Serological investigation of five diseases; Influenza, Newcastle disease, Salmonella, Mycoplasma gallisepticum and Mycoplasma synoviae in native hens of Eghlid, Iran

Aim: The study was conducted to determine seroprevalence of the five diseases influenza, Newcastle, Mycoplasma gallisepticum, Mycoplasma synoviae and salmonella, among around native hens of Eghlid in Iran, on spring 2011. 
 
Materials and Methods: On the basis of native Hens distribution, this region divided into four parts of Eghlid, Doskord, Sedeh and Hasan-abad. Fifty unvaccinated native Hens randomly selected from each part. Blood samples were aseptically collected from the wing veins using 5-ml sterile syringe. Serum from hens was tested for detection and titration for Mycoplasma and Salmonella by the rapid slide agglutination method, and was tested for influenza and Newcastle by the Hemagglutination Inhibition Assay. The data was analyzed completely in randomized design with four treatments, 50 repetitions for each disease. 
 
Results: 34 out of 200 samples (17%) were positive for influenza. There were significant differences between regions (p


Introduction
influenza vary greatly depending on factors such as the strain of virus, the health, age and species of the host, Agents of the disease, influenza, Newcastle disease, other diseases such as coli-bacillus, Newcastle, Salmonella, and Mycoplasma (two of two) are a virus Mycoplasma, and physicochemical factors such as and bacteria respectively.Those diseases are highly dust and ammonia.contagious, dangerous and almost zoonotic.Annual Newcastle is a highly contagious viral disease that great economic losses to the poultry industry have been infects the respiratory and nervous system.Agent of imposed, and may threaten human health.Therefore, this disease is a member of the family Paramyxoviridae.knowledge of disease process and contamination rate That can grow and multiply in egg embryo.Chickens in several regions is needed for control and prevention are highly susceptible to disease, but turkeys, ducks of disease.and other birds do not tend to develop severe signs.Avian influenza is a contagious viral respiratory Newcastle disease virus in humans is mild, and can disease, which is caused by influenza virus A of the cause conjunctivitis.Wild birds are the source of family Orthomyxoviridae.This disease has greater contamination and can spread the virus.The virus may economic importance in the poultry industry.Avian survive a part of its evolution in a wild bird's body.In influenza virus can infect various species of domestic chick mortality is very high, and sometimes reaches and wild birds.Influenza disease in poultry occurs in one hundred percent.In laying hens it decrease egg two forms; highly pathogenic avian influenza (which production.Newcastle disease can also cause softening "Plague Birds" was recognized and causes severe of the shell eggs.Eggs produced by infected birds have systemic disease with high mortality up to 100%) and carried Newcastle virus, so those are not worth to the low virulence avian influenza (which created at incubation.least clinical signs in poultry, and egg production is Salmonella is one of the main microflora of the slightly reduced) [1,2].Signs of infection with gastrointestinal tract, and often is transmitted through experience more severe disease, often accompanied by food to humans, and almost annual one million of swelling of the paranasal (infraorbital) sinus.people infected and died because of salmonella [3].
Conjunctivitis with a frothy ocular exudate is common in turkeys and occurs occasionally in chickens.The disease can be usually transmitted by a very wide Production is lower in infected flocks, with decreased variety of foods and wild animals such as rodents, pets, weight gain, feed efficiency and egg production.The birds, reptiles and insects [3,4].In developed and symptoms of avian mycoplasmosis are typically slow developing countries, infectiousness of salmonella is to develop, and the course of the disease can be one of the most important and widespread of prolonged.However, acute respiratory disease someinfectious food.But the rate of infection is varied times occurs in young birds, particularly turkeys.among countries [3].Foods such as meat, chicken and Mycoplasma synoviae disease is asymptomatic, other poultry products are one of the most important mortality rate is less than ten percent.The disease has a source of ransmission of salmonella to humans.More long incubation period (generally between 11 to 21 than 50% of Salmonella infections can be transmitted days after the transmission).Transmission is also to humans through poultry products.
horizontal or vertical.Survival of Mycoplasma Pullorum disease is almost exclusively a disease synoviae is low outside of the body.But transmission of young chickens and turkeys.Other birds such as though stool is very important [3].quail, pheasants, ducks, peacocks and guinea fowl are The aim of this study was to evaluate the also susceptible.The agent can be recovered from prevalence of influenza, salmonella, New castle almost all organs, tissues and excreta.In older birds Disease virus (NDV), Mycoplama gallisepticum (Mg) that have become carriers, S. Pullorum is most and Mycoplasma synoviae (Ms) in native hens in commonly recovered from the ova and oviduct; and it region of Eghlid (a city in south of Iran). is recovered only occasionally from other organs and tissues, including the alimentary tract.In the acute

