1H NMR spectroscopy as a tool to probe potential biomarkers of the drying-salting process: A proof-of-concept study with the Amazon fish pirarucu.

Dry-salted pirarucu (Arapaima gigas) plays an important cultural role in the Amazon region - South America. In this study, we explored the changes in the chemical composition of pirarucu meat following the drying-salting process via 1H NMR spectroscopy. Combining multivariate and univariate statistical analyses yielded a robust differentiation of metabolites involved in the process. VIP score (>1), p-value (<0.05), and AUC (>0.7) were considered to selecting compounds that had significant fluctuations in their contents along the process. Our results pointed out acetate, lactate, succinate, and creatinine as metabolites undergoing significant changes during the drying-salting process. Creatinine was not detected in fresh samples. The investigation of multiple components delves deeper into the molecular nuances of the salting-drying process's impact on fish meat, providing a more comprehensive understanding of the possible chemical transformations and how the matrix's quality control and nutritional aspects should be addressed.

Yield and Technological Quality of Pirarucu Trimming Surimi According to the Number of Washing Cycles, Rice Flour Content, and Cooking Temperature.

This research aimed to optimize surimi production with innovative raw materials called pirarucu trimmings and broken rice grains, which are by-products from food industries. The effects of three independent variables (number of washing cycles, content, and cooking temperature of rice flour-RF) on surimi's physical, chemical, and technological qualities were investigated through a Box-Behnken design. The number of washing cycles affected yield (77-93%), moisture (55-67%), lipids (18-35%), protein (7.15-11.88%), whiteness (46.73-64.45), chroma (8.86-13.18), hue angle (80.79-93.12°), cohesiveness (0.40-0.61), springiness (0.51-0.99), and freeze stability after 4 weeks (85.16-96.53%). RF concentration affected moisture, lipids, cohesiveness, springiness, and freeze-thaw stability after 4 weeks. RF cooking temperature affected moisture, chroma, cohesiveness, and springiness. The optimal conditions for surimi production with high yield and overall quality were three washing cycles, 6% of RF, and RF cooking temperature at 85 °C. It reveals the promising potential of both by-products to be used as an ingredient in restructured products and contribute to improving agri-industry sustainability.

Molecular Cloning and AlphaFold Modeling of Thyrotropin (ag-TSH) From the Amazonian Fish Pirarucu (Arapaima gigas).

Arapaima gigas, known as Pirarucu in Brazil, is one of the largest freshwater fish in the world. Some individuals could reach 3 m in length and weight up to 200 kg. Due to extinction risks and its economic value, the species has been a focus for preservation and reproduction studies. Thyrotropin (TSH) is a glycoprotein hormone formed by 2 subunits α and ß whose main activity is related to the synthesis of thyroid hormones (THs)-T3 and T4. In this work, we present a combination of bioinformatics tools to identify Arapaima gigas ßTSH (ag-ßTSH), modeling its molecular structure and express the recombinant heterodimer form in mammalian cells. Using the combination of computational biology, based on genome-related information, in silico molecular cloning and modeling led to confirm results of the ag-ßTSH sequence by reverse transcriptase-polymerase chain reaction (RT-PCR) and transient expression in human embryonic kidney (HEK293F) cells. Molecular cloning of ag-ßTSH retrieved 146 amino acids with a signal peptide of 21 amino acid residues and 6 disulfide bonds. The sequence has a similarity to 39 fish species, ranging between 43.1% and 81.6%, whose domains are extremely conserved, such as cystine knot motif and N-glycosylation site. The Arapaima gigas thyrotropin (ag-TSH) model, solved by AlphaFold, was used in molecular dynamics simulations with Scleropages formosus receptor, providing similar values of free energy ΔGbind and ΔGPMF in comparison with Homo sapiens model. The recombinant expression in HEK293F cells reached a yield of 25 mg/L, characterized via chromatographic and physical-chemical techniques. This work shows that other Arapaima gigas proteins could be studied in a similar way, using the combination of these techniques, recovering more information from its genome and improving the reproduction and preservation of this prehistoric fish.

Isolation and Characterization of the Arapaima gigas Growth Hormone (ag-GH) cDNA and Three-Dimensional Modeling of This Hormone in Comparison with the Human Hormone (hGH).

