Research Article

Effects of Oxytetracycline and Flunixin Meglumine Treatments on the Haematological and Serum Biochemistry Profiles of Goats Experimentally Induced with Mannheimia haemolytica Pneumonia

Fathin Faahimaah Abdul Hamid1, Mohd Farhan Hanif Reduan1*, Jasni Sabri1 , Faez Firdaus Jesse Abdullah2,Mohammed Naji Odhah1, Nur Athirah Binti Abdul Manaf1, Mohd Jefri Norsidin2 , Siti Nor Che Yahya1, Intan Noor Aina Kamaruzaman1, Nur Zul Izzati Mohd Rajdi1

1Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, 16100, Pengkalan Chepa, Kota Bharu, Kelantan, Malaysia; 2Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia.

Received | August 19 2022; Accepted | September 15, 2022; Published | February 15, 2023

*Correspondence | Mohd Farhan Hanif Reduan, Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, 16100, Pengkalan Chepa, Kota Bharu, Kelantan, Malaysia; Email: farhan.h@umk.edu.my

Citation | Hamid FFA, Reduan MFH, Sabri J, Abdullah FFJ, Odhah MN, Manaf NABA, Norsidin MJ, Yahya SNC, Kamaruzaman INA, Rajdi IM (2023). Effects of oxytetracycline and flunixin meglumine treatments on the haematological and serum biochemistry profiles of goats experimentally induced with mannheimia haemolytica pneumonia. J. Anim. Health Prod. 11(1): 73-81.

DOI | http://dx.doi.org/10.17582/journal.jahp/2023/11.1.73.81

ISSN | 2308-2801

 

 

Copyright: 2023 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Pneumonia is a common respiratory condition in the tropical region and has been the major contributor to economic losses after diarrhea in small ruminant industries (Dohare et al., 2013). Mannheimia sp. is among the common causative agents contributing to the high prevalence of Mannheimiosis in small ruminants (Chung et al., 2015). These opportunistic pathogens produce diseases when the animal is immunocompromised (Rice et al., 2007). They often disrupt the innate mucosal defense mechanism and lead to acute pulmonary infections characterized by a necrotizing inflammatory response (Sabri et al., 2013).

Haematological and serum biochemistry profiles are valuable diagnostic tools that are helpful in the diagnosis of many organs and systemic diseases supported by history and clinical examination findings. The information assists in diagnosis, surveillance and predicting the prognosis of the animal, planning of the therapeutic options and monitoring response to treatments (Russell and Roussel, 2007; Roland et al., 2014). In this study, oxytetracycline and flunixin meglumine drugs were used to evaluate the effectiveness of the treatment through haematology and serum biochemical changes in goats inoculated with mannheimiosis. Oxytetracycline is a broad spectrum antimicrobial drug commonly applied for gastrointestinal and respiratory tract diseases (Aktas & Yarsan, 2017). Meanwhile, flunixin meglumine is used to alleviate the inflammation response towards respiratory disease or endotoxemia (Weil & Baird, 2021). Until today, the hematological and serum biochemical findings in the post-treatment of mannhemiosis in goats are still lacking. This study aims to evaluate the hematological and serum biochemical changes upon inoculation and post-treatments of mannheimiosis.

Material and Methods

Animals and management

Twenty healthy goats were used in this study and the goats were kept in separate pens and fed with commercial goat pellets. They were provided with ad libitum of Napier grass and drinking water. The experimental procedure was conducted under the approval of the Animal Care and Use Ethics Committee (ACUC), Faculty of Veterinary Medicine, Universiti Malaysia Kelantan. The ethical approval number was (UMK/FPV/ACUC/PG/3/2021).

Bacteria preparation and experimental design

Mannheimia haemolytica growth on the blood agar plate was used as inoculum. The infection dose of the bacteria inoculum was standardized at 107 CFU/ml concentration using 107 McFarland’s standard. Twenty goats were equally divided into five groups (Groups 1, 2, 3, 4 and 5). Group 1 was kept as a negative control and given 1 mL of sterile phosphate-buffered saline (PBS) intranasally and all other groups were inoculated with 107 CFU/1mL of Mannheimia haemolytica intranasally. Goats in the Group 2 were kept as the positive control and no treatment was given in the study period while goats in Group 3 were treated with antimicrobial (Oxytetracycline, 20 mg/kg, SID) on days 6 and 9 post-infection. Group 4 was treated only with an anti-inflammatory drug (Flunixin meglumine, 1.1mg/kg, BID) for 5 days of post-infection and Group 5 received both antimicrobial (Oxytetracycline, 20 mg/kg, SID) on days 6 and 9 and anti-inflammatory drug (Flunixin meglumine, 1.1 mg/kg, BID) treatment for five days on post-infection. Blood samples were collected via jugular vein at 24 hours, days 5, 7, 9, 11, 14, 21, and 28 days post-infection for the analysis.

