استفاده از Nested-PCR در مطالعه انتشار انگل Cryptosporidium spp در پرندگان اهلی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشکده پرستاری، دانشگاه القادسیه، عراق

2 دانشکده بیوتکنولوژی، دانشگاه القادسیه، عراق

3 دانشکده علوم، دانشگاه القادسیه، عراق

چکیده

هدف: ین مطالعه به بررسی تعداد موارد عفونت Cryptosporidium spp. در پرندگان اهلی در استان دیوانیه و همچنین پیامدهای این بیماری و روش های تشخیص آن پرداخت.
مواد و روش‌ها: در این مطالعه، 205 نمونه از گونه های مختلف پرنده بدون در نظر گرفتن جنسیت مورد تجزیه و تحلیل قرار گرفت. برخی از روش‌های مورد استفاده برای تشخیص، روش رنگ آمیزی اسید فست و تکنیک نستد PCR بود. ثابت شد که نستد PCR بهتر از رنگ آمیزی اسید فست AFS است. زیرا می‌تواند کریپتوسپوریدیوم را سریعتر، با بالاترین دقت و حساسیت شناسایی کند.
نتایج: نتایج نشان داد که 90 نمونه از کل نمونه‌ها دارای عفونت کریپتوسپوریدیوم بودند. در بین گونه های پرنده مورد مطالعه، falseco eleonorae  با 16% کمترین بروز کریپتوسپوریدیوزیس را داشت، در حالی که گالوس گالوس داخلی با 3/53 درصد بیشترین بروز را داشت. از 60 نمونه گرفته شده برای هر گونه، 3/48 درصد از گونه Meleagris gallopova و 6/41 درصد از گونه Anas platyrhynchos آلوده بودند. بررسی میکروسکوپی میزان عفونت 9/43 درصد (90 نمونه از 205 نمونه) را در مقایسه با روش دیگر نشان داد. برای استفاده از Nested-PCR برای شناسایی مولکولی، DNA باید از نمونه های آلوده مثبت با استفاده از روش پروتئیناز K استخراج شود. این کار باندهای مثبتی را تولید کرد که معمولاً در محدوده 587 جفت بازی تکثیر می‌شدند. تست Nested-PCR نشان داد که 5/47 درصد نمونه ها (19 از 40 نمونه) آلوده بودند.
نتیجه‌گیری: در نهایت، این مطالعه با نشان دادن شیوع بسیار بالای عفونت کریپتوسپوریدیوم در پرندگان اهلی در استان دیوانیه، نیاز حیاتی به ابزارهای تشخیصی کارآمد و طرح‌های مداخله‌ای را برجسته کرد. به منظور جلوگیری از گسترش کریپتوسپوریدیوزیس در پرندگان، نتایج بر استفاده از روش‌های واکنش زنجیره‌ای پلیمراز نستد (nested-PCR) برای تشخیص عفونت تاکید دارد.

کلیدواژه‌ها

عنوان مقاله [English]

Using Nested-PCR in the Study of Cryptosporidium spp Parasite Spread in Domestic Bird

نویسندگان [English]

  • Saafa Ressan Abdullah Al-Kaeebi 1
  • Rana Saleh Al-Difaie 2
  • Fatima Ibrahim Mohammad 3
1 College of Nursing College, University of Al-Qadisiyah, Iraq
2 Biotechnology College, University of Al-Qadisiyah, Iraq.
3 College of science, University of Al-Qadisiyah, Iraq
چکیده [English]

Objectives
This study examined the number of cases of Cryptosporidium spp. infection in domestic birds in Diwaniyah Governorate, as well as the consequences of this illness and the methods used to diagnose it.
 
Materials and Methods
In the study, 205 samples from different bird species were analyzed without gender being taken into account. Some of the methods used for diagnosis were the Acid Fast Stain method and the DNA nested Polymerization Technique (PCR). Nested - PCR proved out to be better than AFS because it could find Cryptosporidium faster, with highest accuracy, and sensitivity.
 
