Lysin and Lytic Phages Reduce Vibrio Counts in Live Feed and Fish Larvae.

Vibrio species are naturally found in estuarine and marine ecosystems, but are also recognized as significant human enteropathogens, often linked to seafood-related illnesses. In aquaculture settings, Vibrio poses a substantial risk of infectious diseases, resulting in considerable stock losses and prompting the use of antimicrobials. However, this practice contributes to the proliferation of antimicrobial-resistant (AMR) bacteria and resistance genes. Our investigation aimed to explore the potential of biological agents such as bacteriophage CH20 and endolysin LysVPp1 in reducing Vibrio bacterial loads in both rotifer and fish larvae. LysVPp1's lytic activity was assessed by measuring absorbance reduction against various pathogenic Vibrio strains. Phage CH20 exhibited a limited host range, affecting only Vibrio alginolyticus GV09, a highly pathogenic strain. Both CH20 and LysVPp1 were evaluated for their effectiveness in reducing Vibrio load in rotifers or fish larvae through short-setting bioassays. Our results demonstrated the significant lytic effect of endolysin LysVPp1 on strains of Vibrio alginolyticus, Vibrio parahaemolyticus, and Vibrio splendidus. Furthermore, we have showcased the feasibility of reducing the load of pathogenic Vibrio in live feed and fish larvae by using a non-antibiotic-based approach, such as lytic phage and endolysin LysVPp1, thus contributing to the progress of a sustainable aquaculture from a One Health perspective.

Influence of Microalgae Diets on the Biological and Growth Parameters of Oithona nana (Copepoda: Cyclopoida).

Several species of the planktonic free-living genus Oithona have been successfully used in the larviculture of marine fish and shrimp. However, few studies have been published that allow us to estimate the potential of Oithona nana culture under controlled conditions. This work evaluated the effect of the microalgae Isochrysis galbana and Chaetoceros calcitrans as single (200,000 cells/mL) and mixed diets (100,000 + 100,000 cells/mL) on population and individual growth, ingestion rate, number of spawnings, fertility, development time by stage, and sex ratio of O. nana. We cultured this copepod at 28 ± 0.5 °C, 35 PSU salinity, 125 lux, and 12:12 photoperiod. Results showed that diet had no effect on the final population level (6273-7966 ind/L) or on individual growth, nor on sex ratio, with less males than females. With C. calcitrans, O. nana had a higher filtration rate (57 ng C/ind/day). On the other hand, a mixed diet induced a higher number of spawns (0.4 events/day) and nauplii per spawn (23 ind). Similarly, a single or mixed diet, containing I. galbana, accelerated the development rate by 6.33-7.00 days. We concluded that O. nana can be cultured with both microalgae, indicating its potential use in an intensive system for production. However, more research is required to improve the productivity of O. nana rearing.

Marine amphipods (Parhyale hawaiensis) as an alternative feed for the lined seahorse (Hippocampus erectus, Perri 1810): nutritional value and feeding trial.

Finding new alternatives to traditional live preys such as Artemia and rotifers, which do not always promote optimal fish growth and survival, is required for the successful aquaculture of highly specialized predatory species, including seahorses. The present study assessed the nutritional value of an interesting marine amphipod (Parhyale hawaiensis), and evaluates through a feeding trial its potential use as a natural prey for 10-months lined seahorse, Hippocampus erectus. P. hawaiensis showed high levels of valuable lipids (20.4-26.7% on dry matter basis) and polyunsaturated fatty acids (PUFAs) ( 26.4-41% of total FAs), including the long-chain PUFAs (LC-PUFAs) arachidonic acid (ARA) (2.9-7.7%), eicosapentaenoic acid (EPA) (4.3-6.5%) and docosahexaenoic acid (DHA) (2.1-6.2%). A comparison between wild-captured and cultured amphipods revealed a significant improvement of the amphipod FA profile in terms of DHA%, total omega-3 (n3) FAs and n3/n6 ratio when employing both a conventional amphipod culture based on a commercial shrimp diet, and, to a lesser extent, a large (3,500 L) biofloc system. Seahorses fed with frozen/wild amphipods, either singly or in combination with Artemia enriched with Super Selco® (INVE Aquaculture, Belgium) for 57 days, substantially improved seahorse growth and FA profiles in terms of ARA, EPA and DHA%, including indices associated to marine sources, such as Σn3 and n3/n6, compared to a diet based solely on enriched Artemia. These results support the use of marine amphipods as an alternative food organism for juvenile H. erectus and suggest a potential use for general marine aquaculture.