In a previous work, the common gonadotrophic hormone α-subunit (ag-GTHα), the ag-FSH ß- and ag-LH ß-subunit cDNAs, were isolated and characterized by our research group from A. gigas pituitaries, while a preliminary synthesis of ag-FSH was also carried out in human embryonic kidney 293 (HEK293) cells. In the present work, the cDNA sequence encoding the ag-growth hormone (ag-GH) has also been isolated from the same giant Arapaimidae Amazonian fish. The ag-GH consists of 208 amino acids with a putative 23 amino acid signal peptide and a 185 amino acid mature peptide. The highest identity, based on the amino acid sequences, was found with the Elopiformes (82.0%), followed by Anguilliformes (79.7%) and Acipenseriformes (74.5%). The identity with the corresponding human GH (hGH) amino acid sequence is remarkable (44.8%), and the two disulfide bonds present in both sequences were perfectly conserved. Three-dimensional (3D) models of ag-GH, in comparison with hGH, were generated using the threading modeling method followed by molecular dynamics. Our simulations suggest that the two proteins have similar structural properties without major conformational changes under the simulated conditions, even though they are separated from each other by a >100 Myr evolutionary period (1 Myr = 1 million years). The sequence found will be used for the biotechnological synthesis of ag-GH while the ag-GH cDNA obtained will be utilized for preliminary Gene Therapy studies.

Identification of type II interferons and receptors in an osteoglossiform fish, the arapaima Arapaima gigas.

In mammals, type II interferon (IFN; i.e. IFN-γ) signalling transduces through its specific receptors IFN-γR1 and IFN-γR2. In an osteoglossiform fish, the arapaima Arapaima gigas, three type II IFNs, IFN-γ-like, IFN-γ and IFN-γrel, and their four possible receptor subunits IFN-γR1-1, IFN-γR1-2, IFN-γR2-1 and IFN-γR2-2 were identified in this study. The three type II IFN genes are composed of four exons and three introns, and they all contain IFN-γ signature motif and signal peptide, with the presence of potential nuclear localization signal (NLS) in IFN-γ-like and IFN-γ. The IFN-γR1-1, IFN-γR1-2, IFN-γR2-1 and IFN-γR2-2 are composed of seven exons and six introns, with predicted IFN-γR1-1 and IFN-γR1-2 proteins containing JAK1 and STAT1 binding sites, and IFN-γR2-1 and IFN-γR2-2 containing JAK2 binding sites. Gene synteny analysis showed that the type II IFN and their receptor loci are duplicated in arapaima. All these genes were expressed constitutively in all organs/tissues examined, and responded to the stimulation of polyI:C. The prokaryotic recombinant IFN-γ-like, IFN-γ and IFN-γrel proteins can significantly induce the upregulation of immune-related genes in trunk kidney leucocytes. The ligand-receptor relationship analyses revealed that recombinant IFN-γ-like, IFN-γ, and IFN-γrel transduce downstream signalling through IFN-γR1-1/IFN-γR2-1, IFN-γR1-2/IFN-γR2-2, and IFN-γR1-1, respectively, in xenogeneic cells with the overexpression of original or chimeric receptors. In addition, tyrosine (Y) 366 and Y377 in the intracellular region may be essential for the function of IFN-γR1-2 and IFN-γR1-1, respectively. The finding of type II IFN system in A. gigas thus provides different knowledge in understanding the diversity and evolution of type II IFN ligand-receptor relationships in vertebrates.

Extraction and characterization of collagen from the skin of Amazonian freshwater fish pirarucu

The need to fully exploit fishing resources due to increasing production and consequent waste generation requires research to promote the sustainability of the fishing industry. Fish waste from the industry is responsible for relevant environmental contamination. However, these raw materials contain high amounts of collagen and other biomolecules, being attractive due to their industrial and biotechnological applicability. Thus, to reduce the waste from pirarucu (Arapaima gigas) processing, this study aimed to obtain collagen from pirarucu skin tissue. The extraction process used 0.05 M sodium hydroxide, 10% butyl alcohol, and 0.5 M acetic acid, with extraction temperature of 20°C. The obtained yield was 27.8%, and through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), it was determined that the collagen obtained was type I. This study showed that collagen solubility was highest at pH 3 and the lowest solubility was at concentrations of 3% sodium chloride. The denaturation temperature of collagen was 38.1°C, and its intact molecular structure was observed using the Fourier transform infrared spectrophotometry technique with an absorption radius of 1. The results showed that it was possible to obtain collagen from pirarucu skin at 20°C, which has the typical characteristics of commercial type I collagen. In conclusion, the procedures used may be considered to be an interesting alternative for collagen extraction, a new product obtained from the processing of fish waste.