Results
The results showed that 90 of the samples had Cryptosporidium infections. Among the bird species that were studied, falseco eleonorae had the lowest incidence of cryptosporidiosis at 16%, while gallus gallus domesticus had the highest incidence at 53.3%. Out of the 60 samples taken for each species, 48.3% of Meleagris gallopova and 41.6% of Anas platyrhynchos were infected. The microscope examination showed a 43.9% infection rate (90 out of 205 samples) when compared to the other method. To use nested-PCR for molecular identification, DNA had to be taken from positive infected samples using Proteinase K method. This produced positive bands that were usually amplified at around 587 bp. The nested-PCR test showed that 47.5% of the samples (19 out of 40) were infected.
 
Conclusions
Finally, this study highlights the critical need for efficient diagnostic tools and intervention plans by demonstrating the very high incidence of Cryptosporidium infection in domestic birds in Diwaniyah Governorate. In order to prevent the spread of Cryptosporidiosis in birds, the results stress the use of nested polymerase chain reaction (PCR) methods for infection diagnosis.

کلیدواژه‌ها [English]

  • Nested-PCR
  • Microscopic Examination
  • Infection Rate
  • Birds
  • and Cryptosporidium

مراجع

Abbas MM, Abooud KW, Mohammed AQ, et al. (2022) Effects of various irrigation levels and biochar-based fertilizers on peanut production. J Nuts 13(4), 289-300.‏
Abdullah D (2020) A Linear Antenna Array for Wireless Communications. National J Antennas Propag 2(1), 19-24.
ACAR, B.Ç., & Yüksekdağ, Z. (2023). Beta-Glycosidase Activities of Lactobacillus spp. and Bifidobacterium spp. and The Effect of Different Physiological Conditions on Enzyme Activity. Natural and Engineering Sciences, 8(1), 1-17.
Adejinmi JO, Oke M (2011) Gastro-intestinal parasites of domestic duck (Anas platyrhynchos) in Ibadan southwestern Nigeria. Asian J Poult Sci 5(1), 46-50.
Al Nazari AA, Barboza-Arenas LA, Romero-Parra RM, et al. (2023) Investigation and evaluation of the hybrid system of energy storage for renewable energies. Energies 16(5), e2337.‏
Al-dolaimy F, Kzar MH, Hussein SA, et al. (2024) Incorporating of Cobalt into UiO-67 Metal–Organic Framework for Catalysis CO2 Transformations: An Efficient Bi-functional Approach for CO2 Insertion and Photocatalytic Reduction. J Inorg Organomet Polym 34, 864–873.
Al-Hawary SIS, Kadhum WR, Saleh EAM, et al. (2023) Tunneling induced swapping of orbital angular momentum in a quantum dot molecule. Laser Phys 33(9), e096001.‏
Al-Jassani MJ, Sayah MA, Qasim MT, et al. (2022) Isolation and Evaluation of Antibacterial Agents Produced by Soil Bacillus SP. and Study Some of their Immunological Parameters. Revista Electronica de Veterinaria 23(4), 105-111.‏
Al-Safi MT, Qasim MT (2023) Study of some genetic and molecular markers for some rheumatoid arthritis patients in Iraq. Bionatura 3(8), 1-13.
Alseady HH, Kawan MH (2019) Prevalence and molecular identification of Cryptosporidium        spp in cattle in Baghdad province, Iraq. Iraqi J Vet Sci 33(2), 389-394.                                                                                                  
