Unveiling the molecular mechanisms of pigmentation control in Queen Loach, Botia dario (Hamilton, 1822): Insights from sesame seed and marigold-induced antityrosinase effects.

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.

Early Ontogeny of Cichlids Using Selected Species as Examples.

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.

Morphological and Molecular Characterization of Two New Species of Ceratomyxa Thélohan, 1892 (Cnidaria: Myxosporea) from the Marine Ornamental Fish Zanclus cornutus (Linnaeus, 1758) off Lakshadweep Islands, Arabian Sea.

PURPOSE: The present study provides the complete morphological and molecular description of two new species of myxosporeans, Ceratomyxa zancli n. sp. and Ceratomyxa cornuti n. sp. infecting the gallbladder of Zanclus cornutus from the Lakshadweep Islands, Arabian Sea. METHODS: Zanclus cornutus were screened for the presence of myxosporeans, and the recovered myxospores were morphologically characterized using Nomarski Differential Interference Contrast (DIC) optics. The sequences of SSU rDNA were employed for molecular and phylogenetic studies. RESULTS: Both the parasites exhibited a prevalence of 21% each. C. zancli n. sp. is characterized by broadly cresentic myxospores with convex anterior and slightly concave to straight posterior margins and rounded ends. Spore valves two, unequal, measured 9.6 ± 0.7 µm × 25.2 ± 1.3 µm. Polar capsules two, unequal, spherical, measured 4 ± 0.6 µm × 3.5 ± 0.6 µm. Polar filament exceptionally long and arranged irregularly. Myxospores of C. cornuti n. sp. are elongated with convex anterior and slightly concave to straight posterior margins. Spore valves two, unequal, measured 7.00 ± 0.4 µm × 26.56 ± 1.8 µm. Polar capsules spherical, unequal, measured 3.52 ± 0.2 × 3.36 ± 0.35. Molecular analysis of C. zancli n. sp. (ON818297) and C. cornuti n. sp. (ON818298) resulted in 1469 and 1491 bp long SSU rDNA sequences, respectively. Molecularly C. zancli n. sp. is close to C. diplodae and C. barnesi with 91.39% similarity, while C. cornuti n. sp. appears closer to C. robertsthomsoni with 97.46% similarity. In phylogenetic analyses, C. zancli n. sp. branched separately within the Ceratomyxa clade while C. cornuti n. sp. clustered with C. robertsthomsoni and C. thalassomae. CONCLUSION: Based on the differences in morphological, morphometric, molecular, and phylogenetic characteristics, as well as differences in the host and geographic location, the above two species of myxosporeans are considered novel. The study forms the first report of a species of Ceratomyxa from Z. cornutus.

Capture and discard practises associated with an ornamental fishery affect the metabolic rate and aerobic capacity of three-striped dwarf cichlids Apistogramma trifasciata.

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.

Characterization of Aeromonas Isolates from Ornamental Fish: Species, Virulence Genes, and Antimicrobial Susceptibility.

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.

Length-weight relationship for Potamotrygon wallacei (Carvalho, Rosa and Araújo, 2016) caught in the middle Negro River, Barcelos, Brazilian Amazon

Abstract This research aimed to estimate the length-weight ratio (LWR) of the stingray Potamotrygon wallacei, known locally as the cururu, which was caught in streams and lakes in the middle Negro River region, Amazonas, Brazil. The stingrays were captured during the night (from 11 pm to 1 am) near the shores of streams and lakes, through active search using wooden canoes, head lanterns and scoop nets. The samplings were carried out in November 2017 (5 days), February (8 days), March (3 days) and April (2 days) of 2018, totaling 18 days of sampling. The total fresh weight was measured to 0.1 g of accuracy and the disc width to 0.1 cm accuracy. The parameters a and b of the equation W=a.DWb were estimated. This study provides new maximum length data for the cururu stingray.

