Efficacy of Yeast in Ameliorating Deleterious Effects of Coccidiosis in Sonali Chicken

Md. Ariful Alam1, Mst. Misrat Masuma Parvez1, Md. Salauddin2*, Rakibul Islam1 and Md. Bazlar Rashid1

1Department of Physiology and Pharmacology, Faculty of Veterinary and Animal Science, Hajee Mohammad Danesh Science and Technology University, Dinajpur-5200, Bangladesh; 2Department of Microbiology and Public Health, Faculty of Veterinary, Animal and Biomedical Sciences, Khulna Agricultural University, Khulna-9100, Bangladesh.

Editor | Muhammad Abubakar, National Veterinary Laboratories, Park Road, Islamabad, Pakistan.

Received | March 01, 2023; Accepted | November 20, 2023; Published | November 30, 2023

*Correspondence | Md. Salauddin, Department of Microbiology and Public Health, Faculty of Veterinary, Animal and Biomedical Sciences, Khulna Agricultural University, Khulna-9100, Bangladesh; Email: salauddin.dvm@gmail.com

Citation | Alam, M.A., M.M.M. Parvez, M. Salauddin, R. Islam and M.B. Rashid. 2023. Efficacy of yeast in ameliorating deleterious effects of coccidiosis in Sonali chicken. Veterinary Sciences: Research and Reviews, 9(2): 152-160.

DOI | https://dx.doi.org/10.17582/journal.vsrr/2023/9.2.152.160

Keywords | Histopathology, Hematological analysis, Yeast, Amprolium, Coccidiosis in Sonali chicken, Bangladesh

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

The Sonali bird, brought to Northern Bangladesh between 1996 and 2000 via Smallholder and Participatory Livestock Development Project, is a crossbreed of Rhode Island Red (RIR) roosters and Fayoumi hens. It closely resembles the local chickens of Bangladesh in appearance (Uddin et al., 2015). In spite of many positive reasons behind the rearing of Sonali chickens, there are some causes that hamper in optimum production of Sonali chicken by smallholder households in Bangladesh (Biswas et al., 2006). Poultry diseases are one of the major constraints for developing the poultry industry in Bangladesh (Islam and Samad, 2004). However, very few studies have been reported on the prevalence of infectious diseases of chickens in the northern part of Bangladesh. Thus, this study was aimed at investigating the prevalence of various infectious diseases of Sonali chickens in the northern part of Bangladesh.

Avian Coccidiosis is one of the most important diseases of poultry in Bangladesh caused by single-celled protozoan parasites of the genus Eimeria (Lowery, 2023). Coccidiosis represents a severe economic loss in the poultry industry. Control of coccidiosis mostly depends upon chemoprophylaxis by using anticoccidial drugs, but, managerial skills are also important to get the maximum anticoccidial effect of these drugs (Tewari and Maharana, 2011). Chemical anticoccidial additives have been crucial for poultry industry growth, ensuring affordable and high-quality meat. However, their indiscriminate use has led to drug resistance issues and raised concerns about antibiotic presence, contributing to potential antibiotic resistance in bacteria, a worry for consumers (Mak et al., 2022). Due to consumer worries, poultry farmers seek antibiotic-free alternatives for growth. Ideal substitutes should be affordable, widely accessible, and health-conscious. Yeast, with its diverse therapeutic benefits like anti-inflammatory, antimicrobial, and anticoccidial properties, emerges as a promising solution. Yeast cell products are derived from yeast species such as Saccharomyces cerevisiae and Pichia guillermondii (De los Santos et al., 2007; Shanmugasundaram and Selvaraj, 2012). The yeast cell wall contains the polysaccharides mannan oligosaccharides and β-glucans (Thomas et al., 2022). These polysaccharides can have immunomodulatory effects in several species, including poultry (Yitbarek et al., 2012). Yeast products are crucial in poultry farming for their ability to combat coccidial issues, addressing concerns about drug resistance in pathogens (Bilal et al., 2022). Immunomodulation properties of the yeast derivatives have been shown to enhance cellular and humoral immunity in Eimeria challenge models which is critical for the effectiveness of coccidial vaccination. Moreover, yeast nucleotides have been shown to be beneficial in stimulating the healing of the intestinal mucosal surface. Among all the yeast species, S. cerevisiae has been used the most as a probiotic or prebiotic in poultry nutrition (Hooge, 2004; Dhama and Singh, 2010). Because it is readily available and easy to culture, the probiotic yeast Saccharomyces cerevisiae var boulardii is widely used as a low-cost and efficient adjuvant against gastrointestinal tract disorders such as inflammatory bowel disease and treatment of several types of diarrhea, both in humans and animals. Yeast cell surface has also been shown to be an effective oral Eimeria vaccine delivery vehicle. This study aims to compare the impact of amprolium and yeast on coccidiosis-infected Sonali chickens. The evaluation focuses on hematological parameters and histopathological changes.

