Abstracts
This study evaluated the acute inflammatory response induced by carrageenin in the swim bladder of Nile tilapia supplemented with the mixture of natural extracts of propolis and Aloe barbadensis (1:1) at a concentration of 0.5%, 1% and 2% in diet during 15 days. Thirty-six fish were distributed into four treatments with three replicates: fish supplemented with 0.5% of admix of extracts of propolis and Aloe (1:1) injected with 500 µg carrageenin; fish supplemented with 1% of admix of extracts of propolis and Aloe (1:1) injected with 500 µg carrageenin; fish supplemented with 2% of admix of extracts of propolis and Aloe (1:1), injected with 500 µg carrageenin and unsupplemented fish injected with 500 µg carrageenin. Six hours after injection, samples of blood and exudate from the swim bladder of fish were collected. It was observed an increase in the leukocyte count in the swim bladder exudate of fish supplemented with extracts of propolis and Aloe injected with carrageenin. The most frequent cells were macrophages followed by granular leukocytes, thrombocytes and lymphocytes. Supplementation with propolis and Aloe to 0.5% caused a significant increase in the number of cells on the inflammatory focus mainly macrophages, cells responsible for the phagocytic activity in tissues, agent of innate fish immune response.
inflammation; carrageenin; swim bladder; propolis; Aloe
Este estudo avaliou a resposta inflamatória aguda induzida por carragenina na bexiga natatóriade tilápia do Nilo suplementada com a mistura dos extratos naturais de própolis e Aloe barbadensis (1:1), nas concentrações de 0,5%, 1% e 2% na dieta durante o período de 15 dias. Trinta e seis peixes foram distribuídos em quatro tratamentos com três repetições: peixes suplementados com 0,5% da mistura dos extratos de própolis e Aloe (1:1) injetados na bexiga natatória com 500 µg de carragenina; peixes suplementados com 1% da mistura dos extratos de própolis e Aloe (1:1) injetados na bexiga natatória com 500 µg de carragenina; peixes suplementados com 2% da mistura dos extratos de própolis e Aloe (1:1) injetados na bexiga natatória com 500 µg de carragenina e peixes não suplementados injetados na bexiga natatória com 500 µg de carragenina. Seis horas após as injeções foram coletadas amostras de sangue e exsudato da bexiga natatória dos peixes. Foi observado aumento na contagem de leucócitos no exsudato da bexiga natatória de peixes suplementados com os extratos de própolis e Aloe injetados com carragenina. As células mais frequentes foram os macrófagos seguidos pelos leucócitos granulares, trombócitos e linfócitos. A suplementação com própolis e Aloe a 0,5% provocou aumento significativo no número de células no foco inflamatório, principalmente dos macrófagos, células responsáveis pela atividade fagocitária nos tecidos, agente da resposta imune inata nos peixes.
inflamação; carragenina; bexiga natatória; própolis; Aloe
1 Introduction
Inflammation is one of the major host defenses against all forms of aggression. By definition, according to Thomson (1983)THOMSON, RG., 1983. General and veterinary pathology. Philadelphia: W. B. Saunders., inflammation is the vascular response of living tissues and cells to a certain attack, that may result in passive and chemical changes in attacked cells or tissues, which initiate the inflammatory process. In fishes, the inflammatory process may be inducted by biological, chemical or physical agents (Kumar et al., 2004Kumar, V., Abbas, A. and Fausto, N., 2004. Robbins and Cotran: pathologic basis of diseases. 7th ed. Philadelphia: Saunders. 1552 p.).
The role of the inflammatory process is to minimize irritating effects on injured tissues, and the primary response to injury is an accumulation of fluids and cells in the injured area for mobilization of immune cells such as leukocytes, macrophages and lymphocytes to the inflamed site. The rapid migration of leucocytes from microcirculation to inflammatory site is the most important characteristic of this process (Garcia-Leme, 1989Garcia-Leme, J., 1989. Hormones and Inflammation. Boca Raton: CRC Press.). The set of fluids and cells is called exudate, and its purpose is to dilute, find, destroy and remove the irritant, leading to replacement of injured tissue portions.
The exudate is a mean for recognition of inflammatory process in tissues. In fish,
the assessment of inflammatory response depends on three major factors: first is the
difficulty of identification of leukocytes, that depends on the species, the second
is related to ontogenetic variation in blood and inflammatory cells (Tavares-Dias and Moraes, 2004Tavares-Dias, M. and MORAES, FR., 2004. Hematologia de peixes
teleósteos. Ribeirão Preto: Villimpress.), and third
kinetics of inflammatory process, which may be influenced by dietary supplementation
(Moraes et al., 2003MORAES, JR., FREITAS, J., BOZZO, FR., MORAES, FR. and MARTINS, ML.,
2003. A suplementação alimentar com vitamina C acelera a evolução do processo
cicatricial em (Holmberg, 1887). Piaractus mesopotamicusBoletim
do Instituto de Pesca, vol. 29, p. 57-67.; Belo et al., 2005Belo, MAA., Schalch, SHC., Moraes, FR., Soares, VE., Otoboni, AMMB.
and Moraes, JER., 2005. Effect of dietary supplementation with vitamin E and
stocking density on macrophage recruitment and giant cell formation in the
teleost fish, . Piaractus mesopotamicusJournal of Comparative
Pathology, vol. 133, no. 2-3, p. 146-154.
http://dx.doi.org/10.1016/j.jcpa.2005.04.004. PMid:16033696
http://dx.doi.org/10.1016/j.jcpa.2005.04...
