animals

Review
Coccidiosis in Egg-Laying Hens and Potential Nutritional
Strategies to Modulate Performance, Gut Health,
and Immune Response
Milan Kumar Sharma and Woo Kyun Kim *

Department of Poultry Science, University of Georgia, Athens, GA 30602, USA; [email protected]

* Correspondence: [email protected]

Simple Summary: Coccidiosis is one of the most prevalent diseases in poultry production, inflicting
substantial economic losses exceeding USD 15 billion to the poultry industry. Coccidiosis is most
common in young broilers but is not limited to them, and pullets and egg-laying hens are equally
susceptible. Unfortunately, unlike broilers, low emphasis has been given to laying hens. This review
aims to summarize the effect of coccidiosis in laying hens while exploring potential nutritional
interventions to fight coccidiosis.

Abstract: Avian coccidiosis, despite advancements in management, nutrition, genetics, and immunol-
ogy, still remains the most impactful disease, imposing substantial economic losses to the poultry
industry. Coccidiosis may strike any avian species, and it may be mild to severe, depending on the
pathogenicity of Eimeria spp. and the number of oocysts ingested by the bird. Unlike broilers, low
emphasis has been given to laying hens. Coccidiosis in laying hens damages the gastrointestinal tract
and causes physiological changes, including oxidative stress, immunosuppression, and inflammatory
changes, leading to reduced feed intake and a drastic drop in egg production. Several countries
around the world have large numbers of hens raised in cage-free/free-range facilities, and coccidiosis
has already become one of the many problems that producers have to face in the future. However,
limited research has been conducted on egg-laying hens, and our understanding of the physiological
changes following coccidiosis in hens relies heavily on studies conducted on broilers. The aim of
Citation: Sharma, M.K.; Kim, W.K. this review is to summarize the effect of coccidiosis in laying hens to an extent and correlate it with
Coccidiosis in Egg-Laying Hens and the physiological changes that occur in broilers following coccidiosis. Additionally, this review tries
Potential Nutritional Strategies to to explore the nutritional strategies successfully used in broilers to mitigate the negative effects
Modulate Performance, Gut Health, of coccidiosis in improving the gut health and performance of broilers and if they can be used in
and Immune Response. Animals 2024, laying hens.
14, 1015. https://doi.org/10.3390/
ani14071015
Keywords: coccidiosis; laying hens; gut health; oxidative stress; nutritional strategies; immune
Academic Editor: Kenneth Bafundo response; egg production

Received: 6 March 2024

Revised: 21 March 2024
Accepted: 25 March 2024
1. Introduction
Published: 27 March 2024
Avian coccidiosis, caused by various species of the protozoan Eimeria, poses a sig-
nificant threat to the poultry industry. Despite advancements in management, nutrition,
genetics, chemotherapy, and immunology, it remains the most prevalent disease in the
Copyright: © 2024 by the authors. poultry industry, imposing substantial economic losses of more than USD 15 billion [1].
Licensee MDPI, Basel, Switzerland. The economic loss due to coccidiosis is not only due to clinical outbreaks resulting in
This article is an open access article
low production and high mortality but also subclinical coccidiosis without visible clinical
distributed under the terms and
signs; even subclinical coccidiosis contributes to economic losses through reduced nutri-
conditions of the Creative Commons
ent absorption and utilization, thereby compromising optimal performance and the feed
Attribution (CC BY) license (https://
conversion ratio (FCR). Reduced nutrient absorption during subclinical coccidiosis may
creativecommons.org/licenses/by/
result from intestinal enterocyte and morphometric damages, compromised gastrointestinal
4.0/).

Animals 2024, 14, 1015. https://doi.org/10.3390/ani14071015 https://www.mdpi.com/journal/animals

Animals 2024, 14, 1015 2 of 33

integrity, decreased ileal digestibility, and increased susceptibility to secondary diseases

like necrotic enteritis [2–7].
Coccidiosis is common in young broilers or laying hen pullets ranging from 3 to
18 weeks of age but not limited to this age group. Newly hatched chicks have high maternal
antibodies but have little protection against coccidiosis [8]. Chickens usually become infected
when maternal antibodies deplete and the oocyst counts in litter peak at 3–6 weeks of age;
however, immunity develops shortly after mild infections [2,5,8]. Older flocks without
acquired immunity against Eimeria spp. might get the disease [2,8]. In addition, there is no
cross-immunity among Eimeria species; thus, more than one outbreak is possible in the
same flock with different species involved in each episode [2,8]. Generally, anticoccidial
drugs are used in laying hens’ pullets and are withdrawn before they start laying eggs.
Therefore, the acquired immunity against Eimeria during the pullet phase might not be
enough to prevent infection throughout the production period [8–10]. Coccidiosis may
strike any avian species, and it may be mild to severe, depending on the pathogenicity of
the Eimeria spp. and the number of oocysts ingested by the bird [2,5].
Unlike broilers, limited attention has been given to laying hens regarding coccidiosis.
One of the leading causes of laying hen mortality was reported as coccidiosis in cage-free
systems [11–13]. A study conducted in Sweden observed 19% coccidiosis in laying hens
raised on a litter-based cage-free system [14]. Laying hens recovered from coccidiosis
but did not have optimal egg production and had a high FCR. Fossum et al. (2009) [12]
observed higher incidences of coccidiosis in laying hens raised in cage-free systems (aviaries
and free-range) compared with conventional cages in Sweden from 2001 to 2004, after
the ban on conventional cages. Although there are several variations among cage-free
systems (aviaries, free-range, or barn), the litter-based floor is typical in all housing systems
that allow fecal–oral transmission of the oocyst, leading to disease outbreaks, unlike
conventional cages. Therefore, preventing coccidiosis in these systems might become one
of the many challenges for egg producers while transitioning from traditional cages to
cage-free systems.
As the egg industry worldwide is steadily observing an increase in cage-free facilities,
the broad topic of “gut health” associated with “coccidiosis” will have one of the most
significant economic impacts on laying hen health and performance. Many countries in
Europe have already transitioned to cage-free systems, whereas the United States is slowly
transitioning to cage-free systems, and by 2023, more than 35% of laying hens in the U.S.
were raised in cage-free environments, and this is still increasing (UEP, 2023). The evolution
of these new production systems (aviaries; free-range) with large numbers of hens on the
floor/range will influence the occurrence of coccidiosis. Laying hens raised in a litter-
based aviary or free-range systems may provide a perfect environment for transmitting
coccidiosis, and they have a higher chance of getting the disease than those raised in
traditional cage systems. However, laying hens raised in conventional cages are also
susceptible to coccidiosis, which contrasts with the traditional view that raising laying hens
in conventional cages is unlikely to result in coccidiosis infection [10,15]. Hens raised in a
cage system can still ingest enough oocysts from manure belts or edges of cages to induce
clinical coccidiosis; however, the incidences are less [10,15]. Yet, there remains a dearth of
knowledge regarding the effects of coccidiosis on laying hen performance, gastrointestinal
health, oxidative stress, immune response, and skeletal health. However, broilers and
laying hens have been intensively selected for many generations for different purposes
(rapid growth and meat production vs. egg production), which might have changed their
gastrointestinal physiology, immune response, and skeletal development. In this review,
we aim to correlate findings from the broiler studies to best explain what might happen in
laying hens infected with coccidiosis, particularly concerning gastrointestinal physiology,
immune response, and potential mitigation strategies.
Animals 2024, 14, 1015 3 of 33

2. Avian Coccidiosis
Avian coccidiosis is caused by multiple species of the genus Eimeria belonging to the
subkingdom protozoa and phylum Apicomplexa [5]. Eimeria is an intracellular parasite that
infiltrates intestinal enterocytes at different locations, depending on the species. Only seven
have been recognized to infect chickens among the hundreds of Eimeria species including
the following: E. acervulina, E. brunetti, E. maxima, E. necatrix, E. mitis, E. praecox, and
E. tenella [2,5]. Among these Eimeria species known to infect chickens, E. brunetti, E. maxima,
E. necatrix, and E. tenella are considered highly pathogenic, whereas E. acervulina, E. mitis,
and E. praecox exhibit low pathogenicity (Table 1) [2,5]. In the past, there were sporadic
reports of outbreaks of E. hagani and E. mivati in chickens. However, their existence was
doubted, and they are now recognized as nomina dubia [16].

Table 1. List of Eimeria species important in laying hens with their predilection sites, characteristic
lesions, pathogenicity, and immunogenicity.

Predilection Site in the

Species Pathogenicity Characteristic Lesions Immunogenicity
Intestine
Mid-intestine (Meckel’s Mucoid exudate and no
E. mitis Low Moderate
diverticulum to cecal junction) distinct lesions
Mucoid exudate and no
E. praecox Low Anterior intestine (duodenum) Moderate
distinct lesions
E. acervulina Low to moderate Anterior intestine (duodenum) Whitish round lesions Moderate
Mid-intestine (part of jejunum Petechiae hemorrhage and
E. maxima Moderate to high High
and ileum) blood-tinged exudate
Posterior intestine (Meckel’s Blood clot and
E. brunetti Moderate to high High
diverticulum to cecal junction) coagulated necrosis
Mid-intestine (part of jejunum Petechiae hemorrhage and
E. necatrix High Low
and ileum) blood-filled exudate
Hemorrhagic lumen and
E. tenella High Ceca Low
blood clot

