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DETECTION OF PIGEON PARAMYXOVIRUS ANTIBODIES USING HAEMAGGLUTINATON INHIBITION TEST A. A. SHAMAUN*; S. M. SAEED**
*Department of Pathology & Poultry Diseases,
College of Veterinary Medicine, Mosul, Iraq.
** Private Veterinary Clinic.
Abstract
This study was designed to detect the level of Newcastle disease antibody titers of local domestic pigeon in Ninavah governorate in Iraq by using Haemagglutination inhibition test (HI test). Eighty serum Samples were collected from these pigeon. Ninety percent of the positive birds for HI test were clinically affected with digestive, nervous and respiratory signs. The remaining 10% were sub clinically affected with no obvious signs. The nervous signs were the most predominant in affected birds ( 48.75% ) Which were being recovered in Mosul city. The result showed that all serum samples were positive for HI test with titers ranging from 2Log2 to 9Log2 , the highest was 9log2 , while the most predominant was 6log2.
ОБНАРУЖЕНИЕ АНТИТЕЛ ПАРАМИКСОВИРУСА У ГОЛУБЕЙ С ПОМОЩЬЮ РЕАКЦИИ ТОРМОЖЕНИЯ ГЕМАГГЛЮТИНАЦИИ ШАМАУН А .А .*; САИД С. М.**
* Кафедра Патологии и Болезни Птиц,
Колледж Ветеринарной Медицины, Мосул, Ирак. частная Ветеринарная Клиника * *
Резюме
Это исследование было разработано с целью выявления уровня титра антител болезни Ньюкасла у местных домашних голубей в провинции Нинавах в Ираке с помощью реакции Торможения Гемагглютинации (реакции ТГ). Девяносто процентов положительно реагирующих птиц были с клиническими признаками кишечной, нервной и дыхательной форм болезни. Остальные 10% были с субклинической формой болезни, без явно видимых признаков. Наиболее преобладающей оказалась нервная форма болезни (48, 75%),у птиц, взятых в городе Мосул. Результат показал, что все образцы сывороток положительно реагировали в РТГА с
титрами от 2Log2 до 9Log2, самый высокий титр был 9log2, а наиболее преобладающий 6log2.
Introduction
Newcastle Disease (ND) is a highly contagious avian disease which causes severe economic losses in domestic poultry especially in chickens due to mortality and morbidity in high percentage of infected chickens, which depends on virulence of the infecting strain and host susceptibility (1, 2). Newcastle disease is a virus disease of birds characterised by variable combinations of gastroenteritis, respiratory distress and nervous signs (3). All birds are susceptible but the disease is more severe in chickens and turkeys and less severe in ducks and pigeons(4). Over 200 species of birds have been reported to be susceptible to natural and/or experimental infection with ND virus and it seems probable that more are fully susceptible (5). Ducks and geese tend to show few signs even when infected with the most virulent strains of ND virus for chickens (5). Although many species of bird are susceptible to ND it occurs mainly in galliformes such as pheasants, partridges and quails and also in birds of prey, including pigeons and psittaciformes. In most species of bird the young are more susceptible than the adult (6). ND virus (NDV), the only member of the avian paramyxovirus type 1, belongs to the Avulavirus genus of the family Paramyxoviridae (7). The disease normally affects the respiratory, gastrointestinal and nervous systems. The clinical signs reported in birds infected with NDV vary widely but mainly depend on strain of the virus. Other factors determine the outcome of the disease such as the host species, age, immune status, infection with other organisms, and environmental stress (2). In some circumstances, with extremely virulent viruses the disease may result in sudden death (8). The antibody response to NDV occurs rapidly with detectable neutralizing antibodies, usually detected in the serum of birds within 4-6 days after vaccination with live attenuated vaccines (9).
Materials and Methods
Serum samples. Eighty Blood samples were collected in test tubes to get serum from local domestic pigeon in Ninavah governorate in Iraq via the slaughter from May- August 2009.
Chicken red blood cells (RBC). A 1% suspension of washed RBC was prepared for use in haemagglutination (HA) and haemagglutination inhibition (HI) tests (10).