Live Feeds Used in the Larval Culture of Red Cusk Eel, Genypterus chilensis, Carry High Levels of Antimicrobial-Resistant Bacteria and Antibiotic-Resistance Genes (ARGs).

The culture of red cusk eel Genypterus chilensis is currently considered a priority for Chilean aquaculture but low larval survival rates have prompted the need for the continuous use of antibacterials. The main aim of this study was to evaluate the role of live feed as a source of antibacterial-resistant bacteria in a commercial culture of G. chilensis. Samples of rotifer and Artemia cultures used as live feed were collected during the larval growth period and culturable bacterial counts were performed using a spread plate method. Rotifer and Artemia cultures exhibited high levels of resistant bacteria (8.03 × 104 to 1.79 × 107 CFU/g and 1.47 × 106 to 3.50 × 108 CFU/g, respectively). Sixty-five florfenicol-resistant isolates were identified as Vibrio (81.5%) and Pseudoalteromonas (15.4%) using 16S rRNA gene sequence analysis. A high incidence of resistance to streptomycin (93.8%), oxytetracycline (89.2%), co-trimoxazole (84.6%), and kanamycin (73.8%) was exhibited by resistant isolates. A high proportion of isolates (76.9%) carried the florfenicol-resistance encoding genes floR and fexA, as well as plasmid DNA (75.0%). The high prevalence of multiresistant bacteria in live feed increases the incidence of the resistant microbiota in reared fish larvae, thus proper monitoring and management strategies for live feed cultures appear to be a priority for preventing future therapy failures in fish larval cultures.

Stocking density and feed management on larval rearing of Amazonian ornamental fish Heros severus / Manejo alimentar e densidade de estocagem na larvicultura do peixe ornamental amazônico Heros severus

The banded cichlid (Herus severus) is an Amazonian species with potential as ornamental fish, however basic handlings as quantity and feeding frequency as well as stocking density in larviculture are still unknown in order to allow the rational production of this species in captivity. Then the objective was to evaluate the amount of food, feeding frequency and stocking density during the larviculture of Herus severus. Therefore, two experiments were conducted: the first assessed the food concentration (100, 150, 200, 250 and 300 Artemia larvae-1) and feed rate (2 or 4 meals day-1), and the second experiment evaluated different stocking densities (1, 5, 10, 15, 20 larvae L-1) with four replications each. Feeding of 250 nauplii larva day-1 distributed in four daily meals promoted the highest weight, length, specific growth rate, weight gain and survival. In the second experiment, reduction in growth was observed with increased in density due to lower concentrations of dissolved oxygen and elevated total ammonia from 5 larvae L-1. Thus, to promote the rational cultivation of the Banded cichlid larvae in captivity, the species should be reared at 5 larvae L-1 fed with 250nauplii distributed along 4 daily meals.(AU)

