Histological, Bacteriological, and Parasitological Study and Animal Host Origin of Handmade Sausages Marketed in Eight Cities in Iran
Document Type : Original Article
Authors
- Ali Kalantari-Hesari 1
- Mohammad Babaei 1
- Leili Moradi 2
- Mozhdeh Zolfaghari 2
- Arshia Shahbazi-Amraei 3
- Zahra Ghasemi-Teimour 4
1 Department of Basic Sciences, Faculty of Veterinary Medicine, Bu-Ali Sina University, Hamedan, Iran
2 Department of Pathobiology, Faculty of Veterinary Medicine, Bu-Ali Sina University, Hamadan, Iran
3 Department of Food Hygiene, Faculty of Veterinary Medicine, Bu-Ali Sina University, Hamedan, Iran
4 Department of nutrition and food hygiene, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
10.22084/avr.2025.30525.1000
Abstract
The aim of this study was to examine the quality of handmade sausages using histological, bacteriological, and parasitological techniques. During 2023, 19 samples were randomly purchased from the market in eight cities in Iran, i.e., Tehran, Karaj, Tabriz, Hamedan, Borujerd, Mashhad, and Shiraz. Specimens were cut into three parts: i) xed in 10% buffered formalin solution and sectioned and stained for histology according to the Iranian National Standard No. a6103, ii) minced and cultured on Plate Count Agar (PCA) medium and antibiogrammed, iii) used for genomic DNA extraction and conventional PCR to identify the animal host origin of the sausages and examine for coccidian parasites, Toxoplasma gondii, Sarcocystis spp., Neospora caninum DNA. Results showed that 4 samples were suspected of being parasitic cysts. PCR testing revealed that all four samples contained DNA of the coccidian protozoan parasite, and one sample contained DNA of Neospora. Additionally, based on PCR testing, 18 samples contained both chicken and beef meat, while one sample contained lamb meat. In the bacterial analysis, 3 samples contained a signi cant amount of Bacillus subtilis, and one sample contained Bacillus cereus. In the histological examination of the collected samples soybean, cartilage tissue, bone tissue, smooth muscle tissue, and dense connective tissue were observed. Therefore, it can be concluded that since there is no sanitary supervision or quality control in the production of handmade sausages, there is a possibility of using inferior meat products with lower nutritional value.
Keywords
[1] Alikord M, Momtaz H, Yadegarfar G, Keramat J,
Homayooni Rad A. Identi cation in meat prod-
ucts authentication. Food Research Journal. 2017
Dec 22; 27(4): 73-86.
[2] Al-Qassab T, Kamkar A, Shayan P, Khanjari A.
(2019). Mislabeling in Cooked Sausage is a Seri-
ously Increasingly Problem in Food Safety. Iranian
Journal of Veterinary Medicine, 13(1): 101-113.
[3] Asadi MR, Kalantari-Hesari A, Ghaemmaghami
SS, Mosleh N, Ghorbanzadeh B, Abdi P. Meat
products components using histological method
and image analysis software. Veterinary Research
& Biological Products. 2023 Mar 21; 36(1): 102-
12.
[4] Asadi MR, Taghavi M, Hesari AK, Ghorbanzadeh
B. The role of histological test in reducing the use
of unauthorized tissues in meat products between
years of 2014 and 2017. Veterinary Research & Bi-
ological Products. 2020 Sep 22; 33(3): 31-40.
[5] Ballin NZ, Vogensen FK, Karlsson AH. Species
determination-Can we detect and quantify meat
adulteration?. Meat science. 2009 Oct 1; 83(2):
165-74.
[6] Bazyari S, Zamanizadeh HZ, Mizani M, Sharifan
A. Evaluation of presence of soya protein in some
commercial meat products by polymerase chain
reaction assays. Food Sci Nutr. 2015 May; 12: 33-
40.
[7] Bretagne S, Costa JM, Vidaud M, Nhieu JT, Feith
JF. Detection of Toxoplasma gondii by competi-
tive DNA ampli cation of bronchoalveolar lavage
samples. Journal of Infectious Diseases. 1993 Dec
1; 168(6): 1585-8.
[8] Cavin C, Cottenet G, Blancpain C, Bessaire T,
Frank N, Zbinden P. Food adulteration: From vul-
nerability assessment to new analytical solutions.
Chimia. 2016 May 25; 70(5): 329-33.
[9] Ali ME, Razzak MA, Abd Hamid SB, Rahman
MM, Al Amin M, Abd Rashid NR. Multiplex PCR
assay for the detection of ve meat species forbid-
den in Islamic foods. Food chemistry. 2015 Jun 15;
177: 214-24.
