C. alata have numerous bioactive compounds in its leaves with certain alkaloids, flavonoids, saponins, tannins and terpenoids (
Angelina et al., 2021). A recent study by
Elshershaby et al. (2024) indicated that
C. alata extract diminishes oocyst output and alleviates necrotic enteritis and inflammatory alterations caused by
E. papillata, demonstrating its anticoccidial, antioxidant and anti-inflammatory characteristics. In this study, the Folin–Ciocalteu colorimetric analysis assay demonstrated that the total phenolics of CE was 277.4±1.7 mg GAE/g dry weight extract. Besides, the aluminum chloride-based colorimetric method implied that the total flavonoids in CE was 63.6±0.2 mg QE/g dry weight extract. Furthermore, the DPPH radical scavenging activity was 89.9±2% for CE (Table 1).
Coccidiosis is one of the world’s most devastating livestock illnesses, caused by parasite protozoa from the genus
Eimeria following fecal-oral uptake of sporulated oocysts, which affects a variety of animal species and results in significant economic losses (
Al-Quraishy et al., 2011;
Blake and Tomley, 2014). It affects critical animals leading to weight loss and reduced feed efficiency (
Schito et al., 1996). Anticoccidial synthetic chemicals are routinely used to treat coccidiosis, however misuse can consequence in to drug resistance (
Flores et al., 2022). The medication amprolium has a wide spectrum of harmful effects on animal host tissues (
Noack et al., 2019). Diverse animal species have been effectively treated and controlled using various strategies and alternatives globally. Among these choices, various compounds derived from natural herbal medicinal plants (
Lee et al., 2022). Recent previous searches signposted the effective role of plant extracts as anticoccidials as
Persea americana (
Al-Otaibi et al., 2023a),
Krameria lappacea (
Alamari et al., 2024) and
Holothuria polii (
El-Sayed et al., 2024). For this, this study inspected the ameliorative role of CE as a hepatoprotective agent in addition to its anticoccidial, anti-inflammatory and antioxidant properties.
According to the present results,
E. papillata infection induced loss in body weight by -4.67±0.58% and diminish in feed intake (98.00±4.36 grams) on 5
th day post infection in comparison to the control group (Fig 1).
The same results were proved by
Abdel-Gaber et al. (2024a) and
Al-Otaibi et al. (2023a), respectively. Several previous studies illuminated that this weight loss is the result of consuming the nutrients of the intestinal epithelium by the different
Eimeria stages that leads to notable alterations in the intestinal nutrient contents (
Hamid et al., 2019;
Qaid et al., 2022). Also,
Anwar et al. (2008) indicated that this considerable weight loss could be caused by various factors, including decreased daily food and drink intake. On the other hand, the use of CE as a therapeutic natural anticoccidial agent against
E. papillata in this study resulted in a significant enhancement in body weight (-2.33±0.58%) and feed intake (130.33±4.51 grams), in relation to the Inf group (Fig 1). Similarly,
Al-Otaibi et al. (2023a) enhanced the loss of body weight and feed intake that resulted from
E. papillata infection. This significant development may result from the phytochemicals in the plant extract, which improved the nutritional status (
Al-Otaibi et al., 2023a).
C. alata includes innumerable types of phytochemical constituents that denote to its biological activities that possess anti-inflammatory and antioxidant activities such as flavonoids including isoflavones, kaempferol and its derivatives and rutin (
Angelina et al., 2021) as well as anthraquinones, such as phenolic compounds, alkaloids, coumarins, saponins are also present (
Muhammad et al., 2021).
The liver is the first point of interaction with the gut via the portal vein.
E. papillata mainly infects the intestinal tract, predominantly the jejunum and does not usually infiltrate into the liver, however other lymphoid organs including the spleen and lymph nodes might be impacted by similar coccidian protozoa as
E. coecicola parasites (
Renaux et al., 2001). In the current study, light microscopic investigations of hematoxylin and eosin stained liver sections of
E. papillata infected group showed focal hepatic necrosis associated with marked infiltration with mononuclear cells as lymphocytes and macrophages as well as marked periportal hepatic vacuolar changes consistent with marked fatty changes (Fig 2).
These results were in consistent with
Maodaa et al. (2024a), who proved that
E. papillata infection had resulted in a moderate pathological inflammatory reaction in mice’s hepatic tissue that is dominated by an increase in Kupffer cells and leukocytes, mainly lymphocytes, as a typical tissue response to hepatic destruction. Also,
Dkhil and Al-Quraishy (2012) revealed that Eimeria infection caused cell membrane permeability and damage in the hepatocytes. Moreover,
Dkhil et al. (2011) indicated that the structural evidence of mild inflammatory response in the liver correlate with oxidative tissue damage. However, upon treatment with CE in this study, a remarkable decrease in the hepatic vacuolar changes within the cytoplasm of the hepatocytes was observed showing its hepatoprotective effect (Fig 2). Similarly, the use of
Teucrium polium leaves ethanolic extract had the ability to diminish the liver injury induced by
E. papillata and restored the number of hepatic Kupffer cells (
Maodaa et al., 2024a). The hepatoprotective efficacy of CE could be contributed to the anthraquinones and flavonoids contents that considered good anti-inflammatory agents (
Chew et al., 2022).