Antigen. Pigeon paramyxovirus obtained from department of microbiology/ college of veterinary medicine/ university of mosul/ Mosul/ Iraq was used as the antigen for HI test. The HA titres of the Pigeon paramyxovirus antigen were determined as described by (10) and (11) and diluted to contain 4-
HA units. This concentration was used for the HI test. The HI titre for each bird was determined and expressed in log2, and the mean for each species was calculated.
Haemagglutination inhibition test (HI): Two- fold serial dilutions of serum samples were made with normal saline in macro titer plates. 0.25 ml of PBS was dispensed into each well of a macrotitre plate, 0.25 ml of each serum sample placed into the first wells of the plate.Two fold dilutions were made by transferring 0.25 ml of each serum sample from first row of wells to second and so on thus diluting each serum sample as 1:2, 1:4,1:8,1:16: 1:32...1:2048, etc. 0.25 ml of 4 HA units of virus were dispensed in each well with the help of 8-channel micropipette and plates were left for of 30 minutes at room temperature.
0.25 ml of 1% chicken RBCs added to each well and after gentle mixing, the RBCs allowed to settle for about 30 minutes at room temperature (12). The base two logarithmic titer was calculated.
Results
90% of 80 local domestic pigeon suffered from different clinical signs, mainly nervous, digestive and respiratory signs, and the nervous signs revealed the highest percentage representing 48.75% (figure 1).
Figure 1: percentages of clinical signs of sampled pigeon
% No. of serum's samples Clinical signs
48.75% 39 Nervous signs
23.75% 19 Digestive signs
11.25% 9 Respiratory signs
6.25% 5 Another signs
10% 8 Without clinical signs
100% 80 Total
All of the serum samples were collected from local domestic pigeon revealed positive reaction for HI test with different titers of antibodies. Pigeon suffered from nervous and digestive showed highest titer of antibodies 9log2, but 6log2 screened a highest number of 80 samples.
Figure 2: Log of HI paramyxovirus antibody titers of Pigeon
Titer
9log2 8log2 <N g <N 00 'O 5log2 4log2 <N eg 3 2log2
39 5 5 7 11 8 3 Nervous signs
19 4 5 7 3 Digestive signs
9 1 1 2 1 4 Respiratory signs
5 4 1 Another signs
8 3 3 1 1 Without clinical signs
80 9 10 15 18 17 6 5 Total
Discussion
There was serological evidence of paramyxovirus antibodies in local pigeons. This might be due to occasional visits of these birds to the poultry premises where they could have contracted the virus from materials from poultry houses. So, there is the possibility of transmission of paramyxovirus among these specie of birds.
Pigeons are among a group of domesticated birds that were primarily affected by the paramyxovirus (13). The disease spread to all parts of the world, largely as a result of contact between birds at races and shows and the large international trade in such birds (14).
There is serological evidence of paramyxovirus antibodies in pigeons in this work, the spread of paramyxovirus to chickens from pigeons has occurred in several countries including Great Britain where 20 outbreaks in unvaccinated chickens occurred in 1984 as a result of feed that had been contaminated by infected pigeons (15).
The presence of specific antibodies in the serum of a bird gives indication of presence an infection, as well as an immunological response (16) .
In this study, all of the serum samples were collected from local domestic pigeon revealed positive reaction for HI test with different titers of antibodies, and the highest HI paramyxovirus specific antibodies titer "9log2" was recorded
in the pigeon suffered from nervous and digestive signs, and that indicate these birds maybe infected with the virulent strain of virus (17).
References
1. Alexander DJ. Newcastle disease. Br Poult Sci, 2001, 42: 5-22.
2.Alexander DJ. Newcastle disease, other avian paramyxoviruses, and pneumovirus infections, In: Saif, Y.M.; Barnes, H.J.; Glisson, I.R.; Fadly, A.M.; McDougald, J.R. and Swayne, D.E. (Ed.), Disease of Poultry, 11th ed. Iowa State University Press, Ames, 2003, pp: 63-92.
3. Durojaiye OA and Owoade AA. Post - vaccination Newcastle disease in a breeder flock in Nigeria. Nig. J. Immunol, 1990, 2:41 - 42.
4. Munjeri N. An antigen ELISA for the detection of Newcastle disease virus. Zimbabwe Vet J, 1996, Vol. 7, No. 2.
5. Alexander D J. Paramyxoviridae (Newcastle disease and others) In: Poultry Diseases. 4th ed. (Jordan, F.T.W, M. Pattison,m Eds.) W. D. Saunders and Company Limited London. 1990, pp: 139-155.
