Archives of

1. Introduction
The phylum Apicomplexa’s intracellular protozoan Sarcocystis is responsible for the zoonotic disease sarcocystosis [1]. This parasite consists of more than 190 species and affects a variety of domestic animals, including cattle, sheep, goats, and pigs. In these animals, the parasite can lead to weight loss, anorexia, lameness, paralysis, fever, anemia, decreased milk production, abortion, and even death [2-4]. Some Sarcocystis species are important pathogens of humans and domestic and wild animals. These parasites are characterized by an obligatory two-host life cycle, the formation of sarcocysts mainly in the muscles of the intermediate hosts and endogenous sporulation of oocysts in the intestine of the definitive hosts [2]. Due to the condemnation of contaminated carcasses, millions of dollars of damage are caused to the livestock industry every year, and the disease caused by different species of these protozoa is significant both in terms of health and economics [5-8].
By consuming undercooked or raw infected meat, humans can become definite hosts of Sarcocystis hominis and S. suihominis, and they are responsible for intestinal sarcocystosis. Most people with intestinal sarcocystosis remain asymptomatic. Symptoms induced by experimental infections include nausea, abdominal discomfort, and self-limited diarrhea, with symptom severity varying with the amount of meat consumed [9]. Diarrhea usually onset suddenly (in some people 3 to 6 hours after consumption; usually within 48 hours) and symptoms resolve within 36 hours. They can also become intermediate hosts of S. nesbitti and S. lindemanni by consuming sporocysts excreted by reptiles or carnivores, which can cause extraintestinal sarcocystosis and symptoms such as musculoskeletal pain, fever, nausea, abdominal pain, diarrhea, and cardiopathy [9,10]. To date, several Sarcocystis species, including S. medusiformis and S. gigantea (S. ovifelis, which has macroscopic cysts), and S. arieticanis and S. tenella (S. ovicanis, which has microscopic cysts), have been linked to sheep [2,11,12]. S. cruzi, S. hirsute, and S. hominis are the three Sarcocystis species found in cattle, with dogs, cats, people, or primates serving as the three definitive hosts, respectively [13,14].
Some species of Sarcocystis, particularly those having a canid as the definitive host, may cause clinical signs/symptoms including fever, inappetence, abortion, stillbirth, or central nervous signs including acute myopathy, ataxia, paresis, and death in sheep and cattle [15-17].
Those signs have been associated with the release of interleukin-1 (IL-1), prostaglandin E2 (PGE2), and tumor necrosis factor–alpha (TNF-α) by the Sarcocystis infected macrophages. These cytokines can cause inappetence, anemia, and suppression of the release of pituitary growth hormone (GH) as a potential reason for weight loss [18].
Macroscopic and Microscopic Survey of Sarcocystis spp. Infection in Slaughtered Cattle and Sheep in Tabriz, Iran
Nasser Hajipour1*ID
1Department of Food Hygiene and Aquatic, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran
*Corresponding Author: Nasser Hajipour, Email: n.hajipour@tabrizu.ac.ir
Abstract
Background & Aims: Sarcocystis infection is one of the most common protozoan infections between humans and animals, which is caused by different species of Sarcocystis. The study aimed to investigate Sarcocystis infection in cattle and sheep slaughtered in Tabriz slaughterhouse by microscopic and macroscopic methods.
Materials amd Methods: Diaphragmatic muscles of 500 cattle and 800 sheep were randomly selected for macroscopic and microscopic Sarcocystis cysts. A naked eye examination was done for macroscopic sarcocysts, while peptic digestion and Daub smear method were used for the microscopic cysts.
Results: The overall prevalence of Sarcocystis spp. infection was 44.0% and 68.25% in cattle and sheep, respectively. The results showed that rate of infection of cattle with Sarcocystis spp. was 10.4%, 90.0%, and 22.3% by macroscopic, peptic digestion, and Daub smear methods, respectively. Meanwhile, the percentage of infected sheep was determined by macroscopic, peptic digestion, and Daub smear methods as 30.6%, 100.0%, and 44.1%, respectively (P < 0.05).
Conclusion: It is concluded that digestion is a perfect method for diagnosing sarcocysts in cattle and sheep. As well as, the high prevalence of microscopic Sarcocystis spp. in cattle and sheep of Tabriz, is suggested that meat should be cooked sufficiently, the people to be trained not to feed their dogs and cats with uncooked meat.
Keywords: Sarcocystis, Cattle, Sheep, Macroscopic, Tabriz, Iran
Received: January 26, 2023, Accepted: June 20, 2023, ePublished: September 29, 2023
https://jhygiene.muq.ac.ir/
10.34172/AHS.12.3.1.435
Vol. 12, No. 3, 2023, 137-141
Original Article
© 2023 The Author(s); This is an open-access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Hajipour
138 Arch Hyg Sci. Volume 12, Number 3, 2023
The frequency of Sarcocystis spp. in sheep and cattle
in various regions of Iran and other nations has been
the subject of many investigations [19-24]. By using
microscopic and macroscopic techniques, this study’s
goal was to assess the prevalence of Sarcocystis infection
in the diaphragm muscle of cattle and sheep that were
slaughtered for food in Tabriz, Iran.
