Effect of apple cider vinegar and vitamin A on hematological parameters and total immunoglobulin G in sheep: a pilot study

Document Type : Research Article

Authors

1 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.

3 Graduated from Doctor of veterinary Medicine, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.

Abstract

Apple cider vinegar is a fermented compound that contains acetic acid, flavonoids, phenolic compounds, organic acids, minerals, and vitamins. Vitamin A is involved in the development of the immune system and plays regulatory roles in cellular immune responses and humoral immune processes. The aim of this study was to investigate the effect of apple cider vinegar and vitamin A on hematological parameters and immunoglobulin G levels in Gezel lambs. 10 healthy Gazel lambs were used for the study. The lambs were randomly grouped into three groups (control (n = 3), Apple cider vinegar (n = 4), and Vitamin A (n = 3) group). Vitamin A was administrated at a dose of 44,000 IU/kg every ten days for four treatments. ACV was administered orally by drenching (0.5 ml/kg of 6% ACV solution (600 mg ACV), every day, for 40 days). Hematological parameters were determined using standard methods. Total immunoglobulin G concentration was assayed using the turbidimetric immunoassay method. The data obtained before and after drug administration were analyzed by paired T-test and the data of different groups were analyzed using Independent-sample T-test.White blood cells, lymphocytes, and IgG in lambs were significantly increased after administration of Apple cider vinegar (p < 0.05). IgG and lymphocytes were significantly higher in lambs under oral administration of Apple cider vinegar compared to the lambs in the control group (p  < 0.05). White blood cells, neutrophils, and IgG were significantly increased in lambs after vitamin A administration (p < 0.05). Neutrophils and IgG were significantly higher in lambs under vitamin A injection compared to the lambs in the control group (p < 0.05). Administration of vitamin A and apple cider vinegar in sheep is safe. They also improve the immune system.

Keywords

Main Subjects

Abbreviations

APC: apple cider vinegar

RBC: red blood cell

WBC: white blood cell

TP: total protein

PCV: packed cell volume

Hb: Hemoglobin

IgG: immunoglobulin G

Introduction

Apple cider vinegar (ACV) is a fermented compound that contains acetic acid, flavonoids, phenolic compounds, organic acids, minerals, and vitamins [ 1 , 2 ]. ACV is produced through a two-stage fermentation process. First, the sugar is converted into alcohol by yeasts, then the alcohol is converted into acetic acid by acetic acid bacteria [ 3 ].

Animal experiments have reported that ACV has a variety of pharmacological activities, including antioxidant, anti-inflammatory, anti-diabetic, antifungal, antimicrobial, anti-hypertensive, and anti-hyperlipidemic actions [ 4 - 8 ]. A study showed that using ACV in a fish diet improves immunological parameters and gene expression related to immunity, antioxidant system, and growth performance [ 9 ]. Also, a study showed that using ACV in a fish diet increases total immunoglobulin and lysozyme activity [ 10 ]. In addition, a study has reported that dietary supplementation with ACV has beneficial effects on performance and immune response in broiler chickens [ 11 ]. It has been shown that ACV has an anthelmintic impact against gastrointestinal parasites in sheep [ 12 ], and increases the activity of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase [ 13 ].

Vitamin A is known as an anti-inflammation vitamin because of its critical role in enhancing immune function [ 14 ]. Vitamin A is involved in the development of the immune system and plays regulatory roles in cellular immune responses and humoral immune processes [ 14 ].

Thus, the aim of this study was to assess the effect of ACV and vitamin A on hematological parameters and immunoglobulin G in Gezel lambs.

Result

By administering ACV and Vitamin A at the mentioned doses, no disorders and lesions have occurred in the lambs.

ACV

RBC, PCV, Hb, fibrinogen, TP, neutrophils, band neutrophils, eosinophils, and monocytes in lambs before and after administration of ACV were not statistically different.

WBC, lymphocytes, and IgG in lambs were significantly higher after administration of ACV compared to the control group (p < 0.05).

