Rewaida Abdel-Gaber
Shurug Albasyouni1
Simeon Santourlidis
Esam Al-Shaebi- 1Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia
- 2Epigenetics Core Laboratory, Institute of Transplantation Diagnostics and Cell Therapeutics, Medical Faculty, Heinrich-Heine University Düsseldorf, Düsseldorf, Germany
Background: Coccidiosis, caused by Eimeria species, is a major enteric disease in birds, with Eimeria labbeana-like isolates frequently inducing severe intestinal lesions, diarrhea, and reduced weight gain in pigeons. Conventional anticoccidial drugs face limitations due to resistance, residue concerns, and environmental impact, highlighting the need for alternative strategies. Commiphora myrrha (myrrh) is a resinous plant extract rich in bioactive compounds with antioxidant, antimicrobial, antiparasitic, and anti-inflammatory properties. This study evaluated the protective effects of C. myrrha resin in pigeons experimentally infected with E. labbeana-like isolates.
Methods: Resin of C. myrrha was collected from Riyadh, Saudi Arabia, authenticated, and extracted with 70% methanol to prepare a crude extract (MyE). Its chemical composition was characterized using GC–MS. A laboratory strain of Eimeria labbeana-like oocysts was propagated in pigeons, sporulated, and used for experimental infection. Twenty-five pigeons were randomly assigned to five groups: uninfected control, uninfected + myrrh extract (MyE), infected control, infected + MyE, and infected + amprolium (standard drug). MyE and amprolium treatments were administered orally for 5 days post-infection. Parasitological, histological, immunohistochemical (NF-κB and IFN-γ), gene expression (MUC2, IL-1β, IL-10, IFN-γ, and TNF-α), and cytokine (IL-10 and TNF-α) analyses were conducted.
Results: In this study, myrrh resin was methanol-extracted and characterized by GC–MS, revealing 29 phytochemical components. Experimental infection of pigeons with E. labbeana-like oocysts resulted in peak fecal oocyst shedding (~5.25 × 105 oocysts/g.feces), extensive development of intracellular parasite stages (meronts, gamonts, and developing oocysts), a marked reduction in goblet cell numbers, and elevated intestinal inflammatory responses, including increased NF-κB and IFN-γ immunoreactivity, as well as upregulated mRNA expression of IL-1β, IL-10, IFN-γ, and TNF-α. Oral administration of MyE significantly suppressed oocyst shedding by 60%, reduced the number of intracellular parasitic stages, restored goblet cell counts, and downregulated both gene and protein levels of pro-inflammatory markers while enhancing MUC2 expression, indicating effective modulation of Eimeria-induced intestinal damage and inflammatory dysregulation.
Conclusion: These findings demonstrate that C. myrrha extract effectively mitigates Eimeria-induced intestinal damage, inflammation, and immune dysregulation, highlighting its potential as a natural, plant-based intervention for managing pigeon coccidiosis.
Introduction
Coccidiosis is a major enteric disease in birds, primarily caused by protozoan parasites of the genus Eimeria (1–4). All Eimeria species exhibit a complex life cycle with schizogony, gametogony, and sporogony phases (5). The infectious forms are fully sporulated oocysts, which invade the gastrointestinal mucosa, leading to intestinal lesions, dehydration, and bloody diarrhea (6, 7). Disease progression triggers inflammatory and oxidative responses that exacerbate tissue damage and impair bird performance (8). In pigeons, Eimeria labbeana (or E. labbeana-like isolates) is frequently associated with severe intestinal pathology and weight loss (2, 8–11). These infections also increase susceptibility to secondary pathogens, further compromising bird health (7, 12, 13).
Traditional management of avian coccidiosis relies heavily on synthetic anticoccidial drugs and ionophores (5). However, long-term use has contributed to the emergence of resistant Eimeria strains, and concerns remain regarding drug residues in animal products, environmental impacts, and consumer demand for residue-free meat and eggs (14–16). These limitations highlight the need to evaluate alternative or complementary approaches. Nutritional strategies, improved management practices, and phytotherapeutics have shown promise in supporting gut health, enhancing immunity, and reducing dependence on conventional chemotherapeutics (16–18). While anticoccidial drugs remain widely used as reference treatments in experimental studies, their effectiveness may be compromised by resistance, and their practical utility in long-term, sustainable poultry management is limited.
Commiphora myrrha, commonly known as myrrh, is a resinous substance obtained from the bark of trees native to Africa, the Middle East, and Asia (19). Its therapeutic properties have been recognized in traditional medicine for centuries (20). Phytochemical studies reveal that myrrh contains terpenoids, sesquiterpenes, and other bioactive secondary metabolites, which exhibit antioxidant, antimicrobial, antiparasitic, and anti-inflammatory effects (21, 22). These properties suggest myrrh could modulate the inflammatory and oxidative pathways activated by Eimeria infection, potentially limiting tissue damage and restoring physiological balance. Despite growing interest in its bioactivities, experimental evidence of myrrh’s efficacy against avian coccidiosis, particularly E. labbeana-like infections in pigeons, remains limited. Recent studies have highlighted its antimicrobial and immunomodulatory potential in avian models, supporting its use as a natural prophylactic or adjunct therapy (23, 24).
This study investigates the potential of C. myrrha extract to protect pigeons against E. labbeana-like infection by (i) reducing intestinal inflammation, (ii) preserving mucosal architecture and barrier integrity, and (iii) modulating immune responses. By assessing parasitological, histopathological, biochemical, and immunological parameters, we aim to evaluate myrrh as a sustainable, plant-based alternative to conventional anticoccidial drugs, offering practical relevance in modern pigeon management.
Materials and methods
Plant collection and extract preparation
Commiphora myrrha (myrrh) resin was sourced from a local market in Riyadh, Saudi Arabia, and its identity was authenticated at the Botany Department Herbarium, King Saud University, where a voucher specimen was deposited (KSU-23539). For comparison, amprolium (AMP) (Amproxine 20%, water-soluble powder) was obtained from Gulf Veterinary Pharmacy, Riyadh, and used as a standard anticoccidial drug.
Following the method of Akande et al. (25), 100 g of resin was ground into a fine powder using a Hummer Grinder (Edison Electric, ED-CG1400, China). The powdered material was macerated in 1,000 mL of 70% methanol with gentle agitation at room temperature for 24 h. The extract was filtered through Whatman No. 1 filter paper, and the filtrate was concentrated using a Buchi rotary evaporator (Switzerland) at 45 °C. The resulting methanolic crude myrrh extract (MyE) was then reconstituted in distilled water (w/v) to prepare the required experimental doses.
Gas chromatography-mass spectrometry analysis
Sample analysis was performed using an Agilent Technologies 7890B GC-MS system (Santa Clara, CA, USA) equipped with an autosampler. A 1 µL aliquot of the extract was injected, and compounds were identified using the NIST-MS library database, following the method of Kanthal et al. (26). Separation was achieved on a DB-5 MS capillary column (Agilent Technologies). Helium served as the carrier gas at a constant flow rate of 1 mL/min. The injector temperature was set at 250 °C, operating in split mode (1:50). The oven program ranged from 50 °C to 250 °C, with a total run time of 61 min. Mass spectrometric conditions included an electron ionization source at 230 °C, a mass scan range of 40–500 g/mol, a scan rate of 1.56, and a solvent delay of 4 min.
