Detection of Dirofilaria immitis antigen in cats in Mosul city

Detection of Dirofilaria immitis antigen in cats in Mosul city

Wasan Amjad Ahmed¹, Zeena Dhubyan Mohammed Zaki² and Manal Hammadi Hasan¹

¹Department of Microbiology, College of Veterinary Medicine, University of Mosul, ²Department of Dentistry, Al-Noor University College, Mosul, Iraq

[email protected], 0000-0002-3090-9974

[email protected] , 0000-0001-8901-6444

[email protected] , 0000-0002-8351-1230

Abstract

This study is the 1^st trial to detect of Dirofeleria imitis in cats in Mosul city, Iraq, Dirofilariasis is the most dangerous nematodes which infect cats, through this study blood samples was collected from 200 cats (different ages, source and rearing management) to investigate from the mature female worms of D. immitis by using direct ELISA, results showed total percentage of infection was 7% (14/200 animals), the relationship between the infection with age of cats was proportional to the age of cats, as the percentage increases with the age of cats, the high prevalence of infection was recorded in cats more than 2 years old 4%, the kittens less than 1 year old showed low prevalence of infection 0.5%, statistically there is a significant differences (P<0.05) between the oldest animals with other ages, significant differences (P<0.05) between Outdoor cats which recorded high prevalence of infection 4%, while Indoor animals recorded low prevalence of infection 3% with D. imitis, 10 of imported cat was infected with D. imitis with high prevalence of infection (5%) when compare with native cats 2%, without significant differences (P<0.05) between two groups, this study was conducted that the prevalence of D. imitis is high in adult cats, with high prevalence in outdoor rearing and the cats with imported origin.

Keywords: Dirofeleria imitis,cats, Direct ELISA, Mosul

التقصی عن مستضد الخیطیات الدقیقة فی القطط فی مدینة الموصل

وسن أمجد احمد¹ زینة ضبیان محمد زکی² و منال حمادی حسن¹

¹فرع الأحیاء المجهریة، کلیة الطب البیطری، جامعة الموصل، ²فرع طب الأسنان، کلیة النور الجامعة، الموصل، العراق

الخلاصة

تعد هذه الدراسة من الأولى من نوعها للکشف عن طفیلی الخیطیات الدقیقة فی القطط فی مدینة الموصل، یعد داء الخیطیات الدقیقة من أخطر أنواع الدیدان الإسطوانیة التی تصیب القطط، حیث تم من خلال هذه الدراسة جمع 200 عینة دم من قطط (مختلفة الأعمار والمناشی فضلا عن مختلف أسالیب التربیة) وذلک للکشف عن مستضدات الإناث البالغة باستخدام اختبار الممتز المناعی المرتبط الأنزیم من النوع المباشر. بینت النتائج وجود نسبة إصابة کلیة 7%(14/200)، وعند إیجاد العلاقة ما بین نسبة الإصابة وعمر الحیوان تبین وجود علاقة طردیة إذا ارتفعت نسبة الإصابة بتقدم عمر الحیوان، حیث سجلت القطط التی تبلغ أعمارها أکثر من سنتین اعلى نسبة إصابة بالخیطیات الدقیقة 4%, فی حین سجلت اقل نسبة إصابة فی القطط التی یبلغ أعمارها اقل من سنة 0.5% وبفروق معنویة واضحة بین الفئة العمریة الأکبر سنا مع باقی الفئات العمریة. تبین إحصائیاً وجود فروقا بین القطط المربأة فی خارج المنازل والتی سجلت اعلى نسبة إصابة 4% فی حین سجلت القطط المربأة داخل المنازل اقل نسبة إصابة 3%, سجلت 10 قطط مستوردة إصابتها بالخیطیات الدقیقة إذ سجلت اعلى نسبة إصابة 5% مقارنة بالقطط المحلیة التی سجلت نسبة إصابة 2% ومن غیر وجود فوقا معنویا إحصائیا. بینت الدراسة أن اعلى نسبة إصابة بالخیطیات الدقیقة کانت فی القطط الأکبر سنا وکذلک المربأة خارج المنازل فضلا عن القطط المستوردة.