, 2012BELO, MAA., MORAES, JRE., SOARES, VE., MARTINS, ML., BRUM, CD. and
MORAES, FR., 2012. Vitamin C and endogenous cortisol in foreign‑body
inflammatory response in pacus. Pesquisa Agropecuária Brasileira, vol. 47, no.
7, p. 1015-1021,.; Dotta et al.,
2011Dotta, G., MOURIÑO, JLP., JERÔNIMO, GT., JATOBÁ, AMB., MORÁN, REB.
and MARTINS, ML., 2011. Acute inflammatory response in Nile tilapia fed
probiotic . Lactobacillus plantarum in the dietActa
Scientiarum, vol. 33, no. 3, p. 239-246.), by the kind of inflammatory stimulous (Bozzo et al., 2007BOZZO, FR., MORAES, JRE., MORAES, FR., PEREIRA, GT., TAVARES-DIAS,
M. and ONAKA, EM., 2007. Kinetics of cellular component in inflammatory response
induced by different stimuli in the swim bladder of . Piaractus
mesopotamicus, Holmberg, 1887 (Characidae)Journal of the World
Aquaculture Society, vol. 38, no. 2, p. 302-308.
http://dx.doi.org/10.1111/j.1749-7345.2007.00100.x.
http://dx.doi.org/10.1111/j.1749-7345.20...
), by eicosanoids (Claudiano, 2011CLAUDIANO, GS., 2011. Eicosanoides no aumento de permeabilidade
vascular e componente celular da inflamação aguda em . Piaractus
mesopotamicusJaboticabal: Faculdade de Ciências Agrárias e
Veterinárias, Universidade Estadual de São Paulo. Doctoral thesis in Animal
Pathology.), by the ambiental temperature (Dias and Sinhorini, 1991DIAS, JLC. and SINHORINI, IL., 1991. Qualitative evaluation of the
inflammatory response modulated by temperature in tadpoles of Rana
catesbeiana.Ciencia e Cultura, vol. 43, no. 4, p.
304-306.; Jansson Junior and Waaler, 1967JANSSON JUNIOR, CW. and WAALER, E., 1967. Body temperature, antibody
formation and inflammatory response. Acta Pathologica et Microbiologica, vol.
69, no. 4, p. 566-577.) and by
circulating hormones (Martins et al., 2000MARTINS, ML., MORAES, FR., MORAES, JRE. and MALHEIROS, EB., 2000.
Falha na resposta do cortisol ao estresse por captura e por carragenina em
Holmberg, 1887 (Osteichtheyes: Characidae). Piaractus
mesopotamicusActa Scientiarum, vol. 22, no. 2, p.
545-552.;
Belo et al., 2005Belo, MAA., Schalch, SHC., Moraes, FR., Soares, VE., Otoboni, AMMB.
and Moraes, JER., 2005. Effect of dietary supplementation with vitamin E and
stocking density on macrophage recruitment and giant cell formation in the
teleost fish, . Piaractus mesopotamicusJournal of Comparative
Pathology, vol. 133, no. 2-3, p. 146-154.
http://dx.doi.org/10.1016/j.jcpa.2005.04.004. PMid:16033696
http://dx.doi.org/10.1016/j.jcpa.2005.04...
, 2012BELO, MAA., MORAES, JRE., SOARES, VE., MARTINS, ML., BRUM, CD. and
MORAES, FR., 2012. Vitamin C and endogenous cortisol in foreign‑body
inflammatory response in pacus. Pesquisa Agropecuária Brasileira, vol. 47, no.
7, p. 1015-1021,.).
Cellular response can be evaluated by inducing an inflammatory process such as
injections of carrageenin, sulphated polysaccharide, classically used in the acute
inflammation assays. In fish, carrageenin, thioglycolate and inactivated
Aeromonas hydrophila were tested in the swim bladder of
Piaractus mesopotamicus (Martins et al., 2000MARTINS, ML., MORAES, FR., MORAES, JRE. and MALHEIROS, EB., 2000.
Falha na resposta do cortisol ao estresse por captura e por carragenina em
Holmberg, 1887 (Osteichtheyes: Characidae). Piaractus
mesopotamicusActa Scientiarum, vol. 22, no. 2, p.