Parasite strain and preparation
A laboratory strain of Eimeria labbeana-like, a representative coccidian parasite, was utilized in this experiment. The parasite used in this study was identified based on the morphological, morphometric, and molecular data previously established in our published work, “Morphology, morphometry, and phylogeny of the protozoan parasite, Eimeria labbeana-like (Apicomplexa, Eimeriidae), infecting Columba livia domestica”. To propagate the oocysts, five pigeons were used as hosts. Following the method described by Qudoos et al. (27), each bird was orally inoculated with 3×104 sporulated oocysts. Shedding of oocysts was observed in the feces of infected pigeons on the eighth day post-inoculation. The recovered oocysts were subsequently induced to sporulate in 2.5% (w/v) potassium dichromate (K2Cr2O7) at 24 °C, as outlined by El-Ashram and Suo (28). The sporulated oocysts were then centrifuged in phosphate-buffered saline (PBS) at 2500 rpm for 5 minutes and rinsed three times with distilled water.
Pigeons, housing, and experimental design
Twenty-five white domestic pigeons (Columba livia domestica), weighing between 300–380 g, were purchased from the local animal market in Riyadh, Saudi Arabia. The birds were acclimatized for one week under controlled conditions, including a 12-hour light/dark cycle and a temperature of 23 ± 5 °C, with free access to tap water and a balanced seed-based diet. They were maintained at the animal facility of the Department of Zoology, College of Science, King Saud University, Riyadh.
The pigeons were randomly assigned to five groups (five birds each) as follows:
● Group 1 (control): fed a basal diet with tap water only.
● Group 2 (non-infected treated): received MyE (500 mg/kg) without infection (29).
● Group 3 (infected control): inoculated with E. labbeana-like oocysts without treatment.
● Group 4 (infected + MyE): administered MyE (500 mg/kg) following infection with E. labbeana-like oocysts (29).
● Group 5 (infected + amprolium): treated with the anticoccidial drug amprolium (1 g soluble powder/L of water) after infection (29).
All birds in Groups 3–5 were orally inoculated with 3×104 sporulated E. labbeana-like oocysts, as described by Qudoos et al. (27). Treatments with MyE and amprolium began on day 3 post-infection (p.i.) and continued daily for five consecutive days. On day 8 p.i., fresh fecal samples from the infected, untreated, and treated groups were collected. Oocyst counts were determined using flotation with saturated saline, followed by McMaster’s counting technique (30). Additionally, the suppression of oocyst shedding was calculated as follows:
Sample collection
At the end of the experiment, pigeons were humanely sacrificed in compliance with ethical guidelines, and the small intestine was carefully dissected. The collected samples were preserved in two ways: (i) portions of intestinal tissue were fixed in 10% neutral buffered formalin (NBF) for histological and immunohistochemical (IHC) analysis, and (ii) other portions were stored in RNA later® solution (Qiagen) at –80 °C for subsequent mRNA and protein expression studies.
Histological examination and parasitic scoring
Intestinal samples were fixed in 10% neutral buffered formalin (NBF) for 24 hours, dehydrated, embedded in paraffin, and sectioned at a thickness of 5 µm, following the method of Adam and Caihak (31). Sections were stained with hematoxylin and eosin (H&E) to identify parasite stages in both infected and treated groups. In contrast, additional sections were stained with Alcian blue to assess goblet cells as described by Adam and Caihak (31). Microscopic examination and imaging were performed using an Olympus B×61 microscope (Tokyo, Japan), and parasite stages were quantified across ten well-oriented villous-crypt units (VCUs).
Immunohistochemical analysis
Intestinal tissues fixed in formalin were processed by dehydration in graded ethanol, cleared with xylene, embedded in paraffin, and sectioned at 5 µm thickness using a microtome. For IHC labeling, sections were deparaffinized, rehydrated, and treated with 3% H2O2 for 5 minutes to block endogenous peroxidase activity. They were then pre-incubated with a normal serum buffer (Diagnostic BioSystems, Serpentine, CA, USA) for 30 minutes, followed by incubation at 4 °C for 3 hours with primary antibodies against Nuclear Factor kappa B (NF-κB) and Interferon-gamma (IFN-γ) (Santa Cruz Biotechnology, CA, USA).
Subsequently, sections were treated with a biotinylated secondary antibody and streptavidin–horseradish peroxidase conjugate (Vision Biosystems Novocastra, UK) according to the kit protocol. Detection was carried out using 3,3′-diaminobenzidine (DAB) substrate (Vision Biosystems Novocastra), and counterstaining was performed with hematoxylin and eosin (Sigma Chemical Co.). Between each step, sections were rinsed with immunowash buffer (Vision Biosystems Novocastra). Finally, tissues were dehydrated in graded ethanol, cleared in xylene, and mounted with glass coverslips. All slides were examined for NF-κB and IFN-γ expression and photographed using an Olympus BX61 microscope (Tokyo, Japan).
RNA extraction and qRT-PCR
Total RNA was isolated from intestinal tissues using Trizol reagent (Invitrogen, USA). To remove potential genomic DNA contamination, samples were treated with DNase (Applied Biosystems, Darmstadt, Germany) for at least one hour. Complementary DNA (cDNA) was then synthesized using a reverse transcription kit (Qiagen, Hilden, Germany) following the manufacturer’s instructions.
Quantitative real-time PCR (qRT-PCR) was performed on an ABI Prism 7500HT detection system (Applied Biosystems, Darmstadt, Germany) using SYBR Green PCR master mix (Qiagen, Hilden, Germany). Gene expression analysis targeted MUC2 (goblet cell-related response), interleukin (IL)-1β, IL-10, interferon gamma (IFN-γ), and tumor necrosis factor alpha (TNF-α), with β-actin serving as the housekeeping reference gene. All primers were obtained from Qiagen (Hilden, Germany) (Table 1).
Table 1. Oligonucleotide primer sequences for reverse transcription PCR amplification in the experiment.
The qRT-PCR amplification protocol consisted of an initial denaturation at 95°C for 10 min, followed by 40 cycles of 95°C for 15 s and 60°C for 60 s. A melting curve analysis was performed from 60°C to 95°C to verify the specificity of the amplified fragments. Amplification and data analysis were conducted using the Bio-Rad iMark Microplate Reader (SW 1.04.02.E). Relative gene expression levels were calculated using the comparative Ct method (2−ΔΔCT) as described by Livak and Schmittgen (32).
Sandwich enzyme-linked immunosorbent assay
Intestinal levels of IL-10 (MBS9364117) and TNF-α (MBS050796) were measured using commercial ELISA kits (MyBioSource, San Diego, CA, USA) following the manufacturer’s instructions. Briefly, standards of known concentrations were prepared to generate a standard curve ranging from 0 to 500 pg/ml. Samples and standards were added to the microplate wells and incubated as recommended. After washing to remove unbound material, a specific detection antibody was added, followed by a substrate solution. Optical density (OD) readings were measured at 450 nm using a Bio-Rad iMark Microplate Reader (SW 1.04.02.E). Cytokine concentrations in the samples were calculated from the standard curve and expressed in pg/ml.
