Serum Plate Agglutination Test Research Articles

Comparative evaluation of Serum Plate Agglutination Test (SPAT) and Rose Bengal Plate Test (RBPT) for diagnosis of Brucella abortus in sera of cattle and human

Comparative evaluation of Serum Plate Agglutination Test (SPAT) and Rose Bengal Plate Test (RBPT) for diagnosis of Brucella abortus in sera of cattle and human

Development of Rapid Serum Plate Agglutination Test (RSPA) For Screening of Mycoplasma Synoviae Antibodies Using Namakkal Regional Isolate

The aim of the present study was to develop rapid serum plate agglutination test for detecting Mycoplasma synoviae antibodies using Namakkal regional isolates in commercial layer chicken. Mycoplasma synoviae colour antigen was prepared using 0.5 per cent basic fuchsine dye with the antigen concentration of 3x109 organisms/ml for rapid serum plate agglutination test (RSPA). To find out sensitivity and specificity of developed RSPA test in comparison with indirect ELISA 184 serum samples were collected from layer flock and tested by rapid serum plate agglutination test and indirect ELISA. Seropositivity of M. synoviae antibodies in RSPA and indirect ELISA was 51.08 and 54.34 per cent, respectively. The sensitivity and specificity of RSPA with reference to indirect ELISA was found to be 94 and 100 per cent respectively.

Prevalence study of Mycoplasma synoviae in broiler flocks with lameness in Shahrekord and Isfahan, Iran

Identification of Mycoplasma synoviae (M. synoviae), an economically important pathogen in poultry industries worldwide, in clinical samples has critical importance. This study was conducted to determine the prevalence of M. synoviae in broiler flocks with clinical lameness using serological and polymerase chain reaction (PCR) methods. Blood serum samples (200) and joint fluid samples (200) were collected from 20 broiler flocks. All serum samples were studied using the serum plate agglutination (SPA) test while all joint fluid samples were studied using the PCR technique for detection of vIhA gene of M. synoviae. Results showed that 61 serum samples from 11 broiler flocks were positive for SPA test while 42 out of 61 samples were positive using SPA with 1/2 dilution. The PCR method indicated that 102 joint fluid samples of 16 broiler flocks were positive for presence of the vIhA gene of M. synoviae. Our results showed that the SPA and PCR techniques can be used for determination of the prevalence of M. synoviae in clinical samples of poultry.

Exploratory serosurvey for antibodies to avian pathogens in backyard chickens from a satellite community of Jalapa City, Guatemala

An exploratory serosurvey was conducted to determine the presence of circulating antibodies to avian pathogens in backyard chickens from Los Achiotes (LAC), a satellite community of Jalapa City, located in eastern Guatemala. Blood samples from 51 adult chickens belonging to 51 households were taken and investigated for the presence of antibodies to Avian Influenza (AI), Newcastle Disease (ND), Infectious Bronchitis (IB), Infectious Bursal Disease (IBD), Mycoplasma gallisepticum (MG) and M. synoviae (MS). Antibodies for AI, ND, were investigated by Hemagglutination Inhibition, for IB and IBD by ELISA (BioChek®) and for MG and MS by a rapid serum plate agglutination test. The cut-off point for positive titers was 1:4 for AI and ND and a 0.2 S/P ratio for IB and IBD. All sampled chickens were positive for concomitant antibodies to various pathogens. Over half of the chickens were positive reactors to antibodies to all six tested pathogens; about a third carried antibodies to five and the rest to four or three. The frequencies of positive reactors were: AI = 27 (53%); ND = 49 (96.1%); IB = 50 (98%); IBD = 51 (100%); MG = 45 (88%) and MS = 48 (94%). The results show that the dynamic population of backyard chickens in LAC could be a potential threat to backyard poultry, farm poultry, wild birds and human population. The need to develop interventions and policies following the One Health approach (animal health to achieve human health) is stressed.

Molecular identification of Mycoplasma synoviae from seroprevalent commercial breeder farms at Chittagong district, Bangladesh.