Avian coccidiosis is a common and widespread disease of chickens around the world
and is transmitted by the fecal–oral route. Outbreaks of coccidiosis are reported wherever
chickens are raised [2,5]. Eimeria oocysts can remain viable for a long period of time, and
the disease can become endemic to the areas where environmental and managemental con-
ditions favor year-round survival and multiplication [2,5]. The incidences of coccidiosis are
usually higher in litter-based intensive systems, which favor the survival and accumulation
of oocysts [4].
Eimeria’s life cycle is short and has high reproductive potential with self-limiting
infection [2,5]. The life cycle of the Eimeria consists of an exogenous phase involving
the sporulation of the oocyst in the environment and an endogenous phase involving
asexual reproduction (schizogony) followed by sexual reproduction (gametogony) within
the avian intestine. Infection initiates with the ingestion of sporulated oocysts from the
environment [2,5,17]. Each sporulated oocyst contains four sporocysts, each containing
two sporozoites. Upon ingestion, mechanical and biochemical forces break down the
oocyst, releasing the sporozoites in the digestive tract [2,5]. Subsequently, sporozoites
migrate to specific sites depending on the species involved and invade the intestine’s
epithelial enterocytes. Within enterocytes, sporozoites transform into trophozoites and
begin multiple asexual multiplications, forming schizonts, known as schizogony. Mature
schizonts release merozoites, which again invade epithelial enterocytes, repeat the asexual
cycle, and complete the second schizogonous cycle. Depending on the species, they might
complete another schizogonous cycle before forming male (microgametocytes) and female
(macrogametocytes) gametocytes [2,5]. Mature gametocytes release microgametes that
fertilize mature macrogametocytes, resulting in the formation of oocysts. Upon maturation,
oocysts rupture epithelial cells, which are excreted in feces and, under favorable conditions,
Animals 2024, 14, 1015 4 of 33

sporulate within 48–72 h, becoming infective [2,5]. With each successive cycle, the number
of sporulated oocysts increases exponentially in the environment, which retains their
infectivity for extended periods [18]. The development of Eimeria in a host results in the
destruction of the host’s intestinal tissues, leading to clinical manifestation observed in a
disease outbreak.

3. Host–Pathogen Interaction and Immune Responses against Coccidiosis

Eimeria species are highly host-specific and, once infected, can produce protective
immunity against that particular species [19–21]. Notably, no cross-immunity is observed
among Eimeria species. Generally, a large number of Eimeria oocysts are required to
induce complete protective immunity, except for E. maxima, which is considered more
immunogenic than other species [22]. The asexual life cycle of Eimeria is more immunogenic
than the sexual life cycle. However, for the development of complete immunity and
protection, both sexual and asexual phases of the life cycle are equally essential, especially
for adaptive immunity [23,24].
Gut-associated lymphoid tissue (GALT) is an integral component of mucosal-associated
lymphoid tissues and plays a vital role in the immune response as a first line of defense, pre-
venting the progression of diseases and destroying infectious agents at an early stage [18,22].
GALT is a multilayered tissue with antigen-presenting cells, immunoregulatory cells, and
effector cell types different from their systemic counterpart [19]. In poultry, during coc-
cidiosis, GALT serves as antigen recognition and presentation, followed by the release
of intestinal antibodies and activation of the cell-mediated immune response [18,19,25].
Various lymphoid tissues (Peyer’s patches, bursa of Fabricius, and cecal tonsils) have
evolved in the GALT to produce verities of immune cells (epithelial, lymphoid, antigen-
presenting, and natural killer cells) to protect against invading pathogens [18,19,22]. Peyer’s
patches serve as antigen recognition and immune response activation, along with facil-
itating gastrointestinal IgA secretion [19,22,25]. Following this, activated B and T cells
migrate to lamina propria, which serves as the effector site for immune responses, initi-
ating antigen-specific/non-specific responses involving antibody production, leukocyte
accumulation, and locally produced cytokines [19,25]. The activation of antigen-presenting
cells, such as macrophages and dendritic cells, by coccidia upregulates the production of
pro-inflammatory cytokines and chemokines from innate immune cells, which are essential
for the development of the adaptive immune response. T lymphocytes play a crucial role in
the response to coccidia infection, with various T cell subpopulations capable of recognizing
multiple antigens and modulating humoral and cell-mediated immunity [18,19,25].
Cell-mediated immune responses are an important effector mechanism that includes
antigen-specific and non-specific activation of various cell populations, including T lympho-
cytes, CD4+ helper T cells and CD8+ cytotoxic T cells, natural killer cells, and macrophages.
T cell activation is associated with major histocompatibility complex (MHC), with cyto-
toxic T cells recognizing antigens presented in the context of MHC-I and helper T cells
recognizing antigens associated with MHC-II molecules. Antigen-presenting and dendritic
cells help activate naïve CD4+ T cells into different subsets such as Th1, Th2, Treg, and
Th17 [19,26,27]. Activated helper T cells are involved in the humoral immune response, cy-
totoxic activity activation, macrophages, and natural killer cells [21]. Following coccidiosis,
cytotoxic T cells, along with macrophages and natural killer cells, identify coccidia-infected
host cells and modulate mononuclear cells to produce interferon-γ, activating proinflamma-
tory pathways to inhibit the development of intracellular Eimeria within host cells [18,19,28].
Cytokines produced by CD4+ Th1 cells (IL-2, IL-12, IFN-γ, and TNF-α) aid in clearing
the infection by promoting the activation of macrophages and other immune cells capable
of eliminating intracellular coccidia [26,27]. The importance of cytotoxic T cells during
coccidiosis has been demonstrated previously, as evidenced by the presence of intesti-
nal intraepithelial lymphocytes expressing more than 75% CD8+ T cells, the presence of
CD8+ cells in GALT within 24 h of infection, and the presence and activation of mono and
polynuclear cells [19,22,29,30].
Animals 2024, 14, 1015 5 of 33

4. Coccidiosis and Its Effects on Pullets and Laying Hens

4.1. Gut Health and Oxidative Stress
The negative effect of coccidiosis on the parameters associated with gastrointestinal
health, such as increased intestinal lesion scores, gastrointestinal permeability, a breach
in tight junction integrity, damage to intestinal morphology, inflammation, and oxidative
stress, are related to Eimeria multiplication and release in the gut lumen [2,5–7,31]. Con-
tinuous intestinal epithelial cell turnover occurs throughout the lifetime of the chickens.
However, coccidiosis infection increases epithelial cell turnover by two times compared
with noninfected birds [32]. It has been reported that the highest turnover of epithelial cells
was observed between 5 and 7 days post-Eimeria inoculation due to the release of large
numbers of first- and second-generation merozoites into the lumen. As a result, the mRNA
and protein expression of a cell proliferative marker, the proliferating cell nuclear antigen,
in the duodenum epithelium was upregulated, thus increasing crypt depth and resulting
in a lower villus height to crypt depth ratio [33]. The difference between Eimeria-infected
and non-infected birds in terms of villus height might range from 10 to 40% and is also de-
pendent on the Eimeria species and level of infection [7,34–40]. This epithelial cell turnover
during Eimeria infection might be a host defense mechanism to limit the multiplication of
the Eimeria by the expulsion of infected cells [41]. Moreover, the tight junction proteins
maintaining the tight junction integrity of the small intestine are composed of claudin,
occludin, junctional adhesion molecules, and zonula occludens families [42]. These tight
junction proteins were reported to be upregulated in Eimeria-infected birds along with
increased permeability by more than 100%, supporting the observation of a rapid turnover
rate in the intestinal epithelium during Eimeria infection [6,7,37,38,43]. Similar results were
observed in pullets and laying hens of different age groups and at various stages of egg
production [34–36].
During coccidiosis, the equilibrium between reactive oxygen species (ROS) and the
host’s ability to neutralize ROS is disrupted, leading to oxidative stress. These ROS have
a high affinity for the phospholipid bilayer of the cell, initiating lipid peroxidation and
cytotoxic changes, which in turn damages the intestine, altering the gastrointestinal in-
tegrity and causing abnormal changes in the intestinal morphology [44,45]. Under normal
circumstances, the production of ROS and its effects on cellular mechanisms are con-
trolled by the host defense mechanism consisting of enzymes such as glutathione (GSH),
glutathione peroxidases (GPx), and catalase [44]. During Eimeria infection, although ox-
idative defense mechanisms are active, the production of ROS exceeds the production of
enzymes suppressing ROS, aggravating the situation. Previous studies have reported that
Eimeria infection increased the markers associated with oxidative stress, such as reduced
concentration of total antioxidant capacity (TAC) from 15 to 40% [31,46] and reduced
GPx and GSH activity [31,47]. Furthermore, Eimeria infection also increased the markers
of radical-induced damage, such as superoxide dismutase (SOD) and malondialdehyde
(MDA) [34–36,44,48,49].

4.2. Growth Performance

The effect of coccidiosis on the growth performance of birds depends on the Eimeria
species and the severity of the infection. Previous studies have reported that broilers
challenged with 105 oocysts of Eimeria acervulina reduced their body weight gain (BWG) by
10.3%, whereas 106 oocysts per bird decreased BWG by 26.7% [50]. Likewise, similar results
were observed with E. maxima and E. tenella [50]. On average, coccidiosis reduced the body
weight (BW) of the chickens by 10%, irrespective of the species involved in the infection [51].
E. maxima-induced coccidiosis was observed to have the most severe negative impact on the
BW of broilers, ranging from 23% to 37% [6,51], followed by E. tenella, ranging from 15% to
27% [52,53], and E. acervulina, from 16% to 19% [54,55]. However, Choi et al. (2021) [56] did
not observe any reduction in the BW of broilers when they were inoculated with sporulated
oocysts of E. tenella ranging from 6250 to 50,000 [56]. In cases of mixed Eimeria (E. maxima,
E. tenella, and E. acervulina) infections, the reduction in BW can range from 13% to 50%
Animals 2024, 14, 1015 6 of 33

with an increase in Eimeria dosage from 6250 to 50,000 E. maxima, 6250 to 50,000 E. tenella,
and 31,250 to 250,000 E. acervulina [7,37,46]. However, mixed Eimeria infection has been
associated with higher mortality rates, reaching up to 47%, and reductions in BW ranging
from 9% to 28% in laying hen pullets aged two weeks, 8% to 15% in pullets aged 15 weeks,
and 9% to 11% in hens aged 25 weeks during the early phase of infection [34–36]. Most
studies have reported reductions in BW during the early phase of infection (1–8 days post-
Eimeria inoculation) but not during the recovery phase (8–14 days post-Eimeria inoculation).
During the early phase of infection, the reduction in feed intake and damage to the intestinal
linings are more pronounced compared with those in the recovery phase, where feed intake
and intestinal damage recover [7,35]. Additionally, during the early phase, nutritional
redistribution occurs towards the immune response to subside the infection [7,35,36]. This
reduction in feed intake and nutritional redistribution are associated with reduced growth
or body weight loss in pullets and laying hens.

