Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt

Abstract Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Resumo Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis (9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt / Sequenciamento e resistência antimicrobiana múltipla de Pseudomonas fluorescens isoladas de tilápia-do-nilo no Egito

Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis ​​(9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Supervivencia observada en tres familias de tilapia del Nilo (Oreochromis niloticus) infectadas con Streptococcus agalactiae / Survival observed in three families of Nile tilapia (Oreochromis niloticus) infected with Streptococcus agalactiae

RESUMEN La estreptococosis es una de las principales enfermedades en los peces de agua dulce que causa altas tasas de mortalidad. El objetivo de este estudio fue evaluar la respuesta en la supervivencia a la infección por Streptococcus agalactiae en tres familias de tilapia. El experimento se llevó a cabo en el Laboratorio de Enfermedades de los Peces de la Universidad Federal de Lavras. Se utilizaron peces con un peso de 93,7 ± 5,4 g de tres familias diferentes (FA, FB y FC). Se utilizaron 36 peces en cada unidad experimental, inoculados intraperitonealmente con 107 UFC/mL de Streptococcus agalactiae por peces y un grupo control por familia con 9 peces con 1 mL de caldo BHI (Infusión Cerebro Corazón) evaluados durante 15 días. No hubo mortalidad del grupo control. Se observó la presencia de exoftalmia, coloración oscura en todo el cuerpo, letargo y dilatación abdominal antes de la muerte en las tres familias evaluadas expuestas al patógeno. El estimador no paramétrico de Kaplan-Meier se utilizó para observar las curvas de supervivencia. Durante los 15 días del desafío, el tiempo promedio de supervivencia de un individuo en las familias FA, FB y FC fue de 9,4; 6,90 y 8,14 días, respectivamente. Pruebas de Log-rank y Peto & Peto para evaluar la diferencia entre las curvas de supervivencia arrojaron que no hubo diferencias significativas entre las familias evaluadas (P=0,08 y P= 0,09), respectivamente.

ABSTRACT Streptococcosis is one of the main diseases in freshwater fish that causes high mortality rates. The objective of this study was to evaluate the survival response to Streptococcus agalactiae infection in three families of tilapia. The experiment was carried out at the Laboratory of Fish Diseases of the Federal University of Lavras. Fish weighing 93.7 ± 5.4 g from three different families (FA, FB, and FC) were used. 36 fish were used in each experimental unit, intraperitoneally inoculated with 107 CFU/mL of Streptococcus agalactiae per fish and a control group per family with 9 fish with 1 mL of BHI broth (Brain Heart Infusion) evaluated for 15 days. There was no mortality in the control group. The three evaluated families exposed to the pathogen observed the presence of exophthalmia, dark coloration throughout the body, lethargy, and abdominal dilation before death. The Kaplan-Meier nonparametric estimator was used to observe the survival curves. During the 15 days of the challenge, the average survival time of an individual in the FA, FB, and FC families was 9.4, 6.90, and 8.14 days, respectively. Log-rank and Peto & Peto test to evaluate the difference between the survival curves showed no significant differences between the assessed families (P=0.08 and P= 0.09), respectively.

Evaluation of chemical silage on egg quality parameters in ISA Brown line laying hens (Gallus gallus domesticus) / Evaluación del ensilado químico sobre los parámetros de calidad del huevo en gallinas ponedoras de la línea ISA Brown (Gallus gallus domesticus)

ABSTRACT In this study we evaluated the inclusion of chemical silage from red tilapia viscera (Oreochromis sp.) in diets for ISA Brown line laying hens (Gallus gallus domesticus) and its influence on nutritional parameters and egg quality. A total of 56 16-week-old laying hens were randomly divided into two groups (one per diet), which in turn were subdivided into 7 groups, each with 4 birds. Eggs were collected during the first 13 weeks of laying. During this period the egg quality parameters were evaluated until week 11; the bromatological characterization of the egg was carried out in the 2nd and 4th week of laying. The results indicated that the inclusion of chemical silage in 17,18% dry matter does not result in statistically significant differences in egg quality parameters or in their bromatological composition, with respect to the control. This led to the conclusion that chemical silage can be used as an alternative to protein inputs such as fish meal and soybean cake in the diet of laying hens, without modifying the quality of the final product.

RESUMEN En este trabajo se evaluó la inclusion de ensilado químico de vísceras de tilapia roja (Oreochromis sp.) en dietas para gallinas ponedoras (Gallus gallus domesticus) de la raza Isa-Brown, y su influencia sobre parámetros nutricionales y la calidad del huevo. Se utilizó un total de 56 gallinas ponedoras de 16 semanas de edad divididas al azar en dos grupos (uno por dieta), los cuales a su vez se subdividieron en 7 grupos de 4 aves cada uno. Se les recolectó huevos durante las primeras 13 semanas de postura. Durante este periodo se realizó la evaluación de los parámetros de calidad del huevo hasta la semana 11. La caracterización bromatológica del huevo se realizo en las semanas 2 y 4, luego de haber sido puestos. Los resultados indicaron que la inclusion de ensilado químico en un 17,18% materia seca, no genera diferencias estadísticamente significativas (p < 0,05) en los parámetros de calidad del huevo ni en su composición bromatológica, con respecto al control. Esto permitió concluir que el ensilado químico puede ser usado como alternativa a insumos proteicos como harina de pescado y torta de soya en la alimentación de gallinas ponedoras, sin modificar la calidad del producto final.