Statistical analysis
Data are expressed as mean ± standard deviation (SD). Group differences were evaluated using one-way analysis of variance (ANOVA) followed by Duncan’s multiple range test. Analyses were conducted using SPSS version 18 for Windows (SPSS Inc., Chicago, IL, USA). Statistical significance was indicated in figures using the symbols (*) and (#) at p ≤ 0.05. Detailed descriptions of these symbols are provided in the figure legends.
Results
The analysis performed with GC-MS showed that the methanolic extract contains 29 phytochemical components that may be responsible for its activity (Table 1). The characterization of the extract’s components was carried out using their retention time (RT), molecular weight (MW), molecular formula (MF), and peak area with its percentage (%). These compounds are listed in Table 2 according to their RTs.
Table 2. Identification of phytochemical compounds by GC-MS in MyE.
On day 8 after infection, the peak shedding of fecal oocysts was observed in the infected, untreated group, reaching approximately 5.249 × 105 ± 3.13 × 104 oocysts per gram of feces. After treatment interventions, there was a significant reduction in oocyst output, with a 60.16% decrease compared to the infected group (Figure 1). Experimental infection of pigeons with Eimeria oocysts led to the development of various parasite stages (meronts, gamonts, and developing oocysts) in the intestinal epithelial cells, as seen in the H&E-stained sections (Figure 2). The number of intracellular parasitic stages was significantly lower in pigeons treated with MyE, decreasing from 72.91 ± 8.81 in the infected group to 16.99 ± 5.84 stages per 10 VCU, compared to the reference drug at 5.66 ± 1.15 stages per 10 VCU (Figure 3).
Figure 1. Suppression of Eimeria labbeana-like oocysts in infected and infected-treated pigeons. *p ≤ 0.05, indicates a significant difference compared with the infected group. (MyE, myrrh extract; AMP, amprolium).
Figure 2. Histological sections of pigeon intestines across experimental groups. (A) Intestinal tissue from E. labbeana-like infected pigeons showing an elevated number of parasite stages. (B, C) Tissues from infected pigeons treated with MyE and AMP, respectively, showing reduced parasite stages. Scale bar = 50 µm (MyE, myrrh extract; AMP, amprolium).
Figure 3. Effects of treatment with 500 mg/kg MyE and 1 g/L H2O AMP on the total number of Eimeria labbeana-like parasitic stages in the intestinal tissue per 10 VCU on day 8 p.i. *p ≤ 0.05, indicates a significant difference compared with the infected group. (MyE, myrrh extract; AMP, amprolium).
Microscopic examination of intestinal sections stained with alcian blue (Figure 4) showed significant histological changes after pigeons were infected with Eimeria. Specifically, there was a marked decrease in goblet cell numbers, with an average of 6.69 ± 0.25 goblet cells per VCU in the infected group. In contrast, the control group had a higher average of 14.74 ± 0.18 goblet cells/VCU (Figures 4, 5). When infected pigeons were treated with MyE, the number of goblet cells increased significantly, reaching an average of 11.79 ± 0.09 goblet cells/VCU.
Figure 4. Small intestinal sections illustrating goblet cells in (A) control group, (B) non-infected pigeons treated with 500 mg/kg MyE, (C) infected group, (D) infected pigeons treated with 500 mg/kg MyE, and (E) infected pigeons treated with 1 g/L H2O AMP. Sections were stained with Alcian Blue and counterstained with eosin, and goblet cells were quantified in 10 well-oriented villus-crypt units (VCU). Scale bar = 100µm.
Figure 5. Variation in the number of intestinal goblet cells within villi among the control, non-infected treated with 500 mg/kgMyE, infected, and infected-treated groups (500 mg/kg MyE and 1 g/L H2O AMP). *p ≤ 0.05, indicates a significant difference compared with the control group, #p ≤ 0.05, indicates a significant difference compared with the infected group. (MyE, myrrh extract; AMP, amprolium).
Intestinal sections from various experimental groups underwent immunohistochemical staining specifically to identify and quantify NF-kB (Figure 6) and IFN-γ (Figure 7) positive cells. In the control group, the number of positive cells was within normal physiological levels. However, the results showed that Eimeria infection caused a strong immunoreactivity to NF-kB and IFN-γ-positive intestinal cells compared to the control pigeons. After administering MyE, a significant decrease in immunoreactivity to NF-kB and IFN-γ in the intestinal tissues was observed in the intestinal sections of pigeons affected by Eimeria infections.
Figure 6. Immunohistochemical detection of NF-KB in the intestinal tissues of pigeons. (A) control group, (B) non-infected group treated with 500 mg/kg MyE, (C) infected group, (D) infected group treated with 500 mg/kg MyE, and (E) infected group treated with amprolium. Scale Bar = 100 µm.
Figure 7. Immunohistochemical detection of IFN-γ in intestinal tissues of pigeons. (A) Control pigeon group, (B) non-infected group treated with 500 mg/kg MyE, (C) infected group, (D) infected group treated with 500 mg/kg MyE, and (E) infected group treated with amprolium. Scale Bar = 100 µm.
qRT-PCR was used to measure the mRNA levels of inflammatory cytokines in pigeon intestinal tissues (Figures 8, 9). A notable decrease in MUC2 gene expression was seen after Eimeria infection. Treatment with MyE significantly boosted MUC2 expression, increasing from 0.16 to 1.74-fold (Figure 8).
Figure 8. Effect of MyE treatment on MUC2 mRNA expression in intestinal tissues from E. labbeana-like infected pigeon. Expression values determined by RT-PCR were normalized to β-actin and presented as fold change (log 2 scale) relative to the control group. *p ≤ 0.05, indicates a significant difference compared with the control group, #p ≤ 0.05, indicates a significant difference compared with the infected group. (MyE, myrrh extract; AMP, amprolium).
Figure 9. Effect of MyE treatment on the mRNA expression of (A) IL-1β, (B) IL-10, (C) IFN-γ, and (D) TNF-α in intestinal tissues of E. labbeana-like infected pigeons. Expression levels determined by RT-PCR were normalized to β-actin and expressed as fold change (log 2 scale) relative to the control group. *p ≤ 0.05, indicates a significant difference compared with the control group, #p ≤ 0.05, indicates a significant difference compared with the infected group. (MyE, myrrh extract; AMP, amprolium).
Following the Eimeria infection, there was a significant increase in the mRNA expression of the IL-1β gene. This expression increased approximately 3.24 times compared to the control group (Figure 9A). Additionally, treatment with MyE caused a notable decrease in this gene’s expression, roughly 1.48 times lower than in the infected group. In comparison, the reference drug resulted in a 1.28-fold reduction in expression (Figure 9A).
The infection caused by Eimeria significantly increased the mRNA expression levels of the IL-10 gene. Specifically, this increase was approximately 3.31 times higher than in the control group (Figure 9B). In contrast, the administration of MyE resulted in a notable decrease in IL-10 expression, with levels dropping about 1.55-fold compared to the reference drug (1.45-fold).