Novel Microsatellite Markers Used for Determining Genetic Diversity and Tracing of Wild and Farmed Populations of the Amazonian Giant Fish Arapaima gigas.

The Amazonian symbol fish Arapaima gigas is the only living representative of the Arapamidae family. Environmental pressures and illegal fishing threaten the species' survival. To protect wild populations, a national regulation must be developed for the management of A. gigas throughout the Amazon basin. Moreover, the reproductive genetic management and recruitment of additional founders by aquaculture farms are needed to mitigate the damage caused by domestication. To contribute to the sustainable development, we investigated the genetic diversity of wild and cultivated populations of A. gigas and developed a panel composed by 12 microsatellite markers for individual and population genetic tracing. We analyzed 368 samples from three wild and four farmed populations. The results revealed low rates of genetic diversity in all populations, loss of genetic diversity and high inbreeding rates in farmed populations, and genetic structuring among wild and farmed populations. Genetic tracing using the 12 microsatellite markers was effective, and presented a better performance in identifying samples at the population level. The 12-microsatellite panel is appliable to the legal aspects of the trade of the A. gigas, such as origin discrimination, reproductive genetic management by DNA profiling, and evaluation and monitoring of genetic diversity.

Influence of grilling pretreatment and optimization of sous vide processing parameters on the physicochemical and microbiological quality of pirarucu fillet.

The demand for high-quality food products has promoted the study of techniques for its processing and conservation. The present research aimed to evaluate the influence of grilling pretreatment on the physical characteristics of pirarucu fillets and the heat transfer process by a computational modelling, and to optimize the sous vide process parameters. Before and after the sous vide process, the samples were analysed for microbiological, chemical and physical characteristics. There was no significant difference between the total experimental time of grilling and that obtained by computational modelling. Immersion in brine for 300 s in combination with grilling at 200 ℃/120 s was selected because of its water-holding capacity (%) 79.40 ± 0.31, texture (N) 1.91 ± 0.40 and value of L* 74.44 ± 0.38 in the fillets. Cooking at 60 ℃ for 568.8 s were the best sous vide parameters obtained, with highest water-holding capacity (%) 93.60, texture (N) 6.24, ΔE* 7.43, and with microbiological loads below 6 log CFU/g and 7 log MPN/g in the final product. Useful information obtained from this study highlighted the brine and grill pretreatment in combination with sous vide proved it is a potential solution for developing pirarucu products even at an industrial scale.

Effect of density of fingerling and juvenile pirarucu during transportation on water quality and physiological parameters

We assessed the effect of stocking density on physiological parameters (blood lactate, glucose, cortisol, hematocrit), water quality (temperature, dissolved oxygen, pH, unionized ammonia, carbon dioxide), and survival during the transportation of fingerling (24.5 ± 4.7 g) and juvenile (615.8 ± 122.2 g) pirarucu (Arapaima gigas) for six hours in plastic bags. The tested densities were 65, 80, 95, 110 and 125 g L-1 for fingerlings, and 50, 80, 110, 140 and 170 g L-1 for juveniles (three replicates each). Parameters were measured prior to and immediately after transportation, and at 24 and 96 hours recovery after transportation. No mortality was observed, except for fingerlings (< 3%) at densities of 110 and 125 g L-1 during recovery. All the water quality parameters were significantly altered after the transportation of fingerlings and juveniles. Water temperature, dissolved oxygen, carbon dioxide and unionized ammonia increased, but pH decreased. Only carbon dioxide and unionized ammonia differed among densities. Cortisol levels did not increase over time, except for the juveniles at 170 g L-1, which still had high cortisol after 96 hours. Glucose significantly increased after transportation for all the treatments and returned to the initial values during the recovery period. Conversely, the lactate values were still high after 96 hours. Hematocrit was assessed only for juveniles and was significantly lower after transportation. We conclude that fingerling and juvenile pirarucu can be safely transported at densities up to 95 g L-1 and 140 g L-1, respectively. (AU)

Chlorapatite Derived From Fish Scales.