545-552.; Moraes et al.,
2003MORAES, JR., FREITAS, J., BOZZO, FR., MORAES, FR. and MARTINS, ML.,
2003. A suplementação alimentar com vitamina C acelera a evolução do processo
cicatricial em (Holmberg, 1887). Piaractus mesopotamicusBoletim
do Instituto de Pesca, vol. 29, p. 57-67.; Bozzo et al., 2007BOZZO, FR., MORAES, JRE., MORAES, FR., PEREIRA, GT., TAVARES-DIAS,
M. and ONAKA, EM., 2007. Kinetics of cellular component in inflammatory response
induced by different stimuli in the swim bladder of . Piaractus
mesopotamicus, Holmberg, 1887 (Characidae)Journal of the World
Aquaculture Society, vol. 38, no. 2, p. 302-308.
http://dx.doi.org/10.1111/j.1749-7345.2007.00100.x.
http://dx.doi.org/10.1111/j.1749-7345.20...
) and
Oreochromis niloticus (Martins
et al., 2004MARTINS, ML., PILARSKY, F., ONAKA, EM., NOMURA, DT.,
FENERICK-JÚNIOR, J., RIBEIRO, K, MIYAZAKI, DMY., CASTRO, MP., MALHEIROS, EB.,
2004. Haematology and acute inflammatory response of (Osteichthyes: Cichlidae)
submitted to a single and consecutive stress of capture. Oreochromis
niloticusBoletim do Instituto de Pesca, vol. 30, no. 1, p.
71-80.; Dotta et al.,
2011Dotta, G., MOURIÑO, JLP., JERÔNIMO, GT., JATOBÁ, AMB., MORÁN, REB.
and MARTINS, ML., 2011. Acute inflammatory response in Nile tilapia fed
probiotic . Lactobacillus plantarum in the dietActa
Scientiarum, vol. 33, no. 3, p. 239-246.). According to Bozzo et al.
(2007)BOZZO, FR., MORAES, JRE., MORAES, FR., PEREIRA, GT., TAVARES-DIAS,
M. and ONAKA, EM., 2007. Kinetics of cellular component in inflammatory response
induced by different stimuli in the swim bladder of . Piaractus
mesopotamicus, Holmberg, 1887 (Characidae)Journal of the World
Aquaculture Society, vol. 38, no. 2, p. 302-308.
http://dx.doi.org/10.1111/j.1749-7345.2007.00100.x.
http://dx.doi.org/10.1111/j.1749-7345.20...
, thioglycolate and inactivated Aeromonas
hydrophila induced vascular congestion, accumulation of thrombocytes
followed by macrophages, granulocytes and oedema. The choice of swim bladder as
local for assessment of inflammatory response is based on studies of Van Furth (1992)Van-Furth, R., 1992. Development and distribution of mononuclear
phagocytes in normal steady state and inflammation. In GALLIN, JI., GOLDSTEIN,
IM., SNYDERMAN, R. Inflammation: basic principles and clinical correlates. 2nd
ed. New York: Raven Press. p. 325-333., which showed that body
cavities are an experimental model to represent the mechanisms of inflammatory
response that can occur in the other animal tissues.
An efficient method to control diseases in aquaculture is strengthened fish defense
mechanism by prophylactic administration of immunestimulants (Drago et al., 2000Drago, L., Mombelli, B., Vecchi, E., Fassina, MC., Tocalli, L. and
Gismondo, MR., 2000. In vitro antimicrobial activity of propolis dry extract.
Journal of Chemotherapy, vol. 12, no. 5, p. 390-395.
http://dx.doi.org/10.1179/joc.2000.12.5.390. PMid:11128558
http://dx.doi.org/10.1179/joc.2000.12.5....
). Besides, alternatives for the use of
chemicals are needed to prevent and avoid infections (Romano and Mejía, 2003ROMANO, LA. and MEJÍA, J., 2003. Infección por Streptococcus
iniae: Una enfermedad emergente que afecta a peces de cultivo y a
humanos. Revista Aquatic, vol. 18, p. 25-32.). On this view, natural products must
be tested to minimize the environmental degradation by indiscriminate use of
chemicals.
The hypotesis of this assay was to verify whether supplemented diet with natural extracts of propolis and babosa Aloe barbadensis Miller may influence the acute inflammation and hematological response when carrageenin is injected in the swim bladder of Nile tilapia, a proved experimental model.