Althomali RH, Al-Hawary SIS, Gehlot A, et al. (2023) A novel Pt-free counter electrode based on MoSe2 for cost effective dye-sensitized solar cells (DSSCs): Effect of Ni doping. J of Phys Chem Solids 182, e111597.‏
Arif A, Alameri AA, Tariq UB, et al. (2023) The functions and molecular mechanisms of Tribbles homolog 3 (TRIB3) implicated in the pathophysiology of cancer. Int Immunopharmacol 114, e109581.‏
Baroudi D, Khelef D, Goucem R, et al. (2013) Common occurrence of zoonotic pathogen Cryptosporidium meleagridis in broiler chickens and turkey in Algeria. J Vet Parasitol 196(3-4), 334-340.
Barta JR, Thompson RC (2006) What is Cryptosporidium? Reappraising its biology and phylogenetic affinities. Trends Parasitol 22(10), 463- 468.
Bashar SB, Kareem HA, Hasan YM, et al. (2022) Application of novel Fe3O4/Zn-metal organic framework magnetic nanostructures as an antimicrobial agent and magnetic nanocatalyst in the synthesis of heterocyclic compounds. Front Chem 10, e1014731.‏
Blagburn BL, Lindsay DS, Giambrone JJ, et al. (2003) Experimental cryptosporidiosis in broiler chickens. J Poult Sci 128, 442-449.
Cadé D, Blanchet B (2013) From Computationally-Proved Protocol Specifications to Implementations and Application to SSH. J Wireless Mobile Net Ubiquitous Comput Dependable Appl 4(1), 4-31.
Casemore DP (2000) Human Cryptosporidiosis clinical aspect, Epidemiology and control. In: Current Medicine Proc. J Cpll Phys Edin 30, 287-293.
Craddock H, Konudula LP, Kul G (2019) A Survey on the Challenges of Implementing Physical Memory Pools. J Internet Serv Inf Secur 9(2), 57-71.
Gautam AK, Kumar S (2020) Techniques for the Detection, Identification, and Diagnosis of Agricultural Pathogens and Diseases, Editor(s): Chukwuebuka Egbuna, Barbara Sawicka,Natural Remedies for Pest, Disease and Weed Control. Academic Press, pp. 135-142.
Gupta J, Suliman M, Ali R, et al. (2023) Double-edged sword role of miRNA-633 and miRNA-181 in human cancers. Pathol Res Pract 248, e154701.‏
Guyot K, Sarfati  C, Nevez G (2001) Molecular charactrization of Cryptosporidium isolates obtained from humans in France. J Protozoal 39(10), 3472-3480.
Helmy YA (2014) Epidemiological investigations on the public health significance of Cryptosporidium parasites in livestock and people in the Ismailia Canal Zone of Egypt (PhD dissertation). Freie Universität Berlin, Germany. pp. 77.
Hjazi A,  Nasir F, Noor R, et al. (2023) The pathological role of CXC chemokine receptor type 4 (CXCR4) in colorectal cancer (CRC) progression; special focus on molecular mechanisms and possible therapeutics. Pathol Res Pract 248, e154616.‏
Hjazi A, Ahsan M, Alghamdi MI, et al. (2023) Unraveling the Impact of 27-Hydroxycholesterol in Autoimmune Diseases: Exploring Promising Therapeutic Approaches. Pathol Res Pract 248, e154737.‏
Hunter PR, Nichole G (2003) Epidemiological and clinical features of Cryptosporidium infection in immune-compromised patient. J Clin Rev 115(1), 144-154.
Hussein HA, Khudair SA, Alwan M, et al. (2002) Impact of pollution caused by salmon breeding centers on river water quality. Caspian J Environ Sci 20(5), 1039-1045.‏
Jarad NI (2020) Molecular detection of Cryptosporidium parvum in chicken in Al-Diwaniya province. Iraq J Vet Sci 34(2), 441-445.
Jeon S, Lee H, Kim HS, Kim Y (2023) Universal Shift Register: QCA based Novel Technique for Memory Storage Modules. Journal of VLSI Circuits Syst 5(2), 15-21.