Materials and Methods

Animals and records of body mass

Sixty Sonali chickens, aged 23 days, were obtained from Bismillah Farm, Ramsagar, Dinajpur. Upon arrival, they were placed in the experimental shed with proper ventilation and lighting. The birds acclimated for 5 days before the 30-day experiment. Throughout, they experienced a 16-hour photoperiod (natural plus artificial light) in an open-sided house, with additional light from electrical bulbs at night. Weights were measured at the start, every 7 days, and the final weight was recorded on the 30th day.

Collection of feed and yeast

Polli mash commercial feed was collected from Griholokkhi Poultry Feed, Kalitola, Dinajpur from a reputed Sonali feed exporter. Mash and water were provided ad-libitum during the whole experimental period. Yeast (Saccharomyces cerevisiae), company name: Lihama was purchased from the local market of Dinajpur district. Birds were fed with commercial feed and drinking water ad-libitum. Glucose and vitamin C were supplied with drinking water for the first three days to overcome the transportation stress.

Experimental birds grouping

Sonali chickens were uniformly cared for in all groups during the study. Sixty Sonalis were divided into five groups to assess the impact of yeast (Saccharomyces cerevisiae) on Eimeria tenella infection, growth performance, blood parameters, and histopathology. The groups were as follows:

Group T0 (Negative control): Received a normal diet, water ad-libitum, and periodic (10 days interval) weight recordings without Eimeria tenella infection. Blood parameters, and histopathological examination were measured at the times when that of other groups were measured.

Group T1 (Positive control): Induced Eimeria tenella infection without antiprotozoal treatment.

Group T2: Induced Eimeria tenella infection, started treatment with yeast at a dose of 1.5g/kg feed after 72 hours.

Group T3: Sonali’s were supplied with Eimeria tenella protozoa after acclimatization to induce Eimeria tenella infection as like as T1 and T2 group. Induced Eimeria tenella infection, started treatment with yeast at a dose of 1.5g/kg feed after 72 hours.

Group T4: After acclimatization to induce Eimeria tenella infection, Sonalis of this group were supplied with Eimeria tenella protozoa as like as T1, T2 and T3 group. This group left about 72 hours for the establishment of Eimeria tenella infection. After that, we started our treatment with Amprium-Vet (Amprolium hydrocloride 20%-Square Pharmaceuticals Ltd). It was preserved in a dry place at room temperature and was administered at a dose rate of 1g per 3 liters of drinking water for 3-7 days, to compare the antiprotozoal effect with yeast.

Examination of feces for protozoal oocyst investigation

Feces were examined by two different types of qualitative tests; namely direct smear and flotation techniques to identify the morphological features of eggs, cysts, and oocysts (Hendrin and Robinson, 2006; Soulsby, 1982). The direct smear technique was conducted by mixing a drop of water with a few feces using an applicator stick on a glass slide, covered with a coverslip, and examined under a microscope (Figure 1A).

 

Parasite flotation was performed using a specific gravity solution (1.20) to exploit the density of parasite eggs. Five grams of feces were mixed with 5ml of saline, filtered, and centrifuged at 1500 rpm for five minutes. The sediment was mixed with 5ml of saturated sodium chloride, centrifuged at 2500 rpm for one minute, and then suspended with more saturated sodium chloride. After ten minutes without shaking, a coverslip was applied, transferred to a slide, and examined under a microscope.