4.3. Production Performance

Coccidiosis rarely occurs in laying hens because of previous infection and the resulting
immunity [10]. However, pullets reared in cages/on floors with coccidiostat drugs might not
have enough exposure to coccidia to stimulate immunity. Outbreaks can occur after moving to
layer facilities, either conventional cages or alternative housing systems [5,8,10]. Coccidiosis in
laying hens has been reported during the early egg production period, around 23–24 weeks
of age, and is characterized by high morbidity and mortality with a dramatic reduction in
egg production [4,10,57–59]. Temporary cessation of egg production has been observed in
laying hens infected with E. maxima, E. acervulina, and E. tenella. Hegde and Reid (1969) [57]
reported that total egg production dropped to less than 20% when susceptible laying hens
were independently challenged with different Eimeria species (E. acervulina, E. maxima, E.
brunetti, E. necatrix, or E. tenella), with a significant drop observed two weeks post-challenge.
It took 4–6 weeks to recover from infection, which varied among species. The mortality was
more significant with E. necatrix and E. maxima following infection; however, the number
of culled birds was not different from the control unchallenged group. Single-comb white
leghorns infected with E. mitis experienced significant reductions in egg production and
eggshell quality, undergoing complete molting before resuming egg production [58]. Hens
recovered from coccidiosis may not reach their full egg-laying potential [60]. An outbreak
of E. necatrix in a breeder hen facility resulted in significant mortality among breeder hens
and roosters, as well as a 4.3% reduction in egg production following the recovery period
compared with hens of the same age [59]. Recent studies in our lab have observed that
coccidiosis at the pre-lay stage of growth delayed the onset of maturity and egg production
by two weeks, and infection at peak egg production drastically dropped egg production
and temporarily ceased egg production in some hens [34,36]. Reduced egg production during
a coccidiosis outbreak might be because of the malabsorption of nutrients, including amino
acids, carbohydrates, and minerals, which are essential during the laying period [6–8,61]. The
possible mechanism of how coccidiosis affects the performance of laying hens is shown
in Figure 1. Reduced feed intake and damage to intestinal linings following coccidiosis
are evident. This damage caused by Eimeria to the intestinal tract affects the digestion
and absorption of the nutrients required for egg formation [36]. Furthermore, it has been
observed that plasma amino acids are lowered, and amino acid imbalance occurs during
coccidiosis [55]. The limited availability of nutrients and amino acids in laying hens might
reduce the synthesis of albumin, an important component of an egg, thus reducing egg
production. Increased oxidative stress during coccidiosis might damage the developing
oocytes and granulosa cells, leading to follicular atresia, production, and deposition of
yolk precursors in selected ovaries, or occurrence of apoptosis in the oviducts and ovarian
follicles [36,62,63]. These disruptions to normal physiological and reproductive functions
might contribute to a decline in egg production. Additionally, the redistribution of energy
during coccidiosis for tissue repair, inflammation, and maintaining oxidative status might
also be responsible for the decline in egg production [36]. These studies showed that laying
 ovaries, or occurrence of apoptosis in the oviducts and ovarian follicles [36,62,63]. These
disruptions to normal physiological and reproductive functions might contribute to a de-
cline in egg production. Additionally, the redistribution of energy during coccidiosis for
tissue repair, inflammation, and maintaining oxidative status might also be responsible
Animals 2024, 14, 1015 7 of 33
for the decline in egg production [36]. These studies showed that laying hens are suscep-
tible to coccidiosis during the laying phase, with a dramatic reduction in egg production
posing
hens substantial challenges
are susceptible to the
to coccidiosis egg the
during industry. The economic
laying phase, loss toreduction
with a dramatic the industry
in is
mostly from reduced egg production, reduced feed intake, and an increased FCR
egg production posing substantial challenges to the egg industry. The economic loss to the due to
malabsorption of nutrients
industry is mostly and mortality.
from reduced egg production, reduced feed intake, and an increased
FCR due to malabsorption of nutrients and mortality.

Figure 1. Potential mechanism of action showing how coccidiosis affects egg production in laying hens.
Figure 1. Potentialand
5. Prevention mechanism of Coccidiosis
Control of action showing how coccidiosis affects egg production in laying
hens.
5.1. Biosecurity and Management
Biosecurity is an important tool in poultry production to prevent disease outbreaks on
5. Prevention and Control of Coccidiosis
farms. Once a coccidiosis outbreak occurs on a farm and the house becomes contaminated
5.1.with
Biosecurity
oocysts, and
it is Management
virtually impossible to completely decontaminate the environment [2,5].
Strict biosecurity measures
Biosecurity is an important should be implemented
tool between farms
in poultry production and chicken
to prevent houses
disease to
outbreaks
minimize the risk of cross-contamination and outbreaks. In litter-based intensive housing
on farms. Once a coccidiosis outbreak occurs on a farm and the house becomes contami-
systems, Eimeria oocysts accumulate in large amounts and can remain viable for longer
nated with oocysts, it is virtually impossible to completely decontaminate the environ-
periods of time [2]. In those systems, management strategies should focus on maintaining
mentthe[2,5].
optimumStrictlevel
biosecurity measures
of moisture should
in the litter so be
thatimplemented
it can maintain between
the viablefarms and chicken
number of
houses to minimize
sporulated oocysts the risk oftocross-contamination
sufficient and outbreaks.
generate protective immunity but notIn litter-based
high enough tointen-
siveinduce
housing systems,
clinical disease. Eimeria oocysts
Furthermore, accumulate
when employing invaccination
large amounts and can remain
as a preventive viable
measure,
foritlonger
is also periods
critical toofmaintain
time [2]. In thoselitter
optimum systems, management
moisture, strategies
as oocyst recycling should focus
is important in on
inducing a protective
maintaining the optimum immunelevelresponse againstin
of moisture specific Eimeria
the litter so species
that it [64,65]. Additionally,
can maintain the viable
immunosuppression
number of sporulatedfrom stressors
oocysts such asto
sufficient heat, cold, orprotective
generate high stocking density inbut
immunity chicken
not high
houses might increase susceptibility to the disease; therefore, it
enough to induce clinical disease. Furthermore, when employing vaccination as a preven-is critical to maintain
optimum environments in the houses [66].
tive measure, it is also critical to maintain optimum litter moisture, as oocyst recycling is
5.2. Chemotherapy
Traditionally, chemotherapeutic (anticoccidial) drugs have been used in the poultry
industry for years all around the world to control coccidiosis outbreaks in farms; however,
Animals 2024, 14, 1015 8 of 33

the emergence of drug resistance among the zoonotic pathogens and concerns raised by the
World Health Organization and consumers led to a ban of antibiotics use in food-producing
animals including poultry as growth promoters and disease prevention [67,68]. In poultry,
coccidiostat (inhibits the replication of Eimeria) or coccidiocidal (kills and destroys the
Eimeria) drugs are used, which can be either synthetic or ionophores [64,68]. Synthetic
anticoccidial drugs (chemicals) are produced by chemical synthesis from other chemical
compounds that have diverse modes of action in controlling Eimeria, from inhibiting mi-
tochondrial respiration and disrupting energy production (clopidol or decoquinate) to
competitive inhibition of thiamine uptake (amprolium) or inhibiting folic acid pathways
(sulphonamides). Some of the synthesized chemicals approved to be used in the poultry
industry are diclazuril, halofuginone, clopidol, robenidine, decoquinate, nequinate, ampro-
lium, and zoalene [68,69]. These chemicals act on the intracellular stage of Eimeria after they
invade the host intestine and interfere with one or more stages of the life cycle [69]. How-
ever, while using these chemicals, it is important to consider that they are more susceptible
to the development of resistance [68]. Additionally, these chemicals vary in their mode of
action, efficacy, dose range, and susceptibility to resistance; it is important to rotate between
them at certain intervals to minimize the development of resistance [68]. On the other hand,
ionophores are fermented products from bacteria such as Streptomyces spp. and Actino-
madura spp. [64,68]. Ionophores facilitate the transportation of ions across cell membranes,
increasing the concentration of intracellular ions and disrupting the normal ion balance
across cell membranes [70]. The common ionophores used in the poultry industry are
monensin, narasin, salinomycin, maduramicin, semduramicin, and lasalocid. Even though
these ionophores are not of human importance, the ban on antibiotics in animal production
may include these, which have been the backbone of coccidiosis prevention programs for
many years, forcing the poultry industry to look for alternatives [68]. Although the poultry
industry has used shuttle or rotational programs to minimize resistance to these drugs in
the past, most field strains of Eimeria have shown varying levels of resistance to more than
one drug [64,71].