[10] Gjerde B. Phylogenetic relationships among Sar-
cocystis species in cervids, cattle and sheep in-
ferred from the mitochondrial cytochrome c oxi-
dase subunit I gene. International Journal for Par-
asitology. 2013 Jun 1; 43(7): 579-91.
[11] Hajimohammadi B, Fattahi K, Yekta ZK, Sadegh-
inezhad J, Morovvati H, Basti AA. Experimental
Study of the Histological Method for Quantitative
Detection of Meat in Kabab and Cooked Sausage
Model. Journal of Veterinary Research/Majallah-i
Taqqt-i Dmpizishk University. 2020 Jun 1; 75(3).
[12] Ho MS, Barr BC, Tarantal AF, Lai LT, Hendrickx
AG, Marsh AE, Sverlow KW, Packham AE, Con-
rad PA. Detection of Neospora from tissues of ex-
perimentally infected rhesus macaques by PCR
and speci c DNA probe hybridization. Journal of
clinical microbiology. 1997 Jul; 35(7): 1740-5.
[13] Hosseini H, Khaksar R, Shemshadi B. Study of
Sarcocyst in Raw, Ready to Sell Hamburgers in
Tehran. Food Sci Nutr. 2008 Apr. 4(4): 65-71.
[14] Hosseni HS, Tafvizi F, Tajabadi Ebrahimi M,
Sharifan A. Fraud Identi cation in Beef Sausage
in Tehran Province Using Mitochondrial Genes of
Animal Species. Food Hygiene. 2014. 4(1): 81-89.
[15] Khaniki GJ, Kia EB. Detection of Sarcocystis
cysts from meat supplied for hamburger in Iran
by histological method. J Med Sci. 2006 Apr, 6
(1): 18-21.
[16] Khaniki GR, Rokni N. Histological detection of
soya in freezing raw hamburger of Iran. Pajouhesh
and Sazandegi. 2004 Agu, 62: 71-5.
[17] Julini M, Parisi E, Chicco G. Histological aspects
of common frauds in sausage manufacture. Annali
della Facolta di Mediunia Veterinaria di Tirnio.
1979; 26: 231-44.
[18] Karimi S, Bahari A, Nourian A, Azami S, Na-
mavari M, Basso W, Sazmand A, Hemphill A.
Neospora caninum and Toxoplasma gondii infec-
tions in one-humped camels (Camelus dromedar-
ius) in central desert of Iran. Parasitology re-
search. 2023 Mar; 122(3): 847-52.
[19] Khordadmehr M, Sazmand A, Almasi P, Shahbazi
P, Ranjbar V, Otranto D, Hemphill A. Natural
infection with Toxoplasma gondii, Neospora can-
inum and Sarcocystis species in domestic pigeons
(Columba livia domestica) in Iran. Comparative
immunology, microbiology and infectious diseases.
2023 Feb 1; 93: 101946.
[20] Lakzadeh L, Hosseinzadeh S, Shekarforoush SS,
Fazeli M. Quantitative detection of chicken meat
routine mislabeling in emulsion type sausages and
burgers by SYBR green real time PCR assay. Iran
Agricultural Research. 2016 Aug 22; 35(1): 49-54.
[21] Latorre R, Sadeghinezhad J, Hajimohammadi B,
Izadi F, Sheibani MT. Application of morphologi-
cal method for detection of unauthorized tissues in
processed meat products. Journal of food quality
and hazards control. 2015 Jun 10; 2(2): 71-4.
[22] Liscio C, Hopley C. Development of a reference
measurement procedure and certi ed reference
material for the determination of hydroxyproline
in meat. Food Analytical Methods. 2016 Jun; 9:
1461-9.
[23] Mousavi SM, Khaniki GJ, Eskandari S, Rabiei
M, Samiee SM, Mehdizadeh M. Applicability
of species-speci c polymerase chain reaction for
fraud identi cation in raw ground meat commer-
cially sold in Iran. Journal of Food Composition
and Analysis. 2015 Jun 1; 40: 47-51.
[24] Muller N, Zimmermann V, Hentrich B, Gottstein
B. Diagnosis of Neospora caninum and Toxo-
plasma gondii infection by PCR and DNA hy-
bridization immunoassay. Journal of Clinical Mi-
crobiology. 1996 Nov; 34(11): 2850-2.
[25] Nesic K, Stojanovic D, Baltic ZM. Authentication
of meat and meat products vs. detection of ani-
mal species in feedwhat is the difference?. InIOP
Conference Series: Earth and Environmental Sci-
ence 2017 Sep 1 (Vol. 85, No. 1, p. 012043). IOP
Publishing.