2. Materials and Methods
From March 2018 to March 2019, the diaphragmatic
muscles of 500 cattle and 800 sheep were inspected
for macroscopic and microscopic Sarcocystis cysts.
Animals were examined at arrival by veterinarians in
accordance with all the veterinary criteria scheduled for
the antemortem examination included in Iran’s legislation
[25]. The absence of clinical symptoms was established
according to the antemortem veterinary official
examination.
The number of samples was calculated using the
following formula, assuming a prevalence of 20-30% of
Sarcocystis infection in animal meat based on previous
studies, as well as a 96% confidence interval and an
accuracy of 5%.
2
1 /2
2
z p(1 p)
n
d
−α × −
=
Where P: Prevalence, α: Error rate, d: Accuracy
A naked eye examination was done for macroscopic
sarcocysts, while peptic digestion and the Daub smear
method were used for the microscopic cysts [26]. For this
purpose, diaphragmatic muscles that apparently lacked
Sarcocystis cysts resembling rice grains were selected.
About 1 g of diaphragmatic muscles were broken up into
little pieces and firmly compressed between two glass slides
for the Daub smear procedure. Then the samples were
examined under a microscope after staining with Giemsa.
In the digestion procedure, 50 mL of digestion solution
containing 2.5 g pepsin (Sigma-Aldrich, Germany) and
10 mL of HCl (Merck, Germany) in 500 mL of distilled
water, was added to 10 g of crushed muscles and heated
for 4 hours at 40 °C. The solution was filtered through
a strainer, and centrifuged for 2 minutes at 1500 g. The
supernatant was decanted, the sediment was resuspended
in 1 mL of Giemsa saline, prepared a smear, and observed
under a compound microscope at 400 × magnification
[2]. Data were analyzed by SPSS software (version 21) and
chi-square test analysis. A significance level of less than
0.05 was considered.
3. Results
The animals studied in this study were apparently healthy
and had no clinical symptoms. The results of this study
showed that the prevalence of Sarcocystis spp. infection
in sheep (68.25%) was significantly higher than in cattle
(44.0%) (P < 0.05).
The prevalence of microscopic Sarcocystis spp. cysts
in cattle were detected in 90.0% and 22.3% using peptic
digestion and Daub smear, respectively, while 10.4% were
detected in macroscopic examination (Table 1).
Two hundred and forty-five of the 800 sheep (30.6%)
were diagnosed as infected with macroscopic cysts. Out
of 100 diaphragmatic muscles of sheep examined using
the digestive method, all samples were infected with
Sarcocystis cysts. In addition, out of 455 samples tested by
the Daub smear method, 201 samples (44.1%) were found
to be infected.
The results of this study revealed that the prevalence
rate of infection with Sarcocystis spp. in females was
higher than in male animals (P < 0.05). Also, the results
of the comparison of different methods in the diagnosis
of Sarcocystis spp. infection in cattle and/or sheep showed
that the digestive method was better than macroscopic
and/or Daub smear methods.
4. Discussion
One of the most widespread protozoan zoonotic
parasites, Sarcocystis, is found in the striated muscles of
livestock killed for food, such as cattle, sheep, and goats.
It poses serious health and economic risks to both human
and animal civilizations [15,26]. In slaughterhouse
inspections, only macroscopic cysts are detected, and
microscopic cysts remain hidden from the inspectors.
Therefore, the statistics presented by the slaughterhouse
inspection are less than the real value [27].
In the present study, the overall prevalence rate of
Sarcocystis spp. in sheep was higher than that in cattle,
which was consistent with other studies [28-31]. It may
relate to different forage habitats of cattle and sheep,
geographical climates, or high exposure to dogs and/or
cats, which serve as definitive hosts [32].
Table 1. Infection of Sarcocystis spp. in sheep and cattle slaughtered at Tabriz abattoir on the sex by different methods
Method/ Animal
Cattle
P value
Sheep
No. examined No. positive samples (%) P value
Total (%)
No. examined No. positive samples (%)
Total (%)
M F M F M F M F
Macroscopy 250 250 15 (6) 37 (14.8) 52 (10.4)
0.000
400 400 100 (25) 145 (36.2) 245 (30.6)
0.000
Peptic digestion 50 50 45 (90) 45 (90) 90 (90) 50 50 50 (100) 50 (100) 100 (100)
Muscle squeeze 185 163 32 (17.29) 46 (28.22) 78 (22.3) 250 205 88 (35.2) 113 (55.1) 201 (44.1)
Total 250 250 92 (36.8) 128 (51.2) 220 (44) 400 400 238 (59.5) 308 (77) 546 (68.25)
Arch Hyg Sci. Volume 12, Number 3, 2023 139
Sarcocystis spp. in cattle and sheep in Tabriz, Iran
The results of this study showed that the prevalence rate
of infection with Sarcocystis spp. in females was higher
than in male animals (P < 0.05), which was in line with
the studies conducted by Mac et al [22]. However, there
was no significant association between sex and Sarcocystis
spp. infection in cattle [22]. This finding may be due to
the higher mean age of females than males, with females
being slaughtered at an older age [33].