Effect of replacing fish meal in fish diet with shrimp by-product meal on growth performance, feed utilization, length-weight relationship and condition factors of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) / Efecto de reemplazar la harina de pescado en la dieta de pescado por harina de subproducto de camarón sobre el rendimiento del crecimiento, la utilización del alimento, la relación longitud-peso y los factores de condición de la tilapia del Nilo, Oreochromis niloticus (Linnaeus, 1758)

SUMMARY: The present work aimed to study the effect of replacing fish meal (FM) in the fish diet with shrimp by-product meal (SBM) on the growth performance parameters of the Nile Tilapia, Oreochromis niloticus. A total of 200 specimens of monosex O. niloticus fries were obtained from a private fish farm at Kafr El-Sheikh Governorate. They were transported to the fish laboratory at the Animal House of Zoology Department, Faculty of Science, Al-Azhar University. After two weeks for acclimation, the fish were divided into 5 groups, 2 replicates for each group (20 fish in each replicate). The five experimental diets were: C: control group with20 % fish meal (FM) and 0 % shrimp by-product meal (SM); T1, T2, T3, and T4 FM was replaced with SBM as 25 %, 50 %, 75 % and 100 %, respectively. Results indicated that the highest final length, growth in length, length gain, daily length gain and other growth performance parameters including the relationship between length and weight and condition factors of O. niloticus were recorded in T4 group (fed diet in which fish meal was completely replaced with shrimp by-product meal); while, their lowest values were recorded in control group. On the other hand, feed utilization parameters (feed intake, food conversion ratio, the maximum values of feed efficiency ratio and protein efficiency ratio) were recorded in T4 group and the minimum values were recorded in T3 group (fed diet in which 75 % of fish meal was replaced with shrimp by-product meal).

RESUMEN: El presente trabajo tuvo como objetivo estudiar el efecto de sustituir la harina de pescado (HP) en la dieta del pescado por harina de subproductos de camarón (HSC) sobre los parámetros de rendimiento de crecimiento de la Tilapia del Nilo, Oreochromis niloticus. Se obtuvieron un total de 200 especímenes de monosexo de O. niloticus de una piscifactoría privada en la gobernación de Kafr El-Sheikh. Fueron transportados al laboratorio de peces en el Departamento de Zoología de la Facultad de Ciencias, de la Universidad Al-Azhar. Después de dos semanas de aclimatación, los peces se dividieron en 5 grupos: Se realizaron dos repeticiones para cada grupo (20 peces en cada repetición). Las cinco dietas experimentales fueron: C: grupo control con 20 % de harina de pescado (HP) y 0 % de harina de subproductos de camarón (HSC); T1, T2, T3 y T4 FM se reemplazó con HSC con 25 %, 50 %, 75 % y 100 %, respectivamente. Los resultados indicaron que la longitud final más alta, el crecimiento en longitud, la ganancia de longitud, la ganancia de longitud diaria y otros parámetros de rendimiento del crecimiento, como además la relación entre la longitud y el peso, y los factores de condición de O. niloticus, se registraron en el grupo T4 (con una dieta reemplazada con harina de subproducto de camarón); mientras que, sus valores más bajos se registraron en el grupo control. Por otro lado, los parámetros de utilización del alimento (ingesta de alimento, índice de conversión de alimento, los valores máximos de índice de eficiencia alimenticia e índice de eficiencia proteica) se registraron en el grupo T4 y los valores mínimos se registraron en el grupo T3 (alimentación con dieta en la que el 75 % de la harina de pescado fue reemplazada por harina de subproductos de camarón).

Selection of potential lactic acid bacteria from fish intestine of mackerel (Rastrelliger kanagurta) from Lembata Regency, East Nusa Tenggara, Indonesia

Aims@#The aim of this study was to screen lactic acid bacteria (LAB) isolates with probiotic properties for tilapia (Oreochromis niloticus) from the intestines of mackerel and to analyze its ability to produce hydrolase enzymes (amylase, protease, cellulase and lipase).@*Methodology and results@#Ten samples of mackerel were collected from the Fish Auction Place, Lewoleba, Nubatukan District, Lembata Regency, East Nusa Tenggara Province, Indonesia. The process for screening were antimicrobial activity, pH tolerance test of 1, 3 and 5, bile salt concentrations of 2.5%, 5% and 7.5%, and water salinity (0.5%, 3.5% and 6.5%). The autoaggregation and semi-qualitative test of hydrolase enzymes were also carried out. In total, 21 LAB isolates were derived from mackerel intestines. Isolates that have the stronger ability to inhibit the pathogen Aeromonas hydrophila ATCC 7966 are KBP 3.3, KBP 3.3.1 and KBP 6.3, while KBP 1.3, KBP 1.1.1 and KBP 3.3.1 were able to inhibit Streptococcus agalactiae ATCC 13813. The LAB isolates that survived at the tested pH were KBP 3.3.1, KBP 3.3, KBP 1.1.1 and KBP 1.3. Results also suggest that the four isolates were tolerant to bile salt concentrations and water salinity with good survival rates (>94.7%) and had proteolytic, lipolytic and cellulolytic activities. The isolates KBP 3.3, KBP 3.3.1 and KBP 1.1.1 were identified as Weissella confusa 6250, while the isolate KBP 1.3 was identified as W. confusa C5-7.@*Conclusion, significance and impact of study@#The characteristics of four selected isolates indicate their potential as a probiotic. These potential probiotic isolates can be applied directly to fish by utilizing microencapsulation technology or adding to the feed.