5.3. Vaccination
Vaccination against Eimeria species has become one of the most widely used methods
in controlling coccidiosis outbreaks in the United States. The discovery of the self-limiting
infection of Eimeria species and the development of resistance to reinfection by the same
Eimeria species by Beach and Corl. (1925) [72] paved the way for the development and use of
a live vaccine to control coccidiosis in the USA by 1952. In addition to self-limiting infection,
Eimeria species are less susceptible to younger chicks, and the generation of acquired
immunity against Eimeria species with limited to no pathogenesis made vaccination an
efficient strategy to control coccidiosis in commercial poultry production [73]. The objective
of immunization is to expose the birds to Eimeria species to elicit protective immunity
against them, which is adequate to prevent future infections [9,15,65,66,74]. However,
the major disadvantage of vaccination is the absence of cross-immunity among different
Eimeria species; thus, it is more labor-intensive to prepare and has a high cost because
of including multiple species in the vaccine [75]. When vaccinating meat birds against
coccidiosis, the emphasis is placed on E. acervulina, E. maxima, and E. tenella. In contrast,
for egg-laying hens, the focus shifts to E. tenella, E. acervulina, E. maxima, E. brunetti, E.
necatrix, and E. praecox, primarily because of variations in the life spans of the respective
hosts (Table 2) [15]. To ensure birds build strong immunity against Eimeria species, they
must undergo oocyst recycling through three to four successive re-infections so that they
have complete immunity against Eimeria by 3–4 weeks of age when most coccidiosis
outbreaks occur [17,65,75,76]. In the United States, two types of live vaccines are available
based on the pathogenicity of the parasites, including (i) nonattenuated and (ii) attenuated
live oocyst vaccines [65,75]. Nonattenuated vaccines contain laboratory or field strains of
Eimeria oocysts that have their virulence preserved to elicit protective immunity. With a
nonattenuated live oocyst vaccine, immunity against Eimeria is produced by completing the
Animals 2024, 14, 1015 9 of 33

life cycle and boosted by recycling oocysts from the litter for reinfections [9,17,65,73,75–77].
However, the significant disadvantages of nonattenuated vaccines are their short shelf life,
high cost of production due to the inclusion of all Eimeria species, occasional outbreaks
of coccidiosis on farms, halted growth, and a possible lack of compensation for growth
after recovery [9,15,65,75,77]. On the other hand, live attenuated oocyst vaccines are
selected via passage through embryonated hen eggs or by selection for precociousness
and have demonstrated reduced pathogenicity while maintaining the ability to induce
protective immunity [15,65,75,78]. This selection for precocious has decreased pathogenicity
while still having the capacity to stimulate the immune response against selective Eimeria
species [65,73–75,77]. Recombinant or subunit vaccines are composed of immunogenic
antigens capable of producing protective immunity against Eimeria. However, identifying
the antigens that elicit protective immunity and limited to no cross-immunity among
Eimeria species are limiting factors for the development of recombinant vaccines [65,74,77].
Thus, for complete protection against the prevalent Eimeria species, it is essential to identify
antigens against several isolates of the same species and for multiple species to ensure
complete protection [65,77].

Table 2. Commercial vaccines available in the U.S. for use in laying hens; adapted from Price (2014) [15]
and Cervantes (2023) [68].

Eimeria Species Present in Route of Age of Birds at

Trade Name Vaccine Type Manufacturer
Vaccine Administration Vaccination
E. acervulina, E. brunetti, E. Hatchery spray,
Single dose at 1 to
Coccivac-D2 maxima, E. mivati, E. necatrix, Nonattenuated ocular, drinking Merck
4d
and E. tenella water, feed spray
E. acervulina, E. brunetti, E.
Drinking water,
Immucox C maxima, E. necatrix, E. tenella, Nonattenuated Single dose 1 to 4 d Ceva
oral gel
and E. praecox
E. acervulina, E. brunetti, E.
Drinking water, Single dose at 1 to
Paracox maxima, E. mitis, E. necatrix, E. Attenuated Merck
feed spray 9d
praecox, and E. tenella
E. acervulina, E. brunetti, E.
Immunocox5 maxima, E. necatrix, and E. Nonattenuated Gel Single dose 1 to 4 d Ceva
tenella

6. Nutritional Intervention for Coccidiosis

6.1. Role of Vitamins
6.1.1. Vitamin D
Vitamin D is a fat-soluble vitamin, and the active metabolites of vitamin D play an
intrinsic role in calcium and phosphorus homeostasis and bone mineralization [79–82].
More recently, the immunomodulatory roles of vitamin D in improving the host defense
against invading pathogens have been described [83–86]. The absorption of vitamin D is
fat-dependent and undergoes hydroxylation into 25-hydroxyvitamin D3 [25(OH)D3 ] in the
liver [86,87]. Subsequently, 25(OH)D3 is converted into its active form, 1,25-dihydroxy vita-
min D [1,25(OH)2 D3 ], in the kidney by the enzyme 25-hydroxyvitamin D-1α-hydroxylase.
The hydroxylation of 25(OH)D3 to 1,25(OH)2 D3 is not limited only to the kidney but also
occurs in the bone, breast, and thigh muscles, small intestine, and immune cells [88].
The inclusion of 25(OH)D3 at varying levels in either layer pullets or turkey poults
challenged with Eimeria reduced BW loss and fecal oocyst shedding, increased macrophage
nitric oxide (NO) production, stimulated the activity of innate immune cells, and modulated
adaptive immunity (CD4+, CD8+ and CD4+CD25+ T cells) and inflammatory cytokines
(IL-1β and IL-10), as well as maintained tight junction [85,86,89]. Furthermore, supplemen-
tation of either vitamin D or 25(OH)D3 or in combination has been shown to improve the
FCR and BWG in broilers vaccinated against Eimeria spp. at high doses; however, no beneficial
Animals 2024, 14, 1015 10 of 33

effects were observed in reducing intestinal lesions and improving morphometry [90–92].
In broilers challenged with either E. maxima or mixed Eimeria spp. and fed reduced cal-
cium/phosphorus diets, supplementation of 25(OH)D3 at the rate of either 3000 IU/kg or
4000 IU/kg has been shown to improve bone mineralization [83,84,93]. Furthermore, in
laying hens challenged with lipopolysaccharides, 25(OH)D3 supplementation improved
egg production by improving follicular development and oocyte maturation by increasing
the expression of VDR in oocytes, maintaining the plasma estradiol/progesterone and
luteinizing hormone levels [94–97]. Moreover, the lipopolysaccharide challenge increased
the immune response (CD4+CD25+ T cells, proinflammatory cytokines, and plasma IgM
levels) and oxidative stress (SOD, TAC, and GPx) in laying hens, which were normalized
by the addition of 25(OH)D3 [97,98]. Previous studies have reported that inflammation and
oxidative stress associated with coccidiosis are responsible for at least a 10% reduction in
bone volume, bone mineral content and density, and osteoclastic activity [46,48,99]. These
results suggest that vitamin D and its metabolites can potentially reduce the negative
impacts of coccidiosis related to growth performance, egg production, oxidative stress, and
bone mineralization.

6.1.2. Vitamin E
Similar to vitamin D, vitamin E is a fat-soluble vitamin with a potent antioxidant
capacity to protect cells and tissues from lipoperoxidation damage induced by reactive
oxygen species [100–102]. Vitamin E from diets gets absorbed and incorporated into cell
membranes to protect unsaturated fatty acids inside and outside the cells from reactive
oxygen species. Furthermore, vitamin E prevents the excessive generation of reactive
oxygen species from respiratory processes [103,104]. In addition, vitamin E is also involved
in reducing inflammation, production of cytokine (TNF-α and IL-8) protecting cells of
immune responses (lymphocytes, macrophages, and plasma cells), and enhancing the
proliferation and functions of immune cells [100,105].
A previous study by Colnago et al. (1984) [106] observed that supplementation of
100 IU/kg vitamin E (DL-α-tocopheryl acetate) in a broiler diet and subsequent challenge by
E. tenella or E. maxima reduced mortality, improved weight gain and the FCR, and re-
duced lesion scores. Similar results were observed when broiler chicks were challenged
twice at 10 and 38 days of age and supplemented with both vitamin A (8 g/kg) and E
(300 mg/kg) [107]. However, Allen and Fetterer (2002) [101] conducted two consecutive
studies with increasing levels of vitamin E (13–200 IU/kg) in broilers challenged with
E. maxima and did not observe significant differences in performance except for a slight
reduction in lesion score. In broilers fed vitamin E at either 40 or 80 IU/kg alone or
in combination with arginine and challenged with mixed Eimeria spp., heterophil and
monocyte oxidative bursts (ROS production) and NO production decreased at seven
days post-inoculation. They also observed lower lesions in challenged birds fed vita-
min E, and birds fed a high level of vitamin E (80 IU/kg) in combination with arginine
had higher levels of humoral antibodies IgG, IgM, and IGA [108]. Increased humoral
and cell-mediated immune response with lower inflammatory mediators were observed
in laying hens challenged with Salmonella enteritidis and fed diets supplemented with
30 IU/kg vitamin E [109]. Furthermore, da Silva et al. (2011) [110] observed that the
inclusion of vitamin E at the rate of 65 mg/kg of diet increased the cell-mediated immune
response of chickens vaccinated against coccidiosis and New Castle disease as measured
by the cutaneous basophil hypersensitivity test. The antioxidant status (TAC, MDA, GPx
activity, or SOD) and plasma lipid peroxidation of the broilers were improved following
the Eimeria challenge in broilers, but the effects were not enough to improve performance
compared with non-challenged broilers [111,112]. Furthermore, vitamin E has been shown
to enhance the phagocytic activity of macrophages in chickens against pathogens [100].
In the case of egg layers, vitamin E inclusion has increased egg production by enhanc-
ing follicular development by increasing the concentration of reproductive hormones such
as follicle-stimulating hormone, luteinizing hormone, estrogens, and progesterone [102].
Animals 2024, 14, 1015 11 of 33

These results from previous studies showed that vitamin E can be used in laying hens to
improve their performance and boost the immune response; however, further studies are
needed to confirm the effective dose.