Laboratory analyses

Blood samples were analysed using an automatic haematology analyser (Scil Vet abc™, Horiba Medical, Montpellier, France) for the red blood cell (RBC) count, haemoglobin concentration (Hb), hematocrit (HCT), mean corpuscular volume (MCV) and mean corpuscular haemoglobin concentration (MCHC), platelets (PLT), total number of white blood cells (WBC).

An automatic biochemistry analyser (Randox, RX daytona, United Kingdom) was used to determine the concentrations of total protein (TP), albumin (ALB) and globulin (GLOB), blood urea nitrogen (BUN), creatine kinase (CK), gamma-glutamyl transferase (GGT) and lactate dehydrogenase (LDH).

Statistical analysis

The data was analysed using Statistical Package for Social Science (SPSS) software version 26. The values were expressed as mean ± standard deviation (SD) for different parameters. Analysis of variance (ANOVA) tests were done to compare the differences of data between and within groups. Post hoc analysis using Duncan test were used to determine the level of statistical significance which was set at p<0.05.

Results

For the haematological evaluation, within 24 hours of post- inoculation, RBC counts show there was significant (p<0.05) reduction of the RBC levels in all groups compared to the negative control. Levels of the Hb demonstrated no significant difference observed in all groups. The HCT levels showed a significant (p<0.05) decrease in all inoculated groups compared to negative control groups at 24 hours to day 9 of post-inoculation (Table 1). The MCV shows a significant (p<0.05) increase in the levels in all group compared to negative control. Meanwhile, MCHC shows a significant increase (p<0.05) in both groups 2 and 5 compared to negative control (Table 2).

On day 5 of post- inoculation which is the peak of infection, RBC evaluation revealed there was a significant (p<0.05) reduction in RBC level in all groups compared to the negative control. The Hb concentration in group 3 was significantly (p<0.05) decreased compared to the negative control. HCT levels show a significant decrease (p<0.05) in all groups compared to group 1 (negative control)

 

Table 1: Red Blood cells parameters (mean ± SD) of goats experimentally infected with Mannheimiosis

Groups	24 hr	Day 5	Day 7	Day 9		Day 11	Day 14	Day 21	Day 28
Red Blood Cells (RBC) (x106 µl)
Group 1	20.97±2.08c	20.97±2.08d	21.04±2.15b	21.05±2.15c		21.05±2.15c	20.97±2.08c	20.97±2.08b	20.97±2.08b
Group 2	11.59±1.27a	12.17±1.70a	13.15±3.06a	11.38±1.08a		12.18±2.69a	13.29±2.07a	14.15±2.26a	14.06±2.69a
Group 3	12.95±1.29a	13.87±1.75ab	15.50±3.59a	16.16±3.37b		15.27±2.40ab	16.67±2.62ab	16.24±1.89a	17.30±2.39ab
Group 4	14.75±0.95b	18.25±1.06c	19.16±1.03b	18.80±1.41bc		17.23±0.84b	18.66±0.08bc	19.43±1.31b	19.72±3.64b
Group 5	13.31±1.03ab	15.40±1.08b	15.10±1.13a	15.68±1.15b		14.46±1.43ab	15.32±2.75ab	15.95±1.56a	16.97±0.46ab
	Hemoglobin (HGB) (g/dl)
Group 1	9.30±0.87a	9.30±0.87b	9.35±0.88b		9.35±0.88b	9.35±0.88b	9.30±0.87a	9.30±0.87b	9.30±0.87b
Group 2	7.47±0.97a	5.75±1.93ab	7.05±0.70ab		6.30±0.71ab	6.40±0.67a	7.12±0.25a	7.02±0.40ab	7.42± 0.17a
Group 3	5.37±1.33a	4.78±3.26a	7.40±1.22ab		7.67±1.20ab	7.07±0.94ab	8.02±0.99a	7.20±0.48ab	7.87± 0.21a
Group 4	9.16±0.90a	7.83±0.34ab	7.86±0.47a		7.90±0.71a	7.03±0.38a	7.17±0.75a	7.70± 0.50a	7.83 ±1.66a
Group 5	7.02±2.79a	7.65±0.67ab	7.50±0.77ab		7.67±0.68ab	7.05±0.39ab	7.65±1.68a	7.32±0.85ab	8.90± 1.67a
	Haematocrit (HCT) (%)
Group 1	35.55± 3.87b	35.55± 3.87b	35.55±3.87b		35.55±3.88b	35.55±3.88d	35.55±3.87a	35.55 ±3.87a	35.55±3.87ab
Group 2	23.26±0.40a	23.48±0.38a	21.33±2.21a		20.88±1.67a	19.22±1.41b	22.97±3.74a	24.75±2.30a	26.97± 1.06ab
Group 3	22.46±1.03a	22.46±1.03a	21.07±1.10a		20.67±1.79a	25.57±4.16c	28.86±2.34a	25.95±2.21a	28.02±1.72ab
Group 4	21.97±1.12a	21.97±1.12ab	27.68±1.43a		27.68±1.43a	27.68±1.43a	32.10±2.62a	32.70±2.29a	33.55±2.72ab
Group 5	23.05±0.54a	23.05±0.54a	27.42±1.18a		27.42±1.18a	26.45±1.17c	29.75±0.52a	30.52±1.20a	31.10± 1.68a