Crecimiento y supervivencia de Cryphiops caementarius en cocultivo con Oreochromis niloticus a diferentes densidades / Growth and survival of Cryphiops caementarius in coculture with Oreochromis niloticus at different densities

RESUMEN Objetivo. Evaluar el crecimiento y la supervivencia de Cryphiops caementarius en cocultivo con Oreochromis niloticus a diferentes densidades. Materiales y métodos. Se utilizaron camarones machos (5.86 cm y 7.65 g) y alevines revertidos de tilapia (5.65 cm y 2.61 g). Se emplearon nueve acuarios (55 L). En cada acuario se instalaron seis recipientes donde se sembró un camarón por recipiente (32 camarones/m2) y en el agua restante se sembraron tilapias a 100, 200 y 300 alevines/m3. Se empleó alimento balanceado. La ración diaria para camarones fue del 6% y para tilapia fue del 5% de la biomasa total. El experimento duró 90 días. Resultados. En el camarón, la longitud (6.46 cm), peso (9.37 g), las ganancias porcentuales en longitud (10.01% a 10.45%) peso (19.24% a 25.41%), y la supervivencia (88.89% a 94.44%) fueron similares (p<0.05) entre tratamientos. El efecto del síndrome de muerte por muda es discutido. En tilapia, la longitud (9.25 cm), peso (12.90 g), tasa de crecimiento absoluto (0.040 cm/día; 0.114 g/día), tasa de crecimiento específica (0.55% longitud/día; 1.759% peso/día) y la ganancia porcentual (64.21%; 389.48%) fueron mayores (p<0.05) a 100 y 200 alevines/m3. La supervivencia de tilapia fue similar (86.11%) entre tratamientos. Conclusiones. El crecimiento y la supervivencia del camarón fueron afectados por el síndrome de muerte por muda, más no por la presencia de tilapia en el sistema. En cambio, mayor crecimiento de tilapia se obtuvo con 100 alevines/m3 y la supervivencia fue similar entre tratamientos.

ABSTRACT Objective. To evaluate the growth and survival of Cryphiops caementarius in coculture with Oreochromis niloticus at different densities. Materials and methods. Male prawns (5.86 cm and 7.65 g) and reverse tilapia fingerlings (5.65 cm and 2.61 g) were used. Nine aquariums (55 L) were used. Six containers were installed in each aquarium, where one prawn was stocked per container (32 prawn/m2), and in the remaining water, tilapia was stocked at densities of 100, 200 and 300 fish/m3. Balanced feed was used. The daily ration for prawns was 6% and for tilapia, it was 5% of the total biomass. The experiment lasted 90 days. Results. In prawns, the length (6.46 cm), weight (9.37 g), percentage gains in length (10.01% at 10.45%) weight (19.24% a 25.41%), and survival (88.89% to 94.44%) were similar (p<0.05) between treatments. The effect of molting death syndrome is discussed. In tilapia, the length (9.25 cm), weight (12.90 g), absolute growth rate (0.040 cm/day; 0.114 g/day), specific growth rate (0.55% length/day; 1.759% weight/day) and percentage gain (64.21%; 389.48%) were greater (p<0.05) at 100 and 200 fish/m3. Tilapia survival was similar (86.11%) between treatments. Conclusions. Prawn growth and survival were affected by molt death syndrome but not by the presence of tilapia in the system. In contrast, greater growth of tilapia was obtained with 100 fish/m3, although survival was similar between treatments.

Caracterización geométrica de los eritrocitos nucleados de tilapia roja (Oreochromis spp) / Geometric characterization of nucleated erythrocytes of red tilapia (Oreochromis spp)

RESUMEN Introducción. En hematología, el estudio de las alteraciones de la morfología eritrocitaria contribuye con el diagnóstico de la normalidad o anormalidad de estas estructuras, sin embargo, el carácter cualitativo de los criterios diagnósticos dificulta su interpretación y alcance. Objetivo. Caracterizar los eritrocitos nucleados de tilapia roja (Oreochromis spp), en el contexto de la geometría fractal y euclidiana. Metodología. Se tomaron 50 eritrocitos nucleados de 20 extendidos de sangre de tilapia roja. Posteriormente todos los contornos del núcleo y el citoplasma de los eritrocitos fueron delineados, para superponer dos rejillas, una con el doble tamaño que la otra, para calcular mediante el método de Box Counting la dimensión fractal de cada eritrocito delineado. Adicionalmente fue calculada la superficie de estas dos partes del eritrocito. Resultados: Los resultados de este estudio revelaron que los valores de la dimensión fractal no permiten hacer comparaciones entre eritrocitos nucleados. Por su parte, la superposición de rejillas de 5x5 y 10x10 píxeles permitió observar que los valores de ocupación del citoplasma y el núcleo permiten hacer comparaciones entre los eritrocitos nucleados, junto con los valores de la superficie de estas dos partes del eritrocito nucleado. Conclusión: Los eritrocitos nucleados de tilapia roja pueden ser caracterizados mediante la medición de los valores espacios ocupados por su citoplasma y el núcleo, junto con los valores de la superficie de cada una de estas dos partes del eritrocito.

