Generally, the injection method is recommended as the best efficient method for vaccine applications in fish. However, labor-intensive and difficult injection for certain fish sizes is always considered as a limitation to aquatic animals. To demonstrate the effectiveness of a novel oral delivery system for the piscine vaccine with nano-delivery made from nano clay, halloysite nanotubes (HNTs) and their modified forms were loaded with killed vaccines, and we determined the ability of the system in releasing vaccines in a mimic digestive system. The efficaciousness of the oral piscine vaccine nano-delivery system was evaluated for its level of antibody production and for the level of disease prevention in tilapia. Herein, unmodified HNTs (H) and modified HNTs [HNT-Chitosan (HC), HNT-APTES (HA) and HNT-APTES-Chitosan (HAC)] successfully harbored streptococcal bivalent vaccine with inactivated S. agalactiae, designated as HF, HAF, HCF and HACF. The releasing of the loading antigens in the mimic digestive tract demonstrated a diverse pattern of protein releasing depending on the types of HNTs. Remarkably, HCF could properly release loading antigens with relevance to the increasing pH buffer. The oral vaccines revealed the greatest elevation of specific antibodies to S. agalactiae serotype Ia in HCF orally administered fish and to some extent in serotype III. The efficacy of streptococcal disease protection was determined by continually feeding with HF-, HAF-, HCF- and HACF-coated feed pellets for 7 days in the 1st and 3rd week. HCF showed significant RPS (75.00 ± 10.83%) among the other tested groups. Interestingly, the HCF-treated group exhibited noticeable efficacy similar to the bivalent-vaccine-injected group (RPS 81.25 ± 0.00%). This novel nano-delivery system for the fish vaccine was successfully developed and exhibited appropriated immune stimulation and promised disease prevention through oral administration. This delivery system can greatly support animals’ immune stimulation, which conquers the limitation in vaccine applications in aquaculture systems. Moreover, this delivery system can be applied to carrying diverse types of biologics, including DNA, RNA and subunit protein vaccines.

Streptococcosis is one of the major diseases that causes devastation to farmed fish, leading to significant economic losses all around the world. Currently, two serotypes of Streptococcus agalactiae, serotype Ia and III, have been identified as virulent strains and major causative agents of the disease in farmed Nile tilapia (Oreochromis niloticus Linn.) in Thailand. Upon inactivated vaccine development, monovalent inactivated whole-cell vaccines demonstrated high specific antibody production against homologous serotypes and limited production with heterologous serotypes. However, for higher efficacy, a bivalent streptococcal vaccine was designed to maximize protective immunity to both serotypes. Interestingly, our bivalent vaccine could successfully induce specific antibody production against both serotypes with similar levels, and the response could extend over the 8 weeks of the experimental period. Evaluation of vaccines in the laboratory scale revealed relative percent survival (RPS) of vaccinated tilapia to serotype Ia (81.2 ± 9.4%) and serotype III (72.2 ± 4.8%), respectively. The efficacy of the bivalent vaccine showed significant RPS higher than the monovalent vaccine (p < 0.05) at 30 days, and the protection of all those vaccines was reduced thereafter. Evaluation of the vaccine in a farm trial in different locations in Thailand revealed the efficacy of the bivalent vaccine in increasing the production yield by greater than 80% in all tested farms in 2015 and 2021. Taken together, this study affirms the efficacy of the bivalent streptococcal vaccine in the prevention of streptococcus disease in Nile tilapia, which could be used in different areas. This vaccine development could be effectively applied in the tilapia culture industry.

Prior aquatic animal image classification research focused on distinguishing external features in controlled settings, utilizing either digital cameras or webcams. Identifying visually similar species, like Short mackerel (Rastrelliger brachysoma) and Indian mackerel (Rastrelliger kanagurta), is challenging without specialized knowledge. However, advancements in computer technology have paved the way for leveraging machine learning and deep learning systems to address such challenges. In this study, transfer learning techniques were employed, utilizing established pre-trained models such as ResNet50, Xception, InceptionV3, VGG19, VGG16, and MobileNetV3Small. These models were applied to differentiate between the two species using raw images captured by a smartphone under uncontrolled conditions. The core architecture of the pre-trained models remained unchanged, except for the removal of the final fully connected layer. Instead, a global average pooling layer and two dense layers were appended at the end, comprising 1024 units and by a single unit, respectively. To mitigate overfitting concerns, early stopping was implemented. The results revealed that, among the models assessed, the Xception model exhibited the most promising predictive performance. It achieved the highest average accuracy levels of 0.849 and 0.754 during training and validation, surpassing the other models. Furthermore, fine-tuning the Xception model by extending the number of epochs yielded more impressive outcomes. After 30 epochs of fine-tuning, the Xception model demonstrated optimal performance, reaching an accuracy of 0.843 and displaying a 11.508% improvement in predictions compared to the model without fine-tuning. These findings highlight the efficacy of transfer learning, particularly with the Xception model, in accurately distinguishing visually similar aquatic species using smartphone-captured images, even in uncontrolled conditions.

The Krabi mouth-brooding fighting fish, Betta simplex Kottelat, 1994, is a critically endangered and endemic fish species in Krabi province, Southern Thailand. Little information is available on its reproductive ecology and early developmental morphology, which are essential for studying its conservation. Generally, B. simplex is considered an adaptable animal that can tolerate lower alkalinity and higher hardness compared to its natural environment conditions. In this study, wild broodstocks of B. simplex were collected from the reported type localities and bred in captivity under laboratory conditions for size-series collection. Some biological aspects of B. simplex in its natural environmental conditions were determined. We found that its flaring and mating behavior was similar to those of bubble-nesting fighting fish but did not involve bubble-nest building. The fertilized eggs and pre-flexion larvae were nurtured in the mouth cavity of parental males within 11–12 (mode = 11) days after fertilization (DAF). The first-release offspring developed to the post-flexion stage with a body size of 4.39 ± 0.01 mm of standard length (SL; n = 6) and then to the juvenile stage within 30 days after release with 11.72 ± 0.62 mm SL (n = 4). Thus, we propose the following linear regression equation for growth prediction by age (DAF) and body size (SL; mm): age = 0.2425 SL + 1.7036 (r² = 0.9549). The findings of this study will deepen our knowledge of the reproduction and ontogeny of B. simplex and contribute to its future conservation and management.

The family Sciaenidae comprises around 300 species and 70 genera, with the Indo-West Pacific (IWP) region exhibiting the highest species richness. The genus Boesemania Trewavas, 1977 was established as a monotypic group based on the swim bladder and otolith morphology of Johnius microlepis Bleeker. Corvina polykladiskos Bleeker was recognized as the third species of the genus Bahaba. Based on recent morphology and molecular evidence, we assign Bahaba polykladiskos to the genus Boesemania. Boesemania polykladiskos is distinguished from Boesemania microlepis by its distinctly larger scales, the number of lateral-line, upper and lower trans-lateral-line rows, and sagitta and swim-bladder morphology. Phylogenetic analysis of 24 sciaenid species representing 23 genera from the Indo-West Pacific region further confirms that Boesemania is monophyletic and that Boesemania polykladiskos is the sister species of Boesemania microlepis. The two Boesemania species are commercially important fish in their native range and play an essential role in the global luxury dried swim-bladder trade. However, significant knowledge gaps persist regarding their ecology, biology, and population status. Consequently, this paper also addresses the need for further research and conservation measures for these species.