RBC, PCV, Hb, fibrinogen, TP, WBC, neutrophils, band neutrophils, eosinophils, and monocytes were not statistically different between lambs under oral administration of ACV and lambs in the control group.

IgG and lymphocytes were significantly higher in lambs under oral administration of ACV than in lambs in the control group (P<0.05).

The results of the study are summarized in Tables 1 and 2.

Parameter A day before study A day after study P value
Min Max Mean ±SE Min Max Mean ±SE
Case group under vitamin A injection
RBC (104/μl) 1025 1190 1105 ± 47.69 1125 1155 1142 ± 9.06 0.464
PCV (%) 30 35 32.67 ± 1.45 33 34 33.34 ± 0.33 0.635
Hb (g/dl) 10 11.66 10.88 ± 0.48 11 12 11.44 ± 0.29 0.336
TP (g/dl) 6.2 6.66 6.42 ± 0.13 6.4 7.1 6.67 ± 0.21 0.134
Fibrinogen (mg/dl) 100 200 166.7 ± 33.34 100 400 266.7 ± 88.2 0.225
WBC/μl 4446 10349 7248.4 ± 1710 8511 14150 10917.7 ± 1679.3 0.006*
Neutrophil /μl 1269 3895 2421.4 ± 774.9 4514 5382 4898.7 ± 255.3 0.041*
Band neutrophil/μl 30 100 66.7 ± 20.2 112 207 143.7 ± 31.7 0.055
Lymphocyte/μl 2976 5842 4439.4 ± 827.89 3519 7320 5113 ± 1139.2 0.259
Eosinophil /μl 124 307 200.4 ± 54.96 244 724 449.4 ± 142.8 0.105
Monocyte/μl 47 205 120.7 ±45.9 122 517 313 ±114.2 0.107
IgG (g/l) 1.19 1.30 1.24 ± 0.031 1.34 1.40 1.36 ± 0.018 0.020*
Case group under oral administration of ACV
RBC (104/μl) 952 1260 1128 ± 76.03 982 1342 1143 ± 74.95 0.804
PCV (%) 28 37 32.7 ± 1.89 29 40 35.5 ± 2.39 0.102
Hb (g/dl) 9.33 12.33 10.9 ± 0.61 9.6 13.33 8.5 ± 2.4 0.470
TP (g/dl) 5.7 6.5 6.25 ± 0.18 6.1 7.4 6.52 ± 0.29 0.340
Fibrinogen (mg/dl) 100 300 187.5 ± 42.6 100 100 100 ± 0 0.133
WBC/μl 3330 8702 5242 ± 1187.5 5458 12470 8811.5 ± 1459.2 0.01*
Neutrophil /μl 1239 2790 1732.2 ± 356.6 910 4810 3005 ± 813.1 0.105
Band neutrophil/μl 62 80 67.7 ± 4.2 60 100 80 ± 8.16 0.091
Lymphocyte/μl 1909 5460 3217.2 ± 780.1 4250 6900 5337.5 ± 571.3 0.007*
Eosinophil/μl 80 186 121.5 ± 22.9 130 400 232.5 ± 58.36 0.053
Monocyte/μl 40 186 104 ± 30.8 108 260 156.5 ± 35.8 0.063
IgG (g/l) 1 1.25 1.08 ± 0.059 1.55 1.90 1.73 ± 0.074 0.001*
Control group
RBC (104/μl) 1021 1184 1118.4 ± 49.7 1187 1274 1220 ± 27.09 0.096
PCV (%) 30 35 33.4± 1.7 35 38 36 ± 1 0.208
Hb (g/dl) 10 11.66 11 ± 0.54 11.66 12.66 12.1 ± 0.293 0.107
TP (g/dl) 5.8 6.6 6.3 ± 0.25 6.8 7.3 6.97 ± 0.16 0.081
Fibrinogen (mg/dl) 150 300 200 ± 50 200 300 266.7 ± 66.7 0.181
WBC /μl 3851 9960 6722.7 ± 1772.96 5484 7400 6561.4 ± 565.8 0.909
Neutrophil /μl 1692 3840 2677.4 ± 626.3 2557 3040 2765.7 ± 143.2 0.872
