Special Issue:

Emerging and Re-emerging Animal Health Challenges in Low and Middle-Income Countries

A Diagnostic and Immunological Study of the Sarcocystosis in Bovine

Zahraa Sadoon Hadi1*, Mohammed Hassan Flaih2, Hamssa Majid Mustafa3, Ali Esmail Al-Snafi4

1Department of Biology, College of Education for Women, University of Shatrah, Iraq; 2Department of Nursing Techniques, Nasiriyah Technical Institute, Southern Technical University, Nasiriyah, Iraq; 3Department of Medical Laboratory Techniques, Shatra Technical College, Southern Technical University, Shatra, Thi-Qar, Iraq; 4Department of Pharmacology, College of Medicine, University of Thi-Qar, Iraq.

Received | August 20, 2025; Accepted | October 01, 2025; Published | October 16, 2025

*Correspondence | Zahraa Sadoon Hadi, Department of Biology, College of Education for women, University of Shatrah, Iraq; Email: [email protected]

Citation | Hadi ZS, Flaih MH, Mustafa HM, Al-Snafi AE (2025). A diagnostic and immunological study of the Sarcocystosis in bovine. J. Anim. Health Prod. 13(s1): 623-627.

DOI | https://dx.doi.org/10.17582/journal.jahp/2025/13.s1.623.627

ISSN (Online) | 2308-2801

Copyright: 2025 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

Members of the genus Sarcocystis, an intracellular protozoal parasite in the phylum Apicomplexa that is extensively found in mammals, birds, and reptiles, are responsible for causing sarcocystosis (Maky and Mohammed, 2021). Sarcocystis has a number of stages, the oocyst stage contains two sporocysts. Depending on the parasite type, sarcocyst stages vary in size and shape; some are tiny (S. cruzi), while others are gigantic (S. gigantea, S. muris) (Ondrej and David, 2022). These parasites go through two hosts in their life cycle: The definitive host’s intestines create the sexual stage, while the intermediate host’s different tissues host asexual reproduction. Sarcocystis typically finds definite hosts in carnivores and omnivores. A few organisms can develop in more distantly related animals (e.g., cats and dogs), but most parasites are believed to use one or a group of closely related hosts. Mammals, marsupials, birds, reptiles, and perhaps fish can all serve as intermediate hosts. They are frequently omnivores or herbivores. It appears that certain Sarcocystis species are more host-specific than others (Chiesa et al., 2013; Obijiaku et al., 2013). Sarcocystis species occasionally cause myositis, encephalitis, and other illnesses in humans or animals, although the majority of infections appear to be asymptomatic. The dose of the parasites is believed to be one determinant, though it is unclear why some infected people get sick while others stay healthy (Dubey and Rosenthal, 2023; Howe et al., 2018). Worldwide, Sarcocystis infections in cattle are common. The species of Sarcocystis that cause microscopic infection and are spread by the Canidae family are more pathogenic, the parasite toxins lead to fever, anemia, loss of appetite, weight loss, wool loss, decreased milk production, acute myopathy, and central nervous signs such as ataxia, paralysis, muscle tremors that lead to abortion and sometimes death (Rubiola et al., 2021).

The identity of Sarcocystis species in cattle is a topic of recent discussion. Sarcocystis species must be properly diagnosed in order to be evaluated for their significance to the economy and public health (Dubey and Rosenthal, 2023). Our study aimed to evaluate the incidence of bovine sarcocystosis in Nasiriyah province as an effort to provide foundational information for mitigation of the infection.

Materials and methods

Sample collection and processing

This study was carried out in various locations within the Thi-Qar province between 2023 and 2024. A total of 180 blood samples from bovine were chosen at random and 25 blood samples from health bovine. Using anticoagulant-free sterile tubes, jugular vein blood samples were drawn, put on ice, and then brought to the lab. Following a 15-minute centrifugation at 3,000 rpm, the serum was extracted from the coagulated blood and kept at -20 °C for further examination.

Diagnosis of Sarcocystis

Following the manufacturer’s instructions, the Bovine Sarcocystis ELISA kit (SunLong Biotech Co., LTD, China) was used to qualitatively determine the presence of Sarcocystis in all bovine serum samples. Test efficacy for every specimen: Positive control’s average value was greater than 1.00, while negative control’s average value was less than 0.10. Calculating the crucial value (CUT OFF) involves multiplying the average value of negative control by 0.15. Negative samples for bovine Sarcocystis were concluded if the OD value was less than CUT OFF. Positive determination of the sample for bovine Sarcocystis when the OD value is more than or equal to CUT OFF.