6.1.3. Other Vitamins

Vitamin A is a fat-soluble nutrient constituting a broad range of retinoid compounds
and plays a crucial role in various physiological processes, including vision, growth, cell
growth and differentiation, reproduction, immunity, skeletal development, and antioxidant
properties [113–115]. The association between vitamin A deficiency and the increased inci-
dence and severity of coccidiosis was noted as early as 1945 [116]. Dietary supplementation of
vitamin A above the 1960 NRC recommendation (8000 IU/lb) has been shown to enhance
broiler performance and the recovery of chicks infected with E. tenella, acervulina, or necartix
and results in an improved performance after recovery [117,118]. Furthermore, Dalloul et al.
(2002) [113] reported that vitamin A deficiency compromised the intestinal defense mecha-
nism against E. acervulina infection, as evidenced by a reduced population of intraepithelial
lymphocytes (CD4+ and CD8+ T cells) through alterations in concanavalin A-induced spleen
lymphocyte proliferation. Moreover, vitamin A supplementation (12,000 IU/kg) enhanced
intestinal morphometry and tight junction integrity in broiler chickens co-infected with
Clostridium and Eimeria [119].
Unlike vitamins A, D, and E, vitamin C is a water-soluble vitamin with a strong
antioxidant capacity and acts as a cofactor for collagen biosynthesis, thus maintaining
epithelial barrier function and stimulating wound healing [120]. Moreover, it enhances
innate immunity by increasing the phagocytic activity of mononuclear cells and adaptive
immune responses by differentiation and proliferation of B- and T-cells [121]. Supple-
menting vitamin C (110–220 ppm) improved weight gain, increased feed intake, reduced
mortality, and lowered the corticosterone level and heterophil: lymphocyte ratio in the
blood [122,123]. Additionally, concurrent supplementation of vitamin C and protease
improved mucin and NO production [124], and vitamin C and arginine or vitamin C and
vitamin E in different experiments improved oxidative status but were not able to improve
performance in Eimeria-infected broilers [112]. The effect of supplementing different vi-
tamins on minimizing the effect of coccidiosis is summarized in Table 3. A hypothesis
for using vitamins during coccidiosis could posit that both fat-soluble and water-soluble
vitamins may be readily absorbed by the body, potentially exerting beneficial effects on
modulating immune responses despite reduced feed intake and nutrient utilization. While
there is a notable gap in the available literature concerning the utilization of vitamins to
support laying hens during disease conditions, the positive outcomes observed in broilers
suggest that similar benefits might extend to laying hens as well. However, further studies
are necessary to confirm the optimal dosage for this beneficial effect in laying hens.
Animals 2024, 14, 1015 12 of 33

Table 3. Effects of vitamin supplementation on performance and intestinal health of chickens infected with Eimeria species.

Nutritional Interventions Breed Eimeria Infection Dosage Impacts on Performance Impact on Health Reference
• ↓ CD8+ cells.
E. acervulina, E. maxima, and • ↑ CD4+CD25+ cells.
25-hydroxycholecalciferol White Leghorn 4000 IU/kg Improved performance [85]
E. tenella (Inovocox) • ↓ IL-1β expression.
• ↑ IL-10 expression.
7× live vaccine (E. • ↑ Duodenal morphology.
Vitamin D3 + Improved BW, BWG, and
Cobb 700 acervulina, E. maxima, and E. 2000–8000 IU/Kg • ↑ Tibia breaking strength. [91]
25-hydroxycholecalciferol FCR
tenella) • ↑ Tibial bone mineralization.
• ↑ Bone breaking strength.
Vitamin D3 or Improved BW, BWG, and
Ross 308 E. maxima (7000) 4000 IU/Kg • ↑ Bone mineralization. [84]
25-hydroxycholecalciferol FCR
• ↓ Jejunal morphology.
Vitamin D3 or • ↑ Bone breaking strength.
Ross 308 E. maxima (7000) 4000 IU/kg No effect on performance [83]
25-hydroxycholecalciferol • ↑ Ash content and percentage.
• ↓ Intestinal morphology.
Vitamin D3 or Improvement in BW and
Cobb 500 2× live vaccine 1375 or 2750 IU/kg • ↑ Tibia breaking strength. [92]
25-hydroxycholecalciferol FCR
• ↑ Bone ash %.
• ↑ Bone ash.
E. acervulina (62,500), E.
• ↑ Bone mineral content of the birds.
25-hydroxycholecalciferol Cobb 500 tenella (12,500), and E. 3000 IU/kg No effect on performance [93]
• ↑ Tissue and bone volume of the
maxima (12,500)
femur.
• ↓ Mortality.
E. tenella (150,000) or E. Improved BWG, FI, and
DL-alpha tocopheryl acetate Hubbard 100 IU/Kg • ↓ Lesion scores. [106]
maxima (50,000) FCR
• ↑ Packed cell volume.
E. maxima (175,000 or • ↓ Lesion scores.
DL-alpha tocopheryl acetate Ross 13–200 ppm No effect on performance [101]
40,000) • ↑ Plasma NO.
• ↑ Heterophil and monocyte
E. acervulina (100,000), E.
oxidative burst.
DL-alpha tocopheryl acetate Cobb 500 maxima (60,000), and E. 40 or 80 IU/kg - [108]
• ↑ IgG, IgM, and IgA production.
tenella (40,000): Coccivac-B
• ↓ Lesion scores.
D-α-tocopherol Ross 308 Livacox Q vaccination 65 mg/kg - • ↑ Cell-mediated immunity. [110]
D-α-tocopherol Ross 308 E. tenella 100, 316, or 562 mg/kg - • ↓ MDA concentration. [111]
• ↑ Nitric oxide production.
D-α-tocopherol and
Cobb 500 100× Advent vaccine 80 mg/kg VE + 1 g/kg VC Improved BWG • ↑ GPx activity. [112]
L-ascorbic acid
• ↓ Lesion scores.
E. maxima (30,000), E.
Vantress-White Plymouth
L-ascorbic acid brunetti (100,000), E. necatrix 110 mg/kg No effect on performance • Increased mortality. [123]
Rock cross
(30,000), or E. tenella (30,000)
• ↓ Plasma corticosterone.
L-ascorbic acid Hubbard E. tenella (300,000) 150 or 300 mg/kg Improved BW, FI, and FCR • ↓ Heterophil/lymphocyte ratio. [122]
Animals 2024, 14, 1015 13 of 33

Table 3. Cont.

Nutritional Interventions Breed Eimeria Infection Dosage Impacts on Performance Impact on Health Reference
Vitamin A Plymouth Rock 8000 IU/lb Improved BWG • ↑ Recovery. [117]
• ↓ CD4+, CD8+ cells.
• ↑ Oocyst shedding.
Vitamin A Ross 308 E. acervlina (10,000) VE deficiency - [113]
• ↓ IFN-γ.
• ↓ Lymphocyte proliferation.
• ↑ Intestinal morphology.
E. maxima + Clostridium
Vitamin A Broilers 12,000 IU/kg Improved BW, BWG, and FI • ↑ Expression of tight junction [119]
perfringens
proteins.
Lancaster X New
Menadione sodium bisulfite E. tenella (500,000) 1.06 or 9.72 g/ton - • ↓ Mortality. [125]
Hampshire
IU: international unit; BW: body weight; BWG: body weight gain; FI: feed intake; FCR: feed conversion ratio; NO: nitric oxide; IL-1β: Interleukin 1β; IL-10: Interleukin 10; IFN-γ: Interferon γ;
IgA: immunoglobin A; IgG: immunoglobin G; IgM: immunoglobin M; ppm: parts per million; VE: vitamin E; VC: vitamin C; MDA: malondialdehyde; GPx: glutathione peroxidase.
Animals 2024, 14, 1015 14 of 33

6.2. Role of Functional Amino Acids

6.2.1. Arginine
Arginine, an essential functional amino acid, is vital in protein synthesis and accretion
in chickens. Additionally, it contributes to secondary functions such as immune modulation,
wound healing, and antioxidant activity [126–129]. In response to inflammation, arginine
is metabolized to nitric oxide (NO) by nitric oxide synthase, which serves as an immune
modulator, antioxidant defense, and cytotoxic mediator for non-specific host defenses [126–128].
Additionally, metabolites derived from arginine, such as proline, hydroxyproline, and
polyamines, have been demonstrated to promote cell proliferation (Figure 2). Furthermore,
ornithine and proline are primary amino acids found within collagen
Animals 2024, 14, x FOR PEER REVIEW 15 of 35
that are essential for
wound healing and skeletal growth [126–128,130,131].

Figure 2. Effect of L-arginine and its metabolites on gut health and reproductive functions in laying
Figure 2. Effect ofhens.
L-arginine and itshormone;
FSH: follicular-stimulating metabolites
ER: estrogen; on gut health
LH: luteinizing hormone;and reproductive functions in laying
IGF: insulin-like
growth factor 1.
hens. FSH: follicular-stimulating hormone; ER: estrogen; LH: luteinizing hormone; IGF: insulin-like
In the case of laying hens, there is a lack of evidence for arginine supplementation
growth factor 1. and Eimeria challenge; however, in lipopolysaccharide-induced immune suppression, ar-
ginine supplementation above the requirements reduced the expression of inflammatory
cytokines such as IL-1β, IL-10, TLR4, and NF-κB [141]. Furthermore, NO, a metabolite of
The secondary functions
arginine, ofrole
plays a significant arginine during coccidiosis
in the hypothalamus–ovarian axis and plays a have
vital role been extensively investi-
gated in recent years and have shown beneficial effects in chickens. An early study by Allen
in laying hen performance as well [142–144]. Based on previous studies in broilers and
laying hens, supplementation of L-arginine above the requirements can be used in laying
and Fetterer (2000) [132] reported
hens challenged alleviateE.
with Eimeria to that the acervulina
symptoms associatedinfection
with it. reduced the plasma concentra-
tion of arginine,6.2.2.
whereas
Methionine its metabolite NO was significantly increased. The hypothesis
Methionine is considered the first limiting amino acid in corn- and soy-based diets
for using arginine above the requirement could be that additional arginine might aid in
for laying hens and broilers and the first amino acid to be incorporated into the polypep-
balancing aminotioxidant
acidsfunctions,
tide chain in plasma
during and provide
translation [126,145]. a surplus
Aside from protein for secondary
synthesis, methionine has an-
acting as a precursor of cysteine, which is essential for GSH synthesis
functions. However,
supplementation of arginine
[128,146]. (500 protect
Both GSH and cysteine mg/kg) did not
the cells against reverse
ROS [146]. Because ofthe
its rolenegative
in effect associated
balancing oxidative status through GSH and cysteine, its efficacy has been tested in broil-
with E. acervulina, maxima, or tenella infection except for a reduction
ers exposed to coccidiosis. A study by Pourali et al. (2014) [147] reported that supplement- in oocyst shedding
ing 150% of total sulfur amino acids improved BWG, oocyst shedding, and hepatic
of E. tenella [133]. Increasing dietary arginine concentration
malondialdehyde concentration; however, no effect was observed in GPx in broilers chal-above the NRC requirement
lenged with mixed Eimeria species. A study by Lai et al. (2018, 2023) [148,149] reported
improved gut health by increasing villus height and goblet cell count and increasing mTOR
that supplementing extra methionine above the requirement in vaccinated birds did not
pathways for healing [134,135].
have any beneficial The
effect on the intestinal
performance, integrity
immune response, of infected
or oxidative status of birds was improved
Partridge shank broilers. However, reducing the level of methionine in the diets during
when supplementing arginine, ranging from 1.24 to 1.48%, as observed by the upregulation
of tight junction protein, permeability, and intestinal morphology [136,137]. Furthermore,
arginine supplementation also reduced the expression of TLR-4 and IL-1β in broilers
along with modulating the humoral immune response (secretory IgG and IgA) in broilers
challenged with 20× Coccivac-B vaccine [134]. In addition, Yazdanabadi et al. (2020a;
b) [138,139] reported that arginine supplemented 25–50% over the requirement increased
the improved BWG, NO, and proinflammatory concentration along with gut morphometry.
Furthermore, supplementation of arginine above the requirements has been found to im-
prove the oxidative status of chickens following Eimeria challenge by GPx activity, reducing
MDA and maintaining the levels of GSH and TAC [136,137,140].
In the case of laying hens, there is a lack of evidence for arginine supplementation and
Eimeria challenge; however, in lipopolysaccharide-induced immune suppression, arginine
supplementation above the requirements reduced the expression of inflammatory cytokines
such as IL-1β, IL-10, TLR4, and NF-κB [141]. Furthermore, NO, a metabolite of arginine,
Animals 2024, 14, 1015 15 of 33

