Assessment of clinical and hematological changes in sheep pneumonia following vitamin supplement therapy.

Introduction: Pneumonia is a significant health concern in sheep, particularly lambs, characterized by inflammation in the lungs in response to infectious agents affecting bronchioles and alveoli. It is a prevalent condition in sheep populations worldwide (Goodwin et al., 2004). Vitamin C, an essential physiological antioxidant in the immune system, plays a crucial role in protecting host cells from oxidative stress induced by infections. Its high concentration in phagocytes and lymphocytes compared to plasma levels suggests its importance in immune cell function. Vitamin C has been shown to enhance phagocyte activity, T lymphocyte proliferation, interferon production, and decrease viral replication in various experimental settings (Hamila, 2006). Animal studies have demonstrated that vitamin C supplementation can increase resistance to viral and bacterial infections. Furthermore, many infections, including pneumonia, are associated with reduced levels of vitamin C in plasma, leukocytes, and urine (Hamila, 2006). Thiamine is another essential component involved in cellular metabolism, participating in mitochondrial oxidative decarboxylation and glycolytic pathways (Nazir et al., 2019). Patients with septic shock often exhibit thiamine deficiency, which can be attributed to either deficiency or increased loss due to overdose (Donino et., 2010). This study aimed to investigate the efficacy of administering ascorbic acid and thiamine alongside oxytetracycline and tylosin in the treatment of sheep pneumonia as a safe and beneficial approach. The analysis focuses on examining alterations in clinical symptoms and blood parameters to monitor pneumonia improvement.
Materials and Methods:
Twenty 8–12-month-old male sheep with respiratory disease were selected for this study and randomly assigned to four treatment groups: A, B, C, and D. Additionally, five healthy sheep were designated as the control group and received normal saline solution. Group A was administered an antibiotic regimen consisting of oxytetracycline (Oxyvet 10%; hops, Karaj, Alborz, Iran) at a dosage of 11 mg/kg body weight and tylosin (Tylosin 20%; hops, Karaj, Alborz, Iran) intramuscularly at a dose of 18 mg/kg body weight over five days. Group B, in addition to the antibiotics received by Group A, was given thiamine hydrochloride or vitamin B1 (B-ject®; Nasr, Fariman, Khorasan Razavi, Iran) intravenously for five days at a dose of 20 mg/kg body weight (Constable et al., 2016). Group C received ascorbic acid or vitamin C (Vitamin C®; Darou Pakhsh, Tehran, Iran) intravenously for five days at 2.5 g per sheep (Black and Hirdoglu,1996). Group D received a combination of antibiotics, thiamine, and ascorbic acid concurrently. Blood samples were collected from the sheep on the first, third, sixth, and fourteenth days of clinical evaluation for analysis of the blood panel. The effects of the interventions were compared using repeated measures analysis of variance (ANOVA). In cases where the overall ANOVA indicated significant statistical differences, paired comparisons were conducted using the Bonferroni post hoc test. A significance level of P<0.05 was considered for all statistical analyses.
Results and Discussion:
Throughout the treatment period, all groups of animals exhibited clinical improvement characterized by a reduction in respiratory rate, heart rate, body temperature, and other pneumonia-related symptoms. Groups C and D demonstrated a more rapid resolution of shortness of breath and cessation of nasal discharge compared to the other groups. This accelerated response in groups C and D could be attributed to the anti-inflammatory properties of thiamine and vitamin C, which are known to inhibit prooxidative and cytokine activities with anti-inflammatory effects (Mohammadi and Rezaei, 2019; Chen & Wissen, 2020). Additionally, vitamin C has been shown to enhance the clearance of alveolar fluid by preventing active neutrophil accumulation in the alveolar spaces (Fischer et al., 2011). This control of inflammatory processes by thiamine and vitamin C likely contributed to the improvement in clinical symptoms observed in animals with pneumonia, ultimately leading to the resolution of symptoms and reversal of appetite loss.
Following treatment, a significant decrease in the total number of white blood cells, lymphocytes, and neutrophils was observed across all treatment groups (P<0.05), with the D group exhibiting the most pronounced reduction. This study demonstrated that the concurrent administration of vitamin C, thiamine, and antibiotics expedited the healing process in the treated animals, resulting in a faster reduction in blood neutrophil counts. Sheep receiving thiamine supplementation showed more effective neutrophil responses in eliminating infectious agents compared to those without thiamine supplementation (Mohammadi and Rezaee, 2019). Vitamin C, in addition to enhancing the chemotaxis, supported lymphocyte function, facilitated phagocytosis and aided in the intracellular destruction of bacteria (Wilson, 2013).
In terms of hematological parameters, a significant decrease (P<0.05) was observed in the total erythrocyte count, hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular conclusion (MCH), and mean corpuscular hemoglobin concentration (MCHC) values, while the platelet count increased to a normal level in the herd. The initial reduction in erythrocytes, hemoglobin, and hematocrit levels in animals with pneumonia on the first day of the study may be attributed to dehydration, which is common in infected lambs due to decreased appetite, sweating from fever, and fluid loss resulting from lung inflammation (Pesanita et al., 2020). Following treatment, as appetite and hydration were restored in the animals, a decrease in RBC count and subsequent reductions in hemoglobin and hematocrit levels were observed across all treatment groups. In cases of chronic pneumonia, the secretion of inflammatory cytokines such as IL-1β, IL-3, and TNF-α can disrupt the iron cycle, reducing erythrocyte and hemoglobin production by retaining iron in bone marrow macrophages (Trawl et al., 2012). However, in pneumonia-induced hypoxia, increased erythropoietin secretion may contribute to heightened erythrocyte production (Trawl et al., 2012).
Conclusion: The results of this study indicate that the combination of ascorbic acid and thiamine, along with antibiotics, proved to be more effective in treating respiratory diseases in sheep, accelerating the healing process. This combination therapy demonstrated significant reductions in white blood cell counts, lymphocytes, and neutrophils, with the D group showing the most substantial decrease. The administration of ascorbic acid and thiamine in conjunction with antibiotics resulted in a faster resolution of clinical symptoms, particularly in alleviating shortness of breath and nasal discharge. These vitamins' anti-inflammatory properties likely played a crucial role in controlling inflammatory processes and enhancing immune cell function, leading to the improvement in clinical symptoms. Additionally, hematological parameters such as total erythrocyte count, hemoglobin, hematocrit, and red blood cell indices were significantly altered following treatment, reflecting the impact of the therapy on various physiological functions. This study highlights the potential of utilizing ascorbic acid and thiamine in combination with antibiotics as a cost-effective and safe treatment approach for respiratory diseases in sheep.