phase of fowl typhoid the organism is also widely
Out study was conducted between March and distributed, but in carrier birds, the organism is found June 2011.200 apparently healthy native hens in the most often in the liver, spleen and reproductive tract, urban and rural of Eghlid (a city in south of Iran) were and occasionally in the caeca.Clinical signs are typical used in the study.With the distribution of native birds, of a septicaemic condition in poultry and include Eghlid was divided into four regions (Eghlid, Hassanincreasing mortality and poor quality in chicks hatched abad, Doskord and Sedeh).50 unvaccinated hens were from infected eggs.Older birds show signs of anaemia, selected randomly from each region (total 200 hens).A depression, labored breathing and diarrhoea causing total of 200 blood samples were collected from the adherence of excreta to the vent.The highest mortality wing vein of native hens (unvaccinated, mature, and occurs in birds of 2-3 weeks of age.In older birds healthy hens).However, in order to avoid confusion, disease may be mild or unapparent.In breeding flocks before use, the tubes are numbered.To avoid reduced egg production and hatchability may be the corruption of samples, all samples were maintained in only signs, and trans-ovarian infection resulting in the refrigerator and transported to laboratory within infection of the egg and hatched chicks or pullets are 48hrs.If a delay in sample transportation was expected, one of the most important transmission routes for the oc samples were centrifuged and frozen at -20 before disease [5,6].
being submitted to the laboratory.All antigens were Avian mycoplasmosis can be caused by several used from company Golbid, and imported from the species of Mycoplasma (class Mollicutes, order Netherlands (Nobilis® Antigens, Intervet International Mycoplasmatales, family Mycoplasmataceae) Co Netherlands).Antigens of Mg, Ms, and Sp were including Mycoplasma gallisepticum, M. synoviae, M. oc maintained at refrigerator in 4 temperature.Before meleagridis and M. iowae.M. gallisepticum is the most testing, in order to the balance temperature, antigens important pathogen in poultry.Mycoplasma were maintained 30 minutes at room temperature.And gallisepticum is the most economically significant before use, Antigens were stirred.Because of mycoplasmal pathogen of poultry.M. gallisepticum serological test, blood samples were centrifuged and infections are also known as chronic respiratory the serum was separated from it.disease (CRD) of chickens, infectious sinusitis of The detection of antibodies against Mg, Ms, and turkeys and house finch conjunctivitis [7,8].
Salmonella pullorum (Sp) were achieved by the rapid M. gallisepticum infection vary from asymptoslide agglutination method.The first, antigen of Mg, matic to severe, depending on the infecting strain and Ms and salmonella and serum samples without dilution other factors.More severe infections are seen when the are mixed at room temperature (0.03 cc of serum 0.03 birds are infected concurrently with Newcastle disease cc of antigen, proportion 1:1).Then, the plates were virus, infectious bronchitis virus, Escherichia coli or studied under a light source.If agglutination (clot) pathogens other [8].
occurs, the sample was positive.Test was carried out Infected chickens usually develop respiratory for positive samples of Mg and Ms with a dilution onesymptoms that may include rales, coughing, sneezing, eighth (1/8) and for Salmonella with a dilution onenasal discharges and dyspnea.Turkeys typically sixteenth (1/16) (positive serum by phosphate buffer The results showed that 4 out of 200 samples (4%) 0.5M and PH=7.2 was diluted).Therefore, diluted were positive for titers of Mycoplasma synoviae.samples were positive, the test was positive, otherwise Contamination rates from Eghlid, Doskord, Sedeh and it was negative.Of course, if in the first experiment was Hasan-abad were 0, 0, 2, and 6% respectively.The negative, the test was negative and was not required to highest contamination rate was related to the Hasandilute.Antibodies against avian influenza and abad (Table -1).Newcastle virus in the serum were evaluated using the