Note: Different superscript letters on the group means in each column indicate significant difference (p< 0.05).

 

Table 2: Mean hemoglobin concentration and mean corpuscular volume (mean± SD) in goats experimentally infected with Mannheimiosis

Groups

24 Hr

Day 5

Day 7

Day 9

Day 11

Day 14

Day 21

Day 28

Mean hemoglobin concentration (MCHC) (g/dl)

Group 1

40.10± 26.54a

40.10±26.54a

26.35±1.08a

26.35±1.08a

26.35±1.08a

40.10±26.54ab

40.10±26.54ab

40.10± 26.54ab

Group 2

97.05± 8.85bc

65.27±36.98a

58.07±36.82a

58.60±24.21ab

47.17±16.09a

36.05±8.14ab

39.67±23.58ab

46.67± 38.89ab

Group 3

99.05± 25.08bc

56.32±48.27a

73.02±25.66a

45.12±33.70a

27.95±3.94a

52.30±30.25b

42.42±29.34b

31.25± 21.19b

Group 4

60.77± 40.62ab

42.52±52.74a

45.10±41.63a

88.40±58.60ab

44.02±30.86a

15.50±10.70a

17.72± 11.81a

16.85± 12.48a

Group 5

110.25± 14.44c

91.90±43.40a

66.27±43.98a

123.70±64.54b

29.65±1.68a

16.87±19.93a

45.62±15.11b

49.72± 8.02b

Mean corpuscular volume (MCV) (fl)

Group 1

17.91± 1.96a

17.88±1.98a

17.91±1.96a

17.88±1.98a

17.91±1.96a

17.88±1.98a

17.88±1.98a

17.91± 1.96a

Group 2

34.25± 0.96b

34.48± 2.68c

30.37±5.40b

33.75±0.50b

30.45±4.37b

26.51±6.15ab

17.66± 2.45a

33.84± 2.31c

Group 3

33.50± 1.29b

33.25±0.95bc

32.91±2.31b

32.91±2.32b

16.70±0.34a

32.29±2.13b

17.56±1.92a

25.30± 10.07b

Group 4

33.33± 0.47b

29.35± 5.31b

27.53±7.73b

33.33±0.47b

32.33±0.94b

23.25±8.56ab

16.66± 0.09a

31.55± 2.63bc

Group 5

33.25± 0.50b

32.28±2.29bc

29.80±4.37b

29.70±8.60b

16.47±0.15a

25.83±6.80ab

32.60± 2.33b

34.25± 0.50c

Note: Different superscript letters on the group means in each column indicate significant difference (p< 0.05).