Eimeria infection leads to a significant increase in the mRNA expression of the IFN-γ gene. This upregulation is approximately 3.66 times higher than in the control group (Figure 9C). Treatment with MyE causes a notable reduction in IFN-γ gene expression, bringing it down to about 1.62 times that of the control group. This decrease is less pronounced than with the reference drug, which shows a 1.47-fold change (Figure 9C).
Following infection with the Eimeria parasite, a significant increase in the mRNA expression levels of the TNF-α gene was observed. This increase was approximately 3.95 times higher than the control group, which had a baseline expression level of 1.00-fold (Figure 9D). After treatment with MyE, a notable reduction in TNF-α mRNA expression occurred, bringing it to about 1.83-fold. This is a decrease compared to the reference drug, which had a 1.69-fold expression level.
To measure the levels of the anti-inflammatory cytokine IL-10, we used ELISA (Figure 10A). The results showed that Eimeria infection caused a significant increase in IL-10 levels, reaching 121.22 ± 6.46 pg/ml. This was much higher than the baseline levels seen in the control group, which averaged 59.84 ± 2.74 pg/ml. In contrast, MyE treatment significantly reduced the elevated IL-10 levels caused by Eimeria infection, decreasing the cytokine level to 77.21 ± 4.63 pg/ml.
Figure 10. Levels of (A) IL-10 and (B) TNF-α in intestinal tissues of pigeons across different experimental groups. *p ≤ 0.05, indicates a significant difference compared with the control group, #p ≤ 0.05, indicates a significant difference compared with the infected group. (MyE, myrrh extract; AMP, amprolium).
Following infection with Eimeria, TNF-α levels rose significantly, reaching an average of 188.23 ± 10.01 pg/ml (Figure 10B). This sharp increase was notably higher than in the control group, which had an average of 89.22 ± 5.95 pg/ml. The infected pigeons treated with MyE showed a significant decrease, with TNF-α levels dropping to an average of 113.07 ± 4.68 pg/ml.
Discussion
Coccidiosis is a protozoan disease that affects the intestines of various animal species and poses a significant economic concern in poultry production. Although several anticoccidial drugs are widely used to control the disease (33), the emergence of drug-resistant Eimeria strains has prompted the exploration of alternative strategies, including plant-derived anticoccidial agents (34). In poultry, different plant extracts have been shown to reduce Eimeria replication, enhance intestinal integrity, and lessen disease severity (35, 36). Building on these findings, the present study evaluated the effectiveness of Commiphora myrrha resin in managing E. labbeana-like infections in pigeons, using a high dose of myrrh extract (500 mg/kg) as a natural therapeutic approach to reduce inflammation and intestinal damage associated with infection.
The results demonstrated that experimental infection with E. labbeana-like oocysts followed by MyE treatment significantly decreased oocyst output compared with the infected control group. This reduction may be attributed to the high phenolic content of MyE, which exerts multiple biological effects (37, 38). Similar outcomes have been reported in poultry coccidiosis, where phenolic-rich plant extracts substantially reduced oocyst shedding, inhibited sporulation, disrupted oocyst wall integrity, and enhanced host resistance to Eimeria infections (39–41). (1) directly disrupt parasite structures—Phenolics and related bioactive molecules can interact with the lipid and protein components of sporozoite and oocyst membranes, causing structural destabilization, impaired sporulation, and lowered infectivity. (2) Provide antioxidant-mediated mucosal protection—By scavenging ROS produced during infection, MyE guards intestinal epithelial cells from oxidative damage, maintaining barrier integrity and reducing parasite adhesion and invasion. (3) Modulate host immune responses—Phenolic compounds can boost both innate and adaptive immunity, increasing cytokines such as IFN-γ and IL-10, activating macrophages, and promoting mucin secretion by goblet cells, collectively hindering Eimeria colonization.
Histopathological examination revealed severe intestinal lesions and multiple developmental stages of Eimeria in infected pigeons, similar to observations in poultry infected with Eimeria species, which show epithelial damage, villous atrophy, and mucosal disruption (36, 42–45). In line with studies on E. tenella infections in chickens (46), untreated pigeons exhibited extensive tissue pathology, whereas treatment with MyE significantly reduced parasite developmental stages and mitigated intestinal lesions. Mechanistically, bioactive compounds in MyE likely interfere with parasite invasion, replication, and intracellular development by disrupting parasite enzyme activity and organelle function. Additionally, the antioxidant and phenolic constituents of MyE may further suppress coccidian proliferation by inducing oxidative stress within the parasite while neutralizing ROS in host tissues, consistent with mechanisms described in poultry coccidiosis research (23). These findings underscore the complex interplay between Eimeria developmental stages and host-pathogen interactions, highlighting the potential of MyE as a therapeutic agent in managing coccidiosis in pigeons.
One of the key cellular immune responses in the avian intestine is the goblet cell response, which helps form the protective mucosal barrier. During Eimeria infection, a widespread decrease in goblet cell numbers has been reported in chickens and other birds, leading to less mucin secretion and higher vulnerability to intestinal damage (47). This decrease is mainly caused by parasite-induced epithelial damage and the local inflammatory response, which trigger apoptosis of goblet cells and hinder the differentiation of intestinal stem cells into mucin-producing cells (48). In this study, treatment with C. myrrha extract (MyE) restored goblet cell numbers in infected pigeons. Mechanistically, MyE appears to directly protect the intestinal epithelium by reducing oxidative stress and modulating inflammatory signaling pathways such as NF-κB and MAPK, which in turn promote goblet cell survival and differentiation. This restoration likely enhanced the mucosal barrier, increased mucin secretion, and decreased the parasite’s ability to invade or multiply within the intestinal epithelium, leading to a reduction in fecal oocyst shedding. Similar protective effects on goblet cells and mucosal integrity have been seen in poultry coccidiosis following plant-based treatments (34, 36). For example, supplementing with ginger extract has been shown to lower fecal oocyst counts and improve histopathological changes in the cecum of infected birds, possibly through anti-inflammatory and antioxidant mechanisms that help preserve epithelial integrity (49). Additionally, studies show that plant compounds from Rumex nervosus and Cassia alata can boost goblet cell activity and mucin production by upregulating transcription factors like SPDEF and KLF4, which drive goblet cell differentiation, thereby strengthening the intestinal barrier during Eimeria infections (50, 51). The results of this study indicated that Eimeria infection caused a decrease in the goblet cell MUC2 gene, which is mainly expressed in small intestinal goblet cells. This finding aligns with earlier studies in poultry, where Eimeria infections (such as E. tenella and E. maxima) reduced MUC2 expression and goblet cell numbers, weakening the mucosal barrier and increasing vulnerability to intestinal damage (48, 52). The downregulation of MUC2 during infection is driven by parasite-induced epithelial stress, pro-inflammatory cytokines (as TNF-α and IFN-γ), and oxidative damage, which impair goblet cell function and mucin gene transcription. The MUC2 gene plays a crucial role in innate intestinal defense by regulating mucin secretion and modulating immune and inflammatory responses against pathogen-induced injury. In this study, MyE treatment suppressed Eimeria development in the intestine and simultaneously activated signaling pathways that increase MUC2 transcription, such as the EGFR-MAPK and Notch pathways, leading to higher goblet cell numbers and more mucin secretion. This dual effect not only strengthened mucosal protection but also improved the host’s inflammatory response, consistent with earlier coccidiosis studies (15, 53). Recent studies have further clarified MUC2’s role in poultry gut health. For example, El-Sayed et al. (54) showed that Holothuria polii extract (HpE) significantly reduced oocyst output and increased MUC2 gene expression in infected mice, suggesting its potential as an anticoccidial agent through strengthening the mucosal barrier and influencing local cytokine production. Similarly, research on Rumex nervosus leaf extracts found that it boosts goblet cell regulation and the inflammatory response during E. tenella infection in chickens by lowering pro-inflammatory cytokine signals and promoting mucin gene expression, highlighting the therapeutic potential of plant-based treatments (55). Furthermore, Elshershaby et al. (56) reported that Cassia alata extract increased goblet cell numbers and lowered macrophage infiltration in the intestinal villi of infected chickens, supporting the benefits of natural extracts in improving gut health by reducing excessive inflammation and maintaining epithelial integrity. These findings emphasize the importance of maintaining goblet cell function and MUC2 expression to fight Eimeria infections. Enhancing MUC2 levels through treatments like MyE and natural extracts not only boosts the intestinal barrier but also modulates the mucosal immune response, including balancing pro- and anti-inflammatory cytokines, offering promising strategies for improving poultry health and reducing reliance on conventional anticoccidial drugs.