O Acará-severo (Herus severus) é uma espécie amazônica com potencial para a piscicultura ornamental, entretanto manejos básicos como quantidade e frequência de alimentação assim como densidade de estocagem ainda são desconhecidos na larvicultura para permitir a produção racional da espécie em cativeiro. Assim o objetivo foi avaliar quantidade de alimento, frequência alimentar e densidade de estocagem durante a larvicultura de Herus severus. Para tanto, foram realizados dois experimentos: o primeiro avaliando a concentração de alimento (100, 150, 200, 250 e 300 náuplios de Artemia larva-1) e frequência alimentar (2 ou 4 vezes dia-1), e o segundo avaliando diferentes densidades de estocagem (1, 5, 10, 15, 20 larvas L-1) ambos com quatro repetições. A alimentação com 250 náuplios por larva dia-1 em quatro alimentações diárias promoveu maiores valores de peso, comprimento, taxa de crescimento específico, ganho de peso e sobrevivência. No segundo experimento foi observada redução no crescimento com aumento da densidade devido as menores concentrações de oxigênio dissolvido e elevação da amônia total a partir de 5 larva L-1. Assim, para promover o cultivo racional das larvas de acara severo em cativeiro, a espécie deve ser cultivada com 5 larvas L-1, alimentadas com 250 náuplios em 4 alimentações diárias.(AU)

Stocking density and feed management on larval rearing of Amazonian ornamental fish Heros severus / Manejo alimentar e densidade de estocagem na larvicultura do peixe ornamental amazônico Heros severus

The banded cichlid (Herus severus) is an Amazonian species with potential as ornamental fish, however basic handlings as quantity and feeding frequency as well as stocking density in larviculture are still unknown in order to allow the rational production of this species in captivity. Then the objective was to evaluate the amount of food, feeding frequency and stocking density during the larviculture of Herus severus. Therefore, two experiments were conducted: the first assessed the food concentration (100, 150, 200, 250 and 300 Artemia larvae-1) and feed rate (2 or 4 meals day-1), and the second experiment evaluated different stocking densities (1, 5, 10, 15, 20 larvae L-1) with four replications each. Feeding of 250 nauplii larva day-1 distributed in four daily meals promoted the highest weight, length, specific growth rate, weight gain and survival. In the second experiment, reduction in growth was observed with increased in density due to lower concentrations of dissolved oxygen and elevated total ammonia from 5 larvae L-1. Thus, to promote the rational cultivation of the Banded cichlid larvae in captivity, the species should be reared at 5 larvae L-1 fed with 250nauplii distributed along 4 daily meals.

O Acará-severo (Herus severus) é uma espécie amazônica com potencial para a piscicultura ornamental, entretanto manejos básicos como quantidade e frequência de alimentação assim como densidade de estocagem ainda são desconhecidos na larvicultura para permitir a produção racional da espécie em cativeiro. Assim o objetivo foi avaliar quantidade de alimento, frequência alimentar e densidade de estocagem durante a larvicultura de Herus severus. Para tanto, foram realizados dois experimentos: o primeiro avaliando a concentração de alimento (100, 150, 200, 250 e 300 náuplios de Artemia larva-1) e frequência alimentar (2 ou 4 vezes dia-1), e o segundo avaliando diferentes densidades de estocagem (1, 5, 10, 15, 20 larvas L-1) ambos com quatro repetições. A alimentação com 250 náuplios por larva dia-1 em quatro alimentações diárias promoveu maiores valores de peso, comprimento, taxa de crescimento específico, ganho de peso e sobrevivência. No segundo experimento foi observada redução no crescimento com aumento da densidade devido as menores concentrações de oxigênio dissolvido e elevação da amônia total a partir de 5 larva L-1. Assim, para promover o cultivo racional das larvas de acara severo em cativeiro, a espécie deve ser cultivada com 5 larvas L-1, alimentadas com 250 náuplios em 4 alimentações diárias.

Artemia franciscana as a vector for infectious myonecrosis virus (IMNV) to Litopenaeus vannamei juvenile.