Band neutrophil /μl 47 60 51.4 ± 4.3 40 70 53.4 ± 8.8 0.724
Lymphocyte/μl 1909 5460 3623 ± 1026.9 2500 3750 3283.4 ± 394.05 0.675
Eosinophil/μl 141 360 217 ± 71.5 232 340 274 ± 33.4 .278
Monocyte /μl 62 240 154 ± 51.4 155 200 185 ± 15 0.507
IgG (g/l) 1.18 1.25 1.2 ± 0.023 1.20 1.29 1.24 ± 0.026 0.225
*P< 0.05, RBC: red blood cell, PCV: packed cell volume, Hb: hemoglobin, TP: total protein, WBC: white blood cell, IgG: immunoglobulin G, min: minimum, max: maximum, SE: standard error.
 Table 1.Hematological parameters and IgG in lambs before and after administration of vitamin A and ACV
Parameter Case group under vitamin A injection P value Control group P value Case group under oral administration of ACV
Min Max Mean ± SE Min Max Mean ± SE Min Max Mean ± SE
RBC(104/μl) 1125 1155 1142.7 ± 9.06 0.053 1187 1274 1220.4 ± 27.09 0.440 982 1342 1143± 74.9
PCV (%) 33 34 33.4 ± 0.33 0.065 35 38 36 ± 1 0.872 29 40 35.5 ±2.39
Hb (g/dl) 11 12 11.44 ± 0.29 0.186 11.66 12.66 12.1 ± 0.29 0.560 9.6 13.33 11.51± 0.77
TP (g/dl) 6.4 7.1 6.67 ± 0.21 0.413 6.8 7.3 6.8 ± 0.06 0.383 6.8 7.4 6.52 ±0.29
Fibrinogen (mg/dl) 100 400 266.7 ± 88.19 0.742 200 400 233.4 ± 33.4 0.057 100 100 100 ± 0
WBC/μl 8511 14151 10917.7 ± 1679.3 0.070 5484 7400 6561 ± 565.8 0.808 1270 9320 6011.5 ± 1772
Neutrophil/μl 4514 5382 4898.7 ± 255.3 0.002* 2557 3040 2765.7 ± 143.2 0.815 910 4810 3005 ± 813.1
Band neutrophil/μl 112 207 143.7 ± 31.7 0.051 40 70 53.4 ± 8.8 0.080 60 100 80 ±8.1
Lymphocyte/μl 3519 7320 5113 ± 1139.2 0.204 2500 3750 3283 ± 394.05 0.041* 4250 6900 5337.5 ± 571.3
Eosinophil/μl 244 724 449.4 ± 142.8 0.298 232 340 274 ± 33.4 0.601 130 400 232.5 ± 58.3
Monocyte /μl 122 517 313 ± 114.2 0.329 155 2 185 ± 15 0.548 108 260 156.5 ± 35.8
IgG (g/l) 1.34 1.40 1.36 ± 0.018 0.022* 1.25 1.29 1.24 ± 0.026 0.004* 1.55 1.90 1.73± 0.07
*P< 0.05, RBC: red blood cell, PCV: packed cell volume, Hb: hemoglobin, TP: total protein, WBC: white blood cell, IgG: immunoglobulin G, min: minimum, max: maximum, SE: standard error.
 Table 2.Hematological parameters and IgG in lambs after administration of Vitamin A and ACV compared to control group

Vitamin A

RBC, PCV, Hb, fibrinogen, TP, band neutrophils, lymphocytes, eosinophils, and monocytes in lambs before and after administration of vitamin A were not statistically different.

WBC, neutrophils, and IgG were significantly higher in lambs after vitamin A administration compared to the control group (p < 0.05).

