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  3. Detection of Histomonas mele agridis in slaughtered turkeys in Isfahan province

Detection of Histomonas mele agridis in slaughtered turkeys in Isfahan province

Number of pages: 70 File Format: word File Code: 31994
Year: 2014 University Degree: Master's degree Category: Veterinary Medicine
Tags/Keywords: Agridis - Histomonas Mele - Killed turkeys - Meat turkeys - PCR - Trace Histomonas - turkey - Turkey breeding
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  • Summary of Detection of Histomonas mele agridis in slaughtered turkeys in Isfahan province

    Dissertation for receiving a professional doctorate degree in veterinary medicine

    Abstract

    In order to detect Histomonas mele agridis in slaughtered turkeys in Isfahan province, 200 blood samples and 200 liver samples were taken in the slaughterhouse, and after DNA extraction, 550 base pairs related to the gene 18SRNA was amplified. In addition, liver samples were preserved in 10% formalin and after preparation of pathological sections stained with hematoxylin-eosin, they were examined for the presence of parasites. PCR results showed that out of 200 blood samples, 18 samples (9%) and out of 200 liver samples, 32 samples (16%) were positive for Histomonas Melae Agridis. The histopathology results showed that 11.76% of the PCR positive samples were evaluated as positive in histopathology, so it seems that PCR has a high specificity for identifying Histomonas mele agridis. rtl;">- Introduction

    In recent decades, due to the growth of human societies, the need for food resources, especially animal protein items, has increased, and industrial turkey breeding is expanding all over the world in order to meet part of this need. In 2004, the United States ranked first with the production of 2,592,000 tons of turkey meat. In our country, industrial turkey breeding in In recent years, there has been a significant growth, so that its meat production can be sold in various forms or carcasses (5).

    In Iran, the turkey breeding industry started in 1355, and every year the number of turkey farms in the country is added (1). High growth rate (10.74 kg in 16 weeks for female turkeys and 20.39 kg in 20 weeks for male turkeys) low food conversion rate, low percentage of carcass loss and good nutritional value compared to other industrial poultry, industrial breeding of this bird is expanding worldwide to meet some of these needs, so that in 2007, the United States, the European Union, Brazil and Canada in terms of turkey meat production worldwide They were in the first to fourth ranks respectively (7). Histomonas mele agridis[2] is uniflagellate. Parasitic protozoa cause hepatitis of the cecum. The sites of attack of this parasite are the cecum and the liver, and blood circulation disturbance appears in the later stages of this disease and causes a black-headed disease in the bird.

    In recent decades, histomoniasis has caused serious damage to the poultry industry, especially among turkeys. A few cases of disease occurrence in chickens have also been reported, but due to their resistance to the disease, chickens mostly play the role of disease reservoirs and in case of infection with the Hetrakis gallinarum worm[3], they play an important role in the transmission of the disease to turkeys. Turkey chicks get infected by eating infected Hetrakis eggs. This disease has a wide geographical distribution in the world. Clinical observations and necropsy in Iran show the presence of the disease (29). Walking swagger, apathy and a slight increase in mortality. This is the first report of the presence of histomoniasis in the bursa of Fabricius in commercial chickens (10).

    In another study conducted by Hurst (1980) on the topic of investigating histomoniasis in wild turkeys in Mississippi, USA, it was shown that histomoniasis parasites are present in high-density wild turkey populations (19).

    In another study conducted by Lotfi et al. (2012) showed that histomoniasis parasite survives very poorly outside the host body.This investigation was carried out under the same conditions as the farm and at room temperature on several synthetic materials, the results of which include: protozoan survives for up to one hour on wood, rubber and metal, up to 3 hours in egg boxes, egg shells and bricks, up to 6 hours in feathers and wings and turkey feed, and up to 9 hours in non-chlorinated tap water and feces. Therefore, contaminated water and fresh feces or both can play an important role in the transmission of the parasite (24).