The present work demonstrates the production of chlorapatite (ClAp) through thermal decomposition of chemically treated fish scales, originating from an Amazon fish species (Arapaima gigas). The scales were treated with hydrochloric acid (HCl) solution for deproteinization. Afterwards, the solution was neutralized by sodium hydroxide (NaOH) treatment to obtain an apatite-rich slurry. The heat treatment was carried out at different temperatures including 600 °C, 800 °C, and 1000 °C. The powders obtained were characterized through X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray spectroscopy (EDS), and scanning electron microscopy (SEM). The XRD analysis and FTIR spectra confirmed the incorporation of chlorine into the apatite structure. The FTIR results showed absorption bands relative to the OH-, PO43- functional groups which are a characteristic of chlorapatite; moreover, the intensity of the OH-Cl elongation could be observed. Chlorapatite Ca5(PO4)3Cl, NaCl, and NaCaPO4 phases were identified, achieving up to 87.4 wt% for ClAp. From the SEM observations, the ClAp obtained consisted of slightly larger grains and more crystalline with increasing temperature with observed grains ranging in sizes between 1 and 5 µm and 84.27% crystallinity for the ClAp1000 sample. The ClAp and NaCaPO4 can be used in electronics as a phosphor material due to the fact of its luminescence and biomedical applications.

Expression of Acetylcholine- and G protein coupled Muscarinic receptor in the Neuroepithelial cells (NECs) of the obligated air-breathing fish, Arapaima gigas (Arapaimatidae: Teleostei).

The air-breathing specialization has evolved idependently in vertebrates, as many different organs can perfom gas exchange. The largest obligate air-breathing fish from South America Arapaima gigas breathe air using its gas bladder, and its dependence on air breathing increases during its growth. During its development, gill morphology shows a dramatic change, remodeling with a gradual reduction of gill lamellae during the transition from water breathing to air breathing . It has been suggested that in this species the gills remain the main site of O2 and CO2 sensing. Consistent with this, we demonstrate for the first time the occurrence of the neuroepithelial cells (NECs) in the glottis, and in the gill filament epithelia and their distal halves. These cells contain a broader spectrum of neurotransmitters (5-HT, acetylcholine, nNOS), G-protein subunits and the muscarininic receptors that are coupled to G proteins (G-protein coupled receptors). We report also for the first time the presence of G alpha proteins coupled with muscarinic receptors on the NECs, that are thought as receptors that initiate the cardiorespiratory reflexes in aquatic vertebrates. Based on the specific orientation in the epithelia and their closest vicinity to efferent vasculatures, the gill and glottal NECs of A. gigas could be regarded as potential O2 and CO2 sensing receptors. However, future studies are needed to ascertain the neurophysiological characterization of these cells.

Avaliação microbiológica do pirarucu (Arapaima gigas) manejado nas Reservas de Desenvolvimento Sustentável Mamirauá e Amanã (resultados preliminares) / Microbiological evaluation of pirarucu (Arapaima gigas) managed in Mamirauá and Amanã Sustainable Development Reserves (preliminary results)

O pirarucu (Arapaima gigas) é um peixe carnívoro da Amazônia e é considerado um dos peixes mais importantes da região. Ele é manejado de forma sustentável por Acordos de Pesca da Reserva de Desenvolvimento sustentável Mamirauá. Dados microbiológicos são necessários para verificar a qualidade higiênico-sanitária dos peixes manejados nessa região. Devido a essa importância o presente trabalho teve como objetivo avaliar a qualidade microbiológica do pirarucu manejado nas Reservas de Desenvolvimento Sustentável Mamirauá e Amanã. Foram avaliados 16 amostras no total, provenientes de dois Acordos de Pesca diferentes (AP1 e AP2). Os microrganismos analisados foram Mesófilos, Staphylococcus aureus, Coliformes Totais e Escherichia coli. As amostras dos dois Acordos de Pesca tiveram presença para todas as bactérias e grupos de bactérias analisados, com valores mais elevados para o AP1. Tendo assim a partir desses resultados a necessidade de um controle maior das condições higiênico-sanitária no fluxograma de beneficiamento dos locais estudados.