Estimated of IL-6

The Bovine IL-6 (Interleukin 6) ELISA Kit (Reed Biotech Ltd, RE3186B, China) was used to estimate the levels of IL-6.

Calculation of results include following analysis. For each standard and sample, the average duplicate readings were subtracted from the average zero standard optical density. Drawing a logistic curve with four parameters on log-log graph paper, use OD values on the y-axis and standard concentration on the x-axis. Samples were retested using the proper dilution if its optical density (OD) exceeds the upper limit of the standard curve. The computed concentration times the dilution factor is the actual concentration.

Biochemical tests

Following the manufacturer’s instructions, a commercially available kit (Biosystem Laboratory Products LTD, Spain) was used to measure various serum biochemical parameters (Urea, Creatinine, ALT, AST, ALP).

Statistical analysis

The Student’s t test (SPSS, version 26) was used to assess the differences between the groups. The proportions were compared using the Chi-square test. The outcome was deemed significant if the p-value was 0.05 or below.

Results

As shown in Table 1, the incidence of the infection among 180 tested bovine was 26.67%, which include 48 infected animals while the infection was not detected in 132 of the tested animals 73.33%.

 

Table 1: Infection rate in infected bovine with Sarcocystosis using ELISA.

Total samples	Positive sample	Negative sample	%	Sig.
180	48	132	26.67	P ˂ 0.05

 

The results showed an effect of parasitic infection in infected bovine with Sarcocystosis on elevated the level of serum IL-6 (44.31 Pg/ml), and was significantly higher (P ˂ 0.05) compared with its level in control animals (8.36 Pg/ml) Table 2.

 

Table 2: The serum level of IL-6 in infected bovine with Sarcocystosis compared to control samples.

Samples	IL-6 Pg/ml	P-value
Infected(48)	8.36±64.36	0.007
Control(25)	44.31±7.880

 

Examining of the effect of the parasitic infection on the liver and kidney functions in infected bovine with Sarcocystosis, showed that the infection didn’t significantly affected the levels of ALP (72.08±9.375), AST (13.88±7.343), ALT (10.40±5.615), creatinine (0.716±0.1901) and urea (32.94±13.90) compared to its level in control animals Table 3.

Discussion

Animal emaciation caused by the common intracellular protozoan parasite Sarcocystis species has a significant financial impact on livestock. decreased weight gain, decreased quality and quantity of meat, milk, and wool, transboundary restriction, condemnation of infected

 

Table 3: The serum level of ALP, AST, ALT, Creatinine and Urea in the infected bovine with Sarcocystosis compared to control samples.

Samples	ALP (IU/l)	AST (IU/l)	ALT (IU/l)	Creatinine (mg/dl)	Urea (mg/dl)
Infected (48)	72.08±9.375	13.88±7.343	10.40±5.615	0.716±0.1901	32.94±13.90
Control (25)	78.10±14.53	16.12±4.961	12.07±3.731	0.741±0.1908	33.43±11.71
p-value	0.0432	0.1816	0.1906	0.6124	0.8787

 

carcasses, abortion and even death (Hussein et al., 2025; Fayer, 2004; Saleque et al., 1990). In this study, naturally infected bovine samples were used to examine the possible impacts of Sarcocystis infection on inflammation as well as liver and kidney function.

According to estimates, almost 90% of cows worldwide were thought to be infected with Sarcocystis. China and Malaysia both reported the same percentage (Tappe et al., 2014; Yang et al., 2018). Using the tissue expansion method, 94.8% of the bovines under study in Isfahan, Iran, were infected (Fayer, 2004). Infection rates in Tabriz, Iran, were almost same (Shekarforoush et al., 2005). However, almost 90% of the cows analyzed in Australia had Sarcocystis infections, In South Moravia, 75-93% of the examined cows were infected (Pena et al., 2001). Infection was recorded in 78% of the examined cattle in new valley governorate- Egypt (El-Mahdi et al., 2023).