Karthikeyan G, Manoharan A, Swaminathan S (2019) A Scientometric Study on Neuro Science with Special Reference to Growth of Literature. Ind J Inf Sour Serv 9(S1), 77–79.
Khabiri A, Toroghi R, Mohammadabadi M, Tabatabaeizadeh S (2022) Cloning and nucleotide sequencing of the complete matrix protein of Newcastle disease virus subgenotype VII. 1.1 prevalence in broiler flocks of northeastern Iran. Modern Genet J 17(2), 113-125.
Khabiri A, Toroghi R, Mohammadabadi M, Tabatabaeizadeh SE (2023) Introduction of a Newcastle disease virus challenge strain (sub-genotype VII. 1.1) isolated in Iran. Vet Res Forum 14(4), e221.
Khursheed M, Hadi KM, Faraj M, et al. (2023) Methanol extract of Iraqi Kurdistan Region Daphne mucronata as a potent source of antioxidant, antimicrobial, and anticancer agents for the synthesis of novel and bioactive polyvinylpyrrolidone nanofibers. J Front Chem 11, e1287870.  
Laberge I, Griffiths MW (2000) Review article prevalence, detection and control of cryptosporidiosis in food. Int J Food Microbiol 32(1-2), 1-26.
Lafta HA, AbdulHussein AH, Al-Shalah SAJ, et al. (2023) Tumor-Associated Macrophages (TAMs) in Cancer Resistance; Modula-tion by Natural Products. Curr Top Med Chem 23(12), 1104-1122.‏
Lazo AO, Barriga O, Redman DR, Bech-Nielsen S (1986) Identification by transfer blot antigens reactive in the enzyme-linked immunosorbant assay (ELISA) in rabbits immunized and calf infected with Cryptosporidium spp. J Vet Parasitol 21(3), 151-163.  
Lei Z, Alwan M,  Alamir HTA, et al. (2022) Detection of abemaciclib, an anti-breast cancer agent, using a new electrochemical DNA biosensor. Front Chem 10, e980162.‏
Lengmei W, Xue X, Jianqiu L, et al. (2014) Cryptosporidiosis in broiler chickens in Zhejiang Province, China: molecular characterization of oocysts detected in fecal samples. Parasite 21(36), 1-5.
Majewska AC, Graczyk TK, Slodkowicz-Kowalska A, et al. (2009) The role of free-ranging, captive, and domestic birds of Western Poland in environmental contamination with Cryptosporidium parvum oocysts and Giardia lamblia cysts. J Parasitol Res 104(5), 1093-1099.
Margiana R, Alsaikhan F, Al-Awsi GRL, et al. (2022) Functions and therapeutic interventions of non-coding RNAs associated with TLR signaling pathway in atherosclerosis. Cell Signal 100, e110471.‏
Moazeni S, Mohammadabadi MR, Sadeghi M et al. (2016a) Association between UCP Gene Polymorphisms and Growth, Brreeding Value of Growth and Reproductive Traits in Mazandaran Indigenous Chicken. Open J Anim Sci 6, 1-8.
Moazeni SM, Mohammadabadi MR, Sadeghi M et al. (2016b) Association of the melanocortin-3(MC3R) receptor gene with growth and reproductive traits in Mazandaran indigenous chicken. J Livest Sci Technol 4, 51-56.
Mohammadabadi MR, Nikbakhti M, Mirzaee HR, et al. (2010) Genetic variability in three native Iranian chicken populations of the Khorasan province based on microsatellite markers. Russ J Genet 46(4), 505-509
Mohammadifar A, Faghih Imani SA, Mohammadabadi MR, Soflaei M (2014) The effect of TGFb3 gene on phenotypic and breeding values of body weight traits in Fars native fowls. Agric Biotech J 5, 125-136 (In Persian).
Mohammadifar A, Mohammadabadi MR (2017) Effect of Uncoupling Protein Polymorphisms on Growth, Breeding Value of Growth and Reproductive Traits in the Fars Indigenous Chicken. Iran J Appl Anim Sci 7, 679-685.