plays a significant role in the hypothalamus–ovarian axis and plays a vital role in laying
hen performance as well [142–144]. Based on previous studies in broilers and laying
hens, supplementation of L-arginine above the requirements can be used in laying hens
challenged with Eimeria to alleviate the symptoms associated with it.

6.2.2. Methionine
Methionine is considered the first limiting amino acid in corn- and soy-based diets
for laying hens and broilers and the first amino acid to be incorporated into the polypep-
tide chain during translation [126,145]. Aside from protein synthesis, methionine has
antioxidant functions, acting as a precursor of cysteine, which is essential for GSH syn-
thesis [128,146]. Both GSH and cysteine protect the cells against ROS [146]. Because of its
role in balancing oxidative status through GSH and cysteine, its efficacy has been tested
in broilers exposed to coccidiosis. A study by Pourali et al. (2014) [147] reported that
supplementing 150% of total sulfur amino acids improved BWG, oocyst shedding, and
hepatic malondialdehyde concentration; however, no effect was observed in GPx in broilers
challenged with mixed Eimeria species. A study by Lai et al. (2018, 2023) [148,149] reported
that supplementing extra methionine above the requirement in vaccinated birds did not
have any beneficial effect on the performance, immune response, or oxidative status of Par-
tridge shank broilers. However, reducing the level of methionine in the diets during Eimeria
infection significantly affects the performance, oxidative status, and immune responses
in challenged broilers [31,47,150,151], and supplementing 0.75% methionine in low-crude
protein diets increased the detrimental effect of Eimeria infection on broiler health and
performance [6]. In laying hens under heat stress, similar results were observed as those
for Eimeria-challenged broilers, where reducing the methionine level reduced the produc-
tion performance and bone mineralization and increased the oxidative stress of laying
hens [152]. The role of TSAA is evident in maintaining performance, immune response,
and oxidative status during Eimeria infection; however, it is still unclear if supplementing
methionine above the requirement has a beneficial effect on laying hens under coccidiosis.

6.2.3. Other Amino Acids

Glutamine, a conditionally essential amino acid, is the primary fuel for immune and
intestinal epithelial cells and a precursor for GSH synthesis [153]. Supplementation of
glutamine (0.5% or 1%) in coccidia-challenged broilers has been shown to reduce the expres-
sion of inflammatory cytokines IL-10 and IFN-γ, whereas it improves intestinal integrity
and morphometry [154]. Furthermore, supplementing 0.75% glutamine in low-crude pro-
tein diets improved BWG during the recovery phase and maintained the expression of
Claudin-1 compared to that of a normal protein diet in Eimeria-infected broilers [6].
Threonine is another essential amino acid and is essential for mucin and antibody
(IgA, IgM) production in the gastrointestinal tract [155]. The intestinal mucus layer acts
as the first line of defense against invading pathogens [155]. Therefore, factors increas-
ing mucin production or intestinal secretions, as observed in coccidiosis, might increase
threonine requirements during infection [156]. Supplementing 124% of threonine to its
requirement in broilers challenged with mixed Eimeria species improved performance to
the level of non-challenged birds, intestinal morphometry, oocyst shedding, and humoral
immune response as measured by higher antibody production [157]. In contrast, threonine
deficiency worsened the effects of coccidiosis in broilers challenged with coccidiosis by
impairing intestinal morphometry and integrity, inflammatory responses, and lymphocyte
population [158]. The effect of supplementing functional amino acids on the performance
and intestinal health of chickens under coccidiosis infection is summarized in Table 4.
Although these functional amino acids have shown beneficial effects in broilers under
coccidiosis and in laying hens under normal conditions, their beneficial effect in laying
hens under diseased conditions has not been fully explored. These amino acids have shown
potential in mitigating the negative effect of coccidiosis in broilers and might have the same
functions in laying hens, which need to be explored.
Animals 2024, 14, 1015 16 of 33

Table 4. Effects of functional amino acids supplementation on the performance and intestinal health of chickens infected with Eimeria species.

AA Breed Eimeria Challenge AA Dosage Impact on Performance Impact on Health Reference

E. maxima (30,000), or
SexSal E. tenella (50,000), or +500 or +1000 mg/kg No effect on BWG • ↓ Oocyst shedding of E. tenella. [108]
E. acervulina (500,000)
E. acervulina (100,000), • ↑ Heterophil and monocyte
Added 0.3% or 0.6% to make
Cobb 500 E. maxima (60,000), oxidative burst. [108]
1.74% and 2.04% (NRC, 1994)
and E. tenella (40,000): Coccivac-B • ↑ IgG, IgM.
• ↑ Villus height and crypt depth.
Ross 708 20× Coccivac-B 106·4% and 160·8% of (NRC, 1994) Improved BWG and FCR • ↑ Goblet cell counts and density. [141]
• ↓ Expression of TLR4, IL-1β.
• ↑ NO production.
Cobb 500 100× Advent coccidiosis vaccine 1.80% (NRC 1994) No effect on BWG • ↑ GPx activity. [135]
• ↓ Intestinal lesions.
E. acervulina (150,000), E. tenella
105 and 110% (breeder • ↓ Crypt depth and oocyst
Ross 308 (15,000), E. maxima (15,000), and E. No effect on BWG or FCR [135]
recommendation) shedding.
necatrix (15,000)
Arginine
Low arginine (0.74%) Reduced BWG, FI, and FCR • ↓ Plasma Arg: Lys ratio.
[129]
Ross 308 E. acervulina (3.5 × 105 ) • ↑ Plasma Arg: Lys ratio.
High arginine (1.23%) Improved BWG, FI, and FCR
• ↑ NO production.
• ↓ Gut permeability.
• ↑ Tight junction proteins.
E. acervulina (62,500), E. tenella 100%, 108%, and 116% (breeder
Cobb 500 Improved BWG, FI, and FCR • ↑ VH and reduced CD. [136]
(12,500), and E. maxima (12,500) recommendation)
• ↑ NO production.
• ↓ SOD activity.
• ↓ IL-1β, IL-2, IL-6, IFN-γ, and
E. necatrix (15,000), E. maxima
125 and 150% (breeder TNF-α.
Ross 308 (20,000), E. acervulina (15,000), and E. Improved BWG, FI, and FCR [139]
recommendation) • ↑ NO production.
tenella (170,000)
• ↓ Oocyst count.
E. acervulina (62,500), E. tenella • ↓ Gut permeability.
Cobb 500 16% CP plus 0.75% ARG No effect on performance [159]
(12,500), and E. maxima (12,500) • ↑ VH and AA digestibility.
• ↓ Gut permeability.
E. acervulina (62,500), E. tenella • ↑ Tight junction proteins.
Cobb 500 150% (breeder recommendatio) • ↓CD8+:CD4+. [140]
(12,500), and E. maxima (12,500)
• ↑ NO production.
Animals 2024, 14, 1015 17 of 33

Table 4. Cont.