In 2004, the infectious myonecrosis virus (IMNV) was recognized as the main cause of Litopenaeusvannamei shrimp culture's drop in Brazil. In health animal control programs, in order to reduce virus prevalence in production units it is necessary to screen live feed used. Among live diets used in aquaculture, the brine shrimp Artemia sp. is essential in crustacean larviculture and maturation. The aim of the present study was to investigate the susceptibility of Artemiafranciscana to IMNV through an immersion challenge and virus-phytoplankton adhesion route and to elucidate its role as a vector for IMNV transmission to L.vannamei. A. franciscana adults were infected with IMNV through both routes, as demonstrated by PCR-positive reactions. However, infected A. franciscana showed no signs of infection. More than 40% of L. vannamei juveniles fed with IMNV-infected A. franciscana by virus-phytoplankton adhesion route were positive by real-time PCR, whereas only a 10% infection rate was found among shrimp fed with IMNV-infected brine shrimp using the immersion challenge. Significant differences were found in mean viral load between immersion and virus-phytoplankton adhesion shrimp treatments (p ⩽ 0.05). Moreover, the mean viral loads were 1.34 × 10(2) and 1.48 × 10(4) copies/µg(-1) of total RNA for virus-phytoplankton adhesion and IMNV-infected tissue treatments, respectively, and the difference was not significant (p ⩾ 0.05). The results indicated that A. franciscana act as a vector for IMNV transmission under the experimental conditions examined. Although no mass mortalities were detected in L. vannamei fed with IMNV-infected brine shrimp, these infected shrimp should not be disregarded as a source of IMNV in grow-out units.

First feeding of Eugerres brasilianus (Carapeva) larvae with Acartia tonsa (Copepod) nauplii increases survival and resistance to acute stress / Alimentação inicial de larvas de Carapeva Eugerres brasilianus com náuplios de Copepodo Acartia tonsa melhora a sobrevivência e resistência ao estresse agudo

The rotifer Brachionus sp. is commonly used for larval feeding in marine fish hatcheries. The aim of this study was to evaluate whether the inclusion of Acartia tonsa nauplii in the initial diet of carapeva larvae improves their survival, growth and resistance to stress when compared to the regimen containing only rotifers. Adult copepods were collected in the wild and cultured with the microalgae Chaetoceros muelleri, Isochrysis galbana and Nannochloropsis oculata to obtain nauplii. Carapeva larvae were grown for 15 days using four treatments and three replicates: 1) Brachionus plicatilis rotifers (10 to 15/mL); 2) A. tonsa nauplii (0.25 to 0.5/mL); 3) Brachionus plicatilis rotifers (5 to 7.5/mL) + A. tonsa nauplii (0.12 to 0.25/mL), and 4) no supply of live feed. After 15 days, the carapeva larvae were subjected to stress by exposure to air for 10 seconds and then returned to the source tank to evaluate survival after 24 h. Survival and stress resistance were higher in carapeva larvae fed B. plicatilis + A. tonsa nauplii (P<0.05), 20.9 ± 11.2% and 88.9%, respectively. These results confirm the positive effect of the inclusion of copepod nauplii in the diet of fish larvae. However, more research is needed to validate these results...(AU)

O rotífero Brachionus sp. é o alimento vivo mais comumente utilizado na larvicultura de peixes marinhos. O objetivo deste estudo foi avaliar se a inclusão de náuplios de Acartia tonsa na alimentação inicial de larvas de carapeva melhora sua sobrevivência, crescimento e resistência ao estresse, comparado com o regime contendo apenas rotíferos. Copépodos adultos foram coletados no ambiente e cultivados com as microalgas Chaetoceros muelleri, Isochrysis galbana and Nannochloropsis oculata para obtenção dos náuplios. As larvas de carapeva foram cultivadas por 15 dias com quatro tratamentos e três repetições: 1) rotíferos Brachionus plicatilis (10 a 15/mL); 2) náuplios de A. tonsa (0,25 a 0,5/mL); 3) rotíferos Brachionus plicatilis (5 a 7,5/ mL) + náuplios de A. tonsa (0,12 a 0,25/mL) e 4) sem suplementação de alimento vivo. Após 15 dias, as larvas de carapeva foram submetidas a estresse por exposição ao ar, por 10 segundos e retornadas aos tanques para avaliar a sobrevivência após 24 h. As larvas alimentadas com rotífero B. plicatilis + náuplio de A. tonsa apresentaram maiores sobrevivência e resistência ao estresse (P<0,05), 20,9 ± 11,2% e 88,9%, respectivamente. Esses resultados confirmam o efeito positivo da inclusão de náuplios de copépodos na dieta de larvas de peixes. Contudo, são necessárias mais pesquisas para validar esses resultados...(AU)