Resumo Esta pesquisa teve como objetivo estimar a relação peso-comprimento (LWR) da arraia Potamotrygon wallacei, conhecida localmente como cururu, que foi capturada em igarapés e lagos na região do médio Rio Negro, Amazonas, Brasil. As arraias foram capturadas durante a noite (das 23h-1h) nas margens dos igarapés e lagos, por meio de busca ativa utilizando canoas de madeira, lanternas de cabeça e puçás. As coletas foram realizadas nos meses de novembro de 2017 (5 dias), fevereiro (8 dias), março (3 dias) e abril (2 dias) de 2018, totalizando 18 dias de amostragem. Foram medidos o peso fresco total com precisão de 0,01 g e a largura do disco com precisão de 0,1 cm. Os parâmetros a e b da equação W= a.DWb foram estimados. Este estudo prover novos dados de comprimento máximo para a arraia cururu.

Length-weight relationship for Potamotrygon wallacei (Carvalho, Rosa and Araújo, 2016) caught in the middle Negro River, Barcelos, Brazilian Amazon / Relação peso-comprimento de Potamotrygon wallacei (Carvalho, Rosa e Araújo, 2016) capturada no médio Rio Negro, Barcelos, Amazônia Brasileira

Abstract This research aimed to estimate the length-weight ratio (LWR) of the stingray Potamotrygon wallacei, known locally as the cururu, which was caught in streams and lakes in the middle Negro River region, Amazonas, Brazil. The stingrays were captured during the night (from 11 pm to 1 am) near the shores of streams and lakes, through active search using wooden canoes, head lanterns and scoop nets. The samplings were carried out in November 2017 (5 days), February (8 days), March (3 days) and April (2 days) of 2018, totaling 18 days of sampling. The total fresh weight was measured to 0.1 g of accuracy and the disc width to 0.1 cm accuracy. The parameters a and b of the equation W=a.DWb were estimated. This study provides new maximum length data for the cururu stingray.

Resumo Esta pesquisa teve como objetivo estimar a relação peso-comprimento (LWR) da arraia Potamotrygon wallacei, conhecida localmente como cururu, que foi capturada em igarapés e lagos na região do médio Rio Negro, Amazonas, Brasil. As arraias foram capturadas durante a noite (das 23h-1h) nas margens dos igarapés e lagos, por meio de busca ativa utilizando canoas de madeira, lanternas de cabeça e puçás. As coletas foram realizadas nos meses de novembro de 2017 (5 dias), fevereiro (8 dias), março (3 dias) e abril (2 dias) de 2018, totalizando 18 dias de amostragem. Foram medidos o peso fresco total com precisão de 0,01 g e a largura do disco com precisão de 0,1 cm. Os parâmetros a e b da equação W= a.DWb foram estimados. Este estudo prover novos dados de comprimento máximo para a arraia cururu.

A Review of the Industrial Importance, Common Bacterial Diseases, and Zoonotic Risks of Freshwater Aquarium Fish.

Background: The ever-increasing popularity of home aquariums, most often involving freshwater varieties, has exploded in recent years partially due to the Coronavirus pandemic and related to stay-at-home public health precautions for social distancing. With this ever-increasing popularity of aquariums as a hobby, and whether this involves freshwater or marine fish species, a number of important economic, ecological, and public health issues arise for both fish and hobbyists alike. Materials and Methods: This review highlights the history and genesis of aquariums as both a hobby and an important economic factor (industrial, commercial) for many countries on a global basis. Types of aquarium fish are described, and culture conditions leading to homeostasis in aquatic environments are detailed. When these conditions are not met and aquatic systems are out of balance, the disease can result due to stressed fish. Results: Major bacterial diseases associated with freshwater aquarium fish are reviewed, as are potential human infections related to the care and maintenance of home aquaria. Conclusion: Besides, scientific information was also combined with the false facts of hobbyists who tried to identify and treat diseases during an outbreak in the aquarium. Finally, unresolved issues and important misconceptions regarding the field are discussed.

Effects of thermal stress responses in goldfish (Carassius auratus): growth performance, total carotenoids and coloration, hematology, liver histology, and critical thermal maximum.

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.

Molecular epidemiology of Megalocytivirus in freshwater angelfish (Pterophyllum scalare) from Johor, Malaysia.

Background and Aim: 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. Materials and Methods: 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. Results: 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. Conclusion: 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.