AA Breed Eimeria Challenge AA Dosage Impact on Performance Impact on Health Reference

• ↓ Oocyst shedding.
750 E. acervulina, E. tenella, and E. 150% TSAA
Ross 308 Improved BWG • ↓ NO production. [147]
maxima (breeder recommendation)
• ↓ MDA concentration.
• ↓ VH and lesion score.
Partridge Shank 125% and 150% (breeder No effect on BWG; ADFI
50,000 E. tenella • ↓ CD4+ and CD8+ cells. [151]
TSAA broilers recommendation) 150% methionine reduced BWG
• ↓ IFN-γ.
100 × Advent vaccine (E. acervulina, Below the requirement reduced
0.6, 0.8, 0.9, and 1.0% TSAA • ↓ TSAA reduced IgA production. [151]
E. maxima, and E. tenella) BWG, ADFI, and gain: feed
E. acervulina (62,500), E. tenella Negatively affected BWG, FI, and • ↑ Oocyst shedding.
Cobb 500 16% CP + 0.75% Met [159]
(12,500), and E. maxima (12,500) FCR • ↓ Intestinal morphology.
New Hampshire × 125% of threonine (breeder • ↓ IL-1β.
1500 E. maxima Improved BWG, ADFI, and FCR [160]
Colombian recommendation) • ↑ IL-12.
• ↑ Oocyst shedding.
• ↑ Gut permeability, IgA
production.
Ross 708 Coccivac® -B 61.25% (NRC, 1994) Exaggerated performance • ↓ Intestinal morphology. [158]
• ↓ Goblet cell counts.
Threonine • ↓ Lymphocytes expressing CD4+,
CD8+, CD3 cells.
• ↑ Intestinal morphology.
112%, 124%, and 136% (breeder
Cobb 500 Mixed Eimeria oocysts Improved BWG and FCR • ↓ Oocyst shedding. [157]
recommendation)
• ↑ IgG and IgM production.
• ↓ Oocyst shedding.
E. acervulina (62,500), E. tenella Improvement in BWG, FI, and
Cobb 500 16% CP and 0.75% threonine • ↑ Intestinal morphology. [159]
(12,500), and E. maxima (12,500) FCR
• ↓ Claudin-1 expression.
• ↓ Inflammatory and
proinflammatory cytokines (IL-10,
IFN-γ).
Glutamine Cobb 500 20 × dose of Coccivac B 0.5% and 1% glutamine No effect on performance [154]
• ↑ Expression of tight junction
proteins.
• ↓ Intestinal CD and ↑VH.
AA: amino acid; TSAA: total sulfur amino acid; CP: crude protein; BWG: body weight gain; ADFI: average daily feed intake; FCR: feed conversion ratio; NO: nitric oxide; IL-1β:
Interleukin 1 β; MDA: malondialdehyde; GPx: glutathione peroxidase; SOD: superoxide dismutase; TLR-4: toll-like receptor-4; IFN-γ: Interferon γ; IL-2: Interleukin 2; IL-6: Interleukin
6; IL-10: Interleukin 10; IL-12: Interleukin 12; VH: villus height; CD: crypt depth; IgG: immunoglobin G; IgM: immunoglobin M.
Animals 2024, 14, 1015 18 of 33

7. Role of Phytogenic Feed Additives

Phytogenic feed additives (PFAs) are plant-derived natural bioactive compounds
or products that, when fed to animals, have a beneficial effect on performance and
health [161]. Phytogenic feed additives have a wide range of bioactive compounds with
antimicrobial, antioxidant, or anti-inflammatory properties and are used in traditional
human medicines [161,162]. Phytochemicals are generally recognized as safe in the United
States, indicating their safety for consumption. This designation strengthens the poten-
tial utilization of phytochemicals in poultry production for coccidiosis control. In the
poultry industry, PFAs are gaining considerable attention mainly because of improve-
ments in the performance of birds by improving gastrointestinal health alongside antiox-
idative and immunomodulatory effects [161]. The efficacy of various phytogenic feed
additives, including Artemisia annua, curcumin, oregano, thyme, and their essential oils,
has been investigated in broiler chickens infected with coccidiosis, demonstrating some
beneficial effects.
Dietary inclusion of curcumin powder (100–200 mg/kg) has been shown to reduce the
lesion score and improve the oxidative status of broilers challenged with mixed Eimeria
spp. [43]. Furthermore, essential oils of oregano have been shown to improve BWG and
the FCR, reduce intestinal lesions and oxidative stress, and improve the gut morphology
in broilers infected with coccidiosis [163,164]. The beneficial effects of these PFAs include
supporting the host by immunomodulatory effects and providing protection against free radi-
cals by scavenging reactive oxygen species and interfering directly with parasitic metabolism,
reducing oocyst shedding and cecal short-chain fatty acid production [45,163,165]. Further-
more, Felici et al. (2023) [166] reported that bioactive compounds from a PFA can inhibit
the intracellular replication of Eimeria and reduce schizont numbers.
Artemisinin, a bioactive flavonoid found in Artemisia annua leaves (AA), has been
shown to inhibit the growth of several stages of Plasmodium spp. [167,168]. The use of AA
and its extract artemisinin has been shown to demonstrate anticoccidial effects against E.
tenella [169–173] when infected alone. The use of either dried AA leaves or artemisinin
has been shown to improve lesion scores, reduce oocyst shedding and sporulation, and
modulate the humoral and immune response in chickens [169,174–178]. The mechanism
of action of artemisinin is promoting apoptosis of infected host cells, thus neutralizing para-
sites [172]. Furthermore, artemisinin from AA is able to alter the cell wall formation of the
oocysts, leading to the death of developing oocysts and a reduced sporulation rate [170]. The
use of phytogenic feed additives in laying hens infected with coccidiosis has not been studied.
However, results from laying hen studies reported that PFA inclusion in diets improved
the performance, immune response, and antioxidant status of laying hens [179–183]. Since
coccidiosis mainly affects the performance of birds with immune suppression and increased
oxidative stress, PFAs can be helpful in laying hens infected with coccidiosis, as in broilers.

8. Role of Prebiotics, Probiotics, and Symbiotics

8.1. Probiotics
Probiotics are selective nonpathogenic microorganisms that, when administered in
adequate amounts, offer a beneficial advantage to the host and improve gut functions by
altering the gut microflora and reducing pathogenic bacteria colonization in the gastroin-
testinal tract [184–187]. By selectively eliminating pathogenic microorganisms from the GI
tract through competitive exclusion, probiotics promote the growth of beneficial bacteria.
Additionally, their metabolites, including short-chain organic fatty acids and hydrogen
peroxide, exhibit antimicrobial properties, effectively inhibiting the growth of pathogenic
bacteria [188,189]. Probiotic microorganisms help develop immune components in the
GI tract and innate or adaptive immune responses and modulate the phosphorylation
of cytoskeletal and tight junction proteins, improving the intestinal barrier [188,190,191].
Some of the commonly used probiotics in the poultry industry include Bacillus, Lactobacillus,
Enterococcus, Bifidobacterium, and Lactococcus, and yeasts such as Aspergillus, Candida, and
Saccharomyces [188,192].
Animals 2024, 14, 1015 19 of 33

The inclusion of Bacillus spp. in diets has been shown to improve the performance,
tight junction integrity of the intestine, and immune response as well as positively influ-
enced the cecal microbiome of broilers challenged with coccidiosis [193–196]. Similar results
were observed in birds fed lactobacillus-based probiotics, including reduced mortality and
oocyst shedding, upregulation of intestinal integrity, increased intestinal intraepithelial
lymphocytes expressing CD4+ and CD8+ cells, and antibody titers in broilers challenged
with coccidiosis [197–199]. In the case of laying hens, the dietary inclusion of probiotics
(Saccharomyces, Pediococcus, Lactobacillus, or Bacillus), either alone or in combination, has
improved the performance, oxidative status, immune responses, intestinal morphome-
try, and microbial composition of ceca positively in a non-challenge model [197,200,201].
The effectiveness of probiotics (Bacillus amyloliquefaciens, Bacillus licheniformis, and Bacillus
pumilus) against Salmonella Enteritidis in laying hens and observed a significant reduction
in salmonella colonization in ceca of mature laying hens [202,203].

8.2. Prebiotics
Prebiotics are non-digestible or selectively fermentable feed ingredients that, when
incorporated into diets, are utilized by the host intestinal microbiota, selectively promoting
the growth or activity of specific bacterial populations, positively influencing the micro-
biome, and improving the gut health of the host [18,204,205]. While selecting prebiotics,
it is essential to consider (i) digestibility (non-digestible by host enzymes), (ii) absorption
(should not be absorbed directly by host cells), (iii) selective fermentation by intestinal
microbiota, (iv) selective promotion of the growth of beneficial bacterial population, and
(v) stimulation of the immune response of the host [188,204–206]. Some of the prebiotics
commonly used in the poultry industry are oligosaccharides (mannan-, galacto-, and
xylo-oligosaccharides), β-glucan, and fructans [188,206–208].
The efficacy of prebiotics in improving the gut health of poultry has been investigated
using both challenged and unchallenged models [209–213]. In experimentally infected
broilers with coccidiosis, the administration of chitosan oligosaccharide (1 g/kg) improved
various parameters, including BWG, FCR, oocyst shedding, intestinal tight junctions, and mor-
phometry and reduced intestinal inflammation [214]. Similarly, broilers fed mannooligosac-
charides (0.8 g/kg), xylooligosaccharides (0.5 g/kg), galactoglucomannan oligosaccharide
(4%), or yeast cell wall polysaccharides (0.5 g/kg) exhibited improved performance, intestinal
tight junction integrity and nutrient digestibility and mitigation of hostile cecal microbial
populations and fermentation induced by Eimeria infection [209–211,215].
Feeding prebiotics such as dry whey powder (6 g/kg), mannan oligosaccharides
(0.25–2 kg/kg), or fructooligosaccharides (0.25–1 g/kg) to laying hens has shown positive
effects on their intestinal microbial populations. Specifically, it promotes beneficial bac-
teria like Lactobacillus spp. and Olensella spp. while reducing the abundance of harmful
bacteria such as Clostridium perfringens, Escherichia coli, and Salmonella enteritidis [216–218].
Additionally, oligosaccharide supplementation has been linked to improved performance,
digestibility, and upregulation of toll-like receptor-4, interferon-γ, and antibody production
in laying hens [217,218].