ABSTRACT Introduction. In hematology, the study of erythrocyte morphology alterations contributes to the diagnosis of normality or abnormality of these structures. However, the qualitative nature of the diagnostic criteria makes their interpretation and scope difficult. Objective. Characterize the nucleated erythrocytes of red tilapia (Oreochromis spp) in the context of fractal and Euclidean geometry. Methodology. Fifty nucleated erythrocytes were taken from twenty red tilapia blood smears. Subsequently, all the contours of the nucleus and the cytoplasm of the erythrocytes were delineated to superimpose two grids, one twice the size of the other, to calculate the fractal dimension of each delineated erythrocyte using the Box Counting method. Additionally, the surface of these two parts of the erythrocyte was calculated. Results: This study revealed that the fractal dimension values do not allow comparisons between nucleated erythrocytes. The superposition of 5x5 and 10x10 pixel grids allowed us to observe that the occupancy values of the cytoplasm and the nucleus allow comparisons between the nucleated erythrocytes, together with the values of the surface of these two parts of the nucleated erythrocyte. Conclusion: Red tilapia nucleated erythrocytes can be characterized by measuring the values of the spaces occupied by their cytoplasm and nucleus, together with the values of the surface of each of these two parts of the erythrocyte.

Nutritive evaluation, metabolisable energy and digestible amino acid contents of different indigenous feedstuff for Nile tilapia (Oreochromis niloticus) / Avaliação nutricional, energia metabolizável e conteúdo de aminoácidos digestíveis de diferentes alimentos indígenas para tilápia do Nilo (Oreochromis niloticus)

Abstract Three trials were executed to examine the nutritive profile, metabolisable energy and digestible amino acid (AA) contents of four indigenous feed ingredients including wheat (W), wheat middling (WM), canola meal (CM) and rapeseed meal (RSM) in Nile tilapia. Three samples of each test ingredient were collected from three different locations of Multan (MUL) and Sukkar (SKR), of Pakistan. The collected three samples were pooled thereafter to make a homogenous/ representative sample of each test ingredient from a particular study site. Nutrients composition, AA and energy digestibility of these indigenous ingredients were evaluated by using laboratory analyses and fish studies. Proximate analysis indicated variations in some of the nutrients due to location (p < 0.05). Differences were also observed in some AA including arginine, lysine, serine, cysteine, glutamic and aspartic acids, histidine, valine and glycine contents of these ingredients (p < 0.05). Digestibility of leucine, glycine and glutamic acid was higher (p < 0.05) in RSM from MUL. Among W samples from MUL, AA digestibility for lysine, threonine, and aspartic acid was higher (p < 0.05). Crude protein, arginine, alanine, serine, and aspartic acid had higher digestibility (p < 0.05), whereas digestibility was lower (p < 0.05) for threonine, valine and tyrosine in RSM from MUL. Metaboliseable energy contents did not differ among W, WM, CM and RSM regarding their origin (p > 0.05). The results indicated that nutritional profiles and their digestibility indices vary with the location for Nile tilapia.

Resumo Três experimentos foram executados para examinar o perfil nutritivo, a energia metabolizável e o conteúdo de aminoácidos digestíveis (AA) de quatro ingredientes alimentícios, incluindo trigo (W), farelo de trigo (WM), farelo de canola (CM) e farelo de colza (RSM) em tilápia do Nilo. Três amostras de cada ingrediente do teste foram coletadas de dois locais diferentes (Multan (MUL) e Sukkar (SKR), do Paquistão) e assim agrupadas. A composição nutricional, AA e digestibilidade energética desses ingredientes indígenas foram avaliadas por meio de análises laboratoriais e estudos de peixes. A análise imediata indicou variações (p <0,05) em alguns dos nutrientes devido à localização. Variações (p <0,05) também foram observadas em alguns teores de AA desses ingredientes. A digestibilidade da leucina, glicina e ácido glutâmico foi maior (p <0,05) em RSM de MUL. Entre as amostras de W da MUL, a digestibilidade de AA para Lys, Thr e Asp foi maior (p <0,05). Proteína Crud, arginina, alanina, serina e ácido aspártico apresentaram maior digestibilidade (p <0,05), e menor (p <0,05) para treonina, valina e tirosina em MRS. Nenhuma diferença (p> 0,05) relacionada a energia metabolizável foi observada entre esses ingredientes em relação à sua origem. Os resultados indicaram que os perfis nutricionais e sua digestibilidade variam com a localização.