Mohammadifar A, Mohammadabadi MR (2018) Melanocortin-3 receptor (MC3R) gene association with growth and egg production traits in Fars indigenous chicken. Malays Appl Biol 47, 85–90.
Nadham KM, AL-Sadoon IA, Adel T (1996) First report of cryptosporidiosis among Iraq children. J Eastern Mediterranean Health 2(1), 115-119.
Qi M, Wang R, Ning C, et al. (2011) Cryptosporidium spp. in pet birds: genetic diversity and potential public health significance. J Exper Parasitol 128(4), 336-340.
Rhee JJK, Sen YS, Park BK (1991) Isolation and Identification of C. baileyi from Korean chicken. Korean J Parasitol 29, 315-324.
Sane S, Mahoori A, Abdulabbas HS, et al. (2023) Investigating the effect of pregabalin on postoperative pain in non-emergency craniotomy. Clin Neurol Neurosurg 226, e107599.‏
Santin M (2013) Clinical and subclinical infections with cryptosporidium in animals. N Z Vet J 61(1), 1-10.
Shahdadnejad N, Mohammadabadi MR, Shamsadini M (2016) Typing of Clostridium Perfringens Isolated from Broiler Chickens Using Multiplex PCR. Genet Third millennium 14 (4), 4368-4374.
Shukla A, Pandey A, Srivastava S (2013) VEBEK-Working & Functionalities. Int J Commun Comput Technol 1(2), 134-142.
Suljić N (2021) Influence of Roughty of A Rectangular Open Channel on Flow Speeds and Water Flow. Arch Technic Sci 1(24), 57–64.
Tahseen AA, Najlaa OHA (2018) Prevalence Rate of Isolated Cryptosporidium Spp. of Broiler Chicken in Al Qadisiyah Province. Al-Qadisiyah J Agric Sci 8(2), 101–106.
Thantip M, Kalaya A, Suthathip K, et al. (2016) mRNA Expression of Bax, Bcl-2, p53, Cathepsin B, Caspase-3 and Caspase-9 in the HepG2 Cell Line Following Induction by a Novel Monoclonal Ab Hep88 mAb: Cross-Talk for Paraptosis and Apoptosis. Asian Pac J Cancer Prev 17, 703-712.
Valigurova A, Jirku M, Koudela B, et al. (2008) Cryptosporidia: Epicellular parasites embraced by the host cell membrane. Int J Parasitol 38(8-9), 913-922.
Voelz k, May R (2010) Cryptococcal interactions with the host immune system. Eukaryot Cell 9(6), 835-46.
Wang R, Jian F, Sun Y, et al. (2013) Large-scale survey of Cryptosporidium spp. In chickens and Pekin ducks (Anas platyrhynchos) in Henan, China: prevalence and molecular characterization. J Avian Pathol 39(6), 447- 451.
Xiao L, Fayer R, Ryan U, Upton SJ (2004) Cryptosporidium taxonomy: recent advances and implications for public health. Clin Microbiol Rev 17(1), 72 –97.
Yosra AH, Jürgen K, El-Sayed MA, et al. (2017) Molecular diagnosis and characterization of Cryptosporidium spp. in turkeys and chickens in Germany reveals evidence for previously undetected parasite species. PLoS ONE 12(6), e0177150.
Zaman GS, Waleed I, Obeid RA, et al. (2023) Electrochemical determination of zearalenone in agricultural food samples using a flower like nanocomposite-modified electrode. Mat Chem Phys 305, e127986.
Ze L, Al-dolaimy F, Mohammad Sajadi S, et al. (2023) The effect of number of nanoparticles on atomic behavior and aggregation of CuO/water nanofluid flow in microchannels using molecular dynamics simulation. Engin Sci Technol Int J 47, e101556.
مجله بیوتکنولوژی کشاورزی
دوره 16، شماره 2
خرداد 1403
صفحه 281-295

فایل ها

هم رسانی

ارجاع به این مقاله

آمار