[26] Olsen P, Borit M. The components of a food trace-
ability system. Trends in Food Science & Technol-
ogy. 2018 Jul 1; 77: 143-9.
[27] Prayson BE, McMahon JT, Prayson RA. Apply-
ing morphologic techniques to evaluate hotdogs:
what is in the hotdogs we eat?. Annals of diag-
nostic pathology. 2008 Apr 1; 12(2): 98-102.
[28] Sadeghi E, Khazaei M, Almasi A, Shariatifar N,
Bohlouli Oskoii S, Tahvilian R. Recognition of lle-
gal Tissues in the Meat Products from Kerman-
shah Supply Centers during the years 2009-2010.
Internal Medicine Today. 2011 Apr 10; 17(1): 55-
9.
[29] Sarab I. Determination of adulteration and au-
thenticity of meat and meat products using chemi-
cal properties and PCR technique in Tabriz. Jour-
nal of Health. 2020 May; 11(4): 478-88.
[30] Veuthey TV, Herrera MG, Dodero VI. Dyes and
stains: from molecular structure to histologi-
cal application. Frontiers in Bioscience-Landmark.
2014 Apr, 19(1): 91-112.
Homayooni Rad A. Identi cation in meat prod-
ucts authentication. Food Research Journal. 2017
Dec 22; 27(4): 73-86.
[2] Al-Qassab T, Kamkar A, Shayan P, Khanjari A.
(2019). Mislabeling in Cooked Sausage is a Seri-
ously Increasingly Problem in Food Safety. Iranian
Journal of Veterinary Medicine, 13(1): 101-113.
[3] Asadi MR, Kalantari-Hesari A, Ghaemmaghami
SS, Mosleh N, Ghorbanzadeh B, Abdi P. Meat
products components using histological method
and image analysis software. Veterinary Research
& Biological Products. 2023 Mar 21; 36(1): 102-
12.
[4] Asadi MR, Taghavi M, Hesari AK, Ghorbanzadeh
B. The role of histological test in reducing the use
of unauthorized tissues in meat products between
years of 2014 and 2017. Veterinary Research & Bi-
ological Products. 2020 Sep 22; 33(3): 31-40.
[5] Ballin NZ, Vogensen FK, Karlsson AH. Species
determination-Can we detect and quantify meat
adulteration?. Meat science. 2009 Oct 1; 83(2):
165-74.
[6] Bazyari S, Zamanizadeh HZ, Mizani M, Sharifan
A. Evaluation of presence of soya protein in some
commercial meat products by polymerase chain
reaction assays. Food Sci Nutr. 2015 May; 12: 33-
40.
[7] Bretagne S, Costa JM, Vidaud M, Nhieu JT, Feith
JF. Detection of Toxoplasma gondii by competi-
tive DNA ampli cation of bronchoalveolar lavage
samples. Journal of Infectious Diseases. 1993 Dec
1; 168(6): 1585-8.
[8] Cavin C, Cottenet G, Blancpain C, Bessaire T,
Frank N, Zbinden P. Food adulteration: From vul-
nerability assessment to new analytical solutions.
Chimia. 2016 May 25; 70(5): 329-33.
[9] Ali ME, Razzak MA, Abd Hamid SB, Rahman
MM, Al Amin M, Abd Rashid NR. Multiplex PCR
assay for the detection of ve meat species forbid-
den in Islamic foods. Food chemistry. 2015 Jun 15;
177: 214-24.
[10] Gjerde B. Phylogenetic relationships among Sar-
cocystis species in cervids, cattle and sheep in-
ferred from the mitochondrial cytochrome c oxi-
dase subunit I gene. International Journal for Par-
asitology. 2013 Jun 1; 43(7): 579-91.
[11] Hajimohammadi B, Fattahi K, Yekta ZK, Sadegh-
inezhad J, Morovvati H, Basti AA. Experimental
Study of the Histological Method for Quantitative
Detection of Meat in Kabab and Cooked Sausage
Model. Journal of Veterinary Research/Majallah-i
Taqqt-i Dmpizishk University. 2020 Jun 1; 75(3).
[12] Ho MS, Barr BC, Tarantal AF, Lai LT, Hendrickx
AG, Marsh AE, Sverlow KW, Packham AE, Con-
rad PA. Detection of Neospora from tissues of ex-
perimentally infected rhesus macaques by PCR
and speci c DNA probe hybridization. Journal of
clinical microbiology. 1997 Jul; 35(7): 1740-5.
[13] Hosseini H, Khaksar R, Shemshadi B. Study of
Sarcocyst in Raw, Ready to Sell Hamburgers in
Tehran. Food Sci Nutr. 2008 Apr. 4(4): 65-71.