Application of pulsed-vacuum on the salt impregnation process of pirarucu fillet.

The influence of the pulsed-vacuum impregnation (PVI) and periodic pulsed-vacuum impregnation (PPVI), and the temperature, on the salting process of pirarucu fillet was studied. For this, the dorsal region of the fish in the slab-shaped (60 mm × 20 mm × 5 mm) and sodium chloride solution (30 g/100 g NaCl) were used. The process was conducted at different temperatures (10 °C - 40 °C) and the following pulsed-vacuum conditions: initial vacuum pulse (5 min at 10 kPa) and alternating periods of vacuum pulse (5 min at 10 kPa) and atmospheric pressure (101 kPa) for 5 min, 10 min and 15 min intermittently. The solid gain (SG) in the fillet was more pronounced in the first two hours of the salting process. The increase of salt content in the product (0.33 ±â€¯0.02 to 0.43 ±â€¯0.04 g/g db) promoted the gradual reduction of moisture (72.99 ±â€¯1.27 to 57.1 ±â€¯2.4 g/100 g) and water activity (1.00 ±â€¯0.03 to 0.78 ±â€¯0.05). PVI and PPVI processes can be used efficiently at room temperature or under refrigeration in the salting process of pirarucu fillet. On the other hand, at higher temperatures (>30 °C), the effect of temperature prevails over the effect of vacuum. Peleg model satisfactorily explains the salting kinetics of the pirarucu fillet.

Growth, physiological and immune responses of Arapaima gigas (Arapaimidae) to Aeromonas hydrophila challenge and handling stress following feeding with immunostimulant supplemented diets.

The current study tested the efficacy of a dietary immunostimulant additive (Aquate Fish™®) on the growth performance, and on the physiological and immune responses of Arapaima gigas. Two trials were carried out: a feeding trial for 30 days with the experimental diets and a challenge trial for 7 days, in which fish were bacterial challenge (Aeromonas hydrophila) following by 60 s handling stress. During the feeding trial, fingerlings were fed diets supplemented with 0 (control), 6, 9 and 12 g Aquate Fish™®/kg diet. Dietary supplementation did not influence feed intake, feed conversion and condition factor, but increased the final biomass, number of erythrocytes, thrombocytes, leukocytes, lymphocytes, monocytes, hemoglobin, glucose, globulins and plasma triglycerides in fish fed at a concentration of 12 g/kg diet. After bacterial infection, mortality occurred only in fish fed control treatment, whereas respiratory burst of leukocytes, number of leukocytes and lymphocytes increased in fish that received 12 g of dietary supplementation. The results indicated that dietary supplementation with 12 g of Aquate Fish™® improved biomass and immunity performance of A. gigas fingerlings, without negatively affecting blood biochemical parameters.

Cetobacterium Is a Major Component of the Microbiome of Giant Amazonian Fish (Arapaima gigas) in Ecuador.

Arapaima gigas is a large air-breathing fish found in Amazonian rivers, a characteristic that gives this species an advantage in oxygen-deprived waters. It shows high potential for aquaculture in the Amazon region due to its fast growth rate that approaches 10⁻15 kg/year. The aim of this study was to explore the composition of the intestinal bacterial community of Arapaima gigas reared in Ecuador using 16S rRNA gene high-throughput sequencing. The analysis revealed significant differences in alpha diversity indices (p < 0.05) and differential distribution of minor components of the intestinal microbiome between small and large fish. However, components with greater relative abundance, such as Cetobacterium, are found in similar proportions.