2 Material and Methods
2.1 Experimental conditions
Thirty-six juveniles Nile tilapia (22.3 ± 12.7 g mean weight and 12.4 ± 2.7 cm total lenght) from the same spawning were stored into polyethylene water tanks with a capacity of 100 L, provided with a biological filter, heater and constant aeration, maintained under the following conditions: temperature 24.0 ± 2.8 °C and dissolved oxygen 6.0 ± 0.0 mg/L (Hanna, HI 9146), pH 6.51 ± 0.43 (Alfakit, AT-350) and ammonia 0.09 ± 0.33 mg/L (Alfakit, colorimetric method). After seven days of acclimation, feeding with the experimental diets was started and continued for 15 days, to perform induction of inflammatory response to injection of 500 µg carrageenin (Marine Colloids) diluted in 0.5 ml saline solution. Fish were distributed in a completely randomizated factorial design divided in four treatments with three replicates: fish supplemented with 0.5% of admix of extracts of propolis and Aloe (1:1) injected with 500 µg carrageenin; fish supplemented with 1% of admix of extracts of propolis and Aloe (1:1) injected with 500 µg carrageenin; fish supplemented with 2% of admix of extracts of propolis and Aloe (1:1), injected with 500 µg carrageenin and unsupplemented fish injected with 500 µg carrageenin.
2.2 Supplemented diet preparation
After fish biomass calculation fish were fed at 3% body weight with commercial diet supplemented or not with natural extracts. Propolis and A. barbadensis were added to the ration in the amounts of 1:1 to get a final concentration of 0.5; 1 and 2% of the total quantity of offered food per day. The extracts of propolis and babosa were diluted in 50% alcohol before diet mixture.
2.3 Injection and collection of exudate cells
After feeding period of 15 days, fish were anesthetized with clove oil at a concentration of 100 mg.L–1 to be injected with carrageenin in the swim bladder, according to Dotta et al. (2011)Dotta, G., MOURIÑO, JLP., JERÔNIMO, GT., JATOBÁ, AMB., MORÁN, REB. and MARTINS, ML., 2011. Acute inflammatory response in Nile tilapia fed probiotic . Lactobacillus plantarum in the dietActa Scientiarum, vol. 33, no. 3, p. 239-246.. Six hours after injections they were euthanized with clove oil for collection of blood and swim bladder exudate cells (Ethic Committee no 23080.009240/2011-93/CEUA/UFSC). After opening the abdominal cavity, the swim bladder was ruptured and washed with 0.5 ml of phosphate buffer solution (PBS) with a drop of 0.001 ml of 5% EDTA. Briefly, with the aid of pipete the content was collected, maintained in ice for total leucocyte count. Afterthat, the exudate was centrifuged at 150 G for 10 min, the supernatant discharged to use the pellet for smears that were stained with Giemsa for 10 min according to Dotta et al. (2011)Dotta, G., MOURIÑO, JLP., JERÔNIMO, GT., JATOBÁ, AMB., MORÁN, REB. and MARTINS, ML., 2011. Acute inflammatory response in Nile tilapia fed probiotic . Lactobacillus plantarum in the dietActa Scientiarum, vol. 33, no. 3, p. 239-246. to determine differential cell count.
2.4 Hematological parameters
Blood samples were collected from the caudal vein using a syringe containing a drop of 10% EDTA solution (Ethic Committee no 23080.009240/2011-93/CEUA/UFSC) to measure hematocrit (Goldenfarb et al., 1971Goldenfarb, PB., Bowyer, FP., Hall, E. and Brosious, E., 1971. Reproducibility in the hematology laboratory: the microhematocrit determination. American Journal of Clinical Pathology, vol. 56, no. 1, p. 35-39. PMid:5556212.), red blood cells count in a Neubauer chamber, total counts of leukocytes and thrombocytes and differential count of leukocytes. Blood smears in duplicate were stained with a lood combination of Giemsa/May-Grünwald (Rosenfeld, 1947Rosenfeld, G., 1947. Corante pancrômico para hematologia e citologia clínica. Nova combinação dos componentes do May-Grunwald e do Giemsa num só corante de emprego rápido. Memorias do Instituto Butantan, vol. 20, p. 329-335.). The smears were used for differential leukocyte count and the total count of thrombocytes and leukocytes (Martins et al., 2004MARTINS, ML., PILARSKY, F., ONAKA, EM., NOMURA, DT., FENERICK-JÚNIOR, J., RIBEIRO, K, MIYAZAKI, DMY., CASTRO, MP., MALHEIROS, EB., 2004. Haematology and acute inflammatory response of (Osteichthyes: Cichlidae) submitted to a single and consecutive stress of capture. Oreochromis niloticusBoletim do Instituto de Pesca, vol. 30, no. 1, p. 71-80.).
2.5 Statistical analysis
Data was submited to factorial analysis of variance (ANOVA) using Statsoft. Bartlett test was used to verify the homoscedasticity while Tukey test was used to compare means. Data transformations were used according to pertinence.