Coccidiosis frequently affects host cell metabolism (
Al-Quraishy et al., 2014). As well, Eimeria parasite has a noteworthy capability to benefit from the host cells by scavenging their available nutrients (
Hermosilla et al., 2012). In the herein study, a remarkable diminution in both content of carbohydrates (0.37±0.15%) and proteins (7.06±0.67%) of
E. papillata infected group was observed in periodic acid-Schiff and mercuric bromophenol blue stained liver sections, respectively (Fig 3, 4). One possible explanation for the reduced carbohydrate content is the excessive consumption of stored carbohydrates in the jejunal tissues by parasite stages (
Metwaly et al., 2013).
Kouwenhove (1971) stated that intestinal coccidial infections were categorized as protein-losing enteropathy. Many investigations revealed that the infected coccidial tissues had a reduced quantity of total proteins (
Bangoura and Daugschies, 2007). Upon treatment with CE in the present study, the hepatic carbohydrate (6.04±0.45 %) and protein (12.46±1.13%) contents was returned in comparison the infected group (Fig 3, 4).
These reached results were the same that informed by
Abdel-Gaber et al. (2024b), who indicated that the use of
Krameria lappacea root extract had resulted in a significant improvement in the jejunal carbohydrate and protein content.
Al-Otaibi et al. (2023a) revealed that
Persea americana fruit extract could restored its jejunal content of carbohydrates through to its bioactive constituents, which reduced the activity of the glucose-6-phosphatase enzyme that most likely resulted in the reestablishment of tissue carbohydrate content. As well,
Al-Otaibi et al. (2023a) and
Abdel-Gaber et al. (2024b) explained that
Persea americana fruit extract and
Krameria lappacea root extract enhanced jejunal protein content by reducing catalytic pathways of tissue protein and nucleic acid breakdown.
Concerning
Georgieva et al. (2006), Eimeria infection leads to an imbalance in the body’s natural antioxidant defense and release of free radicals. The current findings showed that
E. papillata infected mice exhibited hepatic injury resulting in the elevation of its antioxidant enzymes including NO and MDA (Fig 5).
These results totally in agreement with
Abdel-Gaber et al. (2024a). The higher levels of NO and MDA were due to their imbalanced status. This suggested the presence of oxidative stress resulting from the Eimeria infection and the disruption of the equilibrium between pro-oxidants and antioxidants (
Abdel-Gaber et al., 2024a). in addition,
Koinarski et al. (2005) and
Georgieva et al. (2006) realized that elevated MDA and NO levels were an immune response due to the presence of infectious sporozoite stages, which infiltrate intestinal cells and induce inflammation. Whilst, the usage of CE against
E. papillata infection had resulted in significant reduction in hepatic NO and MDA levels, compared to the infected mice (Fig 5).
Al-Otaibi et al. (2023b) proved that the improvements of NO and MDA levels in the jejuna of
E. papillata coccidian infected mice after medication with
Persea americana extract as a result of the existence of phenolic compounds. This matches with
Al-Otaibi et al. (2023a), who showed that phytochemicals in
Persea americana extract could bind to important macromolecules on parasite membranes, disrupting them and subsequent death of the parasite.
Additionally, infected mice with
E. papillata presented significant reduction in GSH and GPx levels in liver tissues (Fig 5). Similarly,
Alajmi et al. (2023) and
Al-Quraishy et al. (2024) indicated that GPx and GSH levels were decreased, respectively due to
E. papillata infection. During Eimeria infection, GPx and GSH enzymes are crucial for safeguarding the host’s body from damage caused by free radicals (
Al-Sayed et al., 2022).
Dkhil et al. (2015) and
El-Ghareeb et al. (2023) clarified that Eimeria infection produces an imbalance in the antioxidant defense body system, whichdenotes to unfavorable cellular effects. Therapeutic treatment with CE enhanced hepatic levels of GSH and GPx antioxidant enzymes, relative to the infected group (Fig 5). The same results were proved by
Alajmi et al., (2023) and
Al-Quraishy et al. (2024).
Awaad et al. (2016) hypothesize that the antioxidant components found in several different herbals have played a significant role in enhancing the defense against coccidiosis.
C. alata leaves have been reported to contain a wide variety of bioactive phytochemicals (
Veerachari and Bopaiah, 2012). Numerous phytochemical substances, including flavonoids, phenols, alkaloids, anthraquinones, tannins, saponins and cardiac glycosides were reported (
El-Mahmood and Doughari, 2008;
Eliakim-Ikechukwu, 2013). These phytochemicals have a crucial role in pharmacology through their anti-inflammatory, anti-bacterial, antifungal, antioxidant properties as its polyphenols could demonstrate strong scavenging action against free radicals and oxidative agents (
Chew et al., 2022).