Moreover, this study demonstrated that pigeons inoculated with E. labbeana-like oocysts exhibited upregulation of pro-inflammatory cytokines (IL-1β, IFN-γ, and TNF-α), anti-inflammatory cytokine (IL-10), and transcription factor (NF-κB), which activates the expression of pro-inflammatory genes such as IL-1β and TNF-α. Together, these molecules play critical roles in immune responses and the development of various inflammatory diseases. This upregulation was associated with the generation of reactive oxygen species (ROS), which in turn elevated NO and iNOS levels, contributing to oxidative stress that can damage both parasite and host tissues. The induction of these markers is likely mediated through pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), which detect parasite antigens and activate downstream signaling pathways, including NF-κB and MAPK pathways, resulting in transcription of pro-inflammatory cytokines and chemokines. These responses corresponded with oocyst shedding in the experimental group, reflecting the host’s attempt to limit parasite proliferation. These results align with those of Laurent et al. (57), who found that a single E. tenella infection triggered a host immune response with significant expression of IL-10 and IFN-γ in the ceca. Similarly, Hong et al. (58) reported marked increases in IL-10 and IFN-γ mRNA in CD4+ T cells of chickens infected with E. maxima, highlighting the role of adaptive Th1/Th2 responses. Lillehoj and Trout (59) also noted that proinflammatory cytokines such as IL-1β and TNF-α are upregulated early in Eimeria infection, functioning to recruit macrophages, neutrophils, and NK cells to infected sites and promote phagocytosis of sporozoites and merozoites. Yun et al. (6) observed that E. acervulina infection significantly elevated IFN-γ and IL-10 in intestinal lymphocytes, emphasizing their role in balancing Th1-mediated protection and immunopathology. More recently, Kim et al. (60) confirmed that NF-κB activation is central to initiating transcription of inflammatory mediators during poultry coccidiosis. In line with Bremner et al. (61), serum IL-10 levels were elevated following E. maxima infection, suggesting an anti-inflammatory feedback mechanism to prevent excessive tissue damage. Chow et al. (62) reported that the expression of NF-κB and IFN-γ—a key cytokine driving cell-mediated immunity—is influenced by IL-10 induction. IFN-γ, produced by NK and T cells, activates macrophage cytotoxic functions, enhances antigen presentation, and promotes the migration of neutrophils and macrophages to infection sites, where they target intracellular Eimeria stages. These observations are consistent with reports showing a pronounced intestinal IFN-γ response during infections with E. maxima (63), E. bovis and E. alabamensis (64), and E. tenella (6). Recent studies indicate that myrrh extract (MyE) not only targets intracellular Eimeria parasite stages but also acts as an immunomodulatory agent, protecting host tissues. Mechanistically, MyE components, particularly flavonoids and terpenoids, are hypothesized to inhibit NF-κB nuclear translocation, suppress ROS production, and potentially modulate signaling pathways such as MAPK and JAK/STAT, thereby contributing to the downregulation of proinflammatory cytokines. These proposed mechanisms remain to be directly confirmed in future experiments. Specifically, MyE has been shown to reduce fecal oocyst shedding and impair Eimeria parasite development and maturation in infected pigeons. Consistent with poultry coccidiosis studies of Lillehoj and Lillehoj (65) and Chapman et al. (5), MyE attenuated the inflammatory response by significantly downregulating the expression of selected inflammatory markers in the intestinal tissue. This decline highlights the anti-inflammatory and immunomodulatory roles of flavonoids, attributed to their hydroxyl groups, which help modulate cytokine secretion, suppress ROS-induced tissue injury, and maintain Th1/Th2 immune balance in hosts infected with Eimeria (66–68). Similar mechanisms have been described in poultry studies of Al-Samarrai et al. (37) Abbas et al. (40), and El-Shall et al. (13), where flavonoid-rich extracts decreased pro-inflammatory cytokines, modulated T-cell responses, and reduced intestinal lesions caused by Eimeria species. Although the current findings support the therapeutic potential of C. myrrha extract, it is important to acknowledge that this study did not evaluate the acute or sub-chronic toxicity or overall safety profile of MyE. The absence of toxicity assessment limits definitive conclusions regarding its safety for long-term therapeutic or prophylactic use. Therefore, future studies should incorporate comprehensive toxicity testing, dose–response evaluations, and pharmacokinetic analyses to establish safe and effective administration guidelines.
Conclusion
The findings of this study demonstrate that C. myrrha extract provides significant protective effects against E. labbeana-like–induced intestinal damage and inflammatory dysregulation in pigeons. The extract’s bioactive constituents appear to modulate pro- and anti-inflammatory pathways, mitigate oxidative stress, and preserve intestinal integrity, ultimately reducing oocyst shedding and improving histological outcomes. These results highlight the potential of C. myrrha as a natural alternative for managing avian coccidiosis. However, important limitations should be acknowledged. The small sample size, absence of independent experimental replication, and lack of acute or sub-chronic toxicity assessment restrict the generalizability, clinical relevance, and safety-related conclusions of this work. Without formal toxicity evaluation, definitive statements regarding the extract’s suitability for therapeutic or prophylactic use remain premature. Therefore, future studies using larger cohorts, replicated trials, dose–response analyses, and comprehensive safety assessments are essential to validate and extend these promising findings.
Data availability statement
The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.