Antimicrobial activity of characterized Leuconostoc spp. as novel probiotics isolated from Oreochromis spp. (red tilapia) against fish pathogen Edwardsiella tarda

Aims@#This study aimed to isolate and characterize putative new probiotic with antimicrobial properties against common fish pathogens from the gut of Oreochromis spp. (red tilapia). @*Methodology and results@#A total of 28 colonies were isolated from gut of Oreochromis spp. and characterized phenotypically. Eight isolates were selected for probiotic characterization. Temperature, salinity, pH and bile salt tolerance, antibiotic susceptibility and antimicrobial test against selected fish pathogens (Aeromonas hydrophila, Edwardsiella tarda and Staphylococcus aureus subsp. aureus ATCC 25923) were conducted. Characterization studies revealed isolates suited for freshwater environment and exhibited tolerance against wide range of salinity, pH and bile salt. Isolates displayed different antibiotic susceptibility profile, with six exhibited antimicrobial properties against E. tarda. Molecular identification based on 16S rRNA gene sequencing showed 99.44%, 98.59% and 91.21% sequence similarity with Leuconostoc pseudomesenteroides strain 3832T, Leuconostoc lactis strain KCC202369T and Leuconostoc mesenteroides strain 4332T, respectively as compared to known sequence in the GenBank. When identified Leuconostoc spp. were coated on feed pellets, no major decrease in viability over 21 days of storage at 4 °C were observed, with an average of 8 log CFU/mL.@*Conclusion, significance and impact of study@#The characterized species allow further application assessment of the probiotic-supplemented tilapia feed. Host-originated Leuconostoc displayed potential antimicrobial properties against fish pathogen E. tarda. The isolates Leuconostoc is expected to provide protective effect for Oreochromis spp. against edwardsiellosis and to exert beneficial effects more efficiently as compared to commercial probiotics which are not specifically target for Oreochromis spp., thereby indirectly helping fish farmers in achieving economic sustainability and increase affordability of fish.

Bioaccumulation and non-carcinogenic health risk assessment of heavy metals in selected fish species from South Bay of Laguna Lake

Objective@#The study aimed to determine the concentration of three heavy metals namely, lead (Pb), cadmium (Cd) and chromium (Cr) present in head, stomach and muscle parts of Nile Tilapia (Oreochromis niloticu) and Blackchin Tilapia (Sarotherodon melanotheron) collected from different sampling sites in South Bay area of Laguna Lake and evaluate its potential health risk for adult consumers. @*Methods@#Live samples of Nile Tilapia and Blackchin Tilapia were hand-collected from three sites of South Bay of Laguna Lake during wet season. Raw and boiled samples were prepared for analysis. Heavy metal concentrations (mg kg-1) were determined through flame atomic absorption spectrophotometry (AAS). The potential health risk posed by more than one metal was calculated using the Total Hazard Quotient (THQ). @*Results@#The heavy metal detected with the highest concentration was Pb, present in Nile Tilapia. The mean concentrations of Pb, Cd, and Cr in both fish samples were high in the head part. Boiled samples have significantly lower concentrations of heavy metal (p<0.05) compared to raw samples. THQ values for Cd in raw fish samples were higher than the recommended level of exposure (THQ < 1.0). @*Conclusion@#The study revealed that cadmium is of greater concern in terms of possible adverse health effects associated with heavy metal bioaccumulation. However, the presence of other metals may further increase the risk of non-carcinogenic adverse effects. The lower health risk is predicted for boiled fish samples.

Effects of dietary vitamin E on the wound healing induced in Oreochromis niloticus / Efeitos da suplementação com vitamina E na cicatrização de feridas induzidas em Oreochromis niloticus

In the present study, the effect of vitamin E supplementation 450 mg/kg diet was appraised in the process of induced wound healing in Nile tilapias Oreochromis niloticus. Fish were distributed into 18 tanks (10 fish each). Nine tanks were fed the non-supplemented diet and the other 9 tanks were fed 450 mg vitamin E for 60 days. Subsequently, the fish were anesthetized and the epidermis and dermis were surgically removed. The rate of cicatricial retraction and appearance of the wounds, and the histomorphometry of mucous cells, chromatophores, revascularization, inflammatory cells, presence of fibroblasts, collagen fibers, and scales were checked after 3-, 7-, 14-, 21-, and 28-days post-wounding. The retraction rate of the wound was significantly higher in the supplemented fish. The higher concentrations of inflammatory cells, mucous cells, and chromatophores, as well as the production and organization of collagen fibers, resulted in a higher retraction rate. We concluded that a dietary supplementation diet improves specific aspects of the cutaneous healing process in Nile tilapia fish.(AU)

No presente estudo, o efeito da suplementação com vitamina E de 450 mg / kg de dieta foi avaliado no processo de cicatrização induzida de feridas em tilápias do Nilo, Oreochromis niloticus. Os peixes foram distribuídos em 18 tanques (N=10), sendo 9 tanques com dieta não suplementada e os outros 9 tanques suplementados com 450 mg de vitamina E por 60 dias. Posteriormente, os peixes foram anestesiados e a epiderme e derme foram removidas cirurgicamente. Nos tempos pré-determinado de 3, 7, 14, 21 e 28 dias após a ferida foi analisado a taxa de retração cicatricial, a aparência das feridas e a histomorfometria das células mucosas, dos cromatóforos, das células inflamatórias, a revascularização, a presença de fibroblastos, de fibras de colágeno e escamas. A taxa de retração da ferida foi significativamente maior nos peixes suplementados. As maiores concentrações de células inflamatórias, mucosas e cromatóforos, bem como a produção e organização das fibras de colágeno, resultaram em uma maior taxa de retração. Concluímos que a dieta de suplementação melhora aspectos específicos do processo de cicatrização cutânea em peixes de tilápia do Nilo.(AU)

