Daily Egg-Cycle in Japanese Quail: Serum Biochemistry, Bones, and Oviduct Changes

Andrade, KG; Cruz, FK; Kaneko, IN; Nascimento, MC; Iwaki, LCV; Santos, TC

doi:10.1590/1806-9061-2021-1599

KG Andrade

Department of Animal Science, State University of Maringá, Maringá, Paraná, Brazil.

http://orcid.org/0000-0001-6921-0960

FK Cruz

Department of Animal Science, State University of Maringá, Maringá, Paraná, Brazil.

http://orcid.org/0000-0002-1237-8874

IN Kaneko

Department of Animal Science, State University of Maringá, Maringá, Paraná, Brazil.

http://orcid.org/0000-0001-6755-8226

MC Nascimento

Department of Animal Science, State University of Maringá, Maringá, Paraná, Brazil.

http://orcid.org/0000-0002-7461-3577

LCV Iwaki

Department of Odontology, State University of Maringá, Maringá, Paraná, Brazil.

http://orcid.org/0000-0002-1822-3056

TC Santos

Department of Animal Science, State University of Maringá, Maringá, Paraná, Brazil.

http://orcid.org/0000-0003-1463-7785

Corresponding author e-mail address Tatiana C Santos Av. Colombo, 5.790, Maringá, PR, 87020-900, Brazil. Phone: 44 991045100 Email:tcsantos@uem.br

Figures (9)
Tables (6)

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Figure 1
Scheme of group distribution after the oviposition of an egg (e) around 16h00. “X” represents the estimated day period of post-oviposition (POV) when females were analyzed in each group.

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Figure 2
The daily egg-laying formation cycle. At the moment of oviposition (0 hr), follicular hierarchy (F1-F5), preovulatory follicle (F1), and previtellogenic follicles (pv) are visible. At 2 hrs post-oviposition, the magnum (mag) is secreting albumen (al) around the yolk. Between 4 and 20 hrs POV, the formation of membrane fibers and eggshell occurs. At 20 hrs, we can observe eggs in the final period of eggshell formation in some quails, with carbonate calcium deposition (a), pigment deposition (b), and the final egg with cuticle (c).

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Figure 3
Photomicrography of histochemical characteristics of magnum in Japanese quails stained with Alcian Blue (AB) pH 2.5. A) The image represents the 2 hrs treatment when the egg is inside the magnum in the albumen secretion phase. In this period, the cytoplasm of the cells of the duct (white arrows) and the secretory cells from epithelium (epi) are filled with strong AB positives granules (black arrows). The content of the granules is released into the lumen (lu) of the magnum, leaving a secretion of albumen AB+ near the surface of the mucosa. B-C) After 20 hrs of oviposition (group 20 hrs), the egg is inside the uterus and the cytoplasm of secretory cells of the epithelium are filled with AB positive granules again (arrows). Note that cells glands (gl), granules, and secretion are AB negative. C showsa low magnification of a fold of the magnum, evidencing the general characteristic of glands with AB negative contents. Scale bar: A-B) 30 µm, C) 100 µm. Paraffin, Alcian Blue pH 2.5.

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Figure 4
Histology and histochemical reaction to periodic acid of Schiff (PAS) of the magnum of Japanese quails along the egg-cycle. In general, the cytoplasmatic granules of secretory cells (black arrow) in the epithelium and cells of ducts (duc) are PAS+ and do not stain with HE staining. The tubular glands (gl) of submucosa and secretion inside lumen (lu) have PAS+ and eosinophilic granules (*). A-C) In treatment 0hr, female are ready to ovulate and the magnum is filled with albumen secretion inside the tubular glands of the submucosa, and the secretory cells of the epithelium. D-F) In group 2 hrs POV, the epithelium of the ducts and the lumen of the glands have intense secretory activity, and albumen secretion is observed into the lumen. Note the reduction in cell height in the submucosa glands in high magnification. G-I) In the 4 hrs group, the partial emptying of the tubular glands can be easily visualized in low magnification of the folds. In high magnification, the quantity of granules in cytoplasm continues to be reduced. J-L) For females from the 20 hrs group, the folds are filled again and submucosa glands are empty of PAS+ and eosinophilic granules. The secretory cells also show this effect. Stain: HE+PAS: A-B, G-H, J-K; PAS: D-E; HE: C, F, I, L. Scale bar: 100 µm (A, D, G, and J); 30 µm (B, C, E, F, H, I, K, and L)