Histopathological evaluation of formalin toxicity in Arapaima gigas (Arapaimidae), the giant fish from Amazon / Avaliação histopatológica da toxicidade da formalina em Arapaima gigas (Arapaimidae), o peixe gigante da Amazônia

This study aimed to determine the lethal concentration and the structural and ultra-structural effects caused by the formalin exposure on juveniles of Arapaima gigas. Ninety fish (60.1± 2.5g and 20.2±0.9cm) were exposed to 0, 22, 44, 66, 88 and 110mg L-1 in order to determine the lethal concentration (LC50-96h) that was 36.4mg L-1 of formalin. Sublethal effects were evaluated using histopathological analysis on the gills and assessment of behavioral alterations and clinical signs. The LC50 of formalin for 24, 48 and 72h was 88.3, 64.7 and 56.8mg L-1 respectively. Clinical signs and behavioral changes were found: erratic swimming, lethargy, crowding on the water surface, loss of hydrodynamic equilibrium, spasms and agonistic confrontation, which were observed only at 88 and 110mg L-1. The histological alteration index (HAI) showed that 66, 88 and 100mg L-1 presented significant difference (p<0.05) in relation to unexposed fish, indicating that moderate damage to the gills of fish exposed to formalin had occurred. The mean values of alteration (MVA) for 22, 44, 66, 88 and 110mg L-1 were 1.14, 1.29, 1.51, 1.53 and 1.60 respectively, and differences in this index were only observed with 110 mgL-1 of formalin. It is therefore possible to conclude that sublethal concentrations of formalin (22.0mg L-1) did not compromise the health of juveniles of A. gigas. Finally, concentrations greater than to LC50-96h may be carefully used for short-term exposure, since the MVA for all concentrations tested only indicated localized lesions that did not compromise gills functionality of exposed fish.(AU)

O presente estudo teve como objetivo determinar a concentração letal e efeitos estruturais e ultraestruturais causados pela exposição a formalina em juvenis de Arapaima gigas. Noventa peixes (60,1±2,5g e 20,2±0,9cm) foram expostos a 0, 22, 44, 66, 88 e 110mg L-1, para determinar a concentração letal (CL50-96h) de formalina que foi 36,4mg L-1. Os efeitos subletais foram avaliados por análises histopatológicas das brânquias e avaliação das alterações comportamentais e sinais clínicos. A CL50 de formalina para 24, 48 e 72horas foi de 88,3, 64,7 e 56,8; respectivamente. Sinais clínicos e alterações comportamentais encontradas foram: natação errática, letargia, aglomeração de peixes na superfície da água, perda de equilíbrio hidrodinâmico, espasmos e confronto agonísticos, observados apenas nas concentrações de 88 e 110mg L-1. O índice de alteração histológica (IAH) mostrou que as concentrações de 66, 88 e 100mg L-1 apresentaram diferenças significativas (p<0,05) em relação aos controles, indicando a ocorrência de danos moderados nas brânquias dos peixes expostos a formalina. Os valores médios de alteração (VMA) para as concentrações 22, 44, 66, 88 e 110mg L-1 foram 1,14, 1,29, 1,51, 1,53 e 1,60; respectivamente, e as diferenças na composição desse índice foram observados apenas na exposição com 110mg L-1 de formalina. Foi possível concluir que concentrações subletais de formalina (22,0mg L-1) não comprometem a saúde de juvenis de A. gigas. Concentrações de formalina acima da CL50-96h podem ser usadas cuidadosamente para banho de curto tempo, uma vez que o VMA para todas as concentrações testadas indicou apenas lesões localizadas que não comprometem a funcionalidade das brânquias dos peixes expostos.(AU)

Chemical composition of different muscle zones in pirarucu (Arapaima gigas)

Abstract Identifying potential patterns in pirarucu (Arapaima gigas) composition, as a function of the different fish muscle zones (dorse, venter, ventrecha and tail), was the main objective of this research. For such, the different pirarucu muscle zones were evaluated, in order to obtain proximate composition and minerals content. It was also determined amino acids and fatty acids contents in fish muscle. The dorsal, ventral and tail muscle zones presented similar moisture (76.5-78.2%), protein (17.8-18.9%), total lipids (1.0-1.5%) and ash (0.9-1.2%) contents. On the other hand, the ventrecha zone presented 25.8% of protein and the major total lipids content (17.1%). The main minerals found in fish muscle were K (183.5-288.6 mg/100 g muscle) and Na (65.1-175.5 mg/100 g). Glutamic acid (3027.6 mg/100 g muscle) was the main amino acid found in fish muscle, which presented 48% of essential amino acids in the protein fraction. Lipids content showed 57.3% of unsaturated fatty acids and 42.7% of saturated fatty acids.

Combined osmotic dehydration and drying process of pirarucu (Arapaima gigas) fillets.