Hematological parameters

Blood samples were collected from the wing vein of chicken of all groups at the age of 38 days and 58 days. The following parameters were observed:

a. Total Leukocyte Count (TLC) [Thousand/mm3]

b. Hemoglobin estimation (Hb) [mg/dl]

TLC and Hb were performed by Mindray BC-20 Auto Hematology Analyzer at Chatra Bazar, Frakkabad, Birol, Dinajpur.

Postmortem and Histopathological examination

Before and after treatment three chickens from each group were slaughtered to count the number of protozoa (Eimeria tenella) and to see if there were any pathological changes present (Figure 1B, C, D). Histopathological examinations were performed by Rotary Microtome (Tissue sectioning diameter 5 μm, Staining- Hematoxylin and Eosin (H and E), Microscopic examination 100x, 200x) at Veterinary Health Care, Satmatha, Rangpur.

Statistical analysis

Each experiment was repeated at least three times unless otherwise mentioned. The results were expressed as a ratio against each control as mean ± SEM. Single-factor analysis of variance (ANOVA) was used to analyze the statical differences where significances were considered at p<0.05. Furthermore, the Student-Newman-Keul’s test was used to compare the two groups. Differences were considered significant at the level of p<0.05.

Results and Discussion

Body weight

In (Table 1) showed that after post infection body weight of birds in different dietary treatments was statically significant (p<0.01). The effect of dietary supplementation of yeast showed statically significant (p<0.01) all over the experimental period. Among the different treatments of combination maximum body weight (743.17±3.55a gm) was observed under yeast at 1.5g/kg feed (T3) whereas the minimum body weight (595.66±2.76c) was observed under yeast at 1g/kg feed (T2). Meanwhile, the second-highest body weight (734.88±2.36a) was observed under amprolium (T4) with the recommended dose. In addition, the lowest body weight (585.52±2.76c) was found in the T1 group compared to other groups whereas the body weight (627.96±1.82b) of the T0 group was heavier rather than T1 and T2 groups.

 

Table 1: Effect of yeast at a different level and amprolium on body weight of Sonali chicken (10 days to 30 days post-infection).

Treatment	38 day	48 day	58 day
T0	422.40±1.02a	520.48±1.10a	627.96±1.82b
T1	402.70 ± 4.34b	491.24 ± 3.45b	585.52 ± 2.76c
T2	406.36 ± 3.82ab	513.00 ± 1.55a	595.66 ± 2.76c
T3	417.99 ± 1.35ab	525.61 ± 2.89a	743.17 ± 3.55a
T4	415.47 ± 5.41ab	515.11 ± 5.46a	734.88 ± 2.36a
P-value	0.016459*	0.000232***	0.000***

 

All values indicate mean ± standard error of the mean. The mean value with different superscripts within the same column is statistically Significant at a 1% level of significance. ***Significant at the 0.01 level.

 

Hematological parameter

Effect of yeast and amprolium on blood profile of sonali chicken infected with Eimeria oocysts given below:

Estimation of hemoglobin: All four experimentally infected groups revealed statistically highly significant decreases in hemoglobin (Hb) (p<0.01) when compared to the control group (10 days of post-infection, Table 2). Furthermore, chickens of T1, T2, T3, and T4 showed a highly significant reduction (p<0.01) in Hb values from 7.84 ± 0.37 to 9.51±0.30 in comparison to the control group (T0) on 10 days of post-infection. On the other hand, chickens of T0, T1, T2, T3, and T4 showed a statistically significant (p<0.05) increase in Hb values from 10.44±0.24 to 11.15±0.29 at 30 days of post-infection). Among all groups, the highest (11.15±0.29) Hb level was found in the T3 group and the lowest (10.15±0.20) in the T1 group.

Total leucocyte count (TLC)

All four experimentally infected groups revealed statistically highly significant (p<0.01) increases in total leucocyte count when compared to the control group (10 days of post-infection, Table 3). Furthermore, chickens of T1, T2, T3, and T4 showed a highly significant increase in TLC values from 10.64±0.33 to 12.86±0.46 in comparison to the control group (T0) on 10 days of post-infection. On the other hand, chickens of T0, T1, T2, T3, and T4 showed statistically significant (p<0.05) decreases in TLC values from 10.71±0.47 to 9.00±0.36 at 30 days of post-infection). Among all groups, the highest (10.71±0.47) TLC level was found in the T1 group and the lowest (9.00±0.36) in the T3 group.

