Age-associated hemogram and ultrastructural leukocyte morphology in <i>Pygocentrus nattereri</i> (Characiformes: Serrasalmidae) from the Brazilian Pantanal

Siqueira, Mayara S.; Leão, Taynara R. F.; Rodrigues, Robson A.; Farias, Karine N. N.; Silva, André L. N.; Marcondes, Sandriely F.; Veiga, Marcelo L.; Franco-Belussi, Lilian; Fernandes, Carlos E.

doi:10.1590/1982-0224-2020-0136

Mayara S. Siqueira

Laboratório de Patologia Experimental - LAPEx, Instituto de Biociências - INBIO Universidade Federal de Mato Grosso do Sul, UFMS, Av. Costa e Silva, s/n, Universitário, 79002-970 Campo Grande, MS, Brazil. (MSS) mayara_schueroff@hotmail.com; (TRFL) trfleao@gmail.com; (KNNF) kanathiele@gmail.com; (ALNS) andrenascimento31@gmail.com; (SFM) sandy.marcondes@outlook.com; (LFB) lilian.belussi@gmail.com; (CEF) carlos.fernandes@ufms.br (corresponding author).

https://orcid.org/0000-0003-2309-4743

Taynara R. F. Leão

Laboratório de Patologia Experimental - LAPEx, Instituto de Biociências - INBIO Universidade Federal de Mato Grosso do Sul, UFMS, Av. Costa e Silva, s/n, Universitário, 79002-970 Campo Grande, MS, Brazil. (MSS) mayara_schueroff@hotmail.com; (TRFL) trfleao@gmail.com; (KNNF) kanathiele@gmail.com; (ALNS) andrenascimento31@gmail.com; (SFM) sandy.marcondes@outlook.com; (LFB) lilian.belussi@gmail.com; (CEF) carlos.fernandes@ufms.br (corresponding author).

https://orcid.org/0000-0002-7015-4908

Robson A. Rodrigues

Departamento de Aquicultura do Centro de Ciências Agrárias, Universidade Federal de Santa Catarina, Rodovia Admar Gonzaga, 1346, Itacorubi, 88034-001 Florianópolis, SC, Brazil. (RAR) robson.andrade@ufsc.br

https://orcid.org/0000-0003-4807-794X

Karine N. N. Farias

Laboratório de Patologia Experimental - LAPEx, Instituto de Biociências - INBIO Universidade Federal de Mato Grosso do Sul, UFMS, Av. Costa e Silva, s/n, Universitário, 79002-970 Campo Grande, MS, Brazil. (MSS) mayara_schueroff@hotmail.com; (TRFL) trfleao@gmail.com; (KNNF) kanathiele@gmail.com; (ALNS) andrenascimento31@gmail.com; (SFM) sandy.marcondes@outlook.com; (LFB) lilian.belussi@gmail.com; (CEF) carlos.fernandes@ufms.br (corresponding author).

https://orcid.org/0000-0003-3161-055X

André L. N. Silva

Laboratório de Patologia Experimental - LAPEx, Instituto de Biociências - INBIO Universidade Federal de Mato Grosso do Sul, UFMS, Av. Costa e Silva, s/n, Universitário, 79002-970 Campo Grande, MS, Brazil. (MSS) mayara_schueroff@hotmail.com; (TRFL) trfleao@gmail.com; (KNNF) kanathiele@gmail.com; (ALNS) andrenascimento31@gmail.com; (SFM) sandy.marcondes@outlook.com; (LFB) lilian.belussi@gmail.com; (CEF) carlos.fernandes@ufms.br (corresponding author).

https://orcid.org/0000-0002-3772-5748

Sandriely F. Marcondes

Laboratório de Patologia Experimental - LAPEx, Instituto de Biociências - INBIO Universidade Federal de Mato Grosso do Sul, UFMS, Av. Costa e Silva, s/n, Universitário, 79002-970 Campo Grande, MS, Brazil. (MSS) mayara_schueroff@hotmail.com; (TRFL) trfleao@gmail.com; (KNNF) kanathiele@gmail.com; (ALNS) andrenascimento31@gmail.com; (SFM) sandy.marcondes@outlook.com; (LFB) lilian.belussi@gmail.com; (CEF) carlos.fernandes@ufms.br (corresponding author).

https://orcid.org/0000-0002-3439-0022

Marcelo L. Veiga

Centro de Ciências da Saúde, Departamento de Morfologia, Universidade Federal de Santa Maria, Av. Roraima 1000, Cidade Universitária, Camobi, 97105-900 Santa Maria, RS, Brazil. (MLV) marcelolveiga@gmail.com.

https://orcid.org/0000-0002-9303-3324

Lilian Franco-Belussi

Laboratório de Patologia Experimental - LAPEx, Instituto de Biociências - INBIO Universidade Federal de Mato Grosso do Sul, UFMS, Av. Costa e Silva, s/n, Universitário, 79002-970 Campo Grande, MS, Brazil. (MSS) mayara_schueroff@hotmail.com; (TRFL) trfleao@gmail.com; (KNNF) kanathiele@gmail.com; (ALNS) andrenascimento31@gmail.com; (SFM) sandy.marcondes@outlook.com; (LFB) lilian.belussi@gmail.com; (CEF) carlos.fernandes@ufms.br (corresponding author).