8.3. Synbiotics
Synbiotics refer to the combined application of both prebiotics and probiotics, which
exhibit synergistic effects on the host by enhancing gut health, the immune response, and
microbial balance [219]. The rationale behind synbiotic uses lies in the premise that pre-
biotics facilitate the survival and colonization of probiotics in the host’s gastrointestinal
tract, thereby positively influencing the health of the host [188,219]. In broiler chickens,
both in ovo and in vivo studies have demonstrated the beneficial effects of synbiotics on
performance, gut health, and the immune response, irrespective of whether the birds were
challenged with coccidiosis or necrotic enteritis [220–225]. Similarly, in laying hens, sup-
plementation with synbiotics containing both prebiotics and probiotics has been shown to
enhance production performance, decrease levels of inflammatory cytokines, and increase
Animals 2024, 14, 1015 20 of 33

populations of beneficial bacteria in the intestinal tract [226,227]. The efficacy of probiotics,
prebiotics, and synbiotics in chickens infected with coccidiosis is summarized in Table 5.
The effect of probiotics, prebiotics, and synbiotics in laying hens has been investigated in
both normal and diseased (Salmonellosis) conditions and has shown positive impacts on
the performance and wellness of hens. Although they have shown beneficial effects in
broilers under coccidiosis, their effect in laying hens infected with Eimeria spp. hens has
not been tested yet.
Animals 2024, 14, 1015 21 of 33

Table 5. Effects of prebiotics, probiotics, and synbiotics supplementation on performance and intestinal health of chickens infected with Eimeria species.

Nutritional Interventions Breed Eimeria Infection Dosage Impacts on Performance Impact on Health Reference
• ↓ Propionate in ceca.
Galacto-glucomannan
• ↑ IFN-γ, IL-1β, IL-6, IL-12 expression.
oligosaccharide- Ross × Ross E. acervulina (1,000,000) 1, or 2, or 4% Improved FI but not BW [209]
• ↑ Bifidobacterium spp., Lactobacillus spp.
arabinoxylan
in ceca.
Improved BWG, FI, and • ↓ Oocyst shedding.
Mannan-oligosaccharides Ross 308 E. tenella (20,000–30,000) 0.8 g/kg [215]
FCR • ↓ Cecal lesions.
• ↓ Oocyst shedding.
15× Coccivac® -B-52 (E. • ↑ Intestinal morphology.
acervulina, E. maxima, E. Improved BW, BWG, FI, • ↑ Expression of tight junction protein.
Chitosan-oligosaccharide Cobb 500 1 g/kg [214]
maxima MF, E. mivati, and E. and FCR • ↓ Expression of TNF-α, IFN-γ, and
tenella) TLR-4.
• ↑ Expression of IL-6, IL-10, and IL-1β.
Nucleotide-rich yeast E. acervulina (25,000) and E. Improved BW, BWG, and • ↑ Jejunal morphology.
Ross 708 0.5 g/kg [210]
extract maxima (5000) FCR • ↓ Cecal SCFA concentration.
• ↑ Propionic and butyric acid in ceca.
Xylo-oligosaccharides Ross 308 12× Paracox 8 0.025% Improved BWG and FI [228]
• ↓ %G+C profile of cecal bacteria.
E. acervulina (62,500), E. • ↓ Duodenal lesions.
Xylo-oligosaccharides Cobb 500 tenella (12,500), and E. 0.5 or 1 g/kg No effect on performance • ↓ CLDN-1 overexpression. [211,229]
maxima (12,500) • ↓ Branched-chain fatty acids.
Improved BWG, FI, and • ↑ Jejunal morphology.
Yeast cell wall Ross 308 E. tenella (5000) 0.1 or 0.2% [230]
FCR • ↓ Bursa follicle length and area.
• ↓ Oocyst shedding.
Lactobacillus spp. Ross 308 E. acervulina (10,000) 1 g/kg - • ↑ Intraepithelial lymphocytes expressing [199]
CD4, CD8 cells.
E. tenella, E. maxima, and E.
Saccharomyces cerevisiae necatrix (70,000 oocysts 0.1, or 1, or 10 g/kg • ↑ Cellular immune response.
Broilers Improved BWG and FCR [197]
total) • ↑ IgM and IgG concentration.
Lactobacillus salivarius and L. 106 , or107 ,or 108CFU/L • ↓ Oocyst shedding and lesion score.
jhonsonii drinking water
• ↓ Oocyst shedding and lesion score.
Bacillus subtilis 747 Ross 708 E. maxima (10,000) 1.5 × 105 CFU/g feed Improved BWG and FCR • ↓ IL-1β, IL-6, IL-2, and IFN-γ. [195]
• ↑ Expression of tight junction protein.
Bacillus licheniformis-A E. tenella (5000), E. maxima • ↓ Oocyst shedding and lesion score;
Bacillus amyloliquefaciens-B Ross × Ross (5000), and E. acervulina 1.5 × 105 CFU/g of feed Improved performance • ↑ IL-6, IL-8, and IL-10. [196]
Bacillus amyloliquefaciens-D (5000) • ↑ JAM-2.
E. maxima (25,000) and E. 108 CFU/bird/d in No effect on BW, FI, and • ↑ Intestinal morphology.
Bacillus subtilis (BS-9) Ross 708 • ↓ Bursa weight. [231]
acervulina (100,000) drinking water FCR
Animals 2024, 14, 1015 22 of 33

Table 5. Cont.

Nutritional Interventions Breed Eimeria Infection Dosage Impacts on Performance Impact on Health Reference
• ↓ Internal organ growth.
E. acervulina, E. maxima, and
Lactobacillus rhamnosus Yellow broilers 106 , or 108 CFU in ovo - • ↓ Intestinal morphology. [232]
E. tenella (100,000)
• Exaggerated coccidia infection.
106 cfu/mL of Lactobacillus
Lactobacillus plantalum +
Broilers E. tenella (20,000) plantalum + 2% Shallot - • ↓ Cecal lesion score. [220]
Shallot extract
extract in water
Lactobacillus reuteri,
Enterococcus faecium,
E. adenoides and E.
Bifidobacterium animalis, Turkey poult - Improved BWG • ↓ Oocyst shedding and lesion score. [221]
meleagrimitis
Pediococcus acidilactici, and a
fructooligosaccharide
• ↓ Oocyst shedding and lesion score.
Postbiotics (Mantol) Ross 708 E. maxima (10,000) 10 mg/kg No effect on performance [233]
• ↓ IL-1β, IL-6, IL-17, IL-10, and IFN-γ.
BW: body weight; BWG: body weight gain; FI: feed intake; FCR: feed conversion ratio; NO: nitric oxide; IL-1β: Interleukin 1-β; IL-6: Interleukin 6; IL-10: Interleukin 10; IL-12: Interleukin
12; TLR-4: toll-like receptor-4; IFN-γ: Interferon γ; TNF-α: tumor necrosis factor-α; SCFA: short chain fatty acid; CLDN-1: Claudin -1; IL-2: Interleukin 2; IL-8: Interleukin 8; JAM-2:
junction adhesion molecule-2; CFU: colony-forming unit.
Animals 2024, 14, 1015 23 of 33

8.4. Postbiotics
In recent years, it has been established that the beneficial effects of probiotics are not
only limited to when the microorganisms are alive but also after their death, leading to
the development of new antibiotic alternatives, named postbiotics [234,235]. Postbiotics
are non-viable bacterial cells or cell walls or metabolites derived from probiotics that have
biological activity and offer physiological benefits to the host [235]. Different bioactive
metabolites from probiotics with biological activity may include a range of compounds
such as short-chain fatty acids, peptides, enzymes, polysaccharides, vitamins, cell surface
proteins, and organic acids [235]. Recent studies conducted in chickens have shown that
postbiotics confer beneficial effects on the host by positively modulating the intestinal
microbiome, immune responses, and oxidative status [233,236–238]. In laying hen pul-
lets challenged with Salmonella spp., dietary inclusion of postbiotics from Saccharomyces
cerevisiae (1–1.5 kg/MT) or oral inoculation of Lactobacillus spp. postbiotics reduced the in-
testinal colonization of Salmonella [239,240]. Furthermore, Lactobacillus-derived postbiotics
have been shown to enhance the performance, oxidative status, and intestinal morphology
of broilers under heat stress [236,241]. Additionally, in broilers with experimentally induced
necrotic enteritis, postbiotics from Enterococcus spp. Pediococcus spp., Lactobacillus spp., En-
terococcus spp., and Lactiplantibacillus spp. were able to improve performance, lesion scores,
and oxidative status and reduce proinflammatory responses [237,238,242]. Moreover, the
inclusion of maltol (a metabolite of Bacillus subtilis) in a broiler diet infected with E. maxima
was able to reduce intestinal damage and inflammatory response [233]. Although the use of
postbiotics in broilers and laying hens infected with Eimeria spp. has not been extensively
explored, previous studies have demonstrated their efficacy in reducing the symptoms
associated with coccidiosis in other stress conditions.

9. Conclusions
To conclude, this review summarizes the effect of coccidiosis on laying hen health
and performance and its potential to cause a significant economic loss to the egg industry.
Since the ban on antibiotics as growth promoters in animal production, there has been an
increase in the incidences of economically important diseases in poultry, such as coccidiosis
and necrotic enteritis. Several antibiotic alternatives (vitamins, functional amino acids,
phytogenic feed additives, prebiotics, probiotics, synbiotics, and postbiotics) have been
tested in broilers with positive effects on minimizing the effect of coccidiosis, but their
beneficial effects have not been explored in laying hens. These nutritional strategies have
been shown to have the potential to mitigate the negative effects of coccidiosis in laying
hens as well. However, before applying these nutritional strategies either to boost immunity
after vaccination or to mitigate the negative effects of coccidiosis, they need to be tested in
laying hens. Additionally, while implementing these nutritional strategies to mitigate the
adverse effects of coccidiosis, it is important to consider that these strategies are not capable
of curing coccidiosis completely, and poultry producers must evaluate the severity of the
conditions. Future studies should focus on evaluating nutritional strategies either alone
or in combination, focusing on performance, gut health, and immune responses against
coccidiosis in laying hens.

Author Contributions: Conceptualization M.K.S. and W.K.K.; writing—review and editing, M.K.S.
and W.K.K.; supervision, W.K.K.; funding acquisition, W.K.K. All authors have read and agreed to
the published version of the manuscript.
Funding: This research received no external funding.
Conflicts of Interest: The authors declare no conflicts of interest.
Animals 2024, 14, 1015 24 of 33

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