An in-vivo experimental evaluation of the efficacy of fish-derived antimicrobial peptides against multidrug-resistant Pseudomonas aeruginosa.

Introduction: due to the fact that antimicrobial peptides antimicrobial peptides (AMPs) from climbing perch have not been fully explored for their antimicrobial potency, this investigation was undertaken to explore that possibility. Methods: antimicrobial peptides (AMPs) from the mucous secretion of climbing perch were obtained and an in-vivo analysis was conducted using mice. Results: the results showed inhibitory effects on multidrug-resistant multidrug-resistant Pseudomonas aeruginosa with reduced mortality from 100% among the non-treated group to 25%. Similarly, the level of serum transaminase enzymes (AST and ALT), creatinine levels, and pro-inflammatory cytokines (TNF-α and IL-6) were all found to be higher in the non-treatment group compared to the AMP-treatment group. Also, extensive tissue damage in the lung, liver, and spleen of the non-treated control group mice was observed based on the histopathological lesions recorded. As expected, AMPs from climbing perch significantly alleviated multidrug-resistant P. aeruginosa infection in-vivo and produced enhanced therapeutic efficacy superior to the ciprofloxacin treatment. Conclusion: this study provides insight into the potential antimicrobial activity of fish innate immune system-derived peptides that could serve as a candidate for the substitute of antibiotics.

An update on the ornamental fish industry in Malaysia: Aeromonas hydrophila-associated disease and its treatment control.

Malaysia is the world's major producer and exporter of ornamental fish, contributing 9% to the global trade and taking the second position after Singapore. Because of their artistic appeal and tremendous commercial value for international trade, ornamental fish recently gain rapid importance for foreign exchange and as a source of employment. While ornamental fish production is growing, there is an increase in infectious diseases, resulting in high fish mortality with significant economic loss. Bacterial disease is a serious problem for ornamental fish industry. Bacterial species surveillance in diseased freshwater ornamental fish from an aquarium shop reveals that Aeromonas hydrophila is the most dominant bacteria isolated. Consequently, Malaysia is stepping up its efforts by implementing the Economic Transformation Program and other biosecurity steps to address the aquaculture issues and encourage the regrowth of the ornamental fish market. Chemotherapeutic medications, phytobiotics, probiotics, yeast extracts, vaccines, and disinfectants can be used in controlling bacteria. Further studies should be done to find new antibacterial agents from natural sources to combat bacterial fish diseases and reduce fish mortality rate in sustainable aquaculture farms. This review summarizes the literature on ornamental fish industries and aquaculture production in relation to A. hydrophila-associated diseases and ornamental fish health management in Malaysia.

Glyphosate Herbicide Induces Changes in the Growth Pattern and Somatic Indices of Crossbred Red Tilapia (O. niloticus × O. mossambicus).

The development of glyphosate-resistant genetically modified organisms (GMO) has increased the use of herbicide glyphosate by several magnitudes in recent years. It is now the most commonly used pesticide globally that affects aquatic habitats, especially fish. This study aims to add new knowledge on the effect of technical grade glyphosate on several toxicity parameters and to identify the most effective parameter in predicting technical grade glyphosate chronic toxicity (seven weeks) to fish, especially Malaysia's heavily farmed red tilapia. The results show that a relatively high concentration of technical grade glyphosate is needed to induce significant changes in all tested parameters. However, the results also indicate that the bodyweight index is the most sensitive toxicity parameter in that a reduction in body weight was observed at 25 mg/L of glyphosate. Negative correlations between the glyphosate concentration and toxicity parameters such as specific growth rate (SGR), hepato-somatic index (HIS), and gonado-somatic index (GSI) were observed. The fish condition factor and feed conversion ratio were found not to be affected at the highest glyphosate concentration tested (150 mg/L). To conclude, crossbred red tilapia (O. niloticus × O. mossambicus) is one potential species for evaluating the toxic effects of technical grade glyphosate on fish.

Short- and long-term probiotic effects of Enterococcus hirae isolated from fermented vegetable wastes on the growth, immune responses, and disease resistance of hybrid catfish (Clarias gariepinus × Clarias macrocephalus).