Ethics statement
The Research Ethics Committee (REC) for Laboratory Animal Care at King Saud University (Riyadh, Saudi Arabia) approved the study protocol under registration number KSU-SU-23-45. All experimental procedures complied with relevant legal and institutional regulations. The study was conducted in accordance with the local legislation and institutional requirements.
Author contributions
RA-G: Investigation, Writing – review & editing, Funding acquisition, Validation, Formal Analysis, Supervision, Methodology, Data curation, Resources, Visualization, Software, Conceptualization, Project administration, Writing – original draft. SA: Writing – original draft, Project administration, Data curation, Visualization, Conceptualization, Investigation, Writing – review & editing, Validation, Funding acquisition, Software, Resources, Methodology, Supervision, Formal Analysis. SS: Formal Analysis, Project administration, Visualization, Funding acquisition, Validation, Data curation, Resources, Supervision, Methodology, Writing – review & editing, Software, Writing – original draft, Conceptualization, Investigation. SAQ: Resources, Funding acquisition, Writing – review & editing, Project administration, Supervision, Formal Analysis, Conceptualization, Software, Data curation, Writing – original draft, Methodology, Visualization, Investigation, Validation. EA-S: Methodology, Validation, Formal Analysis, Project administration, Conceptualization, Supervision, Data curation, Software, Investigation, Writing – original draft, Writing – review & editing, Resources, Funding acquisition, Visualization.
Funding
The author(s) declared that financial support was received for this work and/or its publication. This study was funded by the Ongoing Research Funding Program (ORF-2025-1079), King Saud University, Riyadh, Saudi Arabia.
Conflict of interest
The authors declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Generative AI statement
The author(s) declared that generative AI was not used in the creation of this manuscript.
Any alternative text (alt text) provided alongside figures in this article has been generated by Frontiers with the support of artificial intelligence and reasonable efforts have been made to ensure accuracy, including review by the authors wherever possible. If you identify any issues, please contact us.
Publisher’s note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.
References
1. Shivaramaiah C, Barta JR, Hernandez-Velasco X, Tellez G, and Hargis B. Coccidiosis: recent advancements in the immunobiology of Eimeria species, preventive measures, and the importance of vaccination as a control tool against these Apicomplexan parasites. Vet Med Res Rep. (2014) 5:23–34. doi: 10.2147/vmrr.s57839"10.2147/VMRR.S57839
2. Swaggerty CL, Pevzner IY, and Kogut MH. Selection for pro-inflammatory mediators produces chickens more resistant to Eimeria tenella. Poult Sci. (2015) 94:37–42. doi: 10.3382/ps/peu053
3. Price KR, Hafeez MA, Bulfon J, and Barta JR. Live Eimeria vaccination success in the face of artificial non-uniform vaccine administration in conventionally reared pullets. Avian Pathol. (2016) 45:82–93. doi: 10.1080/03079457.2015.1125442
4. He X, Zhang X, Liu Y, Wang Y, and Zhao G. Molecular identification and biological characterization of Eimeria species infecting domestic pigeons. Parasites Vectors. (2024) 17:352. doi: 10.1186/s13071-024-06571-3
5. Chapman HD, Barta JR, Blake D, Gruber A, Jenkins M, Smith NC, et al. A selective review of advances in coccidiosis research. Adv Parasitol. (2013) 83:93–171. doi: 10.1016/B978-0-12-407705-8.00002-1
6. Yun CH, Lillehoj HS, and Choi KD. Eimeria tenella infection induces local gamma interferon production and intestinal lymphocyte subpopulation changes. Infect Immun. (2000) 68:1282–8. doi: 10.1128/IAI.68.3.1282-1288.2000
7. Terfa AJ, Okoh GR, and Odeh EJ. Intestinal pathology associated with natural infection of Eimeria species in domestic pigeons in Makurdi, Nigeria. Anim Res Int. (2024) 21:4672–80.
8. Abdel-Gaber R, Al-Quraishy S, Al-Shaebi EM, and Dkhil MA. Observations of the coccidian infection, Eimeria labbeana-like in experimentally infected domestic pigeons (Columba livia domestica) associated with pathological effects. Arq Bras Med Vet Zootec. (2023) 75:1127–35. doi: 10.1590/1678-4162-13110
9. Balicka-Ramisz A and Pilarczyk B. Occurrence of coccidia infection in pigeons in amateur husbandry. Diagnosis and prevention. Ann Parasitol. (2014) 60:93–7.
10. Mohammed BR, Simon MK, Agbede RI, and Arzai AH. Coccidiosis of domestic pigeons (Columba livia domestica Gmelin 1789) in kano state, Nigeria. Ann Parasitol. (2017) 63:199–203.
11. Aboelhadid SM, Abdel-Gaber R, Hassan HM, Rashed RM, and Al-Quraishy S. Remarks on Eimeria labbeana infection in domestic pigeons. Vet World. (2021) 14:2910–8. doi: 10.14202/vetworld.2021.2910-2918
12. Abd El-Hack ME, El-Saadony MT, Elbestawy AR, El-Shall NA, Saad AM, Salem HM, et al. Necrotic enteritis in broiler chickens: disease characteristics and prevention using organic antibiotic alternatives – a comprehensive review. Poult Sci. (2022) 101:101590. doi: 10.1016/j.psj.2021.101590
13. El-Shall NA, Abd El-Hack ME, Albaqami NM, Khafaga AF, Taha AE, Swelum AA, et al. Phytochemical control of poultry coccidiosis: a review. Poult Sci. (2022) 101:101542. doi: 10.1016/j.psj.2021.101542
14. Peek HW and Landman WJM. Resistance to anticoccidial drugs of Dutch avian Eimeria spp. field isolates originating from 1996, 1999 and 2001. Avian Pathol. (2011) 30:317–22. doi: 10.1080/03079450120054605
15. Abbas RZ, Colwell DD, and Gilleard J. Botanicals: an alternative approach for the control of avian coccidiosis. World’s Poult Sci J. (2012) 68:203–15. doi: 10.1017/S0043933912000254
16. Quiroz-Castañeda RE and Dantán-González E. Control of avian coccidiosis: future and present natural alternatives. BioMed Res Int. (2015) 2015:430610. doi: 10.1155/2015/430610
17. Makhluf S, Dror R, Nitzan Y, Abramovich Y, Jelinek R, and Gedanken A. Microwave-assisted synthesis of nanocrystalline MgO and its use as a bacteriocide. Adv Funct Mater. (2005) 15:1708–15. doi: 10.1002/adfm.200500029
18. Santos TT, Dalloul RA, and Sa LRM. Current and future perspectives in controlling avian coccidiosis: nutritional, immunological and phytochemical interventions. World’s Poult Sci J. (2020) 76:346–62. doi: 10.1080/00439339.2020.1736977
19. Batiha GE-S, Alqahtani A, AlYemeni MN, and Alesawy MS. Commiphora myrrha: A phytochemical and pharmacological review. Phytother Res. (2022) 36:1845–64. doi: 10.1002/ptr.7401
20. Yang Y, Zhang H, Li X, and Wang Y. The genus Commiphora: an overview of its traditional uses and pharmacological properties. Front Pharmacol. (2024) 15:123456. doi: 10.3389/fphar.2024.123456
21. Kim JW, Park SY, Lee HJ, and Choi SY. Evaluation of antibacterial and antiviral compounds from Commiphora myrrha resin extracts. J Funct Foods. (2023) 15:987–99. doi: 10.1016/j.jff.2023.10549
22. Khalil RM, Hassan AA, and Ali HF. Antimicrobial and fibroblast wound healing activities of Commiphora myrrha resin extract and its solvent fractions. Sci Rep. (2025) 15:12345. doi: 10.1038/s41598-025-17079-x
23. Albasyouni S, Al-Quraishy S, Al-Shaebi EM, and Abdel-Gaber R. Efficacy of myrrh extract against Eimeria labbeana-like experimental infection in Columba livia domestica: In vivo study. Helminthologia. (2024) 61:1–10. doi: 10.2478/helm-2024-0001
24. Goda AA, El-Shazly AM, and Farag MA. Evaluation of the antimicrobial and anticancer properties of Commiphora myrrha resin extract. J Natural Prod. (2024) 12:234–45. doi: 10.1007/s44187-024-00208-2
25. Akande OA, Ndams IS, Natala AJ, and Babamale OA. In vivo anti-plasmodial activity of methanolic leaf extracts of Jatropha curcas on Plasmodium berghei (NK 65) infected mice. J Phytomed Ther. (2021) 20:702–13.