First feeding of Eugerres brasilianus (Carapeva) larvae with Acartia tonsa (Copepod) nauplii increases survival and resistance to acute stress / Alimentação inicial de larvas de Carapeva Eugerres brasilianus com náuplios de Copepodo Acartia tonsa melhora a sobrevivência e resistência ao estresse agudo

The rotifer Brachionus sp. is commonly used for larval feeding in marine fish hatcheries. The aim of this study was to evaluate whether the inclusion of Acartia tonsa nauplii in the initial diet of carapeva larvae improves their survival, growth and resistance to stress when compared to the regimen containing only rotifers. Adult copepods were collected in the wild and cultured with the microalgae Chaetoceros muelleri, Isochrysis galbana and Nannochloropsis oculata to obtain nauplii. Carapeva larvae were grown for 15 days using four treatments and three replicates: 1) Brachionus plicatilis rotifers (10 to 15/mL); 2) A. tonsa nauplii (0.25 to 0.5/mL); 3) Brachionus plicatilis rotifers (5 to 7.5/mL) + A. tonsa nauplii (0.12 to 0.25/mL), and 4) no supply of live feed. After 15 days, the carapeva larvae were subjected to stress by exposure to air for 10 seconds and then returned to the source tank to evaluate survival after 24 h. Survival and stress resistance were higher in carapeva larvae fed B. plicatilis + A. tonsa nauplii (P<0.05), 20.9 ± 11.2% and 88.9%, respectively. These results confirm the positive effect of the inclusion of copepod nauplii in the diet of fish larvae. However, more research is needed to validate these results...

O rotífero Brachionus sp. é o alimento vivo mais comumente utilizado na larvicultura de peixes marinhos. O objetivo deste estudo foi avaliar se a inclusão de náuplios de Acartia tonsa na alimentação inicial de larvas de carapeva melhora sua sobrevivência, crescimento e resistência ao estresse, comparado com o regime contendo apenas rotíferos. Copépodos adultos foram coletados no ambiente e cultivados com as microalgas Chaetoceros muelleri, Isochrysis galbana and Nannochloropsis oculata para obtenção dos náuplios. As larvas de carapeva foram cultivadas por 15 dias com quatro tratamentos e três repetições: 1) rotíferos Brachionus plicatilis (10 a 15/mL); 2) náuplios de A. tonsa (0,25 a 0,5/mL); 3) rotíferos Brachionus plicatilis (5 a 7,5/ mL) + náuplios de A. tonsa (0,12 a 0,25/mL) e 4) sem suplementação de alimento vivo. Após 15 dias, as larvas de carapeva foram submetidas a estresse por exposição ao ar, por 10 segundos e retornadas aos tanques para avaliar a sobrevivência após 24 h. As larvas alimentadas com rotífero B. plicatilis + náuplio de A. tonsa apresentaram maiores sobrevivência e resistência ao estresse (P<0,05), 20,9 ± 11,2% e 88,9%, respectivamente. Esses resultados confirmam o efeito positivo da inclusão de náuplios de copépodos na dieta de larvas de peixes. Contudo, são necessárias mais pesquisas para validar esses resultados...