3 Results
Hematological parameters as hematocrit and number of red blood cells did not show significant difference (P>0.05) among the treatments (Table1). In the circulating blood, there was a reduction in the number of total leukocytes in fish fed 1% and 2% of the mixtures of extracts. On the other hand, fish fed 0.5% and unsupplemented fish did not differ in the number of total leukocyes (Table 1). The differential leukocyte count in circulating blood (Figure 1) showed changes between treatments. The inflammatory response evaluated from the swim bladder exudate (Table 1) revealed a significant increase in total leukocyte count in fish supplemented with 0.5% extract mixture in the diet, compared to the other treatments, especially in relation to non-supplemented animals.
Mean values and standard deviation of hematological parameters in circulating blood and total leukocytes count in swim bladder exudate of Oreochromis niloticus fed supplemented diet with propolis and Aloe (1:1) in different concentrations.
Mean values of total leukocyte and thrombocyte counts in circulating blood (a) and inflammatory exudate (b) of Oreochromis niloricus fed supplemented diet with propolis and Aloe (1:1) in different concentrations. Lowercase letters indicate a significant difference between treatments (p<0.05). Injected with carrageenin (IC) and non-supplemented (NS).
In differential count of exudate cells, lymphocytes were the most abundant cells, followed by thrombocytes, granulocytes and macrophages (Figures 1 and 2).
Exudate cells of Oreochromis niloticus fed supplemented diet with propolis and Aloe (1:1) in different concentrations. Granulocytes (G), thrombocytes (T), lymphocytes (L) and macrophages (M).
4 Discussion
Carrageenin injection caused accumulation of inflammatory cells into the swim bladder
of Nile tilapia, as reported by Dotta et al.
(2011)Dotta, G., MOURIÑO, JLP., JERÔNIMO, GT., JATOBÁ, AMB., MORÁN, REB.
and MARTINS, ML., 2011. Acute inflammatory response in Nile tilapia fed
probiotic . Lactobacillus plantarum in the dietActa
Scientiarum, vol. 33, no. 3, p. 239-246.. The circulating blood cells that migrate into tissues to
participate in inflammatory response include thrombocytes, lymphocytes, macrophages
and granulocytes, corroborating previous studies. (Bozzo et al., 2007BOZZO, FR., MORAES, JRE., MORAES, FR., PEREIRA, GT., TAVARES-DIAS,
M. and ONAKA, EM., 2007. Kinetics of cellular component in inflammatory response
induced by different stimuli in the swim bladder of . Piaractus
mesopotamicus, Holmberg, 1887 (Characidae)Journal of the World
Aquaculture Society, vol. 38, no. 2, p. 302-308.
http://dx.doi.org/10.1111/j.1749-7345.2007.00100.x.
http://dx.doi.org/10.1111/j.1749-7345.20...
; Reque et al.,
2010REQUE, VR., MORAES, JRE., BELO, MAA. and MORAES, FR., 2010.
Inflammation induced by Aeromonas hydrophila in Nile tilapia
fed diets supplemented with Saccharomyces
cerevisiae.Aquaculture (Amsterdam, Netherlands), vol. 300, no. 1-4, p.
37-42. http://dx.doi.org/10.1016/j.aquaculture.2009.12.014.
http://dx.doi.org/10.1016/j.aquaculture....
; Salvador et al., 2012Salvador, R., Toazza, CS., Moraes, JR. and Moraes, FR., 2012.
Inflammatory responses of Nile tilapia Oreochromis niloticus to
Streptococcus agalactiae: effects of vaccination and yeast
diet supplement Diseases of Aquatic Organisms, vol. 98, no. 3, p. 235-241.
http://dx.doi.org/10.3354/dao02438. PMid:22535873
http://dx.doi.org/10.3354/dao02438...
;
Claudiano, 2011CLAUDIANO, GS., 2011. Eicosanoides no aumento de permeabilidade
vascular e componente celular da inflamação aguda em . Piaractus
mesopotamicusJaboticabal: Faculdade de Ciências Agrárias e
Veterinárias, Universidade Estadual de São Paulo. Doctoral thesis in Animal
Pathology.).
In this assay, macrophages were the most frequent cells, followed by neutrophils,
thrombocytes and lymphocytes in fish supplemented with 0.5% and 2% of extracts
mixture and injected with carrageenin. It should be noted that in studies of Bozzo et al. (2007)BOZZO, FR., MORAES, JRE., MORAES, FR., PEREIRA, GT., TAVARES-DIAS,
M. and ONAKA, EM., 2007. Kinetics of cellular component in inflammatory response
induced by different stimuli in the swim bladder of . Piaractus
mesopotamicus, Holmberg, 1887 (Characidae)Journal of the World
Aquaculture Society, vol. 38, no. 2, p. 302-308.
http://dx.doi.org/10.1111/j.1749-7345.2007.00100.x.
http://dx.doi.org/10.1111/j.1749-7345.20...
the authors utilized
inactivated Aeromonas hydrophila to induce inflammation in
Piaractus mesopotamicus, and the second most frequent cells
were lymphocytes. This may be related to the kind of irritant agent, the bacterium.