By using macroscopy, peptic digestion, and Daub smear
methods in the current investigation, the prevalence of
Sarcocystis spp. in cattle was found to be 10.4%, 90.0%,
and 22.3%, respectively. Similar results were obtained by
Rasouli et al [34], who found an infection rate of 93.33%
by pepsin digestion in Kurdistan, by Nourani et al [35],
who reported an infection rate of 92% by histopathological
in Isfahan.
In the study conducted by Mirzaei and Rezaei [36], the
prevalence of microscopic and macroscopic Sarcocystis
spp. cysts in cattle of Tabriz city, Iran were detected in
100.0% and 8.2%, respectively. In various reports from
different regions of Iran, the infection prevalence rate is
100.0% using the pepsin digestion method, which can be
mentioned in the following studies: Nourollahi-Fard et al
[26] in Kerman, Dalimi Asl et al [37] in Tabriz, Parandin et
al [38] in Hamadan, Rahdar and Salehi [39], Hamidinejat
et al [40] in Ahvaz, and Shekarforoush et al [4] in Shiraz.
The prevalence rate of Sarcocystis spp. by the pepsin
digestion method was estimated to be 59.8% in Bukan
[34] and 26.42% in Saqez [27], which was lower compared
to our results.
However, the prevalence rate obtained in the recent
study was lower compared to some studies conducted by
other researchers. For example; Bonyadian and Meshki
[41] showed an infection rate of 91.0% by Daub smear in
Shahrekord.
Hamidinejat et al [40] reported the prevalence of
Sarcocystis infection by digestive and Daub smear methods
as 100.0% and 94.7% respectively. In another study, of
the 140 inspected cows, 27 (19.28%) were infected with
Sarcocystis spp. by Daub smear [27].
The prevalence rate of Sarcocystis spp. in other countries
has been reported as follows: Hungary (66.0%) [42],
Southern Italy (96.0%) [43], and Nigeria (42.5%) [44].
Our study’s findings, which were in line with other
research, showed that the prevalence of Sarcocystis in
slaughtered sheep was 30.6%, 100.0%, and 44.1% by
macroscopic, digestive, and Daub smear techniques,
respectively (P < 0.05). Daryani et al found the prevalence
of Sarcocystis infection in sheep slaughtered in Ardabil to
be 33.93% by macroscopic method [21], and Arshad et al
[45] in Tabriz and Rahdar [24] in Ahvaz reported 100.0%
by peptic digestion method. The frequency of Sarcocystis
infection was determined to be 6% in sheep slaughtered
in Kerman, according to Mirzaei Dehaghi et al [7].
The prevalence of Sarcocystis spp. in sheep varies in
different parts of the world. In this regard, this rate has
been reported as 92.5% by digestive method [46] and
97.0% and 4.10% by digestive and macroscopic method,
respectively in Iraq [47], 52.51% by digestive method
in China [48] and 100.0% by digestive method in
Lithuania [31].
The reasons for the difference in the level of Sarcocystis
infection in different regions of the world come from the
fact that there are important factors in the transmission
and high frequency of this parasite in ruminants. Some
of these factors include close contact of animals with
carnivores, especially dogs and cats, type of feeding that
is mostly traditional grazing (final hosts infected with this
parasite having access to these pastures and transmitting
this parasite to ruminants), the viability of Sarcocystis
sporocysts for many months in the environment and
weather conditions suitable for the survival of oocysts in
the environment [32].
Since these cysts are of feline origin and in Tabriz
there is more contact between these animals and dogs in
pasture than there is between those animals and cats, the
low prevalence of microscopic sarcocysts in sheep and
cattle in the current study may be the cause [28].
The gold standard method for diagnosing sarcocysts
is pepsin digestion, and research in sheep or cattle
found that it was more sensitive than the muscle squash
method [40].
5. Conclusion
In the present investigation, most cysts in sheep and
cattle are microscopic, and the final host is the dog, and
the contact between the sheep and cattle and the herd
dogs is high and has caused high infection of livestock,
and since it is done macroscopically, the inspection of the
carcasses in slaughterhouses, therefore, it is necessary to
change the method of inspection and control of meat in
slaughterhouses, as well as to decide on special methods
to eliminate meat contamination before consumption to
prevent human infection.
Acknowledgments
I thank Valizadeh for his technical assistance and preparation of
materials and instruments.
Competing Interests
The author declares no conflicts of interest relevant to this study
Ethical Approval
The study entailed recording of the normal meat inspection process
of animals sent to slaughter for human consumption. No ethical
approval was required nor sought as no alteration was required to
the normal processing of these animals.
Funding
This study was supported by Vice-Chancellor for the Research
Affairs University of Tabriz.
Hajipour
140 Arch Hyg Sci. Volume 12, Number 3, 2023
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