https://orcid.org/0000-0003-1938-8423

Carlos E. Fernandes

Laboratório de Patologia Experimental - LAPEx, Instituto de Biociências - INBIO Universidade Federal de Mato Grosso do Sul, UFMS, Av. Costa e Silva, s/n, Universitário, 79002-970 Campo Grande, MS, Brazil. (MSS) mayara_schueroff@hotmail.com; (TRFL) trfleao@gmail.com; (KNNF) kanathiele@gmail.com; (ALNS) andrenascimento31@gmail.com; (SFM) sandy.marcondes@outlook.com; (LFB) lilian.belussi@gmail.com; (CEF) carlos.fernandes@ufms.br (corresponding author).

https://orcid.org/0000-0001-5425-1866

Edited by

Bernardo Baldisserotto

CORRESPONDENCE

Carlos E. Fernandes carlos.fernandes@ufms.br

AUTHOR’S CONTRIBUTION

Mayara S. Siqueira: Conceptualization, Formal analysis, Investigation, Methodology, Writing-original draft. Taynara R. F. Leão: Investigation, Methodology. Robson A. Rodrigues: Conceptualization, Investigation, Methodology, Writing-original draft. Karine N. N. Farias: Investigation, Methodology. André L. N. Silva: Investigation, Methodology. Sandriely F. Marcondes: Investigation, Methodology. Marcelo L. Veiga: Formal analysis, Writing-original draft, Writing-review and editing. Lilian Franco-Belussi: Formal analysis, Writing-original draft, Writing-review and editing. Carlos E. Fernandes: Conceptualization, Formal analysis, Investigation, Methodology, Resources, Writing-original draft, Writing-review and editing.

COMPETING INTERESTS

The authors declare no competing interests.

ETHICAL STATEMENT

The Ethics Committee on Animal Use (CEUA/UFMS, protocol 801/2016) and Biodiversity Information and Authorization System (SISBIO, project no 51270-1) approved the procedures used in this study.

Figures (4)
Tables (2)

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FIGURE 1
| Curve fitting between standard-length (cm) and weight (g) in Pygocentrus nattereri. Age cut off (16 cm) was applied according to Lowe-McConnell (1964Lowe-McConnell RH. The fishes of the Rupununi Savana distinct of British Guyana, South America. Zool J Linn Soc. 1964; 45:103-44.).

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FIGURE 2
| Erythrocytes, thrombocytes and lymphocyte in Pygocentrus nattereri from Brazilian Pantanal in light (A, B, C) and transmission electronic microscopy (D, E, F). A and D. Thrombocytes of elongated shape and scarce cytoplasm rich in canalicular system (arrow). B and E, monocytes of irregular shape, chamfered nucleus, abundant cytoplasm mitochondria (m), vacuoles (v), glycogen granules (g), Golgi apparatus (G), and free ribosomes (r) are also observed. C and F. Small lymphocyte with numerous projections, a large nucleus, and scarce cytoplasm with few mitochondria and organelles. ABC. May Grunwald-Giemsa-Wright stain. Scale bars = A-C = 10 µm; D-F = 2 µm.

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FIGURE 3
| Neutrophil and PAS-positive granular leukocytes in Pygocentrus nattereri from Brazilian Pantanal in light (A and D) and transmission electronic microscopy (B, C, E, F). A-C. Neutrophil with a lumpy nucleus and cytoplasmic vacuoles showing fine azurophilic and neutrophilic granulations. B. Mitochondria (m) and an extensive network of cell synthesis organelles are also observed; nucleus (n); C, vacuoles (v) and granules with varying size and electron density are observed. D-F. PAS-positive granular leukocyte with a displaced euchromatic nucleus; cytoplasm rich in small vacuoles and azurophilic granulation. In E and F, the granules content electrolytic material is observed (g), possibly with internal electrodense areas in an elongated half-moon shape (arrows). Scale bars = A and D = 10 µm (May Grunwald-Giemsa-Wright staining), B and E = 2 µm; C and F = 500 nm.

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FIGURE 4
| Eosinophil and basophil in Pygocentrus nattereri from Brazilian Pantanal in May Grunwald-Giemsa-Wright staining. A. eosinophil showing irregular nucleus, loose chromatin and specific cytoplasmic granules. B. basophil surrounded by erythrocytes presenting cytoplasm filled with basophilic granulations. Scale bars = 10 µm.

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TABLE 1
| Biometrical variables according to age in Pygocentrus nattereri and linear correlation (Pearson’s model, r) with standard-length. Distinct letters represent a significant difference (P < 0.05); SEM = standard error of mean; CI = confidence interval; r = Pearson´s correlation coefficient; K = Fulton’s body condition factor; SI = splenosomatic index; **P < 0.01; *P < 0.05.