Aim:Worldwide, Mycoplasma synoviae (MS) is an important pathogen of poultry, especially for chicken and turkey. It causes respiratory tract infection and infectious sinusitis. The study was conducted to determine the seroprevalence of MS infection with associated risk factors and identification of MS organism in unvaccinated flocks of commercial breeder farms of the Chittagong district, Bangladesh.Materials and Methods:A total of 365 serum samples were collected and tested for MS using serum plate agglutination (SPA) test for determination of MS seroprevalence. On the other hand, tracheal swabs were collected from each seropositive flocks for polymerase chain reaction (PCR) to determine the presence of MS organism.Results:Among the farms, the highest prevalence was found to be 69% and the lowest prevalence was 28% with the average 60%. The seroprevalence of MS infection in breeder farms was highest 70% with the flock size >10,000 birds, whereas it was lowest 57% in the flocks ranging from 4000 to 7000. According to age group, the prevalence was found highest 70% in >60 weeks age group of birds and lowest 42% in 10-19 weeks group. The seroprevalence of MS in winter season was found as highest as 64%, whereas it was found lowest 60% in the summer season. There was a statistically significant difference (p<0.01) among the seroprevalence of MS in different breeder farms, flock size, and age groups, but there was no significant (p>0.05) difference in the winter, summer, and rainy season. To confirm the presence of MS in the samples, PCR test was applied using specific primers to amplify a 214 bp region of the 16S rRNA gene of the organism. In PCR, all seropositive flocks showed a positive result for MS.Conclusion:As the plate agglutination test result showed 100% similar with PCR result, it can be suggested that agglutination test is better than molecular and culture techniques for MS detection and it is also cheaper and less time-consuming method.

Seroprevalence and comparative study of serological tests for detection of mycoplasma gallisepticum infection in commercial layer farms at few districts of Tamil Nadu, India

Avian mycoplasmosis causes great economic losses to the poultry industry, and one of the major agents involved is Mycoplasma gallisepticum (MG). The study was conducted from October 2008 to September 2010 to detect the presence of antibodies against MG in poultry layers. Serum from commercial poultry layers (n = 1350) was tested for MG by using enzyme-linked immunosorbent assay (ELISA) and serum plate agglutination (SPA). A total of 1350 sera from commercial poultry farm were collected and tested in winter, 453 (33.6%) and 350 (26.0%) were positive in SPA test and ELISA, and in summer, 297 (22.0%) and 219 (16.2%) were positive in SPA test and ELISA respectively. In Pearson chi-square test, statistical significant differectnce (p = 0.000) were observed between serological tests (SPA test and ELISA), and it was also demonstrated that seasonal occurrence of infection were higher in zone C. It has been found that MG infection is still an important disease problem in commercial layer farms and there is need for a stringent surveillance and control to combat avian mycoplasmosis in Tamil Nadu and these diagnostic methods (SPA and ELISA) should be only used as screening tools in monitoring programs.

Relationship of Salmonella infection and inflammatory intestinal response with hematological and serum biochemical values in laying hens

There are few studies about the blood serum of laying hens infected with Salmonella. The differential leukocyte count and blood chemistry values are an important aid in the diagnosis of human diseases, but blood parameters in the avian species are not well known. On the other hand, invasive forms of bacterial gastroenteritis, like Salmonella, often cause intestinal inflammation so this study was undertaken to find a biomarker of Salmonella infection and inflammatory intestinal response in the hematological or serum biochemical parameters in laying hens. Furthermore, we evaluated the association of some farm characteristics with Salmonella infection and fecal leukocytes (FL). A fecal sample with at least one fecal leukocyte per field was considered positive for inflammatory intestinal response. False positive serum reactions for Salmonella infection, by serum plate agglutination (SPA) test, were reduced by heating the sample to 56°C for 30min and then diluting it 5-fold. The range of hematological and biochemical parameter values was very wide, in addition, there was a poor agreement between the SPA and FL results. Comparison of the positive and negative samples in SPA and FL showed that 1.3% and 79.8% of the laying hens were positive and negative in both tests, respectively. Hens with a positive SPA result showed a higher percentage of monocytes than those with a negative SPA result. Hens with a positive FL test had a higher percentage of heterophils, ratio of heterophils to lymphocytes and aspartate aminotransferase values, while the percentage of lymphocytes was significantly lower (P<0.05) than those with a negative FL test. The risk of Salmonella infection increased when the age of laying hens and the number of hens per poultry house was greater than or equal to 18 months old and 10,000 laying hens, compared to less than 18 months old and 10,000 laying hens, respectively. On the other hand, the risk of inflammatory intestinal response was higher in laying hens ≥18 months old than in hens <18 months old. Despite the fact that we did not find any specific biomarker of Salmonella infection, this is the first report about the change of Salmonella infection and inflammatory response in hematological/serum biochemical values for laying hens.

Seroprevalence of Mycoplasma gallisepticum antibody by ELISA and serum plate agglutination test of laying chicken.