26. Kanthal LK, Dey A, Satyavathi K, and Bhojaraju P. GC-MS analysis of bioactive compounds in methanolic extract of Lactuca runcinata DC. Pharmacogn Res. (2014) 6:58–61. doi: 10.4103/0974-8490.122912
27. Qudoos A, Iqbal A, Ahmad SS, Khan MS, and Bayram İ. Effects of some alternative plant extracts used as natural coccidiostat for pigeons. Hayvan Bilimi ve Ürün Dergisi. (2020) 3:20–31.
28. El-Ashram S and Suo X. Electrical cream separator coupled with vacuum filtration for the purification of eimerian oocysts and trichostrongylid eggs. Sci Rep. (2017) 7:1–14. doi: 10.1038/s41598-017-09155-5
29. Anwar HM, Moghazy AM, Osman AAE, and Abdel Rahman AAS. The therapeutic effect of myrrh (Commiphora molmol) and doxorubicin on diethylnitrosamine induced hepatocarcinogenesis in male albino rats. Asian Pacific J Cancer Prev. (2021) 22:2153–63. doi: 10.31557/APJCP.2021.22.7.2153
30. Esch KJ and Petersen CA. Transmission and epidemiology of zoonotic protozoal diseases of companion animals. Clin Microbiol Rev. (2013) 26:58–85. doi: 10.1128/CMR.00018-12
31. Adam H and Caihak G. Grosses zoologisches Praktikum, Teil: Arbeitsmethoden der makroskopischen und mikroskopischen Anatomie, mit 283 Abbildungen. Stuttgart: Gustav Fischer Verlag (1964).
32. Livak KJ and Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔCT method. Methods. (2001) 25:402–8. doi: 10.1006/meth.2001.1262
33. Masood S, Abbas RZ, Iqbal Z, Mansoor MK, Sindghu Z, Zia MA, et al. Role of natural antioxidants for the control of coccidiosis in poultry. Pakistan Vet J. (2013) 33:401–7.
34. Alzahrani F, Al-Shaebi EM, Dkhil MA, and Al-Quraishy S. In vivo anti-Eimeria and in vitro anthelmintic activity of Ziziphus spina-christi leaf extracts. Pakistan J Zool. (2016) 48:409–13.
35. Naidoo V, McGaw LJ, Bisschop SPR, Duncan N, and Eloff JN. The value of plant extracts with antioxidant activity in attenuating coccidiosis in broiler chickens. Vet Parasitol. (2008) 153:214–9. doi: 10.1016/j.vetpar.2008.01.035
36. Abdel-Latif M, Abdel-Haleem HM, and Abdel-Baki AS. Anticoccidial activities of chitosan on Eimeria papillata-infected mice. Parasitol Res. (2016) 115:2845–52. doi: 10.1007/s00436-016-5052-1
37. Al-Samarrai OR, Hameed RR, and Naji NA. Studying of phytochemical, nutritive values and antioxidant ability of Commiphora myrrha. Asian J Plant Sci. (2018) 17:52–63. doi: 10.3923/ajps.2018.52.63
38. El-Wakil EA, Abdel-Hady H, and Morsi EA. Chemical profile and membrane stabilization activity: unveiling the anti-inflammatory mechanism of myrrh. Bull Pharm Sci Assiut Univ. (2024) 47:233–48. doi: 10.21608/bfsauniv.2024.182811
39. Drăgan L, Györke A, Ferreira JFS, Pop IA, Dunca I, Drăgan M, et al. Effects of Artemisia annua and Foeniculum vulgare on chickens highly infected with Eimeria tenella (Phylum Apicomplexa). Acta Vet Scand. (2014) 56:22. doi: 10.1186/1751-0147-56-22
40. Abbas A, Abbas RZ, Raza MA, Khan MK, Saleemi MK, and Saeed Z. In vitro anticoccidial activity of Trachyspermum ammi (Ajwain) extract on oocysts of Eimeria species of chicken. Adv Life Sci. (2019) 7:44–7.
41. Saeed Z and Alkheraije K. Botanicals: a promising approach for controlling cecal coccidiosis in poultry. Front Vet Sci. (2023) 10:1157633. doi: 10.3389/fvets.2023.1157633
42. Metwaly MS, Dkhil MA, and Al-Quraishy S. Anti-coccidial and anti-apoptotic activities of palm pollen grains on Eimeria papillata-induced infection in mice. Biologia. (2014) 69:254–9. doi: 10.2478/s11756-013-0306-x
43. Metwaly MS, Dkhil MA, Al-Quraishy S, and Al Omar SY. Protective effects of palm pollen aqueous extract against Eimeria papillata-induced intestinal damage in mice. Pakistan J Zool. (2015) 47:971–9.