Martins et al. (2009)Martins, ML., Vieira, FN., Jerônimo, GT., Mouriño, JLP., Dotta, G.,
Speck, GM., Bezerra, AJM., Pedrotti, FS., Buglione-Neto, CC. and Pereira, G Jr.,
2009. Leukocyte response and phagocytic activity in Nile tilapia experimentally
infected with sp. EnterococcusFish Physiology and Biochemistry,
vol. 35, no. 1, p. 219-222. http://dx.doi.org/10.1007/s10695-008-9262-x.
PMid:18777139
http://dx.doi.org/10.1007/s10695-008-926...
observed that tilapia injected with 1 × 106 colony forming units
Enterococcus/ml showed higher number of thrombocytes.
Thrombocytes are important cells of hemostasis also involved in the organic defense
as reported in previous studies (Ellis et al.,
1976ELLIS, AE., MUNROE, ALS. and ROBERTS, RJ., 1976. Defence mechanisms
in fish. 1. A study of the phagocytic system and the fate of intraperitoneally
injected particulate material in the plaice ( L.). Pleuronectes
platessaJournal of Fish Biology, vol. 8, no. 1. p. 67-78.
http://dx.doi.org/10.1111/j.1095-8649.1976.tb03908.x.
http://dx.doi.org/10.1111/j.1095-8649.19...
; Penha et al., 1996PENHA, ML., DIAS, JLC. and MALUCELLI, BE., 1996. Influence of low
environmental temperature on the phagocytic activity of bullfrog ()
thrombocytes. Rana catesbeianaBrazilian Journal of Veterinary
Research and Animal Science, vol. 33, no. 1, p. 15-18.; Matushima and Mariano, 1996MATUSHIMA, ER. and MARIANO, M., 1996. Kinetics of the inflammatory
reaction induced by carrageenin in the swim bladder of (Nile tilapia).
Oreochromis niloticusBrazilian Journal of Veterinary
Research and Animal Science, vol. 33, p. 5-10.; Martins, et al., 2000MARTINS, ML., MORAES, FR., MORAES, JRE. and MALHEIROS, EB., 2000.
Falha na resposta do cortisol ao estresse por captura e por carragenina em
Holmberg, 1887 (Osteichtheyes: Characidae). Piaractus
mesopotamicusActa Scientiarum, vol. 22, no. 2, p.
545-552.). studies (Ellis et al., 1976ELLIS, AE., MUNROE, ALS. and ROBERTS, RJ., 1976. Defence mechanisms
in fish. 1. A study of the phagocytic system and the fate of intraperitoneally
injected particulate material in the plaice ( L.). Pleuronectes
platessaJournal of Fish Biology, vol. 8, no. 1. p. 67-78.
http://dx.doi.org/10.1111/j.1095-8649.1976.tb03908.x.
http://dx.doi.org/10.1111/j.1095-8649.19...
; Penha et al., 1996PENHA, ML., DIAS, JLC. and MALUCELLI, BE., 1996. Influence of low
environmental temperature on the phagocytic activity of bullfrog ()
thrombocytes. Rana catesbeianaBrazilian Journal of Veterinary
Research and Animal Science, vol. 33, no. 1, p. 15-18.; Matushima
and Mariano, 1996MATUSHIMA, ER. and MARIANO, M., 1996. Kinetics of the inflammatory
reaction induced by carrageenin in the swim bladder of (Nile tilapia).
Oreochromis niloticusBrazilian Journal of Veterinary
Research and Animal Science, vol. 33, p. 5-10.; Martins, et al.,
2000MARTINS, ML., MORAES, FR., MORAES, JRE. and MALHEIROS, EB., 2000.
Falha na resposta do cortisol ao estresse por captura e por carragenina em
Holmberg, 1887 (Osteichtheyes: Characidae). Piaractus
mesopotamicusActa Scientiarum, vol. 22, no. 2, p.
545-552.). The importance of thrombocytes as a phogocytic-cell has been
reported in birds, Gallus gallus domesticus (Grecchi et al., 1980Grecchi, R., Saliba, AM. and Mariano, M., 1980. Morphological
changes, surface receptors and phagocytic potential of fowl mono-nuclear
phagocytes and thrombocytes in vivo and in vitro. The Journal of Pathology, vol.
130, no. 1, p. 23-31. http://dx.doi.org/10.1002/path.1711300104.
PMid:7381624
http://dx.doi.org/10.1002/path.171130010...
; Kajigaya et al., 1985Kajigaya, H., Kamekura, M., Tanahara, N., Otha, A., Suzuki, H.,
Sugiyama, M. and Isoda, M., 1985. Acute and chronic inflammation induced by silk
sutures in chicken mesentery. Bull. Nippon Veterinary Zootechny College, vol.
