimental period: (1) Probiotic 1x108 UFC/g, sodium butyrate 2%, (3) probiotic 1x108 UFC/g + sodium butyrate 2%, (4) control (Base diet). In all treatment we added culture medium in the same doses as in the probiotic treatments. We used 16 experimental units of 60L each, with the bottom Ushaped, stocked with 2,880 post-larvae 5. Fifteen days later, regardless the presence or absence of the probiotic, shrimps from treatment with butyrate had higher survival (p = 0.0039) and lowest individual dry weight (p = 0.0043). No morphological changes were observed in the gut of postlarvae in any treatment. Therefore, the diet supplementation with sodium butyrate increases the survival of shrimp post-larvae of in the pre-nursery phase, without causing morphological changes in its gut.(AU)

Este estudo teve por objetivo avaliar o uso suplementar de probiótico (Lactobacillus plantarum) e butirato de sódio, e a interação deles na dieta de póslarvas de camarão-branco-do-pacífico (Litopenaeus vannamei) em fase de pré-berçário. Durante o experimento foram aplicados nas dietas os tratamentos: (1) probiótico 1x108 UFC/g, (2) butirato de sódio 2%, (3) probiótico 1x108 UFC/g + butirato de sódio 2%, (4) controle (dieta base). Todos os tratamentos tiveram a adição de meio de cultura na mesma dosagem utilizada nos tratamentos com probiótico. Foram utilizadas 16 unidades experimentais de 60 L cada uma, com fundo em formato U, povoadas com 2.880 póslarvas 5. Após quinze dias, independentemente da presença ou não do probiótico, os camarões dos tratamentos com butirato apresentaram maior sobrevivência (p = 0,0039) e menor peso seco individual (p = 0,0043). Não foram observadas alterações morfológicas no intestino das pós-larvas em nenhum dos tratamentos. Portanto, a suplementação com butirato de sódio na ração aumenta a sobrevivência das pós-larvas de camarões na fase de pré-berçário sem causar alteração morfológica em seu intestino.(AU)

Utilização comparativa de enrofloxacino, probiótico (Lactobacilus plantarum) e propionato de sódio na larvicultura de camarão branco do pacífico / Comparative use of enrofloxacin, probiotic (Lactobacillus plantarum) and sodium propionate in pacific white shrimp larvae / Utilización comparativa de enrofloxacino, probiótico (Lactobacillus plantarum) y propionato de sodio en larvicultura de camarón blanco del pacífico

O objetivo do presente trabalho foi avaliar métodos de controle e tratamento de bacterioses na larvicultura de Litopenaeus vannamei, avaliando o uso contínuo de probiótico e o uso pontual de enrofloxacino 15mg.L-1 e propionato de sódio 0,5 mM.L-1 nos momentos de metamorfose sobre os, parâmetros zootécnicos e microbiológicos (larvas e da água). Foram utilizadas 16 unidades de 60L, povoadas na densidade de 325 náuplios.5L-1, divididos em três tratamentos: enrofloxacino, probiótico, propionato de sódio e controle. O enrofloxacino e o propionato foram ministrados em protozoea 3, misis 3 e pós-larva 4 e o probiótico foi ministrado na ração ao longo de todo o experimento. O probiótico aumentou as contagens de bactérias ácido-láticas em relação aos demais tratamentos na água de cultivo (p=0,00001) e em relação ao enrofloxacino e o proprionato nas larvas (p=0,0048). A água do tratamento com probiótico apresentou menor contagem de Vibrio spp. que o enrofloxacino (p=0,0011) e as larvas tratadas com probiótico apresentaram menor contagem de Vibrio spp. que o propionato (p=0,0158). Contudo, não foi observada diferença nos índices zootécnicos avaliados. Assim, os aditivos na dose utilizada não alteram parâmetros zootécnicos da larvicultura do camarão L. vannamei.

