Differential susceptibility and immune response in chicken and Japanese quail towards Avibacterium paragallinarum

Figures & data

Table 1. Experimental set-up showing the number of birds in different group(s), volume of inoculum used, route of administration and sampling time point considered.

Group	No. of birds	Inoculum and volume	Distribution of inoculum (@ 1.8 × 10^8 CFU/mL/bird)	Sampling time point(s) post challenge (h/days)
Naïve Chicken	3	None		0^a
Naïve Japanese quail	3	None		0
Sham-inoculated chicken	9	Sterile PBS (1 mL/bird)		3, 5, 12
Sham-inoculated Japanese quail	9	Sterile PBS (0.5 mL/bird)
Infected Chicken	9 (19^b)	Sterile PBS-washed haemophilus broth + Av. paragallinarum (1 mL/bird)	0.5 mL right intra-sinus	3, 5, 12
			0.5 mL left intra-sinus
Infected Japanese quail	9 (19^b)	Sterile PBS-washed haemophilus broth + Av. Paragallinarum (0.5 mL/bird)	0.25 mL right intra-sinus	3, 5, 12
			0.25 mL left intra-sinus

^a Day 0 was the day before challenge.

^b Total number of infected birds kept in duplicate cages.

Table 2. Criteria set for clinical sign evaluation.

Clinical signs and its description	Intensity	Intensity score given
(A) Nasal discharge
Pressing of nostrils results in flow of scanty nasal secretions	+ (Mild)	1
Partial or intermittent flow of nasal secretions or sticking/drying of previous fresh exudates near nostrils	++ (Moderate)	2
Excessive, free flow of secretions from nasal passages without pressing of nostrils	+++ (Severe)	3
(B) Ocular discharge
Flooding of orbital area with clear ocular secretions	+ (Mild)	1
Partial flow of ocular secretions as evident by sticking/drying of immediate ocular secretions to nearby skin surface	++ (Moderate)	2
Continual or intermittent flow free flow of ocular secretions from ocular orbit	+++ (Severe)	3
(C) Swelling of infra-orbital/ supra-orbital sinus (Facial swelling)
Bilateral or unilateral swelling of triangular fossa (infra-orbital sinus)	+ (Mild)	1
Bilateral or unilateral swelling of sinus with elevation of facial/orbital surface	++ (Moderate)	2
Both side swelling with prominent evidence of raised bumps over facial surface/orbital surface	+++ (Severe)	3
(D) Conjunctival redness
Mild red discolouration of conjunctival surface	+ (Mild)	1
Intense red discolouration of conjunctival surface	++ (Moderate)	2
Prominent dark red discolouration of conjunctival surface	+++ (Severe)	3

Table 3. The clinical signs of infected chicken and Japanese quail inoculated with A. paragallinarum at 4 weeks of age.

Total numbers of birds shown clinical signs^a
Group	Nasal discharge	Ocular discharge	Swelling of infra-orbital sinus (facial swelling)	Redness of conjunctival lining	P-value
Infected Japanese quail	02	02	00	00	.0286^a
Infected chicken	49	14	09	13

^a Cumulative observation(s) made on clinically affected birds for each clinical parameter (s) over a period of 12 days. This observation includes both unilateral and bilateral symptoms, whenever noted.

Table 4. Total clinical signs score in chicken and Japanese quail at different intervals post – Av. Paragallinarum infection.

	Total clinical signs score
	Chicken				Japanese quail
Time interval (days)	Naive control	Sham inoculated	A. paragallinarum infected		Naïve control	Sham inoculated	A. paragallinarum infected
‘0’ Day	Ne	Nd	Nd		Ne	Nd	Nd
			UL	BL			UL	BL
1 DPI	–	Ne	16	8	–	Ne	Ne	Ne
2 DPI	–	Ne	12	4	–	Ne	Ne	Ne
3 DPI	–	Ne	19	2	–	Ne	Ne	Ne
4 DPI	–	Ne	14	Ne	–	Ne	Ne	Ne
5 DPI	–	Ne	13	Ne	–	Ne	Ne	Ne
6 DPI	–	Ne	11	Ne	–	Ne	Ne	Ne
7 DPI	–	Ne	10	Ne	–	Ne	2	Ne
8 DPI	–	Ne	11	Ne	–	Ne	1	Ne
9 DPI	–	Ne	7	Ne	–	Ne	Ne	Ne
10 DPI	–	Ne	7	Ne	–	Ne	Ne	Ne
11 DPI	–	Ne	5	Ne	–	Ne	1	Ne
12 DPI	–	Ne	2	Ne	–	Ne	1	Ne

Note: DPI = days post infection; Nd = not determined; Ne = not evident/no symptoms observed; UL = unilateral; BL = bilateral.

Figure 1. Development of clinical signs following experimentally induced infectious coryza (Av. paragallinarum serovar A) in birds at various days post-infection (DPI). (A) Chicken showing moderate nasal discharge (score 2) on 6 DPI (arrow); (B) Japanese quail showing mild nasal discharge on pressing (score 1) on 8 DPI (arrow); (C) Chicken showing swelling of supra-orbital sinus (star-shaped mark) along with mild nasal discharge (score 1) on 3 DPI (arrow) and (D) Japanese quail showing mild nasal discharge on pressing (score 1) on 12 DPI (arrow).

Table 5. Agglutination titre(s) in chicken and Japanese quail at 3 and 12 DPI.

Group	Inoculum (volume/dose)	Antibody response at days after intra-sinus inoculation (HI titres)^a
		0 DPI	3 DPI	12 DPI
Sham-inoculated Japanese quail	0.5 mL	<1/2	<1/2	<1/2
Sham-inoculated chicken	1.0 mL	<1/2	<1/2	<1/2
Infected Japanese quail	0.5 mL (@ 1.8 × 10^8 CFU/mL)	–	1/8–1/16 (1.94)¤	1/32–1/128 (2.63)¤
Infected chicken	1.0 mL (@ 1.8 × 10^8 CFU/mL)	–	1/8–1/16 (1.85)	1/16–1/256 (2.40)

Note: Geometric mean agglutination titres (GMTs) expressed as log₂X.

^a Number of chicken (n = 3) at each sampling time point.

Figure 2. (A) Effects of experimentally induced infectious coryza (Av. paragallinarum serovar A) on lipid peroxidation in chicken and Japanese quail at various intervals. Each data bar represents the mean ± SEM for three birds from the indicated group. Upper case letters indicate a significant difference with respect to naïve control birds of each species; lower case letters indicate a significant difference between groups as determined by Tukey’s multiple comparison tests. The overall significant difference among groups at various time points as analysed by one way ANOVA is indicated as P < .0001. (B) Effects of experimentally induced infectious coryza infection (Av. paragallinarum serovar A) on reduced glutathione levels in chicken and Japanese quail at various intervals. Each data bar represents the mean ± SEM for three birds from the indicated group. Upper case letters indicate significant difference with respect to naïve control birds of each species. Lower case letters with different case indicate a significant difference between groups as determined by Tukey’s multiple comparison tests. The overall significant difference among groups at various time points as analysed by one way ANOVA is indicated as P < .0001.