34, p. 35-38.), bullfrog, Lithobathes
catesbeianus (Ishida et al.,
1985Ishida, Y., Sugyiama, M., Kajigaya, H., Umedea, M. and Isoda, M.,
1985. Inflammatory reactions on Japanese quails muscular tissue caused by
sutures. Bulletin Nippon Veterinary Zootechny College., vol. 31, p.
39-46.; Dias and Sinhorini, 1991DIAS, JLC. and SINHORINI, IL., 1991. Qualitative evaluation of the
inflammatory response modulated by temperature in tadpoles of Rana
catesbeiana.Ciencia e Cultura, vol. 43, no. 4, p.
304-306.;
Penha et al., 1996PENHA, ML., DIAS, JLC. and MALUCELLI, BE., 1996. Influence of low
environmental temperature on the phagocytic activity of bullfrog ()
thrombocytes. Rana catesbeianaBrazilian Journal of Veterinary
Research and Animal Science, vol. 33, no. 1, p. 15-18.) and tilapia (Suzuki, 1986Suzuki, K., 1986. Morphological and phagocytic characteristics of
peritoneal exudate cells in tilapia, ticus (Trewavas), and carp, L.
Oreochromis niloCyprinus carpioJournal of
Fish Biology, vol. 29, no. 3, p. 349-364.
http://dx.doi.org/10.1111/j.1095-8649.1986.tb04951.x.
http://dx.doi.org/10.1111/j.1095-8649.19...
; Matushima and Mariano, 1996MATUSHIMA, ER. and MARIANO, M., 1996. Kinetics of the inflammatory
reaction induced by carrageenin in the swim bladder of (Nile tilapia).
Oreochromis niloticusBrazilian Journal of Veterinary
Research and Animal Science, vol. 33, p. 5-10.). Similarly to the present results, Reque et al. (2010)REQUE, VR., MORAES, JRE., BELO, MAA. and MORAES, FR., 2010.
Inflammation induced by Aeromonas hydrophila in Nile tilapia
fed diets supplemented with Saccharomyces
cerevisiae.Aquaculture (Amsterdam, Netherlands), vol. 300, no. 1-4, p.
37-42. http://dx.doi.org/10.1016/j.aquaculture.2009.12.014.
http://dx.doi.org/10.1016/j.aquaculture....
observed increased number
of thrombocytes accompanied by lowered number of macrophages, neutrophils and
lymphocytes in the inflammatory exudates of tilapia fed Saccharomyces
cerevisae.
Zanuzzo (2010)ZANUZZO, FS., 2010. Uso da L. no manejo de matrinxã (Brycon
amazonicus). Aloe veraJaboticabal: Faculdade de Ciências
Agrárias e Veterinárias, Universidade Estadual de São Paulo. 86 p. Masters
Dissertation in Aquiculture. evaluated the respiratory
burst of macrophages in Brycon amazonicus, kept in water containing
A. vera, and observed an increase in the respiratory activity
of leucocytes . These findings indicate that A. vera can be reacted
with the neutrophil membrane receptors by stimulating the ROS (reactive oxygen
species) production. According to Jorgensen and
Robertsen (1995)Jørgensen, JB. and Robertsen, B., 1995. Yeast beta-glucan stimulates
respiratory burst activity of Atlantic salmon (. Salmo salar
L.) macrophagesDevelopmental and Comparative Immunology, vol. 19, no. 1, p.
43-57. http://dx.doi.org/10.1016/0145-305X(94)00045-H.
PMid:7615137
http://dx.doi.org/10.1016/0145-305X(94)0...
, an increase in the ROS production could be considered
as indicator of non-specific immune system activation. Zhang et al. (2009)Zhang, G., Gong, S., Yu, D. and Yuan, H., 2009. Propolis and .
Herba Epimedii extracts enhance the non-specific immune
response and disease resistance of Chinese sucker, Myxocyprinus asiaticusFish
& Shellfish Immunology, vol. 26, no. 3, p. 467-472.
http://dx.doi.org/10.1016/j.fsi.2009.01.011. PMid:19185611
http://dx.doi.org/10.1016/j.fsi.2009.01....
, observed significant increase in the
respiratory burst, phagocytosis and lyzozyme activity in O.
niloticus when fed propolis extracts The presence of a great number of
phagocytic cells found in this assay suggests that this fish supplemented with the
extracts of propolis and babosa showed increased acute inflammation . “In vitro”
tests with inflammatory cells from circulating blood are needed, such as conducted
by Zhang et al. (2009)Zhang, G., Gong, S., Yu, D. and Yuan, H., 2009. Propolis and .