RBC, PCV, Hb, fibrinogen, TP, WBC, band neutrophils, lymphocytes, eosinophils, and monocytes were not statistically different between lambs under vitamin A injection and lambs in the control group.

Neutrophils and IgG levels were significantly higher in lambs under vitamin A injection than in the control group (p < 0.05).

The results of the study are summarized in Tables 1 and 2.

Discussion

A study on the effects of ACV on haemato-biochemical parameters in rate showed that there are no significant variations between the groups in post-treatment values of PCV, Hb, RBC, and absolute eosinophil counts [ 15 ]. In our study, these mentioned parameters were not different before and after administration of ACV and also they were not different between the test article groups and the control group. Also, another study found no difference in RBC between control and case groups after adding a combination of onion, garlic, and ACV to broiler chicken diets[ 16 ]. A study showed that there is no significant impact of ACV supplementation on broiler serum total protein levels[ 17 ]. This finding is similar to our finding. Also, a study showed that ACV has no effect on RBC, Hb, and PCV in humans [ 18 ]. The same results are presented in our study.

A study on the effects of an herbal mixture (onion, garlic, and ACV) on hematological parameters of broiler chickens showed that the herbal mixture resulted in significant increases in PCV and Hb values[ 16 ]. This finding is contrary to our results, and probably the results obtained in the mentioned study were not caused by the effect of ACV, or maybe the difference between the results of our study and that study is due to differences between species.

In our study, the significant increase in WBC count after the administration of ACV was mainly due to the substantial increase in lymphocyte count. However, the significant increase in the lymphocyte count in the ACV-administered group compared to the control group did not lead to an increase in WBC count in the ACV-administered group compared to the control group. A study showed that the addition of an herbal mixture (onion, garlic, and ACV) to the diet of broiler chickens increased lymphocyte counts [ 16 ]. The improvement of the immune system due to the administration of ACV has been mentioned in several studies.. Since lymphocytes play a role in humoral immunity, perhaps the improvement of the immune system caused by ACV is responsible for the increase in lymphocytes. However, this requires further study.

In our study, it was shown that immunoglobulin G increased after the administration of vitamin A and ACV. It also increased in the ACV and vitamin A-administered group compared to the control group. These findings show that vitamin A and ACV (as mentioned above) are useful in improving the immune system in lambs. An increase in total immunoglobulin has been reported in fish after administration of ACV [ 10 , 20 , 21 ]. In a study, it was shown that administration of vitamin A leads to an

Materials and Methods

Selection of lambs, grouping and sampling

To perform this study, ten healthy male Gazel lambs were selected. Their health was confirmed by veterinary examinations. The age (6 months) and weight (20 kg) of the selected lambs were similar. Before starting the study, the selected lambs underwent a week of adaptation to the new environment. During the study, lambs were fed alfalfa and hay ad libidum and were observed for any clinical signs of disease throughout the study.

The lambs were randomly grouped into three groups:

1- Control group: 3 lambs were assigned to this group, and no medication was administered to them.

2- Case group under vitamin A injection: 3 lambs were assigned to this group and vitamin A was administered to them (44,000 IU/kg Retinavit®, intramuscularly (IM), every ten days, 4 treatments).

3- Case group under oral administration of ACV: 4 lambs were assigned to this group, and ACV was administered orally by drenching (0.5 ml/kg of 6% ACV solution (600 mg ACV), every day, for 40 days).

Blood samples were taken from the jugular vein of all lambs before and after the study.

Laboratory measurement

Red blood cell (RBC) count, packed cell volume (PCV), hemoglobin (Hb), fibrinogen, total protein (TP), white blood cell (WBC) count, and absolute count of neutrophils, band neutrophils, lymphocytes, eosinophils, and monocytes were determined by routine hematology tests.

Total immunoglobulin G (IgG) concentration was assayed by turbidimetric immunoassay method as previously described [ 25 , 26 ].