Dactylogyrus spp. (Dactylogyridae, Monogenea) from tinfoil barb, Barbonymus schwanenfeldii imported into South Africa: morphometric and molecular characterisation.

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.

Title: Dactylogyrus spp. (Dactylogyridae, Monogenea) de Barbonymus schwanenfeldii importé en Afrique du Sud : caractérisation morphométrique et moléculaire. Abstract: 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.

On a new colorful species of Moenkhausia (Characiformes: Characidae) from the upper rio Madeira basin at the Chapada dos Parecis, Brazil, with comments on its conservation and putative biogeographic history.

A new species, Moenkhausia iris, is described from the upper rio Madeira basin, Mato Grosso State, Brazil. The new species is unique among congeners by its color in life, which is the dorsal half of body blue or green and ventral half red, more intense above the anal fin. Additionally, it can be distinguished by having a single, vertically elongated humeral spot, a lateral surface of body, caudal peduncle, and anal fin without distinct blotches or oblique marks and 18-19 branched anal-fin rays. The new species is endemic from a small tributary of the Rio Guaporé, rio Madeira drainage at Chapada dos Parecis. Its putatively close relatives are endemic to the upper rio Juruena located at the same shield. Both headwaters are very close to each other, sharing other restricted range species, and their hydrogeological topography indicates the upper rio Juruena stretch was captured by the rio Guaporé through a headwater capture event. Moenkhausia iris is much appreciated in the international aquarium trade, as well as many other tetras endemic from the same area. Exportation is based on the capture of wild individuals so sustainable exploitation is necessary for its conservation.

Phylogeography and population genetic structure of the cardinal tetra (Paracheirodon axelrodi) in the Orinoco basin and Negro River (Amazon basin): evaluating connectivity and historical patterns of diversification.

The Neotropics contain one of the most diverse assemblages of freshwater fishes worldwide. Part of this diversity is shared between the Orinoco and Amazon basins. These basins have been separated for a long time due to the Vaupes Arch, rising between 10-11 Ma. Today, there is only one permanent connection between the Orinoco and Negro (Amazon) basins, known as the Casiquiare Canal. However, alternative corridors allowing fish dispersion between both basins have been proposed. The cardinal tetra (Paracheirodon axelrodi), the most important fish in the ornamental world market, is distributed in both basins. Here we investigated P. axelrodi phylogeography, population structure, and potential routes of migration and connectivity between the two basins. A total of 468 bp of the mitochondrial gene (COI), 555 bp of the nuclear gene fragment (MYH6), and eight microsatellite loci were analyzed. As a result, we found two major genetic clusters as the most likely scenario (K = 2), but they were not discreetly distributed between basins. A gradient of genetic admixture was observed in Cucui and São Gabriel da Cachoeira, between the upper Negro River and the upper Orinoco. Samples from the middle-lower Negro River were highly structured. Cucui (Negro basin) was more similar to the Orinoco than to the rest of the Negro basin populations. However, substructure was also observed by the discriminant analysis, fixation indices and other hierarchichal structure analyses (K = 3 - 6), showing three major geographic clusters: Orinoco, Cucui, and the remaining Negro basin. Unidirectional migration patterns were detected between basins: via Cucui toward Orinoco and via the remaining of the Negro basin toward Orinoco. Results from the Relaxed Random Walk analysis support a very recent origin of this species in the headwater Orinoco basin (Western Guiana Shield, at late Pleistocene) with a later rapid colonization of the remaining Orinoco basin and almost simultaneously the Negro River via Cucui, between 0.115 until about 0.001 Ma. Historical biogeography and population genetic patterns observed here for Cardinal tetra, seem to be better explained by river capture, physical, or ecological barriers than due to the geographic distance.

Karyotypic and Molecular Analysis of Pterygoplichthys pardalis (Castelnau 1855) from the Lower Amazon River.