 

Table 3: Total White blood cell count and platelet evaluation (mean± SD) in goats experimentally infected with Mannheimiosis

Groups	24Hr	Day 5	Day 7	Day 9	Day 11	Day 14	Day 21	Day 28
		Total White Blood cell count (cells/µl)
Group 1	31.65± 2.61b	31.65± 2.61a	26.60± 10.46a	26.60± 10.46a	26.60± 10.46a	26.77± 10.13a	26.77± 10.13a	52.47± 41.70b
Group 2	13.30± 4.90a	15.97± 7.47a	19.52± 9.44a	13.30± 2.45a	15.40± 3.82a	19.05± 2.20a	12.27± 8.37a	17.20± 1.26a
Group 3	9.17± 3.63a	13.07± 17.82a	19.25± 10.94a	18.65± 8.05a	17.60± 8.05a	16.85± 6.62a	13.60± 10.91a	29.30± 27.68a
Group 4	10.1± 7.82a	9.025± 26.19a	10.52± 7.29a	8.45± 6.0.5a	9.00± 6.29a	9.67± 7.19a	11.00± 8.17a	9.50± 6.59a
Group 5	8.85± 4.55a	11.97± 2.27a	13.75± 3.78a	13.90± 2.445a	14.52± 4.77a	14.65± 4.01a	15.55± 4.39a	24.72± 18.90a
	Platelet (x103 µl)
Group 1	259.25± 62.55a	259.25± 62.55a	267.00± 54.50a	267.00± 54.49a	267.00± 54.49a	259.25± 62.55a	259.25± 62.55a	259.25± 62.55a
Group 2	267.00± 54.50a	320.75± 261.67a	397.25± 265.63a	406.50± 270.91a	409.25± 270.91a	433.25± 282.52a	435.00± 239.01a	344.25± 215.00a
Group 3	259.25± 62.55a	137.25± 111.19a	414.25± 205.31a	397.50± 203.68a	399.25± 226.44a	459.25± 265.21a	405.00± 279.00a	340± 201.51a
Group 4	267.00± 54.50a	331.00± 290.11a	278.00± 249.44a	328.75± 266.98a	260.00± 218.29a	327.25± 220.83a	257.25± 227.93a	381.25± 402.22a
Group 5	267.00± 54.550a	307.00± 159.95a	282.00± 116.73a	287.00± 140.80a	238.75± 23.71a	538.50± 387.05a	259.75± 91.16a	303.75± 273.15a

Note: Different superscript letters on the group means in each column indicate significant difference (p< 0.05).

 

Table 4: Total proteins, albumin and globulin (mean± SD) level in blood of goats experimentally infected with Mannheimiosis

Groups

24Hr

Day 5

Day 7

Day 9

Day 11

Day 14

Day 21

Day 28

Total protein (TP) (g/dl)

Group 1

6.67 ± 0.68 b

6.70± 0.53a

6.67± 0.53a

6.70± 0.53ab

6.67±0.68a

6.70± 0.53a

6.67±0.68a

6.70±0.53a

Group 2

6.38 ± 0.52 a

6.57 ±0.50a

6.23±0.14 a

6.32±0.37 a

7.37 ± 0.60 ab

7.42± 0.50 ab

7.25 ±0.27b

7.18 ± 0.11b

Group 3

5.90±0.54ab

7.37±0.64a

6.40±0.48a

7.25 ± 0.54b

7.67±0.43b

7.72± 0.54b

7.82 ± 0.51b

7.93±0.50b

Group 4

4.32 ±2.90a

4.92 ±3.40a

6.27±0.05a

6.65±0.63ab

7.62±0.42b

7.57± 0.57ab

7.83±0.41b

7.92±0.40b

Group 5

6.61±0.31b

6.80 ±0.14a

6.70 ±0.08a

6.87±0.40ab

6.87±0.55ab

6.90± 0.57ab

7.55±0.24b

7.65 ±0.27b

Albumin (ALB) (g/dl)

Group 1

3.27 ± 0.30b

3.27 ± 0.30a

3.12 ±0.31ab

3.27 ± 0.31a

3.12 ± 0.31a

3.27 ± 0.30 b

3.27 ± 0.30 a

3.12 ± 0.31b

Group 2

3.17 ± 0.21b

3.02 ±0.27a

3.30 ±0.14b

3.33 ± 0.21a

3.62 ± 0.12b

3.58 ± 0.10 a

3.62 ± 0.08a

3.57 ± 0.05a

Group 3

3.12 ±0.10b

3.37 ± 0.25a

3.17 ± 0.12ab

3.57 ± 0.22a

3.62 ± 0.41b

3.65 ± 0.24 a

3.67 ± 0.19 a

3.64 ±0.17 b

Group 4

2.08 ±1.39a

2.42± 1.63a

2.95 ± 0.21a

3.30 ±0.14a

3.52 ± 0.12b

3.55 ± 0.24 a

3.68 ± 0.20a

3.58 ± 0.09 a

Group 5

3.09 ±0.17b

3.50 ± 0.33 a

3.17 ±0.19ab

3.65 ±0.19a

3.60 ±0.21b

3.70 ± 0.20 a

3.71 ±0.14 a

3.71 ±0.06 a

Globulin (g/dl)