Introduction

Several parasitic diseases affect cats (1,2) and the Dirofilariasis is the disease caused by nematode Dirofilaria immitis known as the heartworm because the site of the worms in the lungs arteries (right ventricle) (3), the disease distributed worldwide and affected both feline and canine species (4). Animals can be infected via female mosquito which can be faded to blood of infected animals, L3 can be developed from microfilariae in feline only (5). Vector adequacy effect of larval development of the mosquito individual (6). Then larvae transcend through the pharynx then gut which stay approximately 1 day (7). They then migrate to the Malpighian tubules and invade the cells then transformation to the so sausage stage L1 mute to the L2 stage then to L3 (8). L3 emerges from the folded labium and rests on the skin of the host immersed in a drop of haemolymph then enters the host (9). Culex and Anopheles is the main vectors of transmission of the disease the clinical signs of infected cats manifested as chronic (subclinical) or acute death form and mainly limited to respiratory signs and the disease called Heart Worm Associated Respiratory Disease (HARD) (10). The direct ELISA, is a perfect test to investigate protein of female worm’s genital tract the number of female heartworms in cats is so low of infection with male worms which high (11). Negative antigen test result in infected animals can caused by two reasons: immaturity female worms or decrease numbers of female worms, while false-positive results is low, so the antigen test is considered to screening test for heartworm infection in cats (12). Detection of D. immitis is attempt by using serological assays, thorax X-ray, and echo-cardiogram, microfilaria is present in circulating 6 months’ post infection, the serological assays are sensitive about 97%, they are considered to have good sensitivity, Therefore, the most effective methods for diagnosis are chest X-ray, echo-cardiogram, and serological assays (13). Because there is no study which include the D. imitis in cats in Iraq, this study was aimed to detection of feline D. immitis antigens in cats using direct-ELISA.

Materials and methods

Animals

Two hundred of house hold and stray cats was examined. The age range of the cats was between 1 and 9 years old, estimated by the teeth, different source, rearing management.

Blood collections

A blood sample 2 ml was collected from cephalic vein of cat after sedation by the intramuscular injection of ketamine 7.5 mg/kg. The collected blood was added in to tube without anticoagulant (11). Serum was separated from clotted blood in a non-additive tube (14).

Direct ELISA

Antigen from the mature female worms of D. immitis was detected by using an ELISA kit (DiroCHEK®, SYNBIOTICS Corporation, San Diego, CA 92127, USA). The ELISA procedures was attempt according the manufacturer’s manual.

Statistical analysis

The difference in the percentages of infection between the various clinical status and ages of cows were assessed by using two-sided Chi-square test in IBM-SPSS statistics version19 program (15).

Results

Table 1 recoded the prevalence of D. imitis antigen in cats with different age, the relationship between the infection with age of cats was proportional to the age of cats, as the percentage increases with the age of cats, the high prevalence of infection was recorded in cats more than 2 years old, the kittens less than 1-year-old showed low prevalence of infection, statically there is a significant difference between the oldest animals with other ages in significantly different (Table 1). Outdoor cats recorded high prevalence of infection, while Indoor animals recorded low prevalence of infection with D. imitis with significant differences between it, in significantly different (Table 2). Ten of imported cat was infected with D. imitis with high prevalence of infection when compare with native cats, without significant differences between two groups (Table 3).

Table 1: The relationship between the infection of D. imitis and age of cats

Values significantly different at P

Table 2: the relationship of percentage of infection of D. imitis with rearing condition of cats

Values significantly different at P

Table 3: The prevalence of infection of D. imitis in Native and imported cats

Values significantly different at P

Discussion

D. imitis is wide spread infected parasite in cats, the parasite is not reported in cats in Iraq, so this study was reported total percentage of infection (7%), this percentage is so low, the reason of that may due to several causes spontaneous elimination of parasite, sudden death of animals, and the detection of microfilieria in peripheral blood is un satisfactory and microifileria is rarely found (16). The results of the present study showed first occurrence of D. imitis in cats in Mosul city. Several species of culicid act as vectors and play important role in D. imitis infection (17). Direct ELISA test give a positive result which indicated active adult worm infection and this result is highly specific, while the sensitivity is decrease when female worm two or fewer (18,19). The study conducted that the antibodies only indicate that an infection occurred and does not provide a guarantee that it still exists. There are variance results of the relationship between D. imitis infection and age of animals (20,21). The infection of D. imitis increase with increase of age because increase of duration of exposure to infection (vectors) (22). The results Showed the infection in all age of animals and revealed the relationship of infection which affected according to health status, immune status and other infectious agents (23). Other reason is the longtime of duration of prepatent period of the D. imitis is 5-6 months (24). The results recorded differences between indoor and outdoor animals, other researchers approve this result of infection with D. immitis in dogs (25). The outdoors animals may increase the chance to exposure of vectors may be bearing infective L3 of D. imitis and live under bad conditions, those animals may have exposed to other infectious agents which may play important role in immunosuppression of outdoor animals (26), indoor animals or house hold animals Puddles a veterinary important care and owners care this decrease the chance of infection and live under quarantine condition this decrease exposed to vectors (24). Animals which imported from different countries reported high prevalence of infection when compare with native animals (27). D. immitis has been reported in some neighborhood countries otherwise countries from which animals are imported, Saudi Arabia (28), Romania (3), Iran (29), Russia (30), Croatia, Syria, Lebanon and Turkey (31), Egypt and Qatar (32), these researchers improve the importation of infected animals to transmission the infection from country to other country as emerging disease.

Conclusions

D. imitis was first investigated in cats in Mosul city, with a higher infection in

Acknowledgment

The authors wish to thank the College of Veterinary Medicine/ University of Mosul for financially supporting this work, laboratory of the veterinary teaching hospital, for their support and the animal’s owners for their cooperation

Conflict of interest

Author declare no conflict of interests of the manuscript.