This study investigated the in vitro antimicrobial and anthelmintic effect of copper nanoparticles (CuNPs) against the bacterium Aeromonas hydrophila, the monogeneans Dactylogyrus minutus, Dactylogyrus extensus, Gyrodactylus cyprini, and the cestode Schyzocotyle acheilognathi, as well as their toxicity to Cyprinus carpio Koi. In the antimicrobial in vitro test, the inhibition zone method and minimum inhibitory concentration (MIC) were performed. In order to determine the time and efficacy of monogenean parasite mortality, the parasites were exposed to CuNP concentrations of 20, 50, 100, 150, 200, and 300 mg L-1, and a control group with tank water and one with copper sulphate pentahydrate (CuSO4.5H2O) at a concentration of 0.3 mg L-1, performed in triplicate. The parasites were observed every 10 min for 300 min, and mortality was recorded. For the cestodes, parasites were immersed in CuNP concentrations of 50, 100, 150, and 300 mg L-1. At the end of the in vitro tests, the anthelmintic efficacy of each treatment was calculated. To assess the tolerance and toxicity in fish, they were exposed to CuNP concentrations of 0.6, 1.25, 2.5, 5, 10, 20, and 50 mg L-1 for 12 h. The MIC demonstrated that CuNPs effectively inhibited the growth of A. hydrophila up to a dilution of 12,500 mg L-1 and showed an inhibition zone of 14.0 ± 1.6 mm for CuNPs. The results of anthelmintic activity showed a dose-dependent effect of concentration for both groups of parasites, with the most effective concentration being 300 mg L-1 in 120 min. In the toxicity test, the carps showed tolerance to lower concentrations. The study indicated that CuNPs were effective against the studied pathogens. However, it proved to be toxic to fish at high concentrations. The use of low concentrations is recommended still requires further investigation.

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety of indigo carmine as a colourant feed additive for cats, dogs and ornamental fish. In its previous opinion, the FEEDAP Panel concluded that indigo carmine is safe for cats and dogs at levels up to 250 mg/kg complete feed and for ornamental fish up to 1000 mg/kg complete feed. These conclusions were limited to indigo carmine with a purity of at least 93% colouring matter. For the current assessment, the applicant submitted additional data on the characterisation and on the toxicological profile to support the safety of the additive with the purity of at least 85% colouring matter. Based on the new data submitted, the FEEDAP Panel concludes that indigo carmine (purity of at least 85% colouring matter) is safe for cats and dogs at levels up to 250 mg/kg complete feed and for ornamental fish up to 1000 mg/kg complete feed.

The aim of this study was to develop a feeding protocol for the larviculture of Apistogramma cacatuoides, using the histological approach to larval nutrition conditions. For this, three experiments were carried out. Experiment 1 was carried out in a randomized design to determine the optimal amount of Artemia nauplii (AN) per larva, and three treatments were evaluated: P1-feeding with 25 A. nauplii per larva (AN/L) during the first 5 days, followed by 50 AN/L from the 6th to the 10th day and 100 AN/L from the 11th to the 20th day; P2 and P3-37 and 50 AN/L during the first 5 days, 75 and 100 AN/L from the 6th to the 10th day and 150 and 200 AN/L from the 11th to the 20th day. Experiment 2 was carried out in a randomized design to determine the daily frequency of feeding and evaluated four feeding frequencies: F1-feeding only once a day (09:00); (F2)-feeding twice a day (09:00 and 17:00); F3-feeding three times a day (09:00, 11:30 and 17:00); and F4-feeding four times a day (09:00, 11: 30, 14:00 and 17:00). Experiment 3 lasted 40 days and was conducted in a randomized design to evaluate three periods for the beginning of the feeding transition: WE10:AN for 10 days, followed by 3 days of co-feeding and commercial feed until the end of the experimental period; WE15:AN for 15 days, followed by 3 days of co-feeding and commercial feed; WE20:AN for 20 days, followed by 3 days of co-feeding and commercial feed. The results of this study showed that, for the best development of the larvae, they should receive the feeding protocol 50-100-200 AN/L (P3) until the 20th day of exogenous feeding. From the 21st day, the transition to inert food should begin with 3 days of co-feeding, and feeding during larviculture should be carried out at a frequency of twice a day; this protocol provided a good nutritional status for the larvae, as shown by the histological approach.