Group 1

3.40± 0.41a

3.42 ± 0.62a

3.55 ± 0.96a

3.42 ± 0.62a

3.55 ± 0.96a

3.42 ± 0.62a

3.40 ±0.82a

3.57 ± 0.64a

Group 2

3.20 ±0.62a

3.55 ±0.26a

2.93±0.25a

3.80 ± 1.44a

3.75 ± 0.62a

3.84 ± 0.55a

3.63 ± 0.30a

3.60 ± 0.12a

Group 3

2.77 ± 0.56a

4.00 ± 0.50a

3.22 ± 0.50a

3.67 ± 0.35a

4.05 ± 0.42a

4.06 ± 0.40a

4.14 ± 0.42a

4.29 ±0.55ab

Group 4

2.24 ± 1.53a

2.50 ± 1.95a

3.32 ± 0.25 a

3.35 ± 0.75a

4.10 ± 0.32a

4.02 ± 0.66a

4.15 ± 0.53a

4.34 ± 0.47b

Group 5

3.52 ± 0.44a

3.30 ± 0.43a

3.52 ±0.25 a

3.22 ± 0.46a

3.27 ± 0.55a

3.20 ± 0.39a

3.84 ± 0.13a

3.94 ±0.21ab

Note: Different superscript letters on the group means in each column indicate significant difference (p< 0.05).

 

Table 5: Blood urea nitrogen and Creatine Kinase (mean± SD) level in goats experimentally infected with Mannheimiosis

Groups	24Hr	Day 5	Day 7	Day 9	Day 11	Day 14	Day 21	Day 28
	Blood Urea Nitrogen (BUN) (mg/dl)
Group 1	12.50 ± 0.82a	12.50 ± 0.82a	12.32 ± 0.87a	12.32 ± 0.87a	12.32 ± 0.87a	12.50 ±0.81a	12.50 ± 0.81a	12.32 ± 0.87a
Group 2	15.82 ± 0.36b	18.95 ± 0.99b	21.25 ± 0.51bc	19.50 ± 1.33bc	19.32 ± 1.67b	19.40 ± 0.77 b	18.77 ±0.55b	18. 40± 1.29b
Group 3	15. 72± 0.50b	19.29± 0.96b	20.35 ± 1.52b	17.55 ± 1.82b	25.37 ± 2.30c	26.55 ± 1.12c	25.47 ± 1.05d	26.55± 1.12c
Group 4	16.00 ± 0.38b	19.15 ±1.32b	22.27 ± 0.29c	20.50 ± 1.58c	30.52 ± 1.90d	26.60 ± 2.30c	26.70 ± 1.60d	26.57± 1.58c
Group 5	16.30 ±0.60b	19.92 ± 1.65b	21.87 ± 0.88c	19.07 ± 0.86bc	20.35 ±1.45b	20.50 ± 0.67 b	21.62± 0.75c	24.37±2.50c
	Creatine Kinase (CK) (U/L)
Group 1	55.75 ± 11.89a	59.00 ± 10.86a	55.75± 11.89 a	55.75 ± 11.89a	59.00 ± 10.86a	55.75 ± 11.89a	59.00 ± 10.86a		55.75 ± 11.89a
Group 2	59.30 ± 1.68ab	78.12 ± 4.51cd	85.30 ± 1.59 d	83.02 ± 15.32b	106.75 ± 12.30b	98.97 ± 12.03bc	94.17 ± 3.58b		95.55 ± 5.28b
Group 3	67.27 ± 3.67bc	72. 25 ± 0.71bc	70.60 ± 1.40 b	82.22 ± 6.32b	126. 45 ± 12.28c	114.17 ± 16.09cd	111.65 ±12.93cd		121.22 ± 13.15c
Group 4	69. 85 ± 2.42c	64. 35 ± 4.79ab	74.55 ± 1.96bc	82. 50 ± 11.48b	131.90 ±4.66c	123.75 ± 4.49d	122.17 ± 5.22d		122.37 ± 3.83c
Group 5	62.12 ± 2.37 abc	86.77 ± 6.12d	80.00 ± 1.36cd	90.00 ±9.61b	94.90 ± 4.38b	94. 45 ±4.68b	104. 02 ± 1.26bc		107.20 ± 5.78b

Note: Different superscript letters on the group means in each column indicate significant difference (p< 0.05).