Primary culture macrophages from fish as potential experimental model in toxicity studies with multiwalled carbon nanotubes

Multiwalled carbon nanotube (MWCNT) has been broadly used in several sectors of society. This material when exposed to the environment might reach the aquatic animals and cause toxic effects. Here, it was evaluated the MWCNTs toxicity in melanomacrophages primary culture that was submitted to 1 µ gm L-1 MWCNTs for 24 hours. After exposition to MWCNT, 48 and 59% liver and spleen melanomacrophages were healthy, respectively. The control group presented 85% viability. Phagocytosis activity of melanomacrophages was observed by presence of black inclusions in cytoplasm. The findings indicate MWCNT was cytotoxic to melanomacrophages, where its release and effect into aquatic environment must be more studied. Finally, the melanomacrophages present large potential as experimental model for evaluation of carbon-based nanomaterial toxicity.

Quality enhancement of frozen Nile tilapia fillets using rosemary and thyme oil / [Melhoria da qualidade de filetes de tilápia do Nilo congelados com óleo de alecrim e tomilho]

The food industry and the frozen fish sector in particular have benefitted greatly from advancements in food processing technologies. This study investigated the effect of adding natural antioxidants such as rosemary and thyme oil to frozen fillets of Nile tilapia (Oreochromis niloticus) in order to preserve their quality for consumers. Fillets were treated with rosemary and thyme at two concentrations (1% and 1.5%) and then were stored at 4°C. Samples were analyzed over 4 days for bacteriological (aerobic plate count, psychotropic count, and coliform count), chemical (determination of pH, thiobarbituric acid reactive substances-TBARS, and total volatile base nitrogen-TVB-N), and sensory quality examination (color, texture, and odor). Significant differences (P<0.05) were observed among different groups in terms of aerobic plate count, psychotropic count, and coliform count during the storage. Moreover, pH, TVB-N, and TBARS mean values in the treated groups were lower than those in the untreated group. The best sensory quality was obtained at the highest concentrations (1.5%) of thyme and rosemary oil.(AU)

A indústria de alimentos e o setor de peixes congelados, em particular, se beneficiaram dos avanços nas tecnologias de processamento de alimentos. Este estudo investigou o efeito da adição de antioxidantes naturais, como óleo de alecrim e tomilho, a filetes congelados de tilápia do Nilo (Oreochromis niloticus), a fim de preservar sua qualidade para os consumidores. Os filés foram tratados com alecrim e tomilho em duas concentrações (1% e 1,5%) e, em seguida, foram armazenados a 4°C. As amostras foram analisadas durante 4 dias para análises bacteriológicas (contagem de placas aeróbicas, psicotrópicas e coliformes), químicas (determinação do pH, substâncias reativas ao ácido tiobarbitúrico-TBARS e nitrogênio base volátil total-TVB-N) e exame da qualidade sensorial (cor, textura e odor). Diferenças significativas (P<0,05) foram observadas entre os diferentes grupos na contagem aeróbia de placas, contagem psicotrópica e contagem de coliformes durante o armazenamento. Além disso, os valores médios de pH, TVB-N e TBARS nos grupos tratados foram inferiores aos do grupo não tratado. A melhor qualidade sensorial foi obtida nas maiores concentrações (1,5%) de tomilho e óleo de alecrim.(AU)

Isolation and characterization of aerobic actinomycetes with probiotic properties in Nile tilapia

This study scoped the isolation of aerobic actinomycetes with probiotic properties against bacterial pathogens in Niletilapia. Eleven rhizosphere soil samples were collected from the agricultural sites in three provinces (Chanthaburi,Nan, and Chachoengsao) of Thailand. A total of 157 actinomycete-like colonies were successfully isolated. Theantibacterial testing against four tested bacteria (Streptococcus agalactiae 2809, Aeromonas jandaei 1929, Aeromonasveronii 1930, and Edwardsiella ictaluri 2234) was carried out by a modified cross-streaked method. The results showedthat 108 strains possessed antibacterial activity against at least one of the bacterial pathogens. Seventeen active isolateswere identified in the actinomycetes by the analysis of the partial 16S ribosomal rRNA gene, and the phylogeneticrelationships of the isolates and their closely related strains were confirmed by the neighbor-joining method. IsolatesLNW002 and YNW004 survived in the liquid cultivation with International Streptomyces Project 2 at pH 2 and thepresence of 0.3% bile salt for 2 hours, which mimics the gastric acidity and bile salt in the gastrointestinal tract of Niletilapia in vitro. In conclusion, these strains might be further investigated for their efficacy as probiotics in Nile tilapia.This study is the first to report on anti-E. ictaluri activity in streptomycetes.