[14] Hosseni HS, Tafvizi F, Tajabadi Ebrahimi M,
Sharifan A. Fraud Identi cation in Beef Sausage
in Tehran Province Using Mitochondrial Genes of
Animal Species. Food Hygiene. 2014. 4(1): 81-89.
[15] Khaniki GJ, Kia EB. Detection of Sarcocystis
cysts from meat supplied for hamburger in Iran
by histological method. J Med Sci. 2006 Apr, 6
(1): 18-21.
[16] Khaniki GR, Rokni N. Histological detection of
soya in freezing raw hamburger of Iran. Pajouhesh
and Sazandegi. 2004 Agu, 62: 71-5.
[17] Julini M, Parisi E, Chicco G. Histological aspects
of common frauds in sausage manufacture. Annali
della Facolta di Mediunia Veterinaria di Tirnio.
1979; 26: 231-44.
[18] Karimi S, Bahari A, Nourian A, Azami S, Na-
mavari M, Basso W, Sazmand A, Hemphill A.
Neospora caninum and Toxoplasma gondii infec-
tions in one-humped camels (Camelus dromedar-
ius) in central desert of Iran. Parasitology re-
search. 2023 Mar; 122(3): 847-52.
[19] Khordadmehr M, Sazmand A, Almasi P, Shahbazi
P, Ranjbar V, Otranto D, Hemphill A. Natural
infection with Toxoplasma gondii, Neospora can-
inum and Sarcocystis species in domestic pigeons
(Columba livia domestica) in Iran. Comparative
immunology, microbiology and infectious diseases.
2023 Feb 1; 93: 101946.
[20] Lakzadeh L, Hosseinzadeh S, Shekarforoush SS,
Fazeli M. Quantitative detection of chicken meat
routine mislabeling in emulsion type sausages and
burgers by SYBR green real time PCR assay. Iran
Agricultural Research. 2016 Aug 22; 35(1): 49-54.
[21] Latorre R, Sadeghinezhad J, Hajimohammadi B,
Izadi F, Sheibani MT. Application of morphologi-
cal method for detection of unauthorized tissues in
processed meat products. Journal of food quality
and hazards control. 2015 Jun 10; 2(2): 71-4.
[22] Liscio C, Hopley C. Development of a reference
measurement procedure and certi ed reference
material for the determination of hydroxyproline
in meat. Food Analytical Methods. 2016 Jun; 9:
1461-9.
[23] Mousavi SM, Khaniki GJ, Eskandari S, Rabiei
M, Samiee SM, Mehdizadeh M. Applicability
of species-speci c polymerase chain reaction for
fraud identi cation in raw ground meat commer-
cially sold in Iran. Journal of Food Composition
and Analysis. 2015 Jun 1; 40: 47-51.
[24] Muller N, Zimmermann V, Hentrich B, Gottstein
B. Diagnosis of Neospora caninum and Toxo-
plasma gondii infection by PCR and DNA hy-
bridization immunoassay. Journal of Clinical Mi-
crobiology. 1996 Nov; 34(11): 2850-2.
[25] Nesic K, Stojanovic D, Baltic ZM. Authentication
of meat and meat products vs. detection of ani-
mal species in feedwhat is the difference?. InIOP
Conference Series: Earth and Environmental Sci-
ence 2017 Sep 1 (Vol. 85, No. 1, p. 012043). IOP
Publishing.
[26] Olsen P, Borit M. The components of a food trace-
ability system. Trends in Food Science & Technol-
ogy. 2018 Jul 1; 77: 143-9.
[27] Prayson BE, McMahon JT, Prayson RA. Apply-
ing morphologic techniques to evaluate hotdogs:
what is in the hotdogs we eat?. Annals of diag-
nostic pathology. 2008 Apr 1; 12(2): 98-102.
[28] Sadeghi E, Khazaei M, Almasi A, Shariatifar N,
Bohlouli Oskoii S, Tahvilian R. Recognition of lle-
gal Tissues in the Meat Products from Kerman-
shah Supply Centers during the years 2009-2010.
Internal Medicine Today. 2011 Apr 10; 17(1): 55-
9.
[29] Sarab I. Determination of adulteration and au-
thenticity of meat and meat products using chemi-
cal properties and PCR technique in Tabriz. Jour-
nal of Health. 2020 May; 11(4): 478-88.
[30] Veuthey TV, Herrera MG, Dodero VI. Dyes and
stains: from molecular structure to histologi-
cal application. Frontiers in Bioscience-Landmark.
2014 Apr, 19(1): 91-112.
)