(Table 1). The MCV shows a significant increase (p<0.05) in all groups except for the negative control. The MCHC showed no significant differences (p>0.05) between groups were evidenced (Table 2).

On days 9 , 14 , 21 and 28 of the post-inoculation (post-treatment), significant decrease of RBC levels were observed in group 2 (positive control) in comparison to group 1 (negative control) and group 4 on day 28. The Hb concentrations at days 9 were significantly decreased (p<0.05) in group 4 compared to the negative control. On day 28 of post-inoculation, the Hb levels declined significantly (p<0.05) in all inoculated groups compared to the negative control (Table 1). On day 14 of post-inoculation, the MCV values in group 3 demonstrated a significant (p<0.05) increase compared to the negative control. On day 21, the levels of MCV in group 5 remained significantly (p<0.05) increased compared to all other groups. On day 28 post-inoculation, all inoculated groups had significantly (p<0.05) higher levels of MCV compared to the negative control. However, the values in group 3 were significantly (p<0.05) lower compared to groups 2 and 5. The MCHC level shows there was significant increase (p<0.05) in group 5 at day 9 in comparison to the negative control and group 3. On day 14, there was a significant (p<0.05) reduction in MCHC levels in groups 4 and 5 compared to the negative control and groups 2 and 3. On days 21 and 28, goats in group 4 demonstrated significantly (p<0.05) reduced levels

 

Table 6: Gamma Glutaryl Transferase and Lactate Dehydrogenase (mean± SD) level of goats blood experimentally infected with Mannheimiosis

Groups	24Hr	Day 5	Day 7	Day 9	Day 11	Day 14	Day 21	Day 28
	Gamma Glutaryl Transferase (GGT) (U/L)
Group 1	78.50 ± 7. 14c	70.75 ± 1.70a	78.5 ± 7.14c	70.75 ± 1.70a	78.50 ± 7.14a	70.75 ± 1.70a	78.50 ± 7.14a	70.75 ± 1.70a
Group 2	61.92 ± 2.08ab	69.87 ±8.09a	63.35 ±2.16b	88.25 ± 7.58 b	103. 82 ± 10.68b	97.00 ± 3.98 b	96.80 ± 2.33b	97.22 ± 2.18b
Group 3	62.62 ± 3.75b	70. 67 ± 7.20a	64.87 ± 3.28b	80.92 ±7.61 b	107.70 ± 8.32b	109.70 ± 4.73c	106.25 ± 6.16c	104. 80 ± 3.16c
Group 4	62.02 ± 3.64ab	65. 37 ± 4.38a	61. 57± 3.90b	88. 72 ± 6.92b	119.02 ± 5.42c	117.57 ± 4.97d	119.17 ±2.10d	117.02 ± 1.54d
Group 5	55.57 ± 2.83a	99.12 ± 4.34b	54.52 ± 0.62a	99. 85 ± 6.42c	100. 37 ± 3.02b	102.55 ± 3.31b	104. 20 ± 1.62c	104.52 ± 2.12c
	Lactate Dehydrogenase (LDH) (U/L)
Group 1	512. 25 ± 73.21a	528.25 ± 58.11a	512.25 ±73.21a	528. 25 ± 58.11a	512.25 ±73.21a	528.25 ± 58.11a	512.25 ± 73.21a	528.25 ± 58.11a
Group 2	1198.75 ±31.61b	1008.75 ± 18. 46b	1129.25 ± 80.03b	1033. 75 ± 58.02b	1105.25 ± 116.36b	1098.00 ± 113.26b	1098.00± 113.26b	1063.75 ± 63.88b
Group 3	1240.75 ± 66.43bc	1170.25 ± 52.82cd	1169.75 ± 92.86b	1162.25 ± 44.97c	1137.75 ± 18.50b	1105.25 ± 62.88b	1105.25 ± 62.88b	1072.50 ± 62.38b
Group 4	1295.25 ± 39.20cd	1075.00 ± 86.26bc	1205.50 ± 67.48b	1124.25 ± 92.65bc	1116.50 ± 86.84b	1092.50 ± 107.46b	1092.50 ± 107.46b	1133.75 ± 122.91b
Group 5	1343.50 ± 31.20d	1225.50 ± 144.15d	1316.25 ±40.59c	1299.25 ± 54.18d	1290.25 ±49.88c	1296.00 ±54.37c	1296.00 ± 54.37c	1267.00 ± 88.97c