 

Table 2: Estimation of hemoglobin (mg/dl) during the experimental period.

Treatment	38 Day	58 Day
T0	10.43 ± 0.34a	11.08 ± 0.28a
T1	7.84 ± 0.37b	10.15 ± 0.20a
T2	9.28 ± 0.23a	10.44 ± 0.24a
T3	9.53 ± 0.25a	11.15 ± 0.29a
T4	9.51 ± 0.30a	10.63 ± 0.29a
P-value	0.000***	0.049*

 

All values indicate mean ± standard error of the mean. The mean value with different superscripts within the same column is statistically Significant at a 1% level of significance. ***Significant at the 0.01 level.

 

Table 3: TLC (Thousand/mm3) during the experimental period.

Treatment	38 Day	58 Day
T0	8.70 ± 0.29c	9.35 ± 0.29ab
T1	12.86 ± 0.46a	10.71 ± 0.47a
T2	10.94 ± 0.34b	9.51 ± 0.33ab
T3	10.72 ± 0.36b	9.00 ± 0.36b
T4	10.64 ± 0.33b	9.35 ± 0.29ab
P-value	0.000***	0.014*

 

All values indicate mean ± standard error of the mean. The mean value with different superscripts within the same column is statistically Significant at a 1% level of significance. ***Significant at the 0.01 level.

 

Histopathology

In histopathological observation of lower intestine, Figure 1E: Intestinal sample obtained from birds based on basal diet (control negative group) showed absence of coccidial oocysts, no inflammatory infiltration and intestinal villi lined by simple columnar epithelium. Figure 1F: Intestinal sample obtained from infected non-treated birds (control positive group) showing thickening of intestinal lamina propia, presence of coccidial oocysts in lamina propia, degeneration, and loss of nuclei. Figure 1G: Intestine of infected birds treated with yeast (1g/kg) chickens, showing mild congestion of sub-mucosal blood vessels, fibrosis and degeneration of sub-mucosal glandular epithelium, and inflammatory infiltration. Figure 1H: Intestine of infected birds treated with yeast (1.5g/kg) showed no hemorrhagic lesions, no inflammatory infiltration, sloughing off intestinal villi slightly and no other distinct changes in the intestinal epithelium. Figure 1I: The infected birds treated with amprolium showed mild inflammatory infiltration of intestinal mucosa and congestion of blood vessels.

In our study, we aimed to highlight the anticoccidial effects of yeast compared to amprolium in inhibiting Eimeria tenella infection. We assessed efficacy based on mean body weight, blood analysis, and histopathological examination. The 35-day study indicates a decrease in body weight in group T1. There was significantly higher at 30 days post-infection bodyweight of sonali chicken than the positive control group. A similar finding was also observed by (Lee et al., 2007; Priya and Babu, 2013; Hana et al., 2015) where significant differences (p<0.05) indicated that feeding yeast to chicks improves body weight gain and feed/gain ratio. The obtained results confirmed the previous findings of several researchers (Zhang et al., 2005; Paryad and Mahmoudi, 2008) that yeast supplementation in broiler ration had a significant effect on body weight gain and feed conversion ratio. Shankar et al. (2017) and Mulatu et al. (2019) also reported that Saccharomyces cerevisiae improved feed/gain ratio and body weight gain. The present study revealed that baker’s yeast supplementation had a positive effect on the bodyweight of sonali chickens in T2 and T3 for the whole trial period which was similar to Islam (2022). Compared with the groups T0, T1, and T4. Kabir (2009) stated that the yeast acts by (i) maintaining normal intestinal microflora by competitive exclusion and antagonism (ii) altering metabolism by increasing digestive enzyme activity and decreasing bacterial enzyme activity and ammonia production (iii) improving digestion, and (iv) stimulating the immune system.