Statistical analysis

The data obtained before and after drug administration were analyzed by paired T-Test in SPSS software. Also, the data of different groups were analyzed using the independent-sample t-test. p &lt; 0.05 was considered significant.

Conclusion

Administering vitamin A and ACV in sheep is safe. They also improve the immune system. Vitamin A and ACV increase total IgG, and also vitamin A increases neutrophils and ACV increases lymphocytes. In general, they can be used to improve the immune system in sheep.

Authors' Contributions

Conceived and designed the experiments: M.T., and E.F. Performed the experiments: M.T., and M.A..Wrote the paper: A .Sh.

Acknowledgements

The authors would to express their gratitude to Research Vice-Chancellor of University of Tabriz for their financial support.

Competing Interests

The authors declare that there is no conflict of interest.

References

  1. Morgan J, Mosawy S. The potential of apple cider vinegar in the management of type 2 diabetes. International Journal of Diabetes Research. 2016; 5(6): 129-34. DOI
  2. Tripathi S, Mazumder PM. Apple cider vinegar (ACV) and their pharmacological approach towards Alzheimer’s disease (AD): A review. Indian J Pharm Educ Res. 2020; 54: s67-s74. DOI
  3. Samad A, Azlan A, Ismail A. Therapeutic effects of vinegar: a review. Current Opinion in Food Science. 2016; 8: 56-61. DOI
  4. Hadi A, Pourmasoumi M, Najafgholizadeh A, Clark CC, Esmaillzadeh A. The effect of apple cider vinegar on lipid profiles and glycemic parameters: a systematic review and meta-analysis of randomized clinical trials. BMC Complementary Medicine and Therapies. 2021; 21(1): 179. DOI
  5. Iman M, Moallem SA, Barahoyee A. Effect of apple cider vinegar on blood glucose level in diabetic mice. Pharmaceutical Sciences. 2014; 20(4): 163-168.
  6. Nazıroğlu M, Güler M, Özgül C, Saydam G, Küçükayaz M, Sözbir E. Apple cider vinegar modulates serum lipid profile, erythrocyte, kidney, and liver membrane oxidative stress in ovariectomized mice fed high cholesterol. The Journal of Membrane Biology. 2014; 247: 667-673. DOI
  7. Shishehbor F, Mansoori A, Sarkaki A, Jalali M, Latifi S. Apple cider vinegar attenuates lipid profile in normal and diabetic rats. Pakistan journal of biological sciences: PJBS. 2008; 11(23):2634-2638. DOI
  8. Mota ACLG, de Castro RD, de Araújo Oliveira J, de Oliveira Lima E. Antifungal activity of apple cider vinegar on Candida species involved in denture stomatitis. Journal of Prosthodontics. 2015; 24(4): 296-302. DOI
  9. Ahmadifar E, Dawood MA, Moghadam MS, Sheikhzadeh N, Hoseinifar SH, Musthafa MS. Modulation of immune parameters and antioxidant defense in zebrafish (Danio rerio) using dietary apple cider vinegar. Aquaculture. 2019; 513: 734412. DOI
  10. Ahmadniaye Motlagh H, Javadmanesh A, Safari O. Improvement of non-specific immunity, growth, and activity of digestive enzymes in Carassius auratus as a result of apple cider vinegar administration to diet. Fish physiology and biochemistry. 2020; 46(4): 1387-1395. DOI
  11. Jahantigh M, Kalantari H, Ayda Davari S, Saadati D. Effects of dietary vinegar on performance, immune response and small intestine histomorphology in 1‐to 28‐day broiler chickens. Veterinary Medicine and Science. 2021; 7(3): 766-772. DOI
  12. Hayajneh FM, Titi HH, Alnimer MA, Irshaid R. Evaluation of anthelmintics resistance against gastrointestinal parasites infection in awassi sheep in Jordan and the use of alternative herbal anthelmentics. Am J Anim Vet Sci. 2019; 14(2):122-126. DOI