Aim:Mycoplasma gallisepticum (MG) is important avian pathogen responsible for chronic respiratory disease of chicken and turkeys, which result in large economic loss for the poultry industry. The objectives of this study were determination of seroprevalence of MG antibody of commercial layer chicken at laying period in selected areas of Bangladesh.Materials and Methods:A total of 563 blood samples were collected randomly from selected commercial layer chickens at laying period during the period from July to December, 2013. Indirect enzyme linked immunosorbent assay (iELISA) and serum plate agglutination (SPA) test were performed to detect the presence of antibodies against MG.Results:Of 563 samples, 64.47% and 56.13% showed an overall prevalence of MG antibodies in iELISA and SPA test respectively. Prevalence of MG was recorded the highest (69.63%) at 50-55 weeks of age compared with lowest (53.26%) at 56-61 weeks of age (p<0.05). Significant (p<0.05) effect of breed were observed in the seroprevalence of MG infection in layer birds in the present study. The overall, 68.77%, 63.74% and 59.37% prevalence were found respectively in sonali, ISA Brown and White leg horn. The prevalence of MG antibodies was the highest (70.13%) in December followed by November (68%), October (65.67%), August (63.46%), September (58.54%) and July (51.78%) month. The seroprevalence of MG antibodies was higher (69.63%) in most of the large flocks and lower (56.82%) in small flocks.Conclusion:Therefore, might be suggested that the commercial layer farms should be routinely checked to monitor MG infection and the reactor birds should be culled since MG organism has the potential to transmit vertically. The correlation between MG antibody in month and flock size was not significant (p=0.359 and p=0.868, respectively).

Prevalence of Mycoplasmosis of Chickens at Kotwali Thana in Chittagong, Bangladesh

A study of prevalence of mycoplasmosis of chicken at kotwali thana in Chittagong was conducted from 23rd November, 2010 to 22nd March, 2011. A total of 455 either dead or sick birds were examined during the period. Diagnosis was based on history, clinical signs, serum plate agglutination (SPA) test and postmortem lesions. The overall prevalence of mycoplasmosis was 15.38%. The prevalence was 73% in broiler and 27% in layer. The result further revealed that highest number of cases were recorded in the age group of 8-21 days (49%) followed by 22-35 days age group (21%), 0-7 days age group (21%), 36-60 days age group (6%) and over 60 days age group (3%). It was also demonstrated that during the study period the highest prevalence of mycoplasmosis was recorded in January (33%) followed by February (30%), March (23%), and December (13%) and lowest in November (1%).

Evaluation of glycerin as preserving agent of chicken serum for plate agglutination test

Serum is widely used for the purpose of monitoring and diagnosis support for most of poultry diseases. In the case of the serum plate agglutination test (SPA), commonly used to detect antibodies for Salmonella Pullorum (SP), Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS), serum cannot be frozen because it may result in false positive. Without freezing, serum can last only for a few days. In this experiment, glycerin was evaluated as a serum preservering agent. About 50 samples for each disease and analyzed by SPA test previously were separated. Glycerin was added to serum from commercial chickens, with and without antibodies for SP, MG and MS, in the proportion of 1:1 (serum:glycerin) and kept at refrigerated conditions (2 to 8 ºC). For four years they were tested by the SPA, initially weekly, afterward monthly and then annually. The results show that serum with glycerin give consistent and valid results according to the kind of antibodies present for the period tested. Sera that glycerin was not added to, the results were valid only for the first week. From the second week on, microbial growth affected the test results of the sera without glycerin. Our investigation shows that glycerin can be used to preserve chicken serum for SPA under refrigerated conditions. It is an easy, simple and cheap procedure that can extend serum shelf life, useful mainly for control sera.

Comparison of Milk Ring Test; Serum Plate Agglutination Test and Polymerase Chain Reaction for the Detection of Bovine Brucellosis

Comparison of Milk Ring Test; Serum Plate Agglutination Test and Polymerase Chain Reaction for the Detection of Bovine Brucellosis

Seroprevalence of Mycoplasma gallisepticum infection in backyard and commercial layer chickens in Bhola district, Bangladesh

This study aimed to determine the seroprevalence of Mycoplasma gallisepticum (MG) infection in the chicken population of Bhola district, Bangladesh, during the period from April 2011 to March 2012. A total of 480 blood samples from chickens were collected from different upazilas (sub-districts) of Bhola district. The sampling considered the types of chicken (backyard and commercial layer), age groups (pullet, adult and old) and seasons (summer and winter). On the basis of the serum plate agglutination test, 55.83% (n=268/480) chickens were found positive for MG. The MG infection was higher (62.5%) in backyard chickens as compared to those being reared in commercial farming systems (53.61%). With respect to age groups, the prevalence was highest in pullets (60.63%) followed by adults (55.63%) and old chickens (51.25%). Moreover, chickens reared in winter showed higher prevalence of MG (60.42%) as compared to those reared in summer (51.25%). In conclusion, MG infection is prevalent in the chicken population of Bhola district, Bangladesh. Appropriate strategies should be taken for successful prevention and control of this disease in Bangladesh.