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Figure 5
Uterus photomicrography in Japanese quails in HE (A-B), AB pH 2.5 stain (C) and PAS (D) stain in the 20 hrs group. The branched character of the uterine folds (fl) was evident. The epithelium of the mucosa shows secretory and ciliated cells (arrows) PAS and AB+. The submucosa glands (gl) had eosinophilic, AB-, and weak PAS+ cytoplasmatic granules. Scale bar: A) 200 µm, B-D) 30 µm.

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Figure 6
Scanning electron microscopy in eggshell at 4 hrs POV in cross-section (A) and outer (C-D) views. Notice the beginning of eggshell formation. The mammillary bodies (mb) are rounded formations arranged randomly on the outer shell membrane fibers (mf), which is still visible at this stage of shell formation. Scale bar: A) 20 µm; B) 50 µm; C) 10 µm.

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Figure 7
Scanning electron microscopy in eggshell at 8 hrs POV in cross-section (A) and outer (B-F) views. At this stage, was possible to observe several stages of eggshell formation. The mammillary bodies (mb) were well developed and fully cover the outer shell membrane (mf) from external vision. The growth (D) of columns (co) through the successive deposition (E) of layers of crystals (black arrows) is visible, and later their junction boundary (C, F) (white arrows) forming random spaces characterizing the gas pores (gp). Scale bar: A) 20 µm, B) 200 µm, C) 50 µm, D-F) 10 µm.

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Figure 8
Scanning electron microscopy in the eggshell at 14 hrs POV in cross-section view (A) and outer (B-C) views. At this stage of eggshell formation, the total junction boundary (white arrows) among columns (co) that were still growing was observed (B), forming the intermediate (A) of the palisade layer (pl) above the mammillary bodies (mb). The outer view of the shell has an irregular spongy appearance (C) and gas pores (gp). Scale bar: A) 50 µm, B) 20 µm, C) 10 µm.

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Figure 9
Scanning electron microscopy in eggshell at 20 hrs post-oviposition in cross-section view (A) and outer (B) views. In this final phase, it was possible to observe all the layers of eggshell formation (A), with distinction of the fibers membrane (mf), mammillary bodies (mb), palisade layer (pl), and the cuticle, which this final layer that gives a smooth appearance to the shell (B), where the presence of sparse gas pores (gp) could be easily distinguished. Scale bar: A) 50 µm, B) 100 µm.

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Table 1
Distribution of quails in the groups based on the period post-oviposition (POV), description of egg in the oviduct, and estimated day-time of each group (dark period 22h30 to 05h30).

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Table 2
Means of the body weight and eggs inside oviduct segments weight (g), relative weights (%) of the ovary + oviduct, magnum, isthmus, uterus liver, and follicular diameter F1 to F4 (mm) in Japanese quails during the daily egg-laying cycle.

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Table 3
Morphometric values of the magnum and uterus in Japanese quail during the daily egg-laying cycle.

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Table 4
Serum biochemistry of total protein (g/dL), albumin (g/dL), phosphorus (mg/L), alkaline phosphatase (U/L), total calcium (mg/dL) andionic calcium (mg/dL, calculated by formula) in Japanese quails during the daily egg-laying cycle.

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Table 5
Bone variables of the femur and tibiotarsus of Japanese quails during the daily egg-laying cycle.

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Table 6
Means of weight, ashes, calcium (Ca), phosphorus (P) and proportions in cortical and medullary bones in femur and tibiotarsus of Japanese quails during the daily egg-laying cycle.