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TABLE 2
| Hematological variables according to age in Pygocentrus nattereri and linear correlation (Pearson’s model, r) with standard-length. Distinct letters represent a significant difference (P < 0.05); SEM = standard error of mean; CI = confidence interval; r = Pearson´s correlation coefficient; RBC = red blood cell; Ht = hematocrit; Hb = hemoglobin; MCV = mean Corpuscular Volume; MCH = mean corpuscular hemoglobin; MCHC = mean corpuscular hemoglobin concentration; ns = not significant; * P < 0.05, ** P < 0.01.

	Juvenile (n = 60)		Adult (n = 75)
Variables	Mean±SEM	CI (95%)	Mean±SEM	CI (95%)	r
Standard length (cm)	14.9±0.30 a	14.40-15.57	19.6±0.25 b	19.08-20.10	-
Weight (g)	160.9±18.03 a	125.11-196.76	363.6±15.7 b	332.47-394.71	0.67**
SI (%)	0.12±0.01 a	0.12-0.15	0.10±0.01 b	0.09-0.11	-0.39**
K-factor	2.7±0.67 a	2.59-2.86	3.0±0.67 b	2.88-3.11	-0.21*

Variables	Juvenile (n = 60)		Adult (n = 75)		r
Variables	Mean ±SEM	(CI (95%))	Mean ±SEM	(CI (95%))	r
Erythrogram
RBC (x10⁶/µl)	1.30 ±0.04 ^a	(1.21 - 1.39)	1.38 ±0.04 ^a	(1.30 - 1.47)	ns
Erythroblasts count (x10³/µl)	48.50 ±3.87 ^a	(40.83 - 56.16)	31.52 ±3.46 ^b	(24.66 - 38.37)	ns
Ht (%)	34.86 ±0.75 ^a	(33.37 - 36.26)	36.61 ±0.67 ^a	(35.27- 37.95)	ns
Hb (g/dL¹)	9.02 ±0.23 ^a	(8.56 - 9.48)	7.71 ±0.21 ^b	(7.30 - 8.12)	-0.29^**
MCV (fL)	282.85 ±7.92 ^a	(267.17-298.53)	273.54 ±7.09 ^a	(259.52-287.57)	ns
MCH (pg)	75.70 ±3.03 ^a	(69.71-81.70)	58.01 ±2.71 ^b	(52.65-63.46)	-0.25^**
MCHC (%)	26.42 ±0.90 ^a	(24.64 - 28.20)	21.90 ±0.80 ^b	(20.31 - 23.50)	-0.22^*
Leukogram
Thrombocytes count (x10³/µl)	47.46 ±4.59 ^a	38.37 - 59.55	60.61 ±4.11 ^b	52.48 - 68.74	0.21^*
Leukocyte count (x10³/µl)	48.66 ±3.54 ^a	41.65 - 55.68	33.65 ±3.17 ^b	27.38 - 39.92	-0.24^**
Lymphocytes (%)	78.13 ±2.07 ^a	74.03 - 82.23	68.93 ±1.85 ^b	65.27 - 72.59	-0.36^**
(x10³/µl)	38.01 ±10.7 ^a	37.48 - 40.01	23.19 ±12.2 ^b	21.96 - 24.42
Neutrophils (%)	9.74 ±1.56 ^a	6.65 - 12.83	18.61 ±1.39 ^b	15.84 - 21.37	0.44^**
(x10³/µl)	4.74 ±0.75 ^a	3.23 - 6.24	6.26 ±0.46 ^a	5.33 - 7.19
Monocytes (%)	8.26 ±0.59 ^a	7.11 - 9.41	6.66 ±0.52 ^b	5.63 - 7.69	ns
(x10³/µl)	2.1 ±.017 ^a	1.71 - 2.50	1.6 ±0.12 ^a	1.21 - 1.93
Leukocyte PAS ⁺ (%)	0.57 ±0.15 ^a	0.26 - 0.88	0.88 ±0.14 ^a	0.60 - 1.16	ns
(x10³/µl)	0.27 ±0.01 ^a	0.12 - 0.42	0.29 ±0.04 ^a	0.20 - 0.39
Eosinophils (%)	0.02 ±0.37 ^a	0.04 - 0.09	0.06 ±0.03 ^a	0.01 - 0.12	0.19^*
(x10³/µl)	0.01 ±0.18 ^a	0.01 - 0.04	2.02 ± 1.00 ^a	0.00 - 4.04
Basophils (%)	0.15 ±0.09 ^a	0.01 - 0.34	0.22 ±0.85 ^a	0.05 - 0.38	ns
(x10³/µl)	0.07 ±0.04 ^a	0.00 - 0.16	0.07 ±0.28 ^a	0.01 - 0.12
Immature leukocyte (%)	3.45 ±0.98 ^a	1.50 - 5.39	4.78 ±0.88 ^a	3.03 - 6.35	ns
(x10³/µl)	1.67 ±0.47 ^a	0.72 - 2.62	1.60 ±0.29 ^a	1.01 - 2.13

[1] Edited by

Bernardo Baldisserotto

Brasil

Brasil

Age-associated hemogram and ultrastructural leukocyte morphology in Pygocentrus nattereri (Characiformes: Serrasalmidae) from the Brazilian Pantanal

ABSTRACT