Hemagglutination Inhibition Assay (HI). The HI assay
In the present study, Avian Influenza Virus (AIV) was performed using 96 'U'-well microtiter plates, 1% antibody titer was found in all the areas and infectious v/v red blood cells and according to present standards lever in four areas weren't uniform.Migrant birds are [7,9,10].Due to unvaccinated hens, samples were one of carriers and agent incidence of AIV.Therefore, considered positive if titers were greater than 2(>2).
existence of more migrant birds and various other The resulting data using software mini tab 15, in a species of animals (such as ducks, goose and so on…) completely randomized design for each sample with in the above mentioned areas is one of the causes of four treatments (regions) and 50 repetitions of each different infectious lever.The absence of clinical signs treatment or region, were analyzed.The average of influenza in native hens, in spite of high antibody infectious rate of each disease was compared with titer in some birds, could be due to persistent and Duncan at 5% level.
acquired resistance of birds to influenza virus in the

Results
environment.The results showed that 34 out of 200 samples Al-Natour (2005) reported that the sero-prevalence (17%) were positive for titers of influenza.Contami-of AIV was 71% among broiler-breeder flocks in nation rate from Eghlid, Doskord, Sedeh and Hassan-Jordan [2].In another study conducted by Nooruddin abad were 8, 10, 28 and 22 percent respectively.The (2006) in Bangladesh, an overall 9.82% serohighest contamination rate was in the Sedeh.There was prevalence of AIV was recorded [11].Dergham (2009) no significant difference between Sedeh and Hassan-reported that sero-prevalence of AIV-H was 54.2 and 9 abad contamination rate (p <0.05), but concerning the 78.3% in broiler and laying hen farms in Jordan, other regions, there were significant differences among respectively [12].those regions.The contamination rate between Eghlid In a serological investigation, Dadras (2006) and Doskord were not significant (p <0.05) (Table 1).
reported that the sero-prevalence of AIV was 53.56%The results of the research showed that 38 out of among the farms around Shiraz city in Iran [13].In a 200 samples (17%) were positive for titer of Newcastle.
The results showed that Newcastle antibody titer The different among the regions was significant (p was found in all areas and prevalence rate of Newcastle <0.05).Difference between Eghlid and Sedeh contain four areas wasn't uniform.Highest contamination mination rate were not significant (p <0.01) .
rate was in the eghlid.Particles suspended in the air, In this study any sample was not positive for titer water and the contaminated food are the most of Salmonella pollurum.
important agent of transmission of Newcastle virus.The results showed that 170 out of 200 samples Prevalence rate was high, where there were maintained (85%) were positive for titers of Mycoplasma pigeon, because contamination can easily expand in the gallisepticum.Contamination rates from Eghlid, area.Lack of vaccination and incorrect vaccination Doskord, Sedeh and Hasan-abad were 88, 80, 90, and could increase prevalence of Newcastle disease.82% respectively.The highest contamination rate was Habibi (2007) reported that the sero-prevalence of related to the Sedeh.There were no significant Newcastle virus was 11.8% among native hens of differences among different regions (p<0.1), and the Kazeron city in Iran [15].Ghaem-meghami (2007), contamination rate was almost the same in four regions reported that the seroprevalence of NDV was 2.7, 19.5 (Table -1).and 0.55% among hens, goose and ducks in Iran,  Dadras (2006), reported that the sero-Conclusion prevalence of salmonella was 34.4% among the farms Contamination of Influenza, Newcastle and around Shiraz city in Iran [13].Fardi-pour (2007) Mycoplasma gallisepticum was high, and the highest reported that the sero-prevalence of salmonella was contamination rate was related to Mycoplasma 36.7% among farms around Isfahan city in Iran.
gallisepticum.It is usually recommended that Contaminated Eggs or foods are one of the main preventive strategies, such as appropriate husbandry transmission agents of the disease, that in these areas, and hygiene, sanitary handling of chicks and eggs, foods aren't brought from other regions.Besides, routine health monitoring, and vaccination of Native housing of cow and poultry are very low, and foods hens should be emphasized.aren't almost entered to these areas [4].
The results showed, that contamination rate by

Table - 1
. Comparison average contamination percent of native hens to several diseases among native hens around Eghlid in Iran.Amol city in Iran, an overall 15.8% serooverall sero-prevalence of the Mg and Ms of the flocks prevalence of NDV was recorded.in Serbia were 9.01 and 47.49% in 2000 and was 11.59The results showed that salmonella antibody titer and 22.17% in 2009 by SPA and Elisa tests respectively was not found in any area, that with results by Habbi and concluded that sero-prevalence of MS was (2007) had different (3.07%).In an investigation decreased ,versus the Mg increased [22].serology,