DETECTION OF TILMICOSIN RESIDUE IN MUSCLES, LIVER AND KIDNEY OF BROILERS AND ITS EFFECT ON LIVER, KIDNEY FUNCTION AND IMMUNITY

One hundred one day day old broiler chickens were fed balanced rations ad libitum with free access to water. At 3 weeks in this study. 10 birds as a random samples were scarified for postmortem and bacteriological examination to detect Pasteurella and to serum plate agglutination test (SPA) to prove that free from Mycoplasma gallisipticum. Other birds were randomly divided into three groups of 30 chickens each. The first group was a control. The second and third groups administered tilmicosin in drinking water at concentrations of 30 and 60 mg/kg bw for 5 days. Six chickens of each group, three males and three females, were slaughtered on days 3, 7, 14 and 21 days after dosing. Theresults reveled significant increase in lymphocytes transformation test (LTT) in high dose administration of tilimicosin at the end of experiment. High dose of tilmicosin (H1,H2) caused decrease albumin levels and ɣ globulin. The result revealed no significant changes in serum creatinine in all period of experiment. Alanine amino transferrase (ALT) showed significant increase in groups treated with low and high dose in all period of experiment. Aspartate amino transeferrase (AST) showed non significant changes on group treated with low dose but high dose treated group revealed significant increase in 1st and 2nd periods only and return to normal level in the 3rd period. Serum alkaline phosphatase (ALP) revealed significant increase in group treated with high dose in all period of experiment. The residue levels in both doses were significantly higher in liver than kidney or muscle at 3, 7, 14 day post treatment and not detected at 21 day post treatment in all examined samples including liver. The results indicated that tilmicosin residue was eliminated from muscle very quickly, but from liver very slowly. The minimum withdrawal time of 7 days for low dose and 14 days for high dose. The chromatograms obtained with this method was validated with limit of tilmicosin detection 10µg/kg.

Mycoplasma gallisepticum and Escherichia coli mixed infection model in broiler chickens for studying valnemulin pharmacokinetics

A Mycoplasma gallisepticum-Escherichia coli mixed infection model was developed in broiler chickens, which was applied to pharmacokinetics of valnemulin in the present experiment. The velogenic M. gallisepticum standard strain S6 was rejuvenated to establish the animal model, and the wild E. coli strain O78 was injected as supplementary inoculum to induce chronic respiratory disease in chickens. The disease model was evaluated based on its clinical signs, histopathological examination, bacteriological assay, and serum plate agglutination test. The pharmacokinetics of valnemulin in infected chickens was determined by intramuscular (i.m.) injection and oral administration (per os, p.o.) of a single dose of 10 mg/kg body weight (BW). Plasma samples were analyzed by liquid chromatography-tandem mass spectrometry. The plasma concentration-time curve of valnemulin was analyzed using the noncompartmental method. After the i.m. administration, the mean values of Cmax , Tmax , AUClast , MRT, CLβ /F, Vz /F, and t1⁄2β , were 27.94 μg/mL, 1.57 h, 171.63 μg·h/mL, 4.51 h, 0.06 L/h/kg, 0.56 L/kg, and 6.50 h, respectively. By contrast, the corresponding values after p.o. administration were 5.93 μg/mL, 7.14 h, 47.60 μg·h/mL, 9.80 h, 0.22 L/h/kg, 3.35 L/kg, and 10.60 h. The disposition of valnemulin was retarded in infected chickens after both modes of extravascular administration as compared to the healthy controls. More attention should be given to monitoring the therapeutic efficacy and adverse effects of mixed infection because of higher required plasma drug concentration and enlarged AUC with valnemulin treatment.