  13. Halima BH, Sarra K, Houda BJ, Sonia G, Abdallah A. Antidiabetic and antioxidant effects of apple cider vinegar on normal and streptozotocin-induced diabetic rats. International Journal for Vitamin and Nutrition Research. 2019. DOI
  14. Huang Z, Liu Y, Qi G, Brand D, Zheng SG. Role of vitamin A in the immune system. Journal of Clinical Medicine. 2018; 7(9): 258.
  15. Ngwu GI, Opara BA, Ngwu MI, Atuogba NC, Ugwuoke WI, Ossai NI, et al. Effects of apple cider vinegar on haemato-biochemical parameters of gestating rats and morphometric indices of their pups at delivery. Comparative Clinical Pathology. 2021; 30(6): 953-960. DOI
  16. Abdulkareem ZA, Mohammed NI, Abdollahi A, Ahmed OR, Ghaffar OR, Khdir HA, et al. Effects of garlic, onion, and apple cider vinegar as a herbal mixture on performance and blood traits of broilers inoculated with chicken infectious anemia virus. Heliyon. 2023; 9(7)DOI
  17. Hayajneh F. Natural feed additives for broiler chickens. South African Journal of Animal Science. 2019; 49(5): 869-875. DOI
  18. Hosseini ZSM, Hosseini J, Nabati S, Hasanshahi G, Mahmoodi M. Survey on the anti-diabetic effects of vinegar on some biochemical factors in type 2 diabetic patients. Clinical Biochemistry-New York. 2011; 44(13): 2. DOI
  19. Pourmozaffar S, Hajimoradloo A, Miandare HK. Dietary effect of apple cider vinegar and propionic acid on immune-related transcriptional responses and growth performance in white shrimp, Litopenaeus vannamei. Fish & shellfish immunology. 2017; 60: 65-71. DOI
  20. Ahmadniaye Motlagh H, Sarkheil M, Safari O, Paolucci M. Supplementation of dietary apple cider vinegar as an organic acidifier on the growth performance, digestive enzymes and mucosal immunity of green terror (Andinoacara rivulatus). Aquaculture Research. 2020; 51(1): 197-205. DOI
  21. Safari R, Hoseinifar SH, Nejadmoghadam S, Khalili M. Apple cider vinegar boosted immunomodulatory and health promoting effects of Lactobacillus casei in common carp (Cyprinus carpio). Fish &shellfish immunology. 2017; 67: 441-448. DOI
  22. Semba R, Scott A, Natadisastra G, West Jr K, Sommer A. Effect of vitamin A supplementation on immunoglobulin G subclass responses to tetanus toxoid in children. Clinical and Diagnostic Laboratory Immunology. 1994; 1(2): 172. DOI
  23. Semba RD. The role of vitamin A and related retinoids in immune function. Nutrition reviews. 1998; 56(1): S38-S48.
  24. Twining SS, Schulte DP, Wilson PM, Fish BL, Moulder JE. Vitamin A deficiency alters rat neutrophil function. The Journal of Nutrition. 1997; 127(4): 558-565.
  25. Davis DG, Schaefer DM, Hinchcliff KW, Wellman ML, Willet VE, Fletcher JM. Measurement of serum IgG in foals by radial immunodiffusion and automated turbidimetric immunoassay. Journal of Veterinary Internal Medicine. 2005; 19(1):93-96. DOI
  26. Ferris RA, McCue PM. How to use a quantitative turbidimetric immunoassay to determine immunoglobulin G concentrations in neonatal foals, Proceedings of the... annual convention. 2009.
Send comment about this article
CAPTCHA Image
Iranian Journal of Veterinary Science and Technology
Volume 16, Issue 3 - Serial Number 36
This issue XML files are being prepared.
September 2024
Pages 85-91
Plum Print visual indicator of research metrics
  • Captures
    • Readers: 2
  • Mentions
    • News Mentions: 1
see details

Files

History

  • Receive Date: 27 September 2023
  • Revise Date: 14 May 2024
  • Accept Date: 10 August 2024

Share

How to cite

Statistics