    In a study conducted by McDougald (2005) on the subject of histomoniasis in birds in Rhode Island, it was shown that blackhead disease causes high mortality in turkeys, sometimes close to 100%, and in chickens, the mortality rate may be as high as 20 to 100 percent be accompanied. In this research, it was shown that histomoniasis spreads rapidly in the turkey flock through direct contact between the birds, and it is probably related to the phenomenon of cloacal drinking. In this research, dependence on Hethrakis gallinarum is mentioned as the source of infection (29).

    In another study conducted by Patra et al. (2013) in India with the aim of identifying the incidence of histomoniasis in meat birds through a molecular method, it showed that out of 4000 birds examined, 40 birds were suffering from histomoniasis Mele agridis, or in other words, the prevalence rate It was 1% (32).

    In a report by Popp et al. (2011) showed that an organic farm in southern Germany for 3 consecutive years among broilers and turkeys, an outbreak of a severe disease caused by the protozoan parasite Histomoniasis Melae agridis occurred and the diagnosis was through DNA (33).

    in another study conducted by Senties-Cue et al. (2009) showed that turkeys aged 7 to 13 weeks had symptoms such as anorexia, depression, diarrhea, weight loss and increased mortality. The signs of its autopsy included severe thickening of the cecum wall, enlargement of the bursa, pancreas, and pale and yellow spleen. Electron microscopy confirmed the presence of histomoniasis in the liver and cecum. This is the first report of the presence of systemic histomoniasis affecting the bursa of Fabricius, kidneys, lungs, pancreas and pre-stomach in turkeys (38). rtl;">1-4-1-1- General objective

    Detection of Histomonas mele agridis parasite in turkey flocks of Isfahan Province by PCR method

    1-4-1-2- Partial objectives

    Estimation of abundance

     

    1-4-2- Research hypotheses

    Histomoniasis exists in turkeys of Isfahan.

     

    1-4-3- Research questions

    Is Is histomoniasis present in the turkeys of Isfahan? Turkey in Isfahan province, referring to the turkey slaughterhouse located in Isfahan province, including Tiran, Koron and Najaf Abad cities, 200 blood samples and 200 liver samples were taken from 20 turkey flocks, and the samples were transferred to the laboratory on ice and kept at -20 degrees Celsius until the time of testing.

     

     

    Abstract

    For detection of Histomonas meleagridis in slaughtered turkeys in Isfahan province, 200 blood and 200 liver samples were collected and after DNA extraction, a 550 bp fragment was amplified. Furthermore, liver samples were collected in formalin 10% and histopathological sections were prepared for the presence of this parasite. PCR results showed 18 out of 200 blood samples (9%) and 32 out of 200 liver samples (16%) were positive for H.meleagridis.

  • Contents & References of Detection of Histomonas mele agridis in slaughtered turkeys in Isfahan province

    List:

    Abstract 1

    Chapter One "Introduction and Research Design"

    1-1- Introduction. 3

    1-2- statement of the problem. 4

    1-3- An overview of research history. 4

    1-4- Objectives, hypotheses and research questions. 6

    1-4-1- Research objectives. 6

    1-4-1-1- general goal. 6

    1-4-1-2- Minor goals. 6

    1-4-2- research assumptions. 6

    1-4-3- Research questions. 6

    1-5- research method. 7

    Chapter Two "Generalities"

    Parasites and diseases of poultry and other birds. 9

    2-1- External parasites. 9

    2-1-1- Bed bugs 9

    2-1-2- Biting lice 9

    2-1-3- Black flies 10

    2-1-4- Poultry mites. 10

    2-1-5-Larumites 10

    2-1-6- The bed bug and its larvae, Alphitubius depaprinus. 11

    2-1-7- Feather mites and mites that cause shedding. 11

    2-1-8- subcutaneous mites. 11

    2-2- internal parasites. 12

    2-2-1- conjunctival sac. 12

    2-2-2- trachea. 13

    2-2-4- pre-stomach 14

    2-2-5- calculus. 14

    2-2-6- small intestine. 15

    2-2-7- cecum 16

    2-2-8- skin. 17

    2-3- Parasitic diseases of poultry 17

    2-3-1- Coccidiosis 17

    2-3-2- Cryptosporidiosis 20

    2-3-3- Hexamithiasis. 24

    2-3-4- Histomoniasis. 27

    2-3-5-Leukocytozoonosis 32

    2-4-Polymerase chain reaction (PCR) 35

    2-4-1- Materials and tools necessary to perform PCR test. 36

    2-4-2- Synthesis. 36

    2-4-3- Detection and analysis of PCR products: 37

    Chapter 3 "Materials and work methods"