Estimated day time¹	Group	N	Description
16h00	0 hr POV	10	Post-oviposition, just before the next ovulation
18h00	2 hrs POV	10	Egg in magnum and albumen secretion
20h00	4 hrs POV	10	Egg in the uterus (inactive phase)
24h00	8 hrs POV	10	Egg in the uterus (beginning of active phase)
06h00 (next day)	14 hrs POV	10	Egg in the uterus (active phase)
12h00 (next day)	20 hrs POV	10	Egg in the uterus (final egg formation)

Variables	Time post-oviposition						Means	SEM	p value
Variables	0 hr	2 hrs	4 hrs	8 hrs	14 hrs	20 hrs
BW (g)	161.40^b	168.30^ab	171.10^ab	165.40^ab	163.10^b	177.70^a	167.96	1.35	0.011
Egg in oviduct (g)	-	-	06.63^d	09.50^c	10.39^b	11.50^a	09.66	0.11	<0.001
Relative weight (%)
Ovary + Oviduct	08.16^d	09.35^c	09.81^c	11.89^b	13.54^a	14.44^a	11.25	0.11	<0.001
Magnum	02.33^a	01.76^b	01.60^b	01.61^b	1.84^b	02.21^a	01.90	0.03	<0.001
Isthmus	00.53^a	00.50^abc	00.47^abc	00.46^bc	0.43^c	00.55^a	00.49	0.01	<0.001
Uterus	01.27	01.32	01.27	01.30	1.39	01.39	01.32	0.03	0.530
Liver	03.71	03.36	03.55	03.54	3.46	03.52	03.53	0.05	0.943
Follicular Diameter (mm)
F1	18.31^a	15.99^c	16.39^bc	16.92^abc	17.67^ab	18.36^a	17.25	0.15	0.012
F2	14.50^ab	11.55^c	12.8^bc	12.39^c	14.26^ab	15.38^a	13.40	0.18	<0.001
F3	10.86^a	07.55^b	08.17^b	08.09^b	10.10^a	10.96^a	09.26	0.17	<0.001
F4	07.26^a	04.69^c	05.02^c	05.54^bc	05.89^abc	06.77^ab	06.03	0.16	<0.001

Variable	Time post-oviposition						Means	SEM	p value
Variable	0 hr	2 hrs	4 hrs	8 hrs	14 hrs	20 hrs
Magnum
Fold height (µm)	1125.07	1006.37	1178.31	1306.24	1251.76	1289.83	1192.93	41.82	0.319
Fold width (µm)	733.01	611.39	706.39	555.65	696.76	767.43	678.27	25.74	0.202
Epithelium height (µm)	12.36^ab	19.45^a	18.45^ab	11.93^ab	9.68^b	19.05^ab	15.15	00.96	0.019
Ciliated cells (%)	50.00	49.67	50.40	50.00	50.00	49.80	50.00	00.57	0.999
Secretory cells (%)	50.00	50.33	49.60	50.00	50.00	50.20	50.00	00.57	0.999
Uterus
Fold height (µm)	1075.84	1324.43	1341.40	977.20	1029.37	1031.58	1126.79	41.00	0.057
Fold width (µm)	179.60	200.51	224.51	194.69	186.88	230.08	203.60	10.15	0.631
Epithelium height (µm)	18.79	20.71	21.83	19.19	20.80	20.11	20.24	00.40	0.293

Variable	Time post-oviposition						Means	SEM	p value
Variable	0 hr	2 hrs	4 hrs	8 hrs	14 hrs	20 hrs
Protein	4.77^b	5.61^ab	5.43^ab	4.90^ab	4.65^b	4.72^b	5.01	0.078	0.002
Albumin	1.22^b	1.65^a	1.42^ab	1.33^b	1.46^ab	1.42^ab	1.42	0.027	0.001
Alkaline phosphatase	1202.89	1311.98	1488.06	1384.58	1280.45	1237.86	1316.87	55.592	0.702
Phosphorus	9.26	8.26	8.80	8.69	8.82	8.59	8.74	0.303	0.963
Total Ca	25.56^abc	27.98^a	26.73^abc	26.20^abc	25.37^b	24.96^bc	26.15	0.316	0.037
Ionic Ca	18.76	19.17	18.88	18.91	18.16	17.94	18.65	0.215	0.537