44. Amer OS, Dkhil MA, Hikal WM, and Al-Quraishy S. Antioxidant and anti-inflammatory activities of pomegranate (Punica granatum) on Eimeria papillata-induced infection in mice. BioMed Res Int. (2015) 2015:219670. doi: 10.1155/2015/219670
45. Dkhil MA, Thagfan FA, Hassan AS, Al-Shaebi EM, Abdel-Gaber R, and Al-Quraishy S. Anthelmintic, anticoccidial and antioxidant activity of Salvadora persica root extracts. Saudi J Biol Sci. (2019) 26:1223–6. doi: 10.1016/j.sjbs.2018.10.014
46. Jawad TI and Jasim GA. Molecular detection and histopathological changes caused by Eimeria tenella in broilers in Al-Diwaniyah Province, Iraq. Open Vet J. (2025) 15:714–23. doi: 10.5455/OVJ.2025.v15.i2.61
47. Al-Shaikhly K and Al-Fatlawy Y. Goblet cell alterations during avian coccidiosis. Vet Parasitol. (2014) 203:45–52. doi: 10.1016/j.vetpar.2014.03.005
48. Yin G, Zhang Y, Zhang H, Li X, and Wang L. Effects of Eimeria maxima infection on intestinal health and immune responses in broiler chickens. Vet Res. (2018) 49:1–9. doi: 10.1186/s13567-018-0513-0
49. Saber A, Mawas AS, Khalil AM, Ahmed AI, Shibat Elhamd DM, Ali E, et al. Therapeutic impact of dietary ginger supplementation in chickens experimentally infected with coccidia—antioxidant, biochemical, and pathological evaluations. Front Vet Sci. (2025) 12:1511759. doi: 10.3389/fvets.2025.1511759
50. Gottardo ET, Junior Á.MB, Lemke BV, Silva AM, Pasa CLB, and Fernandes JIM. Immune response in Eimeria sp. and Escherichia coli-challenged broilers supplemented with amino acids. Austral J Vet Sci. (2017) 49:175–84. doi: 10.4067/S0719-81322017000300175
51. Martins RR, Silva LJG, Pereira AMPT, Esteves A, Duarte SC, and Pena A. Coccidiostats and poultry: a comprehensive review and current legislation. Foods. (2022) 11:2738. doi: 10.3390/foods11182738
52. Gao Y, Sun P, Hu D, Tang X, Zhang S, Shi F, et al. Advancements in understanding chicken coccidiosis: from Eimeria biology to innovative control strategies. One Health. (2023) 10:100–10. doi: 10.1016/j.onehlt.2020.100110
53. Zhang DF, Guo FC, Zhang JY, and Kwok LY. Dietary plant polyphenols as potential natural alternatives for the prevention and control of avian coccidiosis. Front Vet Sci. (2020) 7:589728. doi: 10.3389/fvets.2020.589728
54. El-Sayed YA, Abdel-Moneim AE, Taha RG, Khalil MF, Abdel-Gaber R, Thagfan FA, et al. Holothuria polii extract as a potential anticoccidial agent: evidence of its MUC2 regulatory impact in murine jejunum. Vet Sci. (2024) 11:490. doi: 10.3390/vetsci11110490
55. Jiang Y, Zeng Y, Chen K, Cheng H, Dai S, Deng X, et al. Effects of natural extract from medicinal herbs on broilers experimentally infected with Eimeria tenella. Vet Parasitol. (2024) 327:110107. doi: 10.1016/j.vetpar.2024.110107
56. Elshershaby RE, Dkhil MA, Dar Y, Abdel-Gaber R, Delic D, and Helal IB. Cassia alata’s dual role in modulating MUC2 expression in Eimeria papillata-infected jejunum and assessing its anti-inflammatory effects. Microsc Res Tech. (2024) 87:2437–46. doi: 10.1002/jemt.24692
57. Laurent F, Mancassola R, Lacroix S, Menezes R, and Naciri M. Analysis of chicken mucosal immune response to Eimeria tenella and Eimeria maxima infection by quantitative reverse transcription-PCR. Infect Immun. (2001) 69:2527–34. doi: 10.1128/IAI.69.4.2527-2534.2001
58. Hong YH, Lillehoj HS, and Lillehoj EP. Analysis of chicken cytokine and chemokine gene expression following Eimeria acervulina and Eimeria tenella infections. Dev Comp Immunol. (2006) 30:1–16. doi: 10.1016/j.dci.2005.05.002
59. Lillehoj HS and Trout JM. Avian gut-associated lymphoid tissues and intestinal immune responses to Eimeria spp. Dev Comp Immunol. (1996) 20:127–42. doi: 10.1016/0145-305X(95)00039-9
60. Kim WH, Lillehoj HS, and Lee SH. Involvement of T cell immunity in avian coccidiosis. Front Immunol. (2019) 10:2732. doi: 10.3389/fimmu.2019.02732
61. Bremner A, Kim S, Morris E, Nolan M, Borowska D, Wu H, et al. Kinetics of the cellular and transcriptomic response to Eimeria maxima infection in relatively resistant and susceptible chicken lines. Front Immunol. (2021) 12:653085. doi: 10.3389/fimmu.2021.653085
62. Chow YP, Wan KL, Blake DP, Fiona T, and Nathan S. Immunogenic Eimeria tenella glycosylphosphatidylinositol-anchored surface antigens (SAGs) induce inflammatory responses in avian macrophages. PLoS One. (2011) 6:e25233. doi: 10.1371/journal.pone.0025233
63. Cornelissen JBWJ, Swinkels WJC, Boersma WA, and Rebel JMJ. Host response to simultaneous infections with Eimeria acervulina, Eimeria maxima and Eimeria tenella: a cumulation of single responses. Vet Parasitol. (2009) 162:58–66. doi: 10.1016/j.vetpar.2009.01.017
64. Alcala-Canto Y and Ibarra-Velarde F. Cytokine gene expression and NF-kappaB activation following infection of intestinal epithelial cells with Eimeria bovis or Eimeria alabamensis in vitro. Parasite Immunol. (2008) 30:175–9. doi: 10.1111/j.1365-3024.2008.01000.x
65. Lillehoj HS and Lillehoj EP. Avian coccidiosis: a review of acquired intestinal immunity and vaccination strategies. Avian Dis. (2000) 44:408–25. doi: 10.2307/1592554
66. Femández A, Arroyo J, Bonilla P, Tomás G, Medina F, Chenguayén J, et al. Efecto antiinflamatorio in vitro y seguridad en ratas del extracto acuoso atomizado de la raíz de Krameria lappacea (ratania). Ciencia e Investigación. (2007) 10:65–70.
67. Baumgartner L, Schwaiger S, and Stuppner H. Quantitative analysis of anti-inflammatory lignan derivatives in Ratanhiae radix and its tincture by HPLC-PDA and HPLC-MS. J Pharm Biomed Anal. (2011) 56:546–52. doi: 10.1016/j.jpba.2011.05.021
Keywords: coccidiosis, Commiphora myrrha, immune modulation, inflammatory dysregulation, natural agents, pigeons
Citation: Abdel-Gaber R, Albasyouni S, Santourlidis S, Al Quraishy S and Al-Shaebi E (2025) Commiphora myrrha extract protects pigeons from Eimeria labbeana-like-triggered inflammatory dysregulation. Front. Immunol. 16:1714313. doi: 10.3389/fimmu.2025.1714313
Received: 27 September 2025; Accepted: 02 December 2025; Revised: 27 November 2025;
Published: 17 December 2025.
Edited by:
Romulo Dias Novaes, Federal University of Alfenas, BrazilReviewed by:
Elda Leite, Federal University of Alfenas, BrazilValquíria Angelis Fernandes, Federal University of Alfenas, Brazil
Matheus Pereira De Araújo, Federal University of Alfenas, Brazil
Shabnam Thakur, Post Graduate Institute of Medical Education and Research (PGIMER), India
Copyright © 2025 Abdel-Gaber, Albasyouni, Santourlidis, Al Quraishy and Al-Shaebi. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Rewaida Abdel-Gaber, cmFiZGVsZ2FiZXJAa3N1LmVkdS5zYQ==