Fish pigmentation study can reveal understandings in dermatological research based on functional genomics. Cultured ornamental fish becomes dull coloured and antityrosinase activity through sesame seed may enhance skin colour, which has not been studied. Botia dario is an indigenous fish, having ornamental and aesthetic value and can be studied as a model for fish pigmentation genetics. In this study, fish specimens were fed with 15% marigold petal meal along with 5, 10 and 15% w/w sesame seed in diet. Pigmentation genes, that is, tyr, tyrp1a, asip1, gnaq, kitlga, mc1r, mitf, pax7a, rab38, slc7a11, sox9a, sox10, csf1r, bcdo2 and gsta2 in skin and immunogens, that is, il20, nramp, tlr9 and trail in kidney were studied. Gene expression in tissues revealed enhanced pigmentation and immunity as well as the role of tyr, tyrp1a and asip1 in pigmentation. Immunogenes and blood parameters confirmed the best pigmentation diet. Colorimetric analysis also showed the enhancement of pigmentation. Insights from sesame seed and marigold-induced antityrosinase effects will be applied in aquaculture to develop natural, dietary formulations that will enhance pigmentation in ornamental fish, leading to improved skin colour and market value.

The purpose of this study was to characterize in detail the reproductive strategy, course of embryogenesis, and development of larvae in three species of fishes of the genus Cichlasoma: the green terror (Andinoacara rivulatus), the red discus (Symphysodon discus), and the jaguar cichlid (Parachromis managuensis). Eggs for the study were obtained from five pairs of each species (300 eggs from each female) and incubated at 26 °C. The developing eggs were observed under a microscope (Carl Zeiss Stereo Discovery. V12 and Nikon 2000SE software (NIS-Elements F 4.30.01 64-bit) from fertilization to larval hatching until complete yolk-sac resorption. The largest average number of eggs per female was found in the jaguar cichlid (x¯ = 2991 eggs), a smaller average number of eggs was shown in the green terror (x¯ = 922 eggs), and the red discus showed the smallest average number of eggs (x¯ = 300 eggs). There were significant differences in the sizes of the eggs of the studied species: jaguar cichlid eggs were the smallest (1.060 ± 0.05 mm3), red discus eggs were larger (1.070 ± 0.07 mm3), and green terror eggs were the largest (1.365 ± 0.16 mm3). The embryogenesis time in the red discus was 2132 °H (82 Hpf), in the green terror it was 2158 °H (83 Hpf), and the longest in the jaguar cichlid was 2470 °H (87 Hpf). At the end of embryogenesis, the average size of the larvae after hatching was measured (red discus x¯ = 4.346 mm, green terror x¯ = 5.203 mm, and jaguar cichlid x¯ = 5.301 mm) and the time of yolk-sac resorption from the moment of hatching to the transition from endogenous to exogenous feeding was determined (jaguar cichlid 5 days, green terror 6 days, and red discus 3 days). The results of this study may contribute to the development of reproductive biotechnology for the studied fishes that could be used in aquaculture and, thus, help protect them in their natural habitats.

Fishing causes direct removal of individuals from wild populations but can also cause a physiological disturbance in fish that are released or discarded after capture. While sublethal physiological effects of fish capture have been well studied in commercial and recreational fisheries, this issue has been overlooked for the ornamental fish trade, where it is common to capture fish from the wild and discard non-target species. We examined metabolic responses to capture and discard procedures in the three-striped dwarf cichlid Apistogramma trifasciata, a popular Amazonian aquarium species that nonetheless may be discarded when not a target species. Individuals (n = 34) were tagged and exposed to each of four treatments designed to simulate procedures during the capture and discard process: 1) a non-handling control; 2) netting; 3) netting +30 seconds of air exposure; and 4) netting +60 seconds of air exposure. Metabolic rates were estimated using intermittent-flow respirometry, immediately following each treatment then throughout recovery overnight. Increasing amounts of netting and air exposure caused an acute increase in oxygen uptake and decrease in available aerobic scope. In general, recovery occurred quickly, with rapid decreases in oxygen uptake within the first 30 minutes post-handling. Notably, however, male fish exposed to netting +60 seconds of air exposure showed a delayed response whereby available aerobic scope was constrained <75% of maximum until ~4-6 hours post-stress. Larger fish showed a greater initial increase in oxygen uptake post-stress and slower rates of recovery. The results suggest that in the period following discard, this species may experience a reduced aerobic capacity for additional behavioural/physiological responses including feeding, territory defence and predator avoidance. These results are among the first to examine impacts of discard practises in the ornamental fishery and suggest ecophysiological research can provide valuable insight towards increasing sustainable practises in this global trade.