Note: Different superscript letters on the group means in each column indicate significant difference (p< 0.05).

of MCHC compared to the negative control and groups 2, 3 and 5 (Table 2). No significant changes in the PLT value throughout the 28 days of study were recorded and those were within the normal range. Significantly decreased (p<0.05) total WBC count parameters in all groups except the negative control within 24 hours and on day 28 of post-inoculation was found (Table 3).

Serum biochemistry results of the TP evaluation revealed a significant (p<0.05) decreased in group 4 compared to negative control and group 5 at 24 hours of infection. On day 9, there was a significant (p<0.05) increase in TP levels in group 3 compared to positive control. On day 14, goats in group 3 showed a higher level (p<0.05) of the TP compared to negative control. On days 21 and 28 of post-inoculation, all inoculated groups remained significantly (p<0.05) higher in TP values than the negative control (Table 4).

Results on the BUN showed significant (p<0.05) increase in the levels of BUN in all inoculated groups compared to negative control at 24 hours and day 5 of post inoculation. On day 9 post-inoculation, all inoculated groups had high BUN levels (p<0.05) compared to negative control. On days 11 to 28, all inoculated groups also had significant (p<0.05) increase of BUN levels compared to the negative control group. On days 14 and 21, groups 3 and 4 showed significantly (p<0.05) higher levels of BUN compared to positive control and group 5. On day 28, group 2 has significantly (p<0.05) lower BUN level compared to all treatment groups. However, the value was slightly above the normal range after 9 days of infection (Table 5).

The CK values showed a significant (p<0.05) increase in groups 3 and 4 compared to negative control groups at 24 hours of post-inoculation. On day 5, the CK levels of groups 2, 3 and 5 were significantly (p<0.05) higher than negative control. On day 9, all of the inoculated groups demonstrated a significant (p<0.05) increase in the levels of CK compared to the negative control. Days 11 to 28 showed significantly (p<0.05) higher CK values than the negative control. Groups 3 and 4 had significantly (p<0.05) higher CK levels compared to positive control and treatment group 5 on days 11 and 28. Meanwhile, group 5 had significantly (p<0.05) lower CK levels compared to groups 3 and 4 on day 14 and significantly (p<0.05) lower CK levels compared to group 4 on day 21. However, the values were within the normal range (Table 5).

GGT levels were observed to be decreased in all inoculated groups at 24 hours post-inoculation. On day 5, the GGT levels in group 5 significantly (p<0.05) increased compared to all other groups. On days 9 to 28, all inoculated groups had significantly (p<0.05) higher levels of GGT compared to negative control. Group 4 were significantly (p<0.05) increased compared to groups 2, 3 and 5; and group 3 significantly (p<0.05) higher compared to groups 2 and 5 on day 14. Group 4 has a significantly (p<0.05) higher levels of GGT compared to positive control and treatment groups 3 and 5 on days 21 and 28 (Table 6).

Level of LDH (p<0.05) were observed higher in all inoculated groups from 24 hours until day 28 as compared to the negative control. Among groups, goats in group 5 were significantly (p<0.05) increased in LDH values compared to the positive control group from 24 hours to day 28 of experimental study. Meanwhile, group 3 has significantly (p<0.05) higher level of LDH than the positive control on days 9 and 11 (Table 6).

Discussion

Haematological evaluations contribute information to the diagnosis, surveillance and prediction of the animal’s prognosis (Roland et al., 2014). The variations in the haematological parameters observed in this study indicated that level of red blood cells and haemoglobin were reduced slightly post-infection. A similar pattern of clinicopathological alterations in the blood of sheep and lambs with pneumonia has shown to have lower haemoglobin concentration (Abdalla et al., 2019). Post-infection with Mannheimia haemolytica does reduce the level of haemoglobin in the blood. This reduction of haemoglobin results from the infection through the release of gram-negative endotoxin (LPS), which cause hemolysis and free release of haemoglobin (Brauckmann et al., 2016).