    3-1- Sampling. 40

    3-2- DNA extraction from the liver using a kit. 40

    3-3- DNA extraction from blood using a kit. 41

    3-4- PCR test. 42

    3-4-1- Working method 42

    3-5- Preparation of agarose gel 43

    3-5-1- Necessary materials and tools. 43

    3-5-2- Method of preparation of buffer (1X) TBE for preparation of agarose gel 44

    3-5-3- Method of preparation of agarose gel 44

    3-5-4- Method of performing electrophoresis 44

    3-5-5- PCR product analysis: 45

    3-6- Histopathology tests. 45

    3-6-1- Hematoxylin-eosin (H&E) staining: 45

    3-6-2- Staining slide with color (H&E) method: 46

    3-6-3- Staining interpretation (H&E) 47

    3-6-4- Staining results (H&E) 48

    Chapter Four "Results"

    4-1- Qualitative results related to DNA extraction. 50

    4-2- Results of histopathology tests. 51

     

    Abstract 1

    Chapter One "Introduction and Research Plan"

    1-1- Introduction. 3

    1-2- statement of the problem. 4

    1-3- An overview of research history. 4

    1-4- Objectives, hypotheses and research questions. 6

    1-4-1- Research objectives. 6

    1-4-1-1- general goal. 6

    1-4-1-2- Minor goals. 6

    1-4-2- research assumptions. 6

    1-4-3- Research questions. 6

    1-5- research method. 7

    Chapter Two "Generalities"

    Parasites and diseases of poultry and other birds. 9

    2-1- External parasites. 9

    2-1-1- Bed bugs 9

    2-1-2- Biting lice 9

    2-1-3- Black flies 10

    2-1-4- Poultry mites. 10

    2-1-5-Larumites 10

    2-1-6- The bed bug and its larvae, Alphitubius depaprinus. 11

    2-1-7- Feather mites and mites that cause shedding. 11

    2-1-8- subcutaneous mites. 11

    2-2- internal parasites. 12

    2-2-1- conjunctival sac. 12

    2-2-2- trachea. 13

    2-2-4- pre-stomach 14

    2-2-5- calculus. 14

    2-2-6- small intestine. 15

    2-2-7-Cecum 16

    2-2-8-Skin. 17

    2-3- Parasitic diseases of poultry 17

    2-3-1- Coccidiosis 17

    2-3-2- Cryptosporidiosis 20

    2-3-3- Hexamithiasis. 24

    2-3-4- Histomoniasis. 27

    2-3-5-Leukocytozoonosis 32

    2-4-Polymerase chain reaction (PCR) 35

    2-4-1- Materials and tools necessary to perform PCR test. 36

    2-4-2- Synthesis. 36

    2-4-3- Detection and analysis of PCR products: 37

    Chapter 3 "Materials and work methods"

    3-1- Sampling. 40

    3-2- DNA extraction from the liver using a kit. 40

    3-3- DNA extraction from blood using a kit.41

    3-4- PCR test. 42

    3-4-1- Working method 42

    3-5- Preparation of agarose gel 43

    3-5-1- Necessary materials and tools. 43

    3-5-2- Method of preparation of buffer (1X) TBE for preparation of agarose gel 44

    3-5-3- Method of preparation of agarose gel 44

    3-5-4- Method of performing electrophoresis 44

    3-5-5- PCR product analysis: 45

    3-6- Histopathology tests. 45

    3-6-1- Hematoxylin-eosin (H&E) staining: 45

    3-6-2- Method of staining slides with color (H&E): 46

    3-6-3- Staining interpretation (H&E) 47

    3-6-4- Staining results (H&E) 48

    Chapter Four "Results"

    4-1- Qualitative results related to DNA extraction. 50

    4-2- Results of histopathology tests. 51

    Chapter Five "Discussion and Conclusion"

    5-1- Discussion. 53

    5-2- Suggestions. 56

    Sources 57

    Source:

    Yi, M. (1384). Fundamentals of molecular biology and genetic engineering, Tehran: Mani Publications, pp. 10-35.