Efecto de la sustitución de grasas en salchichas de tilapia roja (Oreochromis sp.) por una mezcla de piel de cerdo y fibra de quinua / Effect of fat replacement in red tilapia sausages (Oreochromis sp.) with a mixture of pork skin and quinoa fiber

RESUMEN La disminución del consumo de grasas saturadas aportadas por los alimentos de origen animal y la búsqueda de alimentos más sanos es una tendencia, a nivel mundial. La producción de tilapia en Colombia ha venido en franco crecimiento en las últimas décadas, lo cual, plantea la necesidad de búsqueda de alternativas para su procesamiento. Se evaluó la sustitución de grasas por una mezcla de piel de cerdo, agua y fibra de quinua (PCFQ) en salchichas de filete de tilapia. Se determinó la composición química de las salchichas elaboradas, el contenido calórico, el pH, las coordenadas de color CIELab, las pérdidas de humedad por cocción, la estabilidad de la emulsión, el perfil de textura y se estableció la apreciación sensorial. La adición de PCFQ conllevó a un aumento en la humedad y en las proteínas totales, gracias a la capacidad de retención de agua de la fibra de quinua. La salchicha con mayor adición de PCFQ (20%) obtuvo un 48,2% menos de grasas, un 32,3% menos de calorías y un 31,1% de pérdida de humedad por cocción, con relación a la salchicha control. Se obtuvieron emulsiones cárnicas más estables, con buenos parámetros de perfil de textura - TPA. Con relación a los parámetros sensoriales, no se apreciaron diferencias significativas entre las salchichas elaboradas y la salchicha control. Por lo anterior, se concluye que, la mezcla, se puede usar como sustituto de grasas en la elaboración de salchichas de filete de tilapia, sin afectar sus propiedades fisicoquímicas y sensoriales.

ABSTRACT The decrease in the consumption of saturated fats provided by animal foods and the search for healthier foods is a worldwide trend. Tilapia production in Colombia has been growing rapidly in recent decades, which raises the need to find alternatives for processing. The substitution of fats for a mixture of pork skin, water and quinoa fiber (PCFQ) in tilapia fillet sausages was evaluated. The chemical composition of the processed sausages, the caloric content, the pH, the CIELab color coordinates, the cooking moisture losses, the emulsion stability, the texture profile and the sensory assessment were determined. The addition of PCFQ led to an increase in moisture and total proteins, thanks to the water retention capacity of quinoa fiber. The sausage with the highest PCFQ addition (20%), obtained 48.2% less fat, 32.3% less calories and 31.1% loss of moisture from cooking compared to the control sausage. More stable meat emulsions were obtained with good texture profile parameters -TPA. Regarding sensory parameters, no significant differences were observed between processed sausages and control sausage. Therefore, it is concluded that the mixture can be used as a fat substitute in the preparation of tilapia fillet sausages, without affecting its physicochemical and sensory properties.

Tratamento de queimaduras de segundo grau profundo em abdômen, coxas e genitália: uso da pele de tilápia como um xenoenxerto / Treatment of deep second-degree burns on the abdomen, thighs, and genitalia: use of tilapia skin as a xenograft

A pele de tilápia possui microbiota não infecciosa e estrutura morfológica semelhante à pele humana. Estudos clínicos fase II, ainda não publicados, mostraram resultados promissores na sua utilização para tratamento de queimaduras. Nos protocolos destes estudos, pacientes com lesões em áreas de dobras de pele, como genitais e região inguinal, foram excluídos, pois achava-se que o biomaterial não aderiria apropriadamente, resultando em um grau de cicatrização inferior. Relato de caso de paciente do sexo feminino, 18 anos, sem comorbidades, com queimaduras de segundo grau profundo em abdômen, região inguinal, parte da genitália e metade superior de ambas as coxas, envolvendo 13,5% da área total da superfície corporal. A pele de tilápia foi aplicada nas lesões levando a uma reepitelização completa com 16 dias de tratamento. Não foram observados efeitos colaterais. A pele de tilápia traz, portanto, a promessa de um produto inovador, de fácil aplicação e alta disponibilidade, que pode se tornar a primeira pele animal nacionalmente estudada e registrada pela Agência Nacional de Vigilância Sanitária, para uso no tratamento de queimaduras. Este relato de caso contribui para reduzir as limitações em relação às áreas anatômicas apropriadas para a aplicação da pele de tilápia, uma vez que, mesmo com a necessidade de reposição de pele, foram obtidos bons resultados com aplicação na genitália e região inguinal.

Tilapia skin has a non-infectious microbiota and a morphological structure similar to human skin. Phase II clinical studies, not yet published, have shown promising results in their use for the treatment of burns. In the protocols of these studies, patients with lesions in areas of skin folds, such as genitals and inguinal regions, were excluded, as it was thought that the biomaterial would not adhere properly, resulting in a lower degree of healing. Case report of a female patient, 18 years old, without comorbidities, with deep second-degree burns in the abdomen, inguinal region, part of the genitalia and upper half of both thighs, involving 13.5% of the total body surface area. Tilapia skin was applied to the lesions leading to a complete re-epithelialization with 16 days of treatment. No side effects were observed. Tilapia skin, therefore, brings the promise of an innovative product, easy to apply, and highly available, which can become the first animal skin nationally studied and registered by the Agência Nacional de Vigilância Sanitária, for use in the treatment of burns. This case report contributes to reduce the limitations concerning the anatomical areas appropriate for the application of tilapia skin, since, even with the need for skin replacement, good results were obtained with application to the genitalia and inguinal region.