Decreasing patterns of haematocrit were observed in all groups at 24 h to day 11 post-infections. A previous study on caprine mycoplasma pneumonia and Pasteurellosis showed a significantly lower hematocrit level (Mondal et al., 2004). Post-infection increases MCV levels in the goats. The levels of MCV reduced and fluctuated throughout the studies in all groups. MCHC levels were slightly increased post-infection, but a pattern of reduction in the MCHC levels was observed on day 11. There was no noticeable treatment effect on MCHC values. A previous study by Saleh and Allam (2014) revealed a significant increase (p<0.05) in the MCV and MCHC values in the pneumonic ewes. Therefore these shows that the treatment does not cause a significant changes to the levels of red blood cells, haemoglobin and haematocrit levels post treatments in goats infected with Mannheimiosis.

Evaluation of total WBC count revealed leukopenia occurred in the goats, which demonstrated the recruitment and mobilisation of white blood cells from the blood to the injury site to combat the infection. These findings contrast with other studies which found that goats affected with common bacteria causing pneumonia, such as Klebsiella pneumoniae, Pasteurella spp., Mannheimia spp., which had an increase in the leucocytic count (Ghanem et al., 2015; Kattimani et al., 2020). Duration and chronicity of the infection also play a role in the recruitment and distribution of the white blood cells, which may cause a decrease in the leucocyte count, as observed in this study. In this study, it shows that treatments do not have a significant effects on the improvement of the white blood cell counts. This could be due to the length of the treatment given to the goats were insufficient in effectively normalize the levels of white blood cells counts in the goat post infections.

The serum biochemistry parameters in the recent study showed that the total protein, albumin and globulin values were within the normal range. Meanwhile, the levels of blood BUN showed a mild increment in post-infection, which was associated with dehydration and injury of the kidney. CK and GGT values were normal in all groups, but their levels had mild increases, which indicates liver injury. The study indicates the following serum biochemistry parameters does not shows a significant reduction nor improvement post-treatment regimes. This could be due to the dose of Mannheimia haemolytica infection in the goats were mild to moderate only.

In contrast to that, a significant increase in LDH was observed post-infection. In relation to respiratory diseases, LDH play role as one of the important indicator which can be used in monitoring the pathological changes related to the lung (Hagadorn et al., 1971). Lactate dehydrogenase is one of the indicators of lung damage as lungs have 500-fold higher LDH compared to their normal concentration in serum. Previous studies demonstrated an increase in the activity of LDH enzymes, particularly (LDH4 and LDH5) in lambs with lung lesions of acute and chronic pneumonia (Milne & Doxey, 1984; Drent et al., 1996; Klein et al., 2020). As observed in this study, Mannheimiosis causes alteration in the serum LDH level leading to their elevation which shows that it is due to the pneumonia development in the goats and the cellular injury that takes place in the lungs. This increment is due to the release of LDH from the epithelial cell of the airways lining that gives indicator of cell damage in the lungs (Drent et al., 1996) and the treatment regime does not significantly reduce the LDH level in the experimental goats. This also shows that the duration of treatment and the progression of disease does interferes with the level of LDH in the study and a longer length of treatment regimes may significantly improved or normalize better the level of LDH post treatment.

Conclusion

The haematological findings in goats experimentally induced with Mannheimia haemolytica demonstrated that post-infection causes a mild reduction in the red blood cell parameters (RBC, Hb, HCT) while decreased the total WBC count. The serum biochemistry findings were found within normal range for TP, ALB and GLOB, with mild to moderate increase in CK, BUN, GGT and LDH level. In conclusion, treatment does not have a great influence on the parameters evaluated except temporary lowering the values of some parameters.

ACKNOWLEDGEMENT

The authors would like to acknowledge the Ministry of Higher Education, Malaysia, for providing the FRGS-RACER grant (R/FRGS/A0600/01391A/001/2019/00668). The author would like to thank Madam Nur Eizzati Badrul Hisham and Miss Nani Izreen Mohd Sani from the Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, for their technical assistance while conducting the research.

conflict of interest

The author declared no conflict of interest.

novelty statement

Treatment does not have a great influence on the parameters evaluated except temporary lowering the values of some parameters.

authors contribution

All author contributed equally to the research work and manuscript.

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