The Natural Reservoirs of Salmonella Enteritidis in Populations of Wild Birds

The aim of our study was to identify possible natural reservoirs of Salmonella Enteritidis among wild birds. Salmonella Enteritidis is dangerous for human due the reason of toxicoinfaction. These pathogen demonstrate high virulence for small children and people with chronic pathologies and can causes people die. The main source of infection to humans is birds (poultry and wild). The study was conducted in populations of wild birds in National Park 'Askania Nova' and peninsula 'Arabat arrow' (the Azov Sea coast). From bird selected samples of blood serum and egg yolks for research in serum plate agglutination test (SPA) and litter samples for bacteriological research. The serological monitoring in populations of wild waterfowl in National Park 'Askania Nova' (Ichthyaetus relictus, Sterna nilotica, Sterna herundo, Casarca ferruginea) has shown the presence of seropozitive individuals in adult birds (average 18%) and egg yolks (avarrage 12%). The bacteriological investigations confirmed circulation of Salmonella in this group of birds. 32.3% of all bacterial pathogens was Salmonella and more then half of them was the reprezentatives of serovar Salmonella Enteritidis.

Identification of avian Mycoplasma species in commercial broilers and layers with respiratory symptoms in Balochistan

Among many avian mycoplasmas, only the Mycoplasmas gallisepticum (MG) andMycoplasmas synoviae (MS) are responsible for causing respiratory disease in commercial poultry. This study reported for the very first time the serological occurrence of M. gallisepticum and M. synoviae in blood samples (n = 600) from sixty flocks (n = 42 broiler and n = 18 layer flocks) with respiratory symptoms in Quetta, Pishin and Kuchlak districts of Balochistan, Pakistan. Sera were tested for MG and MS antibody responses by serum plate agglutination (SPA) test and by enzyme-linked immunosorbant assay (ELISA) Synbiotics kit. It was found that M. gallisepticumantibodies in broiler flocks detected by SPA and by ELISA tests were 10.47 and 19.76%, whereas M. synoviae were 7.86 and 11.19%, respectively. In layer flocks the MG and MS antibodies detected by SPA and ELISA were 19.44, 31.66 and 8.8, 25%, respectively. The overall antibodies of MG and MS in both broiler and layer flocks tested by SPA and ELISA was found to be 13.16, 23.33 and 8.16, 15.33%, respectively. In broiler and layer flocks the presence of antibodies against both M. gallisepticum and M. synoviae were found in tested flocks with respiratory symptoms. Further studies on prevalence and diagnosis of both the M. gallisepticum and M. synoviae in causing respiratory disease in commercial broilers and layers in Balochistan are required. Key words: Mycoplasma, broilers, serum plate agglutination, enzyme-linked immunosorbent assay.

Development and evaluation of a novel subunit vaccine for Mycoplasma gallisepticum

Adhesion proteins from Mycoplasma gallisepticum (MG) encoded by cytadhesion genes mgc1 and mgc2 were cloned into plasmid vectors and transformed into E. coli. Seventeen groups of specific-pathogen free (SPF), birds at four weeks of age were used to inoculate these two proteins (MGC1 and MGC2) mixed into an oil emulsion creating a novel MG vaccine. Six different protein concentrations (50, 100, 200, 400, 800, and 1000µg/bird) were tested with two equal concentration doses at four and seven weeks of age. In addition, many control groups were needed such as bacterin, membrane, no vaccine or challenge, oil emulsion alone, and no vaccine but challenged. Three weeks following the second vaccination, 50% of the birds in each treatment group were challenged with MG strain S6. The remaining birds were left as contacts to verify protection against horizontal transmission. All birds were bled before vaccinations, challenge and euthanasia. Birds were negative for MG at the first vaccination, as shown by serum plate agglutination test. At necropsy, tissue samples (trachea, lungs, and air sacs) were collected for histopathological examination. Swabs from trachea were used for PCR analysis. ELISA results showed a strong immune response to both protein preparations and almost the same response level for different doses tested, proving the immunogenic features of MGC1 and MGC2. However, humoral responses failed to prevent MG infection and disease when challenged as demonstrated by PCR and histopathology. MGC1 contact birds showed some degree of infection by PCR analysis. In addition, histopathological and ELISA results suggest that contact birds did not have enough time to develop lesions and to mount an immune response.