This study aimed to characterize 300 Aeromonas spp. strains isolated from 123 ornamental fish of 32 different species presenting with septicemia, skin lesions, and/or eye lesions. Within the 300 strains, 53.0% were identified as A. veronii, 41.3% as A. hydrophila, and 5.7% as A. caviae. Among the six virulence genes investigated, the most frequent were act (90.3%) and aer (79.3%). More than 50% of A. hydrophila strains were positive for all the studied genes. A total of 30 virulence profiles were identified, with the five main profiles identified comprising 75% of strains. Only five strains were negative for all genes and were identified as A. caviae and A. veronii. The antimicrobial susceptibility profile was performed for 234 strains, with sulfonamides presenting more than 50% of the resistance rates. Susceptibility was observed mainly for cephalosporins, aminoglycosides, chloramphenicol and piperacillin-tazobactam. Multidrug resistance was detected in 82.5% of the studied strains, including A. caviae with 100% multidrug resistance, and A. hydrophila with 90.9% multidrug resistance. The SE-AFLP analysis resulted in 66 genotypes of A. hydrophila, 118 genotypes of A. veronii, and 14 genotypes of A. caviae, demonstrating the greater heterogeneity of A. veronii and A. caviae. However, no direct correlation was observed between the genotypes and the strains\' origins or virulence and resistance profiles.

UNASSIGNED: Malaysia has more than 630 culturists who are involved in the ornamental fish industry and culture 250 species, including local and exotic species. Among these viruses, megalocytiviruses have been associated with severe systemic diseases and economic losses in ornamental fish. The intensity of Megalocytivirus infection in Pterophyllum scalare in Malaysia remains unknown. Thus, this study aimed to investigate the occurrence of Megalocytivirus while discovering its associated risk factors and the genotypes of its causative agents in an ornamental fish farm in Malaysia.
UNASSIGNED: Seven broodstock pairs of P. scalare were used in this study to follow the life stages of fish, from egg to market size. Water samples and other samples, such as mucus swabs, gill swabs, P. scalare eggs, fries, juveniles, snails, snail eggs, live feed (Tubifex worms and Moina spp.), sediment samples, and wild fish, were collected periodically for initial environmental sampling from day 0 to day 60. Nested polymerase chain reaction amplifications were performed for megalocytivirus-related sequences. The phylogenetic tree, including the sampled causative agents of megalocytiviruses, was inferred from the major capsid protein genes of all known Iridoviridae species. Pearson\'s correlation coefficients were calculated to determine the strength of the correlation between the presence of megalocytiviruses in P. scalare samples and the associated risk factors.
UNASSIGNED: A total of 312 out of 935 pooled and individual samples tested positive for the presence of Megalocytivirus-related sequences, except snail eggs and wild fish (Poecilia reticulata). No clinical symptoms were observed in any fish samples. Megalocytivirus-associated viruses detected in water samples indicate horizontal transmission of the virus. All the nucleotide sequences found in this study had high nucleotide identities of 95%-99 % and were closely related to Megalocytivirus genotype I infectious spleen and kidney necrosis virus. Risk factors associated with Megalocytivirus include water temperature, dissolved oxygen (DO), pH, ammonia, nitrate, nitrite, and the life stages of P. scalare. High Megalocytivirus infection was detected when the water temperature, DO, and pH were high in P. scalare, high water temperature and nitrate in the water samples, and the same rate of Megalocytivirus infection in P. scalare fry and juveniles.
UNASSIGNED: This is the first study to confirm the existence of different possible routes of megalocytivirus distribution in ornamental fish farms in Malaysia. Nevertheless, the connection between the mode of transmission and the risk factors for this virus needs to be explored further to recognize the evolution and potential new host species.