Alimento vivo enriquecido con ácidos grasos para el desarrollo larvario de peces / Alimento vivo e seu enriquecimento com ácidos graxos no desenvolvimento larvário em peixes / Live feed enriched with fatty acids for larval fish development

La nutrición de las larvas representa uno de los principales problemas en la producción de peces a granescala; el objetivo principal de esta revisión es profundizar en el conocimiento de los factores que intervienen enel desarrollo orgánico y digestivo de los peces y en cómo éstos, pueden afectar su desarrollo y sobrevivencia. Eldesarrollo del tracto digestivo en las larvas de peces está condicionado por aspectos de tipo anatomo-fisiológico,que permiten a la poslarva adaptarse bioquímica e histológicamente a los períodos de transición entre lafinalización de la reabsorción del vitelo y el inicio del consumo de alimento vivo, y en la transición de alimentovivo a dieta comercial balanceada. Las especies empleadas en la primera alimentación de la poslarva, afectansu sobrevivencia y desarrollo, debido a que un suministro inadecuado, podría ocasionar una alta mortalidad,ya que las especies zooplanctónicas que proliferan, no siempre satisfacen los requerimientos nutricionales dela poslarva, se producen en un volumen inferior al requerido y/o un mal manejo en su producción, favorece laproliferación de especies planctónicas depredadoras. Entre los principales nutrientes que aporta el alimentovivo, están los ácidos grasos insaturados y poliinsaturados. Así mismo deficiencias nutricionales en el alimentovivo pueden ser complementadas mediante enriquecimiento con ácidos grasos, esenciales en los procesos depigmentación, producción de prostaglandinas, respuesta inmunológica, desarrollo retinal entre otras.

The larvae nutrition represents one of the main problems in the great scale production of fish. Theprimary objective of this review is to enlarge the knowledge of the factors that take part in the organic and digestive development of fish and the way these factors can affect their growth and survival. Thedevelopment of digestive tract in the larvae of fish is conditioned by anatomo-physiological aspects thatallow pos-larvae to adapt biochemical and histologically to periods of transition between the end of thereabsorption of the yolk sac and the beginning of consumption of live feed, and from the consumptionof live feed to a balanced commercial diet. The species used in the first feeding of post-larvae affect theirsurvival and development. An inadequate provision could cause a high mortality due to the fact that thezooplankter species that proliferate everywhere not always satisfy the growing fish nutritional requirements,their volume growth is inferior to the required volume and, on top of that, a wrong production handlingfavors the proliferation of depredating planktonic species. Bearing in mind that one of the main nutrientst0hat live feed contributes, are the non-saturated and polyunsaturated fatty acids, nutritional deficienciesin the live feed can be complemented with the enrichment of fatty acids, which are essential componentsin the processes of pigmentation, production of prostaglandins and immunological defenses, plus retinaldevelopment required to improve visual capacity and sharpness, among others.

A nutrição das larvas representa um das principais problemas na produção de peixes a grandeescala; o objetivo principal de esta revisão é afundar no conhecimento dos fatores que influenciam odesenvolvimento orgânico e digestivo dos peixes e como estes fatores podem afetar o seu desenvolvimentoe sua sobrevivência. O desenvolvimento do trato digestivo nas larvas dos peixes está condicionado aosaspectos do tipo anatomo-fisiológico que permitem à póslarva adaptar-se bioquímica e histologicamenteaos períodos de transição entre a finalização da reabsorção do vitelo e o inicio do consumo de alimento vivoe na transição do alimento vivo na dieta comercial balanceada. As espécies usadas na primeira alimentaçãopóslarva afetam sua sobrevivência e desenvolvimento devido a que uma inadequada alimentação poderiaocasionar uma alta mortalidade, já que as espécies zooplanctônicas que proliferam, não sempre satisfazemos requerimentos nutricionais da póslarva, as quais se produzem em volume inferior ao requerido e/ouuma mal manejo na sua produção, favorece a proliferação de espécies planctônicas depredadoras. Entreos principais nutrientes que aporta o alimento vivo, estão os ácidos graxos insaturados e poliinsaturados.Também as deficiências nutricionais no alimento vivo podem ser complementadas mediante enriquecimentocom ácidos graxos essenciais no processo de pigmentação, produção de prostaglandinas, respostasimunológicas, desenvolvimento retinal entre outras.