Variables	Time post-oviposition						Means	SEM	p value
Variables	0 hr	2 hrs	4 hrs	8 hrs	14 hrs	20 hrs
Femur
Hardness (kgf)	2.61	2.97	2.69	2.15	3.14	2.46	2.66	0.11	0.144
Fractureness (kg)	2.49	2.85	2.39	2.13	3.12	2.44	2.56	0.13	0.269
Time (s)	7.20	8.08	7.47	6.18	7.25	5.84	6.97	0.26	0.136
Density (mmAl)	0.99^abc	0.35^c	0.57^abc	0.71^abc	1.06^a	0.99^ab	0.79	0.07	0.012
Seedor (mg/mm)	14.43	14.56	15.23	15.16	14.29	14.82	14.77	0.28	0.863
Tibiotarsus
Hardness (kgf)	2.51	2.57	2.37	2.47	2.34	2.55	2.47	0,08	0.948
Fractureness (kg)	2.37	2.57	2.24	2.47	2.34	2.51	2.42	0.09	0.088
Time (s)	7.56	10.42	8.54	10.66	9.18	10.50	9.65	0.31	0.064
Density (mmAl)	0.92^ab	0.43^b	0.51^ab	0.61^ab	1.02^a	0.84^ab	0.73	0.06	0.039
Seedor (mg/mm)	11.99	12.12	12.74	11.52	11.73	11.34	11.90	0.20	0.390

Variables	Time post-oviposition						Means	SEM	p value
Variables	0 hr	2 hrs	4 hrs	8 hrs	14 hrs	20 hrs
Femur
Weight (g)	0.52	0.57	0.56	0.55	0.52	0.52	0.540	0.01	0.480
Cortical (%)	80.60	79.99	76.31	77.69	79.25	76.99	78.42	0.60	0.269
Medullar (%)	19.40	20.01	23.69	22.31	20.75	23.01	21.58	0.60	0.269
Ash cortical (%)	67.01	63.65	63.53	63.63	61.54	64.90	64.06	0.80	0.516
Ash medullar (%)	56.09	52.17	42.09	46.38	40.47	44.68	46.80	1.91	0.184
Ca cortical (%)	12.00	12.21	13.34	11.81	10.71	9.38	11.55	0.85	0.812
Ca medullar (%)	16.43^a	13.48^ab	11.83^ab	11.48^ab	9.42^b	13.20^ab	12.61	0.57	0.034
P cortical (%)	8.09	7.23	7.91	7.73	7.78	7.42	07.71	0.52	0.997
P medullar (%)	8.15^a	6.92^ab	6.27^ab	5.60^ab	4.35^b	7.45^a	06.44	0.29	0.010
Tibiotarsus
Weight (g)	0.55	0.56	0.59	0.53	0.55	0.52	00.55	0.01	0.359
Cortical (%)	82.81	79.49	81.23	77.82	81.28	80.59	80.54	0.50	0.117
Medullar (%)	17.19	20.51	18.77	22.18	18.72	19.41	19.46	0.50	0.117
Ash cortical (%)	72.22	70.42	69.54	73.21	69.89	70.29	70.93	0.55	0.346
Ash medular (%)	50.79	47.53	45.50	46.44	42.07	44.13	46.08	1.18	0.406
Ca cortical (%)	16.93	16.14	15.86	15.41	17.29	17.97	16.63	0.71	0.903
Ca medullar (%)	13.30^a	9.92^ab	11.83^ab	9.22^ab	6.45^b	10.41^ab	10.20	0.57	0.036
P cortical (%)	9.77^ab	8.06^b	9.78^ab	9.87^ab	11.86^a	11.48^a	10.08	0.32	0.032
P medullar (%)	7.56	6.87	5.90	7.27	5.19	6.20	06.50	0.26	0.124

[1] Corresponding author e-mail address Tatiana C Santos Av. Colombo, 5.790, Maringá, PR, 87020-900, Brazil. Phone: 44 991045100 Email:tcsantos@uem.br

Brasil

Brasil

Daily Egg-Cycle in Japanese Quail: Serum Biochemistry, Bones, and Oviduct Changes

ABSTRACT