Pterygoplichthys pardalis is an armored catfish native to South America and an important resource for the ornamental fish industry. Recently, several exotic populations have been introduced into rivers on five continents. Despite its commercial and environmental importance, P. pardalis is poorly studied from a genetic perspective. In this study, we analyzed the karyotype of P. pardalis from the Amazon River and molecular variations in the mitochondrial gene Cytochrome oxidase I (COI) between native and exotic populations. The karyotype presented diploid number 2n = 52 and NF = 100 without cytogenetic variation between males and females. Nucleolus organizer regions (Ag-NOR) in the distal region of the long arm of pair 12 coincided with the 18S hybridization signal, whereas 5S was syntenic to this chromosome but localized in the short arm. The constitutive heterochromatin was restricted in the distal regions of pairs 4, 12, 25, and 26. Telomeric probes showed only distal hybridization signals. The karyotype of P. pardalis diverged from that of its congeners, and COI molecular variation revealed four haplotypes. The Philippine population revealed the greatest diversity with three haplotypes, while haplotype H1 was the most abundant and observed in both native and exotic populations. This new genetic data contributes to species management and provides useful information from an aquaculture perspective.

Enhancement of body coloration of sword-tail fish (Xiphophorus helleri): Plant-derived bio-resources could be converted into a potential dietary carotenoid supplement.

The effects of natural plant-derived carotenoid sources on body coloration and growth performance of Sword-Tail Fish (Xiphophorus helleri) were investigated using four different dietary treatments; Commercial Feed (CF), China Rose Flower (CRF), Marigold Flower (MFF), and Carrot (CAF). CF was used as the control treatment without a natural source of carotenoids. The sources were mixed with feed (15 g/100 g) during the diet preparation. Fish were fed with four treatments twice a day for eight weeks, and then sampling was done weekly. A total of 96 fish were used for the experiment, and the mean initial weights were 3.314 g. MFF-treated fishes showed better growth performance in terms of mean weight, length and Specific Growth Rate (SGR), and carotenoid gain than the other treatments. The lowest Food Conversion Ratio (FCR) was also observed in the MFF-treated fish. The study suggested that marigold flower mixed feed could increase the body coloration and boost the growth performance of sword-tail fish and may be manufactured.

Cutaneous angiomatosis-like presentation in koi carp (Cyprinus carpio koi): Clinical-pathological investigations.

The skin represents an indicator of an animal's health status. Causes of cutaneous diseases in fish most often trace back to biological agents. However, fish skin diseases can also arise from a complex interaction of infectious and non-infectious causes, making it more difficult to identify a specific aetiology. In the period between April and September of the years 2019-2022, four koi carp (Cyprinus carpio koi) from two European countries presented with multifocal, irregularly round, few mm to 1 cm, variably raised cutaneous reddened areas. The fish displayed good general condition. Cutaneous samples, investigated by microbiological and molecular methods and microscopy, did not indicate a primary pathogenic agent. Gross and histological findings of the cutaneous biopsies were consistent with a multifocal/reactive process centred on dermal vessels. The histological features were reminiscent of angiomatosis, a benign proliferative condition affecting the dermal vessels of mammals, including human patients. The clinical-pathological presentation and the dermatologic condition that affected the koi carp are discussed and compared with the veterinary and human literature.

Generation of Albino Phenotype in Ornamental Fish by CRISPR/Cas9-Mediated Genome Editing of slc45a2 Gene.

Albinism is the most common color variation described in fish and is a fascinating trait of some ornamental fish species. Albino mutants can be generated by knocking out core genes affecting melanin synthesis like slc45a2 in several fish species. However, genetic mutation remains challenging for species with unknown genome information. In this study, we generated albino mutants in two selected ornamental fish species, royal farlowella (Sturisoma panamense), and redhead cichlid (Vieja melanura). For this purpose, we carried out phylogenetic analyses of fish slc45a2 sequences and identified a highly conserved region among different fish species. A pair of degenerate primers spanning this region was designed and used to amplify a conserved slc45a2 fragment of 340 bp from the two fish species. Based on the amplified sequences, a target site in the 6th exon was used for designing guide RNA and this targeted site was first verified by the CRISPR/Cas9 system in the zebrafish (Danio rerio) model for the effectiveness. Then, specific guide RNAs were designed for the two ornamental fish species and tested. Most of the injected larvae completely lost black pigment over the whole body and eyes. DNA sequencing confirmed a high degree of mutation at the targeted site. Overall, we described a fast and efficient method to generate albino phenotype in fish species by targeting the conserved 6th exon of slc45a2 gene for genome editing via CRISPR/Cas9 and this approach could be a new genetic tool to generate desirable albino ornamental fish.

Early detection of Pseudocapillaria tomentosa by qPCR in four lines of zebrafish, Danio rerio (Hamilton 1882).

The intestinal nematode Pseudocapillaria tomentosa in zebrafish (Danio rerio) causes profound intestinal lesions, emaciation and death and is a promoter of a common intestinal cancer in zebrafish. This nematode has been detected in zebrafish from about 15% of the laboratories. Adult worms are readily detected about 3 weeks after exposure by either histology or wet mount preparations of the intestine, and larval worms are inconsistently observed in fish before this time. A quantitative PCR (qPCR) test was recently developed to detect the worm in fish and water, and here we determined that the test on zebrafish intestines was effective for earlier detection. Four lines of zebrafish (AB, TU, 5D and Casper) were experimentally infected and evaluated by wet mounts and qPCR at 8, 15-, 22-, 31- and 44-day post-exposure (dpe). At the first two time points, only 8% of the wet mounts from exposed fish were identified as infected, while the same intestines screened by qPCR showed 78% positivity, with low and consistent cycle threshold (Ct) values at these times. Wet mounts at later time points showed a high prevalence of infection, but this was still surpassed by qPCR.

Spawning, egg and larval description of the brazilian basslet Gramma brasiliensis Sazima, Gasparini & Moura, 1998 in captivity.

Brazilian basslet Gramma brasiliensis is a fish highly appreciated by the marine ornamental industry. There is an increasing interest in the development of a breeding protocol for this species. However, descriptions of the reproductive biology, eggs and larval development are scarce. This study was the first to describe the spawning, eggs and larvae of G. brasiliensis in captivity, including mouth size information. Six spawning events produced egg masses with 27, 127, 600, 750, 850, and 950 eggs. Larger egg masses showed embryos with at least two different developmental stages. Eggs are spherical (∼1.0 mm diameter), held together by filaments entangling chorionic projections. Larvae with fewer than 12 hph (hours post-hatch) presented 3.55 mm standard-length, well developed eyes, fully absorbed yolk sac, an inflated swim bladder and mouth opened. Exogenous feeding on rotifers began within 12 hph. The average mouth width at first feeding was 0.38 mm. The first settled larva was noted by day 21. This information should help to determine appropriate diets and prey-shift time during the larviculture of the species.

Feeding and locomotor rhythm of the ornamental cichlid fish Amatitlania sp. in different social groups.

The aim of this study was to evaluate the feeding activity and the rhythm of daily locomotor activity of the convict cichlid (Amatitlania sp.) kept in different social groups under a self-feeding system. A total of 120 animals was distributed among six repetitions of four social groups, as follows: group 1 with one male and one female per tank; group 2 with three males and three females per tank; group 3 with six males per tank; and group 4 with six females per tank. Feeding activity (FA) and locomotor activity (LA) were evaluated using photoelectric presence-sensors connected to automatic feeders. The fish were fed a commercial extruded diet (46% crude protein and 3600 kcal kg-1 of digestible energy). Animal growth was evaluated for all groups. After 30 days of experimentation, the fish stabilized their demands by adjusting their consumption. Amatitlania sp. showed predominantly diurnal FA and LA. All groups showed a peak of activity when the light was turned on and when it was turned off. In summary, FA and LA of Amatitlania sp. are predominantly diurnal and independent of social group. Pairs and groups of males and females together consume less food in relation to groups of one sex or the other due to reproductive behaviour. On the other hand, groups of only males or females consume more food because they lack reproductive stimuli and thus prioritize growth. These results may support good feeding management practices for this ornamental cichlid. Studies relating feeding behaviour with different social groups are of great importance for determining effective feeding strategies for this species in captivity. Thus, such a study assists in a more efficient production of Amatitlania sp.