The osmotic dehydration (OD) and complementary drying of pirarucu (Arapaima gigas) fillets were studied. Pieces of the dorsal portion of pirarucu (60 mm × 20 mm × 10 mm) underwent OD in a binary solution (NaCl-water) with the application of vacuum pulse following a central rotatable composite design. The effect of the following process variables was assessed: temperature (20-40 °C), osmotic solution concentration (15-25% NaCl), and vacuum pulse pressure (7-101 kPa) on water loss (WL), solid gain (SG), and water activity (aw). OD kinetics was obtained and the Peleg model was fitted to WL and SG data. The osmotically dehydrated pirarucu was dried (40-70 °C) in a fixed-bed dryer and mathematical models were fitted to the drying data. The optimal operational condition for the OD process was 35 °C, solution with 25% NaCl, and atmospheric pressure, which yielded WL of 14.87 ± 1.46%, SG of 8.56 ± 0.45%, and aw of 0.87 ± 0.02. The Peleg model efficiently predicted the WL and SG kinetics. The increase in the water loss in drying was more evident at low temperatures (40-50 °C) with effective diffusivity ranging from 10.85 × 10-9 to 12.30 × 10-9 m2/s. The Midilli and Page models efficiently predicted the drying kinetics.

Characterization of microbiota in Arapaima gigas intestine and isolation of potential probiotic bacteria.

AIMS: The aim of this study was to determine the intestinal microbiota of pirarucu (Arapaima gigas) in different growth stages (adult and fingerlings) and to isolate and identify potential probiotic bacteria. METHODS AND RESULTS: High-throughput sequencing analysis of the intestinal contents revealed that the majority of sequences belonged to the Proteobacteria, Fusobacteria and Firmicutes phyla. At the genus level, the greatest number of sequences belonged to Bradyrhizobium in adult fish, while Cetobacterium was the most abundant in juvenile fish. Twenty-three lactic-acid bacteria (LABs) were isolated on MRS agar from healthy juvenile fish. The isolates were tested in vitro for probiotic properties. Two isolates (identified as strains of Lactococcus lactis subsp. lactis and Enterococcus faecium) displayed antagonism against all 10 pathogens tested, were nonhaemolytic and maintained good viability for at least 3 weeks when supplemented to fish diets. The presence of a number of antibiotic resistance genes (ARGs), conferring resistance to erythromycin, tetracycline and chloramphenicol, was investigated by PCR. CONCLUSIONS: The absence of ARGs investigated the potential to antagonize pathogens, and favourable growth and survival characteristics indicate that these autochthonous isolates have the potential to be considered probiotics, which will be studied in future in vivo experiments. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has demonstrated, for the first time, the normal microbiota in the A. gigas intestine during different life stages and the presence of LAB strains. It also demonstrated LAB antibiotic resistance and antagonistic behaviour against pathogens isolated from the same fish.

Dawestrema cycloancistrium (Monogenea) from the head pores of arapaimas.

Arapaima gigas is one of the main cultured fish species in South America, and monogenean parasites of this species cause large economic losses to fish farmers. During surveys of the parasites of cultured arapaimas from Mexiana Island in the Amazon River Delta, Rio Branco, in northwestern Brazilian Amazonia, and Yurimaguas, Peru, the monogenean Dawestrema cycloancistrium was found in the gills of A. gigas as well as in previously unreported sites, i.e. the head pores and chambers. The aim of this study was to investigate the transmission route of this parasite and its geographical distribution as well as to describe its morphology as observed by light and confocal imaging. Phalloidin labeling confirmed the presence of 2 prostatic reservoirs and showed muscular branches of fibers supporting haptoral sclerites. In arapaimas, the head connects to the gill chambers via 2 perforated scales located at the dorsolateral sides of the distal part of the head. The scales connect to thin channels and chambers situated in the proximal part of the head. These chambers are filled with cephalic mucus, which flows out to the environment through terminal pores. Adults and egg masses of monogeneans were found on the gills and inside the head pores and cavities of fish along with cephalic mucus. This indicates a specialized method of parasite transmission from adult fish to fingerlings during parental care (holding offspring in the mouth) or via head secretions, providing evidence of a new adapted mechanism of dispersion.