Development of Immunochromatographic Assay for the Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae Antibodies

Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP), which is a highly significant respiratory disease in goats leading to significant economic losses in Africa and Asia. Currently available procedures for the diagnosis of CCPP have some limitations in sensitivity, specificity, operation time, requirement of sophisticated equipment or skilled personnel, and cost. In this study, we developed a rapid, sensitive, and specific colloidal gold-based immunochromatographic assay (GICA) strip for the efficient on-site detection of antibodies against Mccp in the serum within 10 min. For the preparation of this colloidal GICA strip, recombinant P20 protein, the membrane protein of Mccp, was expressed by Escherichia coli prokaryotic expression system after purification was used as the binding antigen in the test. The rabbit anti-goat immunoglobulin G labeled with the colloidal gold was used as the detection probe, whereas the goat anti-rabbit immunoglobulin G was coated on the nitrocellulose membrane as the control line. The concentration of the coating antibody was optimized, and the effectiveness of this colloidal GICA strip was evaluated. Our results proved that the detection limit of the test strip was up to 1:64 dilutions for the Mccp antibody-positive serum samples with no cross-reactivity with other pathogens commonly infecting small ruminants,including goat pox virus, peste des petits ruminants virus, foot-and-mouth disease virus type A, or other mycoplasmas. Moreover, the colloidal GICA strip was more sensitive and specific than the indirect hemagglutination assay for the detection of Mccp antibodies. The 106 clinical serum samples were detected by the colloidal GICA strip compared with the complement fixation test, demonstrating an 87.74% concordance with the complement fixation test. This novel colloidal GICA strip would be an effective tool for the cost-effective and rapid diagnosis of CCPP in the field.

PREVALENCE OF TOXOPLASMOSIS AMONG SHEEP AND GOATS IN BAGHDAD AREA

The complement fixation test (CFT) and the direct fluorescent antibody test were used for detecting anti Toxoplasma gondii antibodies in sera obtained from 143 sheep and 44 goats. Complement fixing antibodies were detected in 38 (26.2%) for sheep sera and 24 (54.5%) of goat sera tested by CFT. On the other hand, 26(18.2%) of sheep sera were positive by the FAT. The combined use of CFT and FAT allows the differentiation between an acute or latent T.gondii infection.

Detection of Antibodies Brucella abortus in dairy cattle in Puspo District, Pasuruan using Rose Bengal Test and Complement Fixation Test

Brucellosis was an infectious disease caused by the genus brucella. Brucellosis in dairy cattle was caused by Brucella abortus that impaction an abortion. The aim of this study was to determine the presence of antibodies of the Brucella abortus in adult female dairy cattle in the Puspo district Pasuruan using Rose Bengal Test (RBT) and Complement Fixation Test (CFT). A descriptive analysis method with quantitative approach was used in this study. Meanwhile, interviews were conducted to get supporting information from the farmer. Used 100 samples. The results of the study showed 2% samples considered positive brucellosis tested by RBT. To confidence false-positive result, RBT test was followed by CFT. After tested by CFT, in this study there were no brucella antibodies in adult female dairy cattle in Puspo district, Pasuruan.

Seroprevalensi Brucellosis pada Sapi Potong di Papua Barat

Brucellosis merupakan penyakit bakterial yang disebabkan oleh Brucella abortus, yang menga-kibatkan kerugian ekonomi akibat terjadinya keguguran pada sapi betina yang bunting pada triwulan ketiga kebuntingan. Surveilans brucellosis dalam mendukung pengembangan peternakan sapi di Propinsi Papua Barat telah dilakukan oleh Balai Besar Veteriner Maros. Tujuan pelaksanaan surveilans ini adalah untuk mengetahui seroprevalensi brucellosis pada sapi potong di Papua Barat. Surveilans dilakukan di 4 kabupaten/kota, 14 kecamatan dan 33 desa. Sebanyak 684 sampel serum sapi telah diambil guna pengujian terhadap brucellosis. Sampling dilakukan pada lokasi kabupaten, kecamatan dan desa terpilih yang padat ternak. Metode pengujian dilakukan secara seri, dengan melakukan screening test menggunakan Rose Bengal Test, yang bila hasilnya positif diuji lebih lanjut dengan Complement Fixation Test. Hasil pengujian menunjukkan adanya reaktor brucellosis pada 5 ekor dari 684 ekor sapi yang diambil dan diuji serumnya. Seroprevalensi brucellosis pada sapi sebesar 0,73% di desa Remu Utara, kecamatan Sorong, kota Sorong sebanyak 1 ekor serta di desa KaliMerah, kecamatan Masni, kabupaten Manokwari. Untuk mencegah penularan dan penyebaran brucellosis pada sapi maka sapi reaktor brucellosis segera dipotong dengan pengawasan dari petugas.

