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, [Tesis de maestría])

"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."

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

How a holobiome perspective could promote intensification, biosecurity and eco-efficiency in the shrimp aquaculture industry

Eric Daniel Gutiérrez Pérez RICARDO VAZQUEZ JUAREZ FRANCISCO JAVIER MAGALLON BARAJAS MIGUEL ANGEL MARTINEZ MERCADO GRISEL ALEJANDRA ESCOBAR ZEPEDA Paola Magallón Servín (2022, [Artículo])

"The aquaculture industry faces many challenges regarding the intensification of shrimp rearing systems. One of these challenges is the release of excessive amounts of nitrogen and phosphorus into coastal areas, causing disruption in nutrient cycling and microbial equilibrium, which are important for coastal productivity. Biosecurity within the shrimp rearing systems can also be compromised by disruption to the nutrient fluxes, and as consequence the microbiome of the system. In certain conditions, these changes could lead to the blooming of potentially pathogenic bacteria. These changes in the external microbiome of the system and the constant fluctuations of nutrients can affect the intestinal microbiome of shrimp, which is involved in the growth and development of the host, affecting nutrient absorption, regulating metabolic processes, synthesising vitamins, modulating the immune response and preventing growth of pathogenic bacteria. It has been suggested that specific changes in the intestinal microbiome of Litopenaeus vannamei may be an avenue through which to overcome some of the problems that this industry faces, in terms of health, growth and waste. Recent research, however, has focussed mainly on changes in the intestinal microbiome. Researchers have overlooked the relevance of other aspects of the system, such as the microbiome from the benthic biofilms; zooplankton, plankton and bacterioplankton; and other sources of microorganisms that can directly affect the microbial status of the intestinal and epiphytic communities, especially in rearing systems that are based on intensification and microbial maturation processes, such as a biofloc system. It is therefore necessary to place holobiome studies into context, including the ‘holobiome of the aquaculture system’ (microbiomes that make up the culture system and their interactions) and not only the intestinal microbiome. Thus, we describe factors that affect the shrimp microbiome, the methodology of study, from sampling to bioinformatic workflows, and introduce the concept of the ‘holobiome of the aquaculture system’ and how this enables us to promote the intensification, biosafety and eco-efficiency of shrimp farming. The holobiome perspective implies a greater investment of resources and time for research, but it will accelerate the development of technology that will benefit the development and sustainability of the aquaculture industry."

litopenaeus vannamei, microbiome, intensification, biofloc, holobiome of aquaculture systems CIENCIAS AGROPECUARIAS Y BIOTECNOLOGÍA CIENCIAS AGRARIAS PRODUCCIÓN ANIMAL NUTRICIÓN NUTRICIÓN

Uso del cártamo (Carthamus tinctorius L.) como ingrediente en alimentos para juveniles del camarón Litopenaeus vannamei

ALFONSO GALICIA GONZALEZ (2009, [Tesis de doctorado])

Debido a los altos costos e incertidumbre en el abasto de la harina de pescado y pasta de soya, en los últimos años se han buscado alternativas que puedan sustituirlos sin tener un efecto negativo en el crecimiento de los camarones en el cultivo. En el presente trabajo se realizó un estudio para evaluar el valor nutricional del cártamo (Carthamus tinctorius) como fuente proteica en alimentos para juveniles de camarón blanco del Pacífico Litopenaeus vannamei. El primer capítulo comprende el estudio de la caracterización de tres productos de cártamo (HIC; Harina Integral de Cártamo, PCB; Pasta de cártamo baja en proteína y PCA; Pasta de cártamo alta en proteína) en base a su composición de nutrientes y algunos factores antinutricionales (hemaglutininas, saponinas, actividad uréasica, aflatoxinas e inhibidor de tripsina). Los productos de cártamo tuvieron un contenido de proteína entre 20.6 y 36.8%. La harina integral fue la que presentó el mayor nivel de extracto etéreo 31%, mientras que las pastas tuvieron un nivel bajo (1.8-1.0%). El contenido de fibra en el cártamo fue alto (17-23%). Los aminoácidos más abundantes en los productos de cártamo fueron el ácido aspártico y glutámico, en contraste los menos abundantes fueron lisina y metionina. Los ácidos grasos más abundantes fueron el ácido oleico, el linoleico, el palmítico y el estéarico. El contenido de calcio de los productos de cártamo varió de 0.25 a 0.42%, mientras que el contenido de fósforo varió del 0.23 a 0.25%. No se encontraron los factores antinutricionales buscados, a excepción del inhibidor de tripsina que tuvo valores bajos (7.56 UTI/mg de muestra). No se encontraron diferencias significativas en los coeficientes de utilización digestiva aparente (CUDA) de materia seca y carbohidratos de los productos de cártamo; sin embargo, sí se encontraron diferencias en los CUDA de proteína, lípidos (CUDAl) y de energía digestible (CUDAe). La actividad de las enzimas digestivas de camarones que fueron alimentados con productos de cártamo mostró un incremento en proteinasas generales y quimotripsina. La atractabilidad y el consumo de los alimentos con un 30% de inclusión de los productos de cártamo se vieron afectadas negativamente en los camarones [...]

In recent years, alternatives to fish and soybean meals in aquacultural feeds that do not have negative effects on growth of cultivated shrimp have been intensively studied in an effort to reduce the high costs and uncertainty in supplies. In this study, the nutritional value of safflower (Carthamus tinctorius) meal as a protein source in diets for juveniles Pacific whiteleg shrimp (Litopenaeus vannamei) was investigated. Initially, characterization of three safflower meals (i.e. WSM; whole safflower meal, LPSM; low-protein safflower meal and HPSM; high-protein safflower meal), focusing on chemical composition and antinutritional factors (hemaglutinine, saponine, ureasic activity, aflatoxin, and trypsin inhibitor) was conducted. Protein content ranged from 20.6 to 36.8%. Whole safflower meal contained the highest level of lipids (31%); lipids in the other safflower meals were as low as 1.0–1.8%. Fiber content was high (17–23%). The most abundant amino acids were aspartic and glutamic acids and lysine and methionine were less abundant. Oleic, linoleic, palmitic, and stearic fatty acids were present in greatest amounts. Calcium content ranged from 0.25–0.42% and phosphorus content ranged from 0.23–0.25%. Anti-nutritional factors were not found in the analyses, except for trypsin inhibitor, with values as low as 7.56 UTI mg–1 per sample. No significant differences between the apparent digestibility coefficients (ADC) for dry matter and carbohydrates in safflower meals were detected; however, protein ADC, lipid ADC, and energy ADC were different. Total proteinase and chymotrypsin increased significantly in diets where safflower meals were included. Atractability and consumption of feed with 30% safflower meals appeared to have negative effects on the juveniles [...]

atractabilidad; camarón; cártamo; Carthamus tinctorius; digestibilidad; enzimas digestivas; factores antinutricionales; Litopenaeus vannamei CIENCIAS FÍSICO MATEMÁTICAS Y CIENCIAS DE LA TIERRA CIENCIAS DE LA TIERRA Y DEL ESPACIO OCEANOGRAFÍA OCEANOGRAFÍA ACUICULTURA MARINA OCEANOGRAFÍA ACUICULTURA MARINA

Gene expression in primary hemocyte culture of the Pacific white shrimp Penaeus vannamei infected with different white spot syndrome virus (WSSV) strains

Delia Patricia Parrilla Taylor REGINA ELIZONDO GONZALEZ Jesús Neftalí Gutiérrez Rivera SILVIA ALEJANDRA GARCIA GASCA NORBERTO VIBANCO PEREZ MA. DE JESUS DURAN AVELAR RICARDO VAZQUEZ JUAREZ (2022, [Artículo])

"Five previously analyzed white spot syndrome virus (WSSV) strains from northwest Mexico,differing in their genome architecture as well as in virulence, were selected (high virulence JP and LG strains;moderate virulence GVE and DIV strains; and low virulence LC10 strain) to evaluate pathogenesis response in vitro. Expression of phagocytosis-activating protein PAP, manganese superoxide dismutase MnSOD and peroxiredoxin PRX, and two genes of immediate-early expression (IE1 and WSSV304) were measured by qPCR in a primary hemocyte cell culture from Penaeus vannamei at 1, 3, 6, 12, and 24 h post-infection (hpi). PAP expression was significantly higher at 1 and 3 hpi, and JP and LC10 strains induced the highest expression. The response of MnSOD was high at 1 hpi, and a significant increase in PRX expression was detected at 3 hpi, probably due to the occurrence of an oxidative burst; expression levels of MnSOD and PRX were significantly higher at 1 and 3 hpi, respectively, induced by the LG strain (high virulence), suggesting an acute response. In general, expression of most immune-related - genes decreased after the initial hours of infection. Expression levels of IE1 and WSSV304 were exceptionally high at 1 hpi in almost all five WSSV analyzed strains,confirming their efficient mechanism for replication and viral fitness. The results of this study do not show an accurate link between the genome size and WSSV virulence of the strains, albeit the strain with the smallest genome showed the highest virulence. All strains induced an early immune response in heterogeneous ways."

Penaeus vannamei, virulence, gene expression, viral fitness, viral pathogenesis, immune response BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA BIOLOGÍA ANIMAL (ZOOLOGÍA) PATOLOGÍA ANIMAL PATOLOGÍA ANIMAL

Chronic toxicity of shrimp feed added with silver nanoparticles (Argovit-4®) in Litopenaeus vannamei and immune response to white spot syndrome virus infection

Carlos Rosendo Romo Quiñonez Píndaro Álvarez Ruiz CLAUDIO HUMBERTO MEJIA RUIZ NINA BOGDANCHIKOVA Alexey Pestryakov CARINA GAMEZ JIMENEZ WENCESLAO VALENZUELA QUIÑONEZ Magnolia Montoya Mejía EUSEBIO NAVA PEREZ (2022, [Artículo])

"In recent years, the application of silver nanoparticles (AgNPs) as antibacterial compounds has been widely used in human and veterinary medicine. In this work, we investigated the effects of AgNPs (Argovit-4 R ) as feed additives (feed-AgNPs) on shrimp (Litopenaeus vannamei) using three different methods: 1) chronic toxicity after 28 days of feeding, 2) Effects against white spot syndrome virus (WSSV) challenged by oral route, and 3) transcriptional responses of immune-related genes (PAP, ProPO, CTL-3, Crustin, PEN3, and PEN4) following WSSV infection. The results showed that the feed-AgNPs did not interfere with the growth and survival of shrimp. Also, mild lesions in the hepatopancreas were recorded, proportional to the frequency of the feed-AgNP supply. Challenge test versus WSSV showed that feeding every 7 days with feed-AgNPs reduced mortality, reaching a survival rate of 53%, compared to the survival rates observed in groups fed every 4 days, daily and control groups of feed-AgNPs for the 30%, 10%, and 7% groups, respectively. Feed-AgNPs negatively regulated the expression of PAP, ProPO, and Crustin genes after 28 days of treatment and altered the transcriptional responses of PAP, ProPO, CTL-3, and Crustin after WSSV exposure. The results showed that weekly feeding-AgNPs could partially prevent WSSV infection in shrimp culture. However, whether or not transcriptional responses against pathogens are advantageous remains to be elucidated."

Silver nanoparticles, Shrimp, Aquaculture, Chronic toxicity, WSSV, AgNP, Argovit, Litopenaeus vannamei , Silver fed, White spot syndrome virus BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA INMUNOLOGÍA INMUNIZACIÓN INMUNIZACIÓN