The diversity of ursodeoxycholic acid precursors from bile waste of commercially available fishes, poultry and livestock in Indonesia

Ursodeoxycholic acid (UDCA), a secondary bile acid (BA), has been used as a drug to treat various liver diseases. UDCA is synthesised from cholic or chenodeoxycholic acid (CA/CDCA), two primary BAs frequently used as the starting materials. Nowadays, swine, cattle, and poultry bile are the main sources of those BAs. However, other commercial animals could be promising sources as well. We identified two livestock, two poultries, and eight fishes that are commercially cultivated in Indonesia. Four free BAs including CA, CDCA, deoxycholic acid (DCA), and lithocholic acid (LA) were identified for their occurrences using thin-layer chromatography and high-performance liquid chromatography. CA was detected in cow, duck, red tilapia, gourami, the common carp, and grouper, whereas CDCA was only detected in two poultries and the common carp. The occurrence of DCA was common and abundant in most tested animals. In contrast, the presence of LA was found to be very low in all samples. The biliary bile of tilapia has been found to contain a high abundance of free CA (43% of the total bile). A simple extraction was able to purify CA from biliary bile of tilapia. This is a new promising and competitive source of CA.

Uso da pele de tilápia do Nilo em medicina regenerativa: Status atual e perspectivas futuras / Nile tilapia fish skin use in regenerative medicine: Current status and future perspectives

OBJETIVO: O presente estudo tem como objetivo apresentar a trajetória do desenvolvimento da pele de tilápia como curativo biológico. MÉTODO: Descrição do processo de desenvolvimento da pele de tilápia como curativo biológico, desde a concepção da ideia para utilização no tratamento de queimaduras, até as perspectivas de sua utilização como scaffold em múltiplas especialidades médicas. RESULTADOS: Com o objetivo de oferecer uma alternativa segura, eficaz e de baixo custo para o tratamento de queimaduras, pesquisadores brasileiros desenvolveram um curativo biológico derivado da pele de tilápia. CONCLUSÃO: A pele de tilápia vem demonstrando um notável potencial como curativo biológico no tratamento de queimaduras e na medicina regenerativa.

OBJECTIVE: The present study aims to present the development trajectory of the tilapia skin as a biological dressing. METHODS: To describe the development process of tilapia skin as biological dressing, from the conception to the burn wounds treatment to the perspectives of its use as a scaffold in multiple medical specialties. RESULTS: In order to provide a safe, effective and low-cost alternative for the treatment of burn injuries, Brazilian researchers have recently developed a biological dressing derived from tilapia skin. CONCLUSION: The tilapia skin have been demonstrating the remarkable potential as a biological dressing in burn injuries treatment and in regenerative medicine.

Elaboração, desenvolvimento e instalação do primeiro banco de pele animal no Brasil para o tratamento de queimaduras e feridas / Elaboration, development, and installation of the first animal skin bank in Brazil for the treatment of burns and wounds

Introdução: A produção e a distribuição de pele humana pelos Bancos de Pele do Brasil é insuficiente para atender a demanda do país, suprindo menos de 1% da necessidade para tratar as vítimas de queimaduras. O objetivo do trabalho foi apresentar a elaboração e o desenvolvimento da implantação do primeiro Banco de Pele Animal do Brasil para tratamento da queimadura. Métodos: Trata-se de um estudo metodológico. O Banco de Pele Animal Aquático foi elaborado, desenvolvido e instalado a partir do desenvolvimento do processamento de pele da tilápia, da revisão sistemática de estudos referentes a banco de pele animal, de visitas à pscicultura em Jaguaribara-CE, da visita técnica aos bancos de pele humana no Brasil, de consulta técnica e de treinamento no Banco de Pele de Recife, da observação de todas as fases de processamento da pele de tilápia e da identificação da estrutura física da área onde ocorrem todos os processos. Resultados: Além da produção e da distribuição da pele da tilápia para os estudos em vítima de queimaduras, o banco está processando mais de 5000 peles de tilápia e encontra-se em fase de distribuição desta pele para estudos multicêntricos em outros estados e outras especialidades, que desenvolvem pesquisas nas áreas de ginecologia, ortopedia, endoscopia, estomaterapia, cirurgia vascular, odontologia e veterinária. Conclusão: este trabalho possibilitou a elaboração, o desenvolvimento e a implementação do primeiro Banco de Pele Animal do país e o primeiro Banco de Pele Aquática do mundo.

Introduction: The production and distribution of human skin by Brazilian skin banks is insufficient to meet the country's demand, supplying less than 1% of the need for treating burn victims. The objective of this work was to present the elaboration and development of the first animal skin bank of Brazil for the treatment of burns. Methods: This methodological study elaborated, developed, and installed The Aquatic Animal Skin Bank in terms of the development of tilapia skin processing after a systematic review of studies referring to animal skin banks based on visits to the pisciculture center in Jaguaribara-CE, technical visits to human skin banks in Brazil, technical consultation and training in the Recife Skin Bank, the observation of all phases of tilapia skin processing, and the identification of the physical structure of the area where the processes occur. Results: In addition to the production and distribution of tilapia skin for studies on burn victims, the bank is processing more than 5000 tilapia skin samples and is in the distribution phase of this skin for multicentric studies in other states and specialties including gynecology, orthopedics, endoscopy, stomatherapy, vascular surgery, dentistry, and veterinary medicine. Conclusion: This work enabled the elaboration, development, and implementation of Brazil's first animal skin bank and the world's first aquatic skin bank.