Herba Epimedii extracts enhance the non-specific immune
response and disease resistance of Chinese sucker, Myxocyprinus asiaticusFish
& Shellfish Immunology, vol. 26, no. 3, p. 467-472.
http://dx.doi.org/10.1016/j.fsi.2009.01.011. PMid:19185611
http://dx.doi.org/10.1016/j.fsi.2009.01....
and Zanuzzo (2010)ZANUZZO, FS., 2010. Uso da L. no manejo de matrinxã (Brycon
amazonicus). Aloe veraJaboticabal: Faculdade de Ciências
Agrárias e Veterinárias, Universidade Estadual de São Paulo. 86 p. Masters
Dissertation in Aquiculture. to evaluate the phagocytic
activity of these cells.
In agreement with the findings of Reque et
al. (2010)REQUE, VR., MORAES, JRE., BELO, MAA. and MORAES, FR., 2010.
Inflammation induced by Aeromonas hydrophila in Nile tilapia
fed diets supplemented with Saccharomyces
cerevisiae.Aquaculture (Amsterdam, Netherlands), vol. 300, no. 1-4, p.
37-42. http://dx.doi.org/10.1016/j.aquaculture.2009.12.014.
http://dx.doi.org/10.1016/j.aquaculture....
in tilapia fed supplemented diet with
Saccaromyces cerevisiae, the acute inflammation induced by
A. hydrophila did not show alterations in
hematocrit.
Hematological parameters assessments may be important indicators of fish health
status (Martins et al., 2008Martins, ML., Mouriño, JL., Amaral, GV., Vieira, FN., Dotta, G.,
Jatobá, AM., Pedrotti, FS., Jerônimo, GT., Buglione-Neto, CC. and
Pereira-JUNIOr, G., 2008. Haematological changes in Nile tilapia experimentally
infected with sp. EnterococcusBrazilian Journal of Biology =
Revista Brasileira de Biologia, vol. 68, no. 3, p. 657-661.
http://dx.doi.org/10.1590/S1519-69842008000300025.
PMid:18833489
http://dx.doi.org/10.1590/S1519-69842008...
). Similarly,
Cuesta et al. (2005)Cuesta, A., Rodríguez, A., Esteban, MA. and Meseguer, J., 2005. In
vivo effects of propolis, a honeybee product, on gilthead seabream innate immune
responses. Fish & Shellfish Immunology, vol. 18, no. 1, p. 71-80.
http://dx.doi.org/10.1016/j.fsi.2004.06.002. PMid:15450970
http://dx.doi.org/10.1016/j.fsi.2004.06....
have related
increased number of monocyte-macrophages and granulocytes phagocytosis from the
blood of Sparus aurata fed supplemented diet with propolis. On the
other hand, a gradated increase in leucocyte accumulation on the inflammatory site
induced by Escherichia coli injection was reported for tilapia with
the highest migration activity 24 hours after injection (Matsuyama and Iida, 1999Matsuyama, T. and IIDA, T., 1999. Degranulation of eosinophilic
granular cells with possible involvement in neutrophil migration to site of
inflammation in tilapia. Developmental and Comparative Immunology, vol. 23, no.
6, p. 451-457. http://dx.doi.org/10.1016/S0145-305X(99)00027-0.
PMid:10512456
http://dx.doi.org/10.1016/S0145-305X(99)...
).
The use of vegetal extracts on fish food has been responsible for enhancement of the
immune system not only in the specific response but also in the non-specific as
shown in Catla catla fed diet supplemented with Achyranthes
aspera (Rao Y and Chakrabarti,
2005Rao Y, V. and Chakrabarti, R., 2005. Stimulation of immunity in
Indian major carp Catla catla with herbal feed ingredients. Fish & Shellfish
Immunology, vol. 18, no. 4, p. 327-334.
http://dx.doi.org/10.1016/j.fsi.2004.08.005. PMid:15561562
http://dx.doi.org/10.1016/j.fsi.2004.08....
).
To state precisely the real effectiveness of immunestimulants supplementation in
fish, careful analysis is needed (Zhang et al.,
2009Zhang, G., Gong, S., Yu, D. and Yuan, H., 2009. Propolis and .
Herba Epimedii extracts enhance the non-specific immune
response and disease resistance of Chinese sucker, Myxocyprinus asiaticusFish
& Shellfish Immunology, vol. 26, no. 3, p. 467-472.
http://dx.doi.org/10.1016/j.fsi.2009.01.011. PMid:19185611
http://dx.doi.org/10.1016/j.fsi.2009.01....
). With the evaluation of inflammatory response in the swim bladder
of tilapia, there was a significant increase in cell accumulation to the site of
injection of carrageenin, and increased number of macrophages in fish supplemented
with 0.5% of propolis and Aloe extracts mixture.
In addition, studies must be encouraged to analysis the feasibility of the use of natural extracts in fish farm to minimize the economic losses caused by infectious or parasitic diseases.
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» http://dx.doi.org/10.1016/j.fsi.2009.01.011
-
(With 2 Figures)
Publication Dates
-
Publication in this collection
May 2015
History
-
Received
07 Feb 2014 -
Accepted
17 Apr 2014