Kondisi Brucellosis setelah Vaksinasi di Kecamatan Majauleng Kabupaten Wajo Sulawesi Selatan

Vaksinasi brucellosis telah dilakukan secara massal di Kabupaten Majauleng, Kabupaten Wajo, Sulawesi Selatan tahun 2013 dan 2014 menggunakan vaksin brucellosis RB51. Vaksinasi dimaksudkan untuk mengendalikan dan menekan angka prevalensi yang tinggi di Kabupaten Wajo yang mencapai angka 30%. Surveilans telah dilakukan oleh Balai Besar Veteriner Maros untuk melihat dan mengevaluasi prevalensi brucellosis yang terjadi beberapa tahun kemudian untuk melihat penurunan prevalensi brucellosis pasca vaksinasi. Pada tahun 2016 telah dilakukan pengujian sampel serum yang berasal dari Kabupaten Wajo. Sebanyak 118 serum telah diuji dengan Rose Bengal Test (RBT) yang diteruskan dengan Complement Fixation Test (CFT). Hasilnya menunjukkan bahwa brucellosis masih ditemukan di Kabupaten Wajo dengan angka prevalensi sebesar 25,42%. Prevalensi terbesar ditemukan di Kecamatan Takalalla yang mencapai 73,68%, sedangkan di Kecamatan Majauleng sebesar 3.13%. Sampling lanjutan dilakukan pada tahun 2020 di kecamatan terpadat populasi sapinya yaitu Kecamatan Majauleng. Prevalensi brucellosis pada tahun 2020 di dua desa di Kecamatan Majauleng meningkat menjadi sebesar 6 %.

Complement Fixation Test for Specific Antibody Detection against Bovine Brucellosis in Selected Peasant Association of Guto Gida District, East Wollega Zone, Oromia, Ethiopia

Brucellosis is one of the major zoonotic diseases yet prevalent in Ethiopia. It is contagious and has harmful effects on free animal movement and export. A cross-sectional study was conducted from November 2016 to April 2017 in the Loko and Uke peasant association of Guto Gida District. The goal of this study is to determine the seroprevalence and associated risk factors of bovine brucellosis. The questionnaire survey was conducted on 200 respondents to collect the livestock owner’s perception of this disease. Eighty (80) blood samples were collected from grazing cattle above six months of age. Serum was tested by complement fixation test (CFT) for Brucella antibody detection. Pearson chi-square is used to assess the relation of independent variables such as sex, site, and age with seroprevalence in a 95% confidence interval if P value is less than 0.05, recoded as significant. Seroprevalence of bovine brucellosis in the present study is 22.5%. Bovine brucellosis prevalence recorded in Uke (28.6%) was higher than that in Loko (21.2%). Again bovine brucellosis is higher in males (30.4%) than females (19.2%). There is prevalence variation among site, sex, and age which is statistically insignificant ( P value>0.05). Survey findings revealed that 52% (104/200) of respondents did not know the causes of abortion, while 48% (96/200) of them confirmed abortion in their herd and 64% (124/200) of respondents removed retained fetal membrane by themselves. Brucellosis is a contagious reproductive disease of cattle with zoonotic implications and needs to design and implement control measures aiming at preventing further spread of the disease.

Detection of Brucellosis in Dairy Cattle in Turen District Malang Regency using Rose Bengal Test (RBT) and Complement Fixation Test (CFT) Methods

The purpose of this research was to detect Brucellosis in female dairy cattle in Turen District, Malang Regency. A total of 78 samples were used in this research taken from 351 total population of female dairy cattle that have been calculated based on the slovin formula. The methods used in this research were Rose Bengal Test (RBT) for the screening test and continued with Complement Fixation Test (CFT) for the confirmation test. The results RBT of this research clearly showed that positive results from 10 samples were confirmed as a final diagnostic test of Brucellosis using CFT. CFT showed a positive result from 9 samples and negative in 1 sample. The percentage of Brucellosis occurrence in the Turen District was recorded at 11.5%. Moreover, Turen District also became an area with a high Brucellosis occurrence (>2%).

Problems and prospects of bovine tuberculosis serological diagnosis

For the purposes of tuberculosis eradication on any tuberculosis-infected farm, it is necessary to identify tuberculin anergic animals, being a potential source of the infection. The purpose of this study was to analyze the role of complement fixing and haemagglutinating antibodies for the detection cattle infected with bovine tuberculosis (TB). 977 cattle of different sex and age groups on two tuberculosis-infected farms were tested thrice over time. After 35 days all tuberculin reactive cattle (132 animals; 13.5%) were subjected to complex testing using allergy and serology methods. After 40 days, (Stage 3) animals demonstrating apparent specific antibody activity and low cell immunity were tested. Allergy