Supervivencia de Artemia franciscana en cocultivo con Vibrio campbellii y Nitzschia thermalis f. kurta

Survival of Artemia franciscana in coculture with Vibrio campbellii and Nitzschia thermalis f. kurta

Dalia Azucena Verde Inzunza (2023)

La acuicultura es uno de los sectores más importantes y de mayor producción alimentaria. Sin embargo, se ve afectada por enfermedades bacterianas como la vibriosis, la cual puede ocasionar pérdidas totales en los cultivos. El método más utilizado para controlar la vibriosis es el uso de antibióticos, lo cual ha contribuido al desarrollo de cepas bacterianas resistentes. Por ello, se buscan medios de control naturales para inhibir el crecimiento de Vibrio, como el uso de aguas verdes. El objetivo de este trabajo fue evaluar el efecto inhibitorio de Nitzschia thermalis f. kurta en la concentración de V. campbellii y en la supervivencia de nauplios de Artemia franciscana al estar en cocultivo. Se realizó un bioensayo de inhibición de V. campbellii en cocultivo con N. thermalis y nauplios de A. franciscana. Se mantuvieron cultivos monoespecíficos, no axénicos y por triplicado en medio “f” con una densidad celular de 250,000 cél. mL-1 de N. thermalis. Los cultivos fueron inoculados con una concentración inicial de 1 x105 V. campbellii mL-1 y una densidad de 1 nauplio de A. franciscana mL-1. Para cada tratamiento se evaluó la densidad celular de N. thermalis, conteo de bacterias heterotróficas, cuantificación de la concentración de V. campbellii y la supervivencia de nauplios de A. franciscana durante 5 días. Se obtuvo una supervivencia promedio de 55% de nauplios de A. franciscana al estar en cocultivo con N. thermalis. Mientras que el tratamiento control sin la presencia de N. thermalis obtuvo una supervivencia de nauplios de A. franciscana del 0%. El contenido de bacterias heterotróficas se presentó en todos los tratamientos en un orden de magnitud de 106 UFC mL-1, lo cual nos indica que el efecto inhibitorio de N. thermalis es específico para V. campbellii. La concentración de V. campbellii disminuyó a 53 V. campbellii mL-1 al día 5 en el cocultivo de V. campbellii con N. thermalis y nauplios de A. franciscana. Se concluye que N. thermalis tiene un efecto inhibitorio contra V. campbellii y brinda protección a los nauplios de A. franciscana en cocultivo con V. campbellii.

Aquaculture is one of the most important sectors with the highest food production. However, it is affected by bacterial diseases such as vibriosis, which can cause total crop losses. The most used method to control vibriosis is antibiotics, which have contributed to the developing of resistant bacterial strains. For this reason, natural control means are sought to inhibit Vibrio growth, such as using green water. This work aimed to evaluate the inhibitory effect of Nitzschia thermalis f. kurta in the concentration of V. campbellii and the survival of Artemia franciscana nauplii when in coculture. An inhibition bioassay of V. campbellii was carried out in coculture with N. thermalis and nauplii of A. franciscana. Monospecific, non-axenic cultures were maintained in triplicate in “f” medium with a cell density of 250,000 cells mL-1 of N. thermalis. The cultures were inoculated with an initial concentration of 1 x105 V. campbellii mL-1 and a density of 1 nauplius of A. franciscana mL-1. For each treatment, the cell density of N. thermalis, count of heterotrophic bacteria, quantification of the concentration of V. campbellii, and the survival of A. franciscana nauplii were evaluated for 5 days. Was obtained an average survival of 55% of A. franciscana nauplii when cocultured with N. thermalis. In contrast, the control treatment without the presence of N. thermalis obtained a survival of A. franciscana nauplii of 0%. The content of heterotrophic bacteria was present in all treatments in order of magnitude of 106 CFU mL-1, which indicates that the inhibitory effect of N. thermalis is specific to V. campbellii. The concentration of V. campbellii decreased to 53 V. campbellii mL-1 on day 5 in the co-culture of V. campbellii with N. thermalis and A. franciscana nauplii. It is concluded that N. thermalis has an inhibitory effect against V. campbellii and provides protection to A. franciscana nauplii in coculture with V. campbellii.