A CROSS-SECTIONAL STUDY FOR &lt;i&gt; MYCOPLASMA GALLISEPTICUM &lt;/i&gt; ANTIBODIES IN NON-VACCINATED COMMERCIAL LAYER BIRDS IN KHULNA DISTRICT

A cross-sectional study was carried out on 96 commercial layer farms of six upazilas of Khulna district to know the seroprevalence of Mycoplasma gallisepticum (MG) infection during the period from August 2009 to July 2010. A total of 1268 sera samples were collected from randomly selected layer birds of different age groups and tested by serum plate agglutination (SPA) test by using commercial MG antigen (Nobilis® MG, Intervet International) to detect the presence of antibodies against MG. The overall prevalence of MG infection was recorded as 67.4%. Age-wise analysis revealed that there was significant relationship (p&lt;0.001) between MG infection and age of birds. The highest prevalence of MG infection was 71.2% in 21-56 weeks age group followed by 66.8% in 8-20 weeks age group. There was significant (p&lt;0.001) variation in the seroprevalence in terms of areas of study (upazilas). Significant (p&lt;0.001) effect of seasons in the seroprevalence of MG infection in layer birds was observed in the present study. The prevalence of MG infection was the highest (75.6%) in winter season followed by summer (66.3%) and rainy (60.8%) seasons. The seroprevalence of MG infection was higher in most of the large flocks and lower in small flocks with some exceptions. It is evident from the present study that above 50% layer birds are infected with MG organism. Therefore, it may be suggested that the layer farms should be routinely checked to monitor MG infection and the reactor birds should be culled since MG organism has the potential to transmit vertically.DOI = http://dx.doi.org/10.3329/bjvm.v8i2.9620Bangl. J. Vet. Med. (2010). 8 (2) : 93-96

SEROLOGICAL SURVEY OF SALMONELLA INFECTION IN NON-VACCINATED COMMERCIAL LAYER BIRDS IN KHULNA DISTRICT OF BANGLADESH

A cross sectional study was conducted on 164120 non-vaccinated layer birds of 96 farms in six upazilas (Sadar, Batiaghata, Dumuria, Dighulia, Rupsha and Fultala) of Khulna district to determine the seroprevalence of Salmonella infection (S. pullorum and S. gallinarum) during the period from August 2009 to July 2010. Sera samples were collected from 1268 layer birds of different ages and the birds were selected through a disproportionate stratified random sampling technique based on the flock size of each farm. Sera samples were tested by Serum plate agglutination (SPA) test applying commercial Salmonella antigen (Nobilis® SA, Intervet International B.V. Boxmeer- Holland) to detect the presence of antibodies against Salmonella. The overall seroprevalence of Salmonella infection was recorded as 65.9%. The significantly higher seroprevalence (76.6%) of Salmonella infection was recorded in layer birds of &gt;56 weeks of age than those of other age groups. Seasons had significant influence on the seroprevalence of Salmonella infection. The seroprevalence was significantly higher in summer (82%) than that in rainy (66.8%) and winter (50%) seasons. The location of farms, i.e. upazilas also had significant association with the occurrence of Salmonella infection. The seroprevalence significantly differed between the different categories of flock size. The flock size of 5001 and above had the highest seroprevalence (81.4%) among other categories. It may be concluded that above 60% layer birds in 92 out of 96 farms are infected with Salmonella organism, which requires keeping of vigilant eye of the poultry farmers and the hatchery owners in the control of Salmonella infection in poultry farms.DOI = http://dx.doi.org/10.3329/bjvm.v9i1.11206Bangl. J. Vet. Med. (2011). 9(1): 27-31

Detection of Mycoplasma gallisepticum in experimentally infected broiler chickens using Culture, SPA, ELISA, and PCR methods

Avian mycoplasmosis is caused by several pathogenic mycoplasmas of which Mycoplasma gallisepticum and Mycoplasma synoviae are the most important. This experiment was designed to study the diagnosis of M. gallisepticum infection in broiler chicken receiving M. gallisepticum. One hundred and twenty one-day-old chickens were raised in separate isolated rooms and equally allocated into two groups and renamed groups A and B were 121-day-old chickens. At 3 weeks old, group A was considered as the control group while group B was challenged with M. gallisepticum containing 1 × 106 CFU/bird via an eye drop. In this study, culture, serum plate agglutination (SPA) test, the commercially available enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR) tests were compared for the detection of M. gallisepticum infection during the 1–7 weeks of age. Results revealed that M. gallisepticum was found in group B using culture and PCR during the 4–7-week-old period. The SPA results showed positive reactors in group B during the 5–7-week-old period. The positive reactors were detected by commercial ELISA test kit from 6 to 7 weeks of age. In this study, sensitivity of the PCR method was higher as compared to culture and serological tests. Its high sensitivity and fast turnaround time is making it the most frequently used test to monitor M. gallisepticum infection beside other tests.