In our study, 48 bovines were infected out of 180 (26.67%) tested, the reason of the low percent of the infection cold be attributed to using of Sarcocystis ELISA which is more accurate diagnostic test. Furthermore, the majority of these studies were carried out on the slaughtered bovine with high contamination rate reaching to 75-93% by the digestion method in bovine (Pena et al., 2001).

The serum level of IL-6 was significantly elevated in bovine infected with Sarcocystis. In response to infections and tissue damage, the soluble cytokine mediator interleukin 6 (IL-6) is quickly and temporarily produced. In the early stages of inflammation, IL-6 is generated in a local lesion and then circulated to the liver, where it rapidly initiates the production of a wide range of acute phase proteins, such as fibrinogen, haptoglobin, serum amyloid A (SAA), C-reactive protein (CRP), and α1-antichymotrypsin. It supports host defense by encouraging hematopoiesis, immunological responses, and acute phase responses (Tanaka et al., 2014). 

Many studies showed elevated IL-6 in Trypanosoma, Trichomonas and Toxoplasma infections and suggested that IL-6 mediates anti-parasite protective responses (Al- Hadraawy et al., 2014; Gao and Pereira, 2002). An infection with Sarcocystis is a major contributing factor to a number of serious inflammatory conditions, including intestinal infections, hepatitis, encephalitis, and encephalomyelitis (Fayer, 2004; AbuBakar et al., 2013). Infection with Sarcocystis camelicanis increased the expression of IL-6 in many organs of the infected animals (Metwally et al., 2021).

Fresh Sarcocystis extract significantly raised the levels of several cytokines, including IL-1β, IL-18, IL-15, IL-12, IL-10, and IL-13, in rats used in experimental infection (Badr et al., 2020).

During protozoan infections, the immune system generates a number of defense mechanisms, such as innate immune responses to extracellular protozoan parasites by immune cells like natural killer (NK) cells, neutrophils, and macrophages. Both innate and adaptive immune responses to intracellular parasites depend on the cytokines generated by natural killer cells and activated macrophages (Melby et al., 2019).

Our result showed that Sarcocystis didn’t significantly change hepatic and renal functions. However, the effect of the parasite on the values of these tests depend on the severity and distribution of the parasite, When the parasite invaded liver, it cased hepatocellular injury represented by hepatocytic and sinusoidal necrosis and alterations. The hepatocyte nuclei showed a pleomorphic appearance with dilatation of the core veins and pale-violet staining. The border architecture of the central vein and sinusoids was uneven and often looked torn. When it entered the kidney, a part of the renal cortex showed highly damaged renal corpuscles with a destroyed glomerulus, dilated lumen, proximal and distal convoluted tubules that were twisted and dilated, and leukocyte infiltration. These histological changes associated with alterations of liver and kidney function tests (El-Mahdi et al., 2023).

Conclusions and Recommendations

This study evaluated the incidence of bovine sarcocystosis in Thi-Qar province. The incidence of the infection reached 26.67%, most of the cases were subclinical. Sarcocystosis species must be properly diagnosed in order to be evaluated for their significance to the economy and public health. Researchers recommend conducting more molecular studies and studying the histological effects on animals infected with the Sarcocystis parasite.

ACKNOWLEDGEMENT

The authors sincerely thank the Research Laboratories, Department of Biology, College of Education for Women, University of Al-Shatrah, Iraq, for their infrastructure and technical support. We also thank the staff of the Veterinary Hospital in Shatrah and Nasiriyah, Iraq, for their assistance in sample collection.

NOVELTY STATEMENT

The novelty of our research is that it includes studying the effect of parasitic infection in bovine, which are considered an important source of human food, with few studies in Thi-Qar Governorate that include the effect of this infection on Immunological and biochemical parameters in infected bovine.

AUTHOR’S CONTRIBUTION

ZSH: Preparing the research idea, collecting samples, parasites diagnosed and writing the research. Immunological and biochemical parameters work.

MHF: Preparing and providing the necessary materials to complete the immunological parameters work.

HMM: Preparing and providing the necessary materials to complete the biochemical parameters work.

AEA-S: Preparing the research idea, writing the research, supervising it and analyze the results.

Generative AI and AI-assisted technology statement

In this manuscript, AI was not used.

Conflict of interest

The authors have declared no conflict of interest.

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