Decreased mean hemoglobin (Hb) concentration had been recorded in infected groups T1, T2, T3, and T4 in comparison with the control group on 10 days post-infection. The research findings are in accordance with Ogbe et al. (2010) and Matthew et al. (2022) who were reported a decrease in the Hb counts in E. tennela infected chickens. This could be attributed to the reduction of blood components due to hemorrhage caused by the disease. These findings are similar to Razzaq et al. (2003). A significant increase in the mean Hb values was observed in all treated groups when compared with the control group on 30 days post-infection. A similar finding was also observed by Mulatu et al. (2019) and Mares et al. (2023). This indicates that there was a dietary effect of yeast on Hb.

Increased mean total leucocyte count (TLC) concentration had been recorded in infected groupsT1, T2, T3, T4 in comparison with the control group on 10 days post-infection. The study findings are in accordance with Hirani et al. (2018) who reported an increase in the TLC counts in E. tennela infected chickens. A significant increase in TLC has also been reported by Padmavathi and Muralidharan (1986), Panda et al. (1997), Kumar and Padmavathi (2000), Jaipurkar et al. (2004), and Ogbe et al. (2010). Adamu et al. (2013) and Shekhar et al. (2020) also reported similar findings of higher TLC with increased numbers of lymphocytes, eosinophils, and heterophils in E. tenella and E. brunetti infected broiler. In this study chickens (Sonali) of T1, T2, T3, and T4 showed a highly significant increase in TLC values from 10.64 ± 0.33 to 12.86 ± 0.46 in comparison to the control group (T0) on 10 days of post-infection. The increased total leucocytes count in coccidia-affected birds might be due to suppressed body immune system from the infection (Hirani et al., 2018; Stephen, 1965). This increase was suggestive of increased leucopoiesis as the first step of defense mechanism to infection (Padmavathi and Muralidharan, 1986). A significant decrease in the mean TLC values was observed in all treated groups when compared with the control group on 30-day post-infection. These findings clearly indicated the ability of yeast to produce immunity against coccidiosis. Yitbarek et al. (2012), Shanmugasundaram and Selvaraj (2012) stated that the whole yeast cell is a good source of mannan oligosaccharides and β-glucans known for their immune-modulatory effects. Lee et al. (2007), also stated that Saccharomyces-based probiotic, when included in the diet may improve the resistance against E. tenella and E. acervulina by enhancing humoral immune response as well as reducing oocyst production in birds.

Conclusions and Recommendations

In conclusion, the current experimental findings strongly support the notion that yeasts exhibit a notable anticoccidial effect against Eimeria tenella in Sonali chickens, concurrently influencing the growth performance of the poultry. Given the observed drug resistance and adverse effects associated with amprolium, yeast emerges as a preferable alternative. The absence of adverse effects at dosage rates of 1g/kg and 1.5g/kg feed further underscores the safety profile of yeast during this study. This suggests that yeast could offer a promising therapeutic approach against E. tenella, considering its accessibility and efficacy. The potential of yeast as a viable agent, particularly in the face of multidrug-resistant species like Eimeria tenella, warrants further consideration and exploration in future research endeavors.

Acknowledgments

The authors are really thankful to the Department of Physiology and Pharmacology, HSTU, Bangladesh, for the laboratory support to this work.

Novelty Statement

This helps to combat against multidrug-resistant Eimeria tenella. Through this work we tried to focus on Eimeria tenella infection by using yeast and amprolium as a potential anticoccidial and evaluate their effeteness. Finally, this research helps to reduce coccidial treatment cost in sonali chicken as well as creating public awareness against coccidiosis.

Author’s Contribution

Conceptualization: MMMP, RI, MS. Research work: MAA, MMMP. Draft writing: MAA, MMMP, MS, RI, MBR. Data arrangement, reviewing and final manuscript writing: MS. All authors reviewed the final version of the manuscript.

Ethical statement

This research work was approved based on University laws and National Law especially following the Bangladesh Veterinary Council (BVC) laws. In this study ethical approval for the use of Pigeon was granted by the Institute of Research and Training (IRT), and Department of Physiology and Pharmacology (Memo No: HSTU/VAS/PPH/1352), Faculty of Veterinary and Animal Science Hajee Mohammad Danesh Science and Technology University, Dinajpur-5200, Bangladesh.

Funding

This research did not receive any specific grant from funding agencies.

Conflict of interest

The authors have no potential conflicts of interest.

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