The purpose of this study was to assess control and treatment methods for bacterial diseases in Litopenaeus vannamei, evaluating the continuous use of probiotics and occasional use of 15mg-L-1 enrofloxacin and 0.5-mM.L-1 sodium propionate at the morphological change moments on the performance and microbiological parameters of larvae and water. A total of 16 60-L units were used, stocked with 325 nauplii/5L-1, divided into three treatments and one control. Enrofloxacin and propionate were administered into protozoea 3, misis 3 and 4, and post-larvae 4, while the probiotic was administered in the feed throughout the experiment. The probiotic increased the counts of lactic acid bacteria in relation to the other treatments in the culture water (p = 0.00001) and in relation to enrofloxacin and propionate in larvae (p = 0.0048). The treatment water with probiotic had lower counts of Vibrio ssp. than enrofloxacin (p = 0.0011) and larvae treated with probiotic showed lower counts of Vibrio ssp. that propionate (p = 0.0158). However, no difference was observed in the performance indexes assessed. Thus, it can be concluded that additives in the assessed doses did not influence the performance parameters of L. vannamei.

El objetivo del presente trabajo fue evaluar métodos de control y tratamiento de enfermedades bacterianas en larvicultura de Litopenaeus vannamei, evaluando el uso continuo de probiótico y el uso puntual de 15 mg.L-1 de enrofloxacino y 0,5 mM.L-1 de propionato de sodio en momentos de metamorfosis sobre los parámetros zootécnicos y microbiológicos (larvas y del agua). Se han utilizado 16 unidades de 60L, pobladas en la densidad de 325 nauplios.5L-1, divididos en tres tratamientos: enrofloxacino, probiótico, propionato de sodio y control. El enrofloxacino y el propionato fueron suministrados en protozoea 3, misis 3 y postlarva 4 y el probiótico suministrado en alimento durante el transcurso del experimento. El probiótico aumentó el contaje de bacterias ácido-lácticas en correlación a los demás tratamientos en agua de cultivo (p=0,0001) y las larvas en relación al enrofloxacino y el propionato en las larvas (p=0,0048). El agua del tratamiento con probiótico presentó menor contaje de Vibrio spp. que el enrofloxacino (p=0,0011) y las larvas tratadas con probiótico presentaron menor contaje de Vibrio spp. que el propionato (p=0,0158). Sin embargo, no se ha observado diferencia en los índices zootécnicos de larvicultura de camarón L. vannamei.

Utilização comparativa de enrofloxacino, probiótico (Lactobacilus plantarum) e propionato de sódio na larvicultura de camarão branco do pacífico / Comparative use of enrofloxacin, probiotic (Lactobacillus plantarum) and sodium propionate in pacific white shrimp larvae / Utilización comparativa de enrofloxacino, probiótico (Lactobacillus plantarum) y propionato de sodio en larvicultura de camarón blanco del pacífico

O objetivo do presente trabalho foi avaliar métodos de controle e tratamento de bacterioses na larvicultura de Litopenaeus vannamei, avaliando o uso contínuo de probiótico e o uso pontual de enrofloxacino 15mg.L-1 e propionato de sódio 0,5 mM.L-1 nos momentos de metamorfose sobre os, parâmetros zootécnicos e microbiológicos (larvas e da água). Foram utilizadas 16 unidades de 60L, povoadas na densidade de 325 náuplios.5L-1, divididos em três tratamentos: enrofloxacino, probiótico, propionato de sódio e controle. O enrofloxacino e o propionato foram ministrados em protozoea 3, misis 3 e pós-larva 4 e o probiótico foi ministrado na ração ao longo de todo o experimento. O probiótico aumentou as contagens de bactérias ácido-láticas em relação aos demais tratamentos na água de cultivo (p=0,00001) e em relação ao enrofloxacino e o proprionato nas larvas (p=0,0048). A água do tratamento com probiótico apresentou menor contagem de Vibrio spp. que o enrofloxacino (p=0,0011) e as larvas tratadas com probiótico apresentaram menor contagem de Vibrio spp. que o propionato (p=0,0158). Contudo, não foi observada diferença nos índices zootécnicos avaliados. Assim, os aditivos na dose utilizada não alteram parâmetros zootécnicos da larvicultura do camarão L. vannamei.(AU)

The purpose of this study was to assess control and treatment methods for bacterial diseases in Litopenaeus vannamei, evaluating the continuous use of probiotics and occasional use of 15mg-L-1 enrofloxacin and 0.5-mM.L-1 sodium propionate at the morphological change moments on the performance and microbiological parameters of larvae and water. A total of 16 60-L units were used, stocked with 325 nauplii/5L-1, divided into three treatments and one control. Enrofloxacin and propionate were administered into protozoea 3, misis 3 and 4, and post-larvae 4, while the probiotic was administered in the feed throughout the experiment. The probiotic increased the counts of lactic acid bacteria in relation to the other treatments in the culture water (p = 0.00001) and in relation to enrofloxacin and propionate in larvae (p = 0.0048). The treatment water with probiotic had lower counts of Vibrio ssp. than enrofloxacin (p = 0.0011) and larvae treated with probiotic showed lower counts of Vibrio ssp. that propionate (p = 0.0158). However, no difference was observed in the performance indexes assessed. Thus, it can be concluded that additives in the assessed doses did not influence the performance parameters of L. vannamei.(AU)

El objetivo del presente trabajo fue evaluar métodos de control y tratamiento de enfermedades bacterianas en larvicultura de Litopenaeus vannamei, evaluando el uso continuo de probiótico y el uso puntual de 15 mg.L-1 de enrofloxacino y 0,5 mM.L-1 de propionato de sodio en momentos de metamorfosis sobre los parámetros zootécnicos y microbiológicos (larvas y del agua). Se han utilizado 16 unidades de 60L, pobladas en la densidad de 325 nauplios.5L-1, divididos en tres tratamientos: enrofloxacino, probiótico, propionato de sodio y control. El enrofloxacino y el propionato fueron suministrados en protozoea 3, misis 3 y postlarva 4 y el probiótico suministrado en alimento durante el transcurso del experimento. El probiótico aumentó el contaje de bacterias ácido-lácticas en correlación a los demás tratamientos en agua de cultivo (p=0,0001) y las larvas en relación al enrofloxacino y el propionato en las larvas (p=0,0048). El agua del tratamiento con probiótico presentó menor contaje de Vibrio spp. que el enrofloxacino (p=0,0011) y las larvas tratadas con probiótico presentaron menor contaje de Vibrio spp. que el propionato (p=0,0158). Sin embargo, no se ha observado diferencia en los índices zootécnicos de larvicultura de camarón L. vannamei.(AU)

Characterization and pathogenicity of Vibrio splendidus strains associated with massive mortalities of commercial hatchery-reared larvae of scallop Argopecten purpuratus (Lamarck, 1819).

Three strains (VPAP16, VPAP18 and VPAP23 strains) were isolated as the most predominant organisms from 3 different episodes of massive mortalities of larval cultures of the Chilean scallop Argopecten purpuratus occurred in different commercial hatcheries located in northern Chile. The main aims of this study were to identify the pathogenic strains and investigate their pathogenic activity. Based on selected phenotypic features and sequence identity of the 16S rRNA gene and the housekeeping gene, RNA polymerase α-chain rpoA, all pathogenic strains were identified as Vibrio splendidus. Healthy 10-day-old scallop larvae cultures exhibited mortality percentages of 69.61±3.35%, 79.78±6.11% and 61.73±3.71% after 48 h when were inoculated with 1×10(6) CFU (colony forming units)mL(-1) of VPAP16, VPAP18 and VPAP23 strains, respectively, and evidenced that concentrations ⩾10(4) CFU mL(-1) would probably be detrimental for the larval culture. The main clinical signs observed in challenged larvae for 24h were bacterial swarms on the margins of the larvae, extension and disruption of the velum, detachment of velum cilia cells and digestive tissue necrosis. Otherwise, challenge assays using pathogenic strains stained with 5-([4,6-dichlorotriazin-2-yl]amino)fluorescein hydrochloride (5-DTAF)evidenced that after 1h stained bacteria were detected in high density in the digestive gland and the margin of the shell. When larval cultures were inoculated with cell-free extracellular products (ECP) of V. splendidus strains, exhibited larval mortalities higher than 70% (VPAP16), 80% (VPAP18) and 50% (VPAP23) after 24 h, even when ECP were treated with proteinase K or heat, indicating that extracellular pathogenic activity is mainly mediated by non-proteic thermostable compounds. In this study all Koch's postulates were fulfilled and it was demonstrated for the first time the pathogenic activity of V. splendidus strains on reared-larvae of scallop A. purpuratus and prompt the necessity to maintain this species at concentrations lower than 10(4) CFU mL(-1) to avoid episodes of mass mortalities in scallop hatcheries.