6. Arnall L and Keymer I P. Bird Diseases. Baillere and Tindall, 8 Henrietta Street London. 1975, pp: 451-459.
7. Wise MG, Sellers HS, Alvarez R and Seal BS. RNA-dependent RNA polymerase gene analysis of worldwide Newcastle disease virus isolates representing different virulence types and their phylogenetic relationship with other members of Paramyxoviridae. Virus Res, 2004, 104:71-80.
8. Brown C, King DJ and Seal B. Pathogenesis of Newcastle disease in chickens experimentally infected with viruses of different virulence. Vet Pathol, 1999, 36:125-132.
9. Alexander DJ. Newcastle disease and other Paramyxoviridae infections. In: Calnek, B.W.; Barnes, H.J.; Beard, C.W. and McDougald, L., Saif Y.M., (Ed.), Diseases of poultry, 10th ed., Iowa State University Press, Ames, Iowa, 1997, pp: 541-570.
10. Allan, W. H., R. E. A. Gough (1974): Standard haemagglutination inhibition test for Newcastle disease: A comparison of macro and micro methods. Vet. Rec. 95, 120-123.
11. Allan WH, and Lancaster JE and Toth B. Newcastle disease vaccines. Their Production and Use: FAO Animal Production and Health Series, 1978, No. 106, pp:50-69.
12. Gould EA and Clegg JCS. Growth. Titration and purification of Alphaviruses and flavi virusis. In: virology. A practical approach. Mahy BW,(Ed.), IrI press, Oxford, 1987, pp:65-67.
13. Kaleta EF, Alexander DJ and Russell PH. The first isolation of the PMV-1 virus responsible for the current panzootic in pigeons. Avian Pathol, 1985, 14: 553 - 557.
14. Biancifiori F and Fioroni A. An occurrence of Newcastle disease in pigeons: Virological and serological studies on the isolates. Comp Immunol Microbiol Infect Dis, 1983, 6:247-252.
15. Alexander DJ, Wilson GWC, Rusell PH, Lister SA and Parsons G. Newcastledisease outbreaks in fowl in Great Britain during 1984. Vet Rec, 1985, 117:429-434.
16. Shuaib M, Khan H, Sajid-ur-Rehman and Ashfaque M. .Humoral Immune Response to Newcastle Disease Vaccine (Lasota Strain ) in Broilers . International Journal of Poultry Science, 2006, (5): 411-414.
17. Hassanzadeh M and Bozorgmeri fard MH. A serological study of Newcastle Disease in pre-and post-vaccinated Village chickens in north of Iran. International journal of poultry science, 2004, 3:658-661.
УДК 619:616.9-036.2+612.017.11/.12+576.8
СОВРЕМЕННЫЕ ПРОБЛЕМЫ ЭПИЗООТОЛОГИИ, ИММУНОЛОГИИ И МИКРОБИОЛОГИИ Юсупова И.Т.
Буинский ветеринарный техникум
Ключевые слова: проблема, эпизоотология, иммунология,
микробиология.
Key words: problem, Epizootology, immunology, microbiology.
Хусаин Галеевич Г изатуллин - заслуженный деятель науки РСФСР и ТАССР, доктор ветеринарных наук, профессор, награждён многими правительственными наградами, орденами Ленина, Октябрьской
Революции, Отечественной войны 2 степени, Красной Звезды, Трудового Красного Знамени, «Знак Почёта» и многими медалями.
В сентябре 2010 года вся передовая общественность ветеринарии отмечает 100 лет со дня рождения Х. Г. Гизатуллина.
В октябре 1960 года Хусаин Галеевич назначен проректором по научной работе института, а в 1963 - 1975 годах возглавил институт в должности ректора. Казанский ветеринарный институт под его руководством стал комплексным учебным и научно - исследовательским центром.
Х.Г.Гизатуллин возглавлял кафедру эпизоотологии и одну из ведущих лабораторий, которая занималась научными исследованиями в области иммунологии и вирусологии. Им, совместно с научными сотрудниками разработаны ускоренные методы диагностики инфекционных