    2- Ohadinia, H. (2011). Industrial breeding and diseases of turkeys, Tehran: Alam and Qalam Publications. P. 161.

    3- Haj Sadegh, N. (1387). A review of poultry diseases, Tehran: Perto Vacheh Publications, pp. 76-110.

    4- Razmi, G., Bassami, M.H. and Maleki, M. (1384). Report of a case of histomoniasis in turkeys of Mashhad city, Tehran University Veterinary Faculty Journal. 60 (2): 145-143.

    5- Sohani, A., Khairkhah, A. and Masoudian, A. (1385). Management of turkey breeding, Tehran: Parto Vacheh Publications, p. 19. 6- Madervar, B. P. (1387). Homeopathic treatment of poultry diseases, translated by Hamid Khanghahi Abianeh, Tehran: Rosenha Publications, p. 60. 7- Waez, J. (1390). Investigating the prevalence of Ornithobacterium rhinotracheal infection in turkey flocks of Isfahan province by PCR method. In order to obtain the veterinary medicine thesis of the Islamic Azad University, Shahrekord branch.

    8- Whiteman, C.A. and Bickford, A.E. (1375). Guide to poultry diseases, translated by Mohammad Hassan Bozormehri Fard and collaboration. Tehran: Publications of the Education and Research Unit of the Ministry of Agriculture, pp. 229-268.

    9- Calnek, BW., John Barnes, H., Beard, CW., Reid, WM. and Yoder, HW. (1991) Disease of Poultry ninth ed.chapt33.Pp779-814.

    10- Cortes, P., Chin, RP., Bland, MC., Crespo, R. and Shivaprasad, HL. (2004). Histomoniasis in the Bursa of Fabricius of Chickens. Avian Diseases.48 (3):711-715.

    11- Curtice, C. (1907). further experiments in connection with the blackhead disease in turkeys. R. I. Agr. Exp. Sta. Bull. 124:67-105.

    12- Dearstyne, RS. (1945). When and why do they die? Turkey World.14:62-63.

    13- Desowitz, RS. (1951). Age as a factor influencing fatal infections of histomoniasis in chickens. J. Com. Path. And Therap. 61:231-236.

    14- Edgar, SA. (1986). Coccidiosis in turkeys: Biology and incidence. Proc Georgia Coccidiosis Conf, University of Georgia, Athens.116-123.

    15- Grabensteiner, E., Liebhart, D., Weissenbock, H. and Hess, M. (2006). Broad dissemination of Histomonas meleagridis determined by the detection of nucleic acid in different organs after experimental infection of turkeys and specified pathogen-free chickens using a mono-eukaryotic culture of the parasite. Parasitology International.65(4):317-322.

    16- Glisson, JR., Brown, TP., Brugh, M., Page, RK., Kleven, SH. and Davis, RB. (1984). Sinusitis in turkey associated with respiratory cryptosporidiosis. Avian Disease. 28:783-790.

    17- Hishaw, WR. (1937). Disease of turkeys Bulletin. Berkeley, CA: University of California Agricultural Experiment Station.

    18- Hoerr, FJ., Ranck, FM. and Hastings, TF. (1978). Respiratory cryptosporidiosis in turkeys. J Am Vet Med Assoc.173:1591-1593.

    19- Hurst, G.A. (1980). Journal of wildlife diseases 16(3).

    20- Karaman, M., Ozen, H. and Ozcan, K. (2009). Histomoniasis in turkeys: pathological observations and PCR detection. Dtsch Tierarztl Wochenschr, 116(6):214-219.

    21- Kendall, SB. (1957). Some factors influencing resistance to histomoniasis in turkeys. Brit.Vet. J. 133:435-439.

    22- Liu, W., Peng, J., Li, F., Sun, H., Ding, Y.

Detection of Histomonas mele agridis in slaughtered turkeys in Isfahan province
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