The present study aimed to investigate the effect of thermal stress on growth, feed utilization, coloration, hematology, liver histology, and critical thermal maximum (CTmax) in goldfish (Carassius auratus) cultured at three different acclimation temperatures including 27 °C, 30 °C, and 34 °C for 10 weeks. Goldfish were assigned randomly to tanks with a quadruplicate setup, accommodating 20 fish per tank. The result showed that fish acclimated to different temperatures did not significantly differ in weight gain (WG) and specific growth rate (SGR). However, increasing temperature significantly decreased feed efficiency ratio (FER), protein efficiency ratio (PER), and protein productive value (PPV), but significantly increased feed conversion ratio (FCR) (P < 0.05). The coloration parameters significantly decreased by high temperature in the trunk region with increasing temperature (L* and a* at week 5; L*, a*, and b* at week 10; P < 0.05). Total carotenoid contents in serum, fin, muscle, and skin also significantly decreased with increasing temperature (P < 0.05). Total protein, albumin, and globulin levels exhibited a notable decrease, while the albumin: globulin ratio showed a slight insignificant increase, with increasing temperature. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total cholesterol, and triglycerides significantly increased with increasing temperature (P < 0.05). While, high-density lipoprotein cholesterol (HDL-c) decreased linearly (P < 0.05). Glucose and cortisol levels linearly increased with increasing temperature, the highest levels being observed in the 34 °C group. Liver histology showed swollen hepatocytes, nuclei displacement, and infiltration of inflammation in fish cultured at 34 °C. Goldfish acclimated to 34 °C displayed a higher CTmax of 43.83 °C compared to other groups. The present study showed that temperature should be kept below 34 °C for goldfish culture to prevent high FCR, fading coloration, and liver damages.

This study reports on three species of Dactylogyrus Diesing, 1850 (Dactylogyridae) collected from tinfoil barb, Barbonymus schwanenfeldii (Bleeker) which were imported into South Africa as ornamental fish from Sri Lanka and Thailand. Supplementary morphometric characterisation and molecular data (partial 18S and 28S rDNA, and ITS1 region sequences) are presented for Dactylogyrus lampam (Lim & Furtado, 1986), Dactylogyrus tapienensis Chinabut & Lim, 1993 and Dactylogyrus viticulus Chinabut & Lim, 1993. Prevalence of Dactylogyrus spp. infection was 87% and 80% for fish from Sri Lanka and Thailand, respectively. Composition of the parasites between the fish of each origin differed. All three species were found to infect fish from Thailand, but only D. lampam was present on the fish received from Sri Lanka. Phylogenetic analysis revealed the position of studied species, with D. lampam clustering within the lineages of varicorhini-type species, while D. tapienensis and D. viticulus form a sister lineage to Dactylogyrus spp. associated with Cyprinus carpio L. and Carassius spp., species parasitising central African large cyprinids (Labeo Cuvier), and species parasitising African and Middle Eastern Carasobarbus spp.
UNASSIGNED: Dactylogyrus spp. (Dactylogyridae, Monogenea) de Barbonymus schwanenfeldii importé en Afrique du Sud : caractérisation morphométrique et moléculaire.
UNASSIGNED: Cette étude porte sur trois espèces de Dactylogyrus Diesing, 1850 (Dactylogyridae), prélevées sur des Barbonymus schwanenfeldii (Bleeker) qui ont été importés en Afrique du Sud comme poissons d’ornement depuis le Sri Lanka et la Thaïlande. Une caractérisation morphométrique et des données moléculaires supplémentaires (ADNr 18S et 28S partiels et séquences de la région ITS1) sont présentées pour Dactylogyrus lampam (Lim & Furtado, 1986), Dactylogyrus tapienensis Chinabut & Lim, 1993 et Dactylogyrus viticulus Chinabut & Lim, 1993. La prévalence de l’infection par les Dactylogyrus spp. était respectivement de 87 % et 80 % pour les poissons du Sri Lanka et de Thaïlande. La composition des parasites entre les poissons des deux origines différait. Les trois espèces infectaient les poissons de Thaïlande, mais seul D. lampam était présent sur les poissons du Sri Lanka. L’analyse phylogénétique a révélé la position des espèces étudiées, D. lampam se regroupant dans les lignées d’espèces de type varicorhini, tandis que D. tapienensis et D. viticulus forment une lignée sœur des Dactylogyrus spp. associés à Cyprinus carpio L. et Carassius spp., espèces parasitant les grands cyprinidés d’Afrique centrale (Labeo Cuvier), et espèces parasitant les Carasobarbus spp. d’Afrique et du Moyen-Orient.