This study evaluated the short- and long-term effects of dietary supplementation with Enterococcus hirae strain UPM02 on the growth performance, immunity, and disease resistance of hybrid catfish (Clarias gariepinus × Clarias macrocephalus) against Aeromonas hydrophila infection. In the long-term trial, fingerling fish were fed diets containing 0 (control), 2 × 105, or 2 × 107 CFU/g E. hirae UPM02 for 120 days. Administration of E. hirae UPM02 had significant effects on the specific growth rate (SGR), feed utilization efficiency, body indices (P < 0.05), and gut villus physiology of the catfish. E. hirae UPM02 application also significantly increased the complete blood cell counts, phagocytic activity, respiratory burst, lysozyme activity, and alternative complement pathway hemolytic (ACH50) activity in tested catfish throughout the experimental periods (P < 0.05). Dietary E. hirae UPM02 at both concentrations significantly increased the expression levels of the alpha-2-macroglobulin (α2M), CC chemokines, CXC chemokines, lysozyme c (LYZC), myeloperoxidase (MYE), NF-kappa-B1 p105 subunit (NF-K), and bactericidal permeability-increasing protein (BPIP) genes in the head kidney, liver, and spleen (P < 0.05) at days 80, 100 and 120 after application. However, heat shock protein 70 (HSP70) gene expression was slightly downregulated in these organs. Interestingly, fish fed the diets containing 2 × 105 and 2 × 107 CFU/g E. hirae UPM02 exhibited a significantly lower (P < 0.05) postchallenge mortality rates (32% and 30%, respectively) after 14 days of A. hydrophila challenge than the control fish (58%). In short-term (28 days) application to juvenile catfish, the two concentrations of E. hirae did not affect all growth parameters. Nevertheless, these concentrations markedly elevated all tested immune parameters, similarly to long-term application. Immune-related gene expression was significantly upregulated at day 28 in the head kidney, at day 14 in the liver, and at day 7 in the spleen in fish treated with the two concentrations of the probiotics (P < 0.05). Mortality at 14 days after challenge with A. hydrophila in the groups receiving the two concentrations of the probiotic was significantly lower than that in the control group, at 28, 24, and 48%, respectively (P < 0.05). These results collectively suggest that dietary supplementation with E. hirae UPM02 at 2 × 105 and 2 × 107 CFU/g effectively influenced immune responses, enhanced disease protection, and stimulated immunity-related gene expression in hybrid catfish under both short- and long-term application. However, growth enhancement was significantly evidenced with long-term application only.

Mucosal responses of brown-marbled grouper Epinephelus fuscoguttatus (Forsskål, 1775) following intraperitoneal infection with Vibrio harveyi.

Groupers are popular aquaculture species in South-East Asia, but their cultivation is affected by infectious disease outbreaks. Mucosa-associated lymphoid tissues provide a first-line defence against pathogens; however, few studies are available relating to cellular or proteomic responses of mucosal immunity in grouper. Skin, gill and intestine were sampled from brown-marbled grouper Epinephelus fuscoguttatus (Forsskål, 1775) at 4 and 96 hr post-infection (hpi) and 7 days post-infection (dpi) following intraperitoneal infection with Vibrio harveyi, and stained with haematoxylin/eosin and Alcian Blue/periodic acid-Schiff. Skin mucus was analysed by 2D-gel electrophoresis, and proteins modulated by the bacterial infection identified. In the infected fish, significant increases in sacciform cells in skin and increased levels of nucleoside diphosphate kinase in mucus were detected at 4 hpi. At 96 hpi, goblet cells containing acidic mucins significantly increased in the intestine, while those containing mixed mucins increased in skin and gills of infected fish. Proteasome subunit alpha type-I and extracellular Cu/Zn superoxide dismutase levels also increased in mucus. Rodlet and mast cells did not appear to respond to the infection. Mucosal tissues of grouper appeared actively involved in response to Vibrio infection. This information may help future research on improving grouper health, production and vaccine development.

In vivo and in vitro effects on cholinesterase of blood of Oreochromis mossambicus by copper.

The present study is aimed to evaluate the effects of sub-acute toxicity testing of copper sulphate (CuSO4), on behavioural, histological and biochemical changes of the Oreochromis mossambicus (black tilapia) blood tissues. The effects were assessed according to the previous results on sub-acute toxicity test after exposing fish to several concentrations (0.0, 2.5, 5.0, and 10.0 mg/L). The observations of scanning electron microscope, and transmission electron microscope studies revealed severe histopathological changes on the surface and the cellular changes in blood tissues, respectively. The morphological alterations in blood involved irregular structure of red blood cell and blood clot formation. CuSO4 affected the biochemical alteration of the blood cholinesterase also known as serum cholinesterase (ChE). Blood ChE inhibited up to 80% of activity when exposed to 10.0 mg/L CuSO4. The findings from this study can further improve the quality standards of aquaculture industry and the fundamental basis in selecting suitable strains among freshwater fish species to be used as bioindicator.

Live vaccines against bacterial fish diseases: A review.

Fish diseases are often caused either by bacteria, viruses, fungi, parasites, or a combination of these pathogens. Of these, bacterial fish diseases are considered to be a major problem in the aquaculture industry. Hence, the prevention of such diseases by proper vaccination is one of the integral strategies in fish health management, aimed at reducing the fish mortality rate in the aquaculture farms. Vaccination offers an effective yet low-cost solution to combat the risk of disease in fish farming. An appropriate vaccination regime to prevent bacterial diseases offers a solution against the harmful effects of antibiotic applications. This review discusses the role of live-attenuated vaccine in controlling bacterial diseases and the development of such vaccines and their vaccination strategy. The current achievements and potential applications of live-attenuated and combined vaccines are also highlighted. Vaccine development is concluded to be a demanding process, as it must satisfy the requirements of the aquaculture industry.

Toxic effects of copper on liver and cholinesterase of Clarias gariepinus.

The release of pollutants, especially heavy metals, into the aquatic environment is known to have detrimental effects on such an environment and on living organisms including humans when those pollutants are allowed to enter the food chain. The aim of this study is to analyse the damage to Clarias gariepinus' liver caused by exposure to different concentrations of copper. In the present study, samples of C. gariepinus were exposed to sub-lethal copper sulphate (CuSO4) concentrations (from 0.2 to 20.0 mg/L) for 96 h. Physiological and behavioural alterations were observed with respect to their swimming pattern, mucus secretion and skin colour. Mortality was also observed at high concentrations of copper. Histopathological alterations of the liver were analysed under light, transmission and scanning electron microscopies. The liver of the untreated group showed normal tissue structures, while histopathological abnormalities were observed in the treated fish under light and electron microscopes with increased copper concentrations. Histopathological abnormalities include necrosis, melanomacrophage, hepatic fibrosis and congested blood vessels. In addition, the enzyme activity of liver cholinesterase (ChE) was also found to be affected by copper sulphate, as 100% of cholinesterase activity was inhibited at 20.0 mg/L. Thus, liver enzyme activity and histopathological changes are proven to be alternative sources for biomarkers of metal toxicity.

Histological features of the gastrointestinal tract of wild Indonesian shortfin eel, Anguilla bicolor bicolor (McClelland, 1844), captured in Peninsular Malaysia.

This study was conducted to record the histological features of the gastrointestinal tract of wild Indonesian shortfin eel, Anguilla bicolor bicolor (McClelland, 1844), captured in Peninsular Malaysia. The gastrointestinal tract was segmented into the oesophagus, stomach, and intestine. Then, the oesophagus was divided into five (first to fifth), the stomach into two (cardiac and pyloric), and the intestine into four segments (anterior, intermediate, posterior, and rectum) for histological examinations. The stomach had significantly taller villi and thicker inner circular muscles compared to the intestine and oesophagus. The lamina propria was thickest in stomach, significantly when compared with oesophagus, but not with the intestine. However, the intestine showed significantly thicker outer longitudinal muscle while gastric glands were observed only in the stomach. The histological features were closely associated with the functions of the different segments of the gastrointestinal tract. In conclusion, the histological features of the gastrointestinal tract of A. b. bicolor are consistent with the feeding habit of a carnivorous fish.

Detection and phylogenetic profiling of nodavirus associated with white tail disease in Malaysian Macrobrachium rosenbergii de Man.

White tail disease (WTD) is a serious viral disease in the hatcheries and nursery ponds of Macrobrachium rosenbergii in many parts of the world. A new disease similar to WTD was observed in larvae and post larvae of M. rosenbergii cultured in Malaysia. In the present study, RT-PCR assay was used to detect the causative agents of WTD, M. rosenbergii nodavirus (MrNV) and extra small virus (XSV) using specific primers for MrNV RNA2 and XSV. The results showed the presence of MrNV in the samples with or without signs of WTD. However, XSV was only detected in some of the MrNV-positive samples. Phylogenetic analysis showed that the RNA2 of our Malaysian isolates were significantly different from the other isolates. Histopathological studies revealed myofiber degeneration of the tail muscles and liquefactive myopathy in the infected prawns. This was the first report on the occurrence of MrNV in the Malaysian freshwater prawn.