Master thesis

Artemia franciscana, Vibrio campbellii, inhibición, Nitzschia thermalis Artemia franciscana, Vibrio campbellii, inhibition, Nitzschia thermalis CIENCIAS AGROPECUARIAS Y BIOTECNOLOGÍA CIENCIAS AGRARIAS PECES Y FAUNA SILVESTRE PISCICULTURA PISCICULTURA

Evaluación del uso de vibriófagos encapsulados en el alimento para el control de Vibrio parahaemolitycus causante de AHPND en el camarón Penaeus vannamei

Esther Imelda Ponce García (2023)

"Las enfermedades causadas por bacterias del género Vibrio en el cultivo de camarón son de gran importancia debido a su impacto negativo en la industria acuícola. Estas bacterias pueden causar diversas enfermedades en camarones, como la necrosis hepatopancreática aguda (AHPND). La fagoterapia es una alternativa para el biocontrol de estas bacterias patógenas que presenta importantes ventajas, como la alta especificidad a sus huéspedes, la replicación en el sitio de infección, y la capacidad de infectar a bacterias resistentes a antibióticos. Sin embargo, la estabilidad de los fagos en el sistema de cultivo es una preocupación latente, ya que los fagos presentan una estabilidad limitada en solución y experimentan una caída significativa en el título de fagos durante el procesamiento y almacenamiento. El método de encapsulación de bacteriófagos, un área que aún no se estudia del todo, se presenta como una potencial alternativa para atender esta problemática. En el presente estudio se evaluó la eficacia terapéutica de un cóctel de bacteriófagos (vB_Vp_PvVp05, vB_Vp_PvVp07 y vB_Vp_PvVp11), encapsulados en alginato de sodio comercial y de mediana viscosidad, pectina, carboxilmetilcelulosa, liposomas y liofilizado en pectina como agente de biocontrol de Vibrio parahaemolyticus para su uso en terapias fagicas pasiva y activa. Se observó que el encapsulado en alginato de sodio de mediana viscosidad presentó la mayor liberación de fagos activos y al momento de ser incorporado a alimento para camarón y horneados a 80 – 100°C, se mantuvo una concentración de 1.3 x105 UFP/g después de 100 días de almacenamiento. Para la evaluación de las terapias pasiva y activa se realizó una infección experimental en juveniles de Penaeus vannamei para conocer la efectividad de los encapsulados, utilizando para la terapia pasiva la cepa Vp M0904 y para la activa la cepa Vp M0605. El experimento de terapia pasiva mostró una mortalidad del 80.6% en los camarones a las 48 hpi (p > 0.05) y una concentración de 5 x102 UFC/mL de Vibrio spp. no fermentativos en TCBS. En el caso de la terapia activa no se presentó mortalidades, pero hubo una disminución de la concentración de Vibrio spp. no fermentativos en TCBS con 5.56 x101 UFC/mL a las 48 hpi (p < 0.05) y aumentó la densidad de vibriófagos en 1.4 x102 UFP/mL en el agua de los acuarios (p < 0.05)..."

"Diseases caused by bacteria of the Vibrio genus in shrimp farming are of great importance due to their negative impact on the aquaculture industry. These bacteria can cause various diseases in shrimp, such as acute hepatopancreatic necrosis disease (AHPND). Phage therapy is an alternative for the biocontrol of these pathogenic bacteria, offering significant advantages, including high specificity to their hosts, replication at the infection site, and the ability to infect antibiotic-resistant bacteria. However, the stability of phages in the farming system is a persistent concern, as phages have limited stability in solution and undergo a significant drop in phage titer during processing and storage. The method of bacteriophage encapsulation, an area that is not yet fully explored, emerges as a potential alternative to address this issue. In this study, the therapeutic efficacy of a cocktail of bacteriophages (vB_Vp_PvVp05, vB_Vp_PvVp07, and vB_Vp_PvVp11), encapsulated in commercial medium-viscosity sodium alginate, pectin, carboxymethylcellulose, liposomes, and lyophilized pectin, was evaluated as a biocontrol agent against Vibrio parahaemolyticus for use in passive and active phage therapies. It was observed that encapsulation in medium-viscosity sodium alginate showed the highest release of active phages, maintaining a concentration of 1.3 x 105 PFU/g after 100 days of storage when incorporated into shrimp feed and baked at 80–100°C.For the evaluation of passive and active therapies, an experimental infection was conducted on Penaeus vannamei juveniles to assess the effectiveness of the encapsulated phages. The passive therapy experiment showed a mortality rate of 80.6% in shrimp at 48 hpi (p > 0.05) and a concentration of 5 x 102 CFU/mL of non-fermentative Vibrio spp. on TCBS agar. In the case of active therapy, no mortalities were observed, but there was a decrease in the concentration of non-fermentative Vibrio sp. on TCBS agar to 5.56 x 101 CFU/mL at 48 hpi (p < 0.05), and the density of vibriophages increased to 1.4 x 102 PFU/mL in the aquarium water (p < 0.05). It was determined that passive therapy offered no advantage for the control of Vibrio parahaemolyticus, unlike active therapy, which proved to be effective in controlling this bacterium."

Master thesis

vibriófagos, encapsulado, Penaeus vannamei, terapia activa, terapia pasiva vibriophages, encapsulated, active therapy, passive therapy BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA MICROBIOLOGÍA BACTERIÓFAGOS BACTERIÓFAGOS

Whole-genome comparison between reference sequences and oyster Vibrio vulnificus C-genotype strains

CARLOS ABRAHAM GUERRERO RUIZ (2019)

Whole-genome sequences of Vibrio vulnificus clinical genotype (C-genotype) from the CICESE Culture Collection, isolated from oysters, were compared with reference sequences of CMCP6 and YJ016 V. vulnificus C-genotype strains of clinical origin. The RAST web server estimated the whole genome to be ~4.8 Mb in CICESE strain 316 and ~4.7 Mb in CICESE strain 325. No plasmids were detected in the CICESE strains. Based on a phylogenetic tree that was constructed with the whole-genome results, we observed high similarity between the reference sequences and oyster C-genotype isolates and a sharp contrast with environmental genotype (E-genotype) reference sequences, indicating that the differences between the C- and E-genotypes do not necessarily correspond to their isolation origin. The CICESE strains share 3488 genes (63.2%) with the YJ016 strain and 3500 genes (63.9%) with the CMCP6 strain. A total of 237 pathogenicity associated genes were selected from reference clinical strains, where—92 genes were from CMCP6, 126 genes from YJ016, and 19 from MO6-24/ O; the presence or absence of these genes was recorded for the CICESE strains. Of the 92 genes that were selected for CMCP6, 67 were present in both CICESE strains, as were as 86 of the 126 YJ016 genes and 13 of the 19 MO6-24/O genes. The detection of elements that are related to virulence in CICESE strains—such as the RTX gene cluster, vvhA and vvpE, the type IV pili cluster, the XII genomic island, and the viuB genes, suggests that environmental isolates with the C-genotype, have significant potential for infection. © 2019 Guerrero et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Article

Article, bacterial gene, bacterial strain, bacterial virulence, comparative study, controlled study, gene cluster, gene identification, genomic island, genotype, nonhuman, phylogenetic tree, sequence analysis, strain identification, Vibrio vulnificus BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA GENÉTICA GENÉTICA

Phylogenetic relationships of Pseudo-nitzschia subpacifica (Bacillariophyceae) from the Mexican Pacific, and its production of domoic acid in culture

Sonia Quijano (2020)

Pseudo-nitzschia is a cosmopolitan genus, some species of which can produce domoic acid (DA), a neurotoxin responsible for the Amnesic Shellfish Poisoning (ASP). In this study, we identified P. subpacifica for the first time in Todos Santos Bay and Manzanillo Bay, in the Mexican Pacific using SEM and molecular methods. Isolates from Todos Santos Bay were cultivated under conditions of phosphate sufficiency and deficiency at 16°C and 22°C to evaluate the production of DA. This toxin was detected in the particulate (DAp) and dissolved (DAd) fractions of the cultures during the exponential and stationary phases of growth of the cultures. The highest DA concentration was detected during the exponential phase grown in cells maintained in P-deficient medium at 16°C (1.14 ± 0.08 ng mL-1 DAd and 4.71 ± 1.11 × 10−5 ng cell-1 of DAp). In P-sufficient cultures DA was higher in cells maintained at 16°C (0.25 ± 0.05 ng mL-1 DAd and 9.41 ± 1.23 × 10−7 ng cell-1 of DAp) than in cells cultured at 22°C. Therefore, we confirm that P. subpacifica can produce DA, especially under P-limited conditions that could be associated with extraordinary oceanographic events such as the 2013–2016 "Blob" in the northeastern Pacific Ocean. This event altered local oceanographic conditions and possibly generated the presence of potential harmful species in areas with economic importance on the Mexican Pacific coast. © 2020 Quijano-Scheggia et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Article

domoic acid, domoic acid, kainic acid, Article, cell growth, controlled study, diatom, Mexico, morphology, nonhuman, Pacific Ocean, phylogeny, plant cell, plant growth, Pseudo nitzschia, toxin analysis, cell culture technique, classification, diatom, CIENCIAS FÍSICO MATEMÁTICAS Y CIENCIAS DE LA TIERRA CIENCIAS DE LA TIERRA Y DEL ESPACIO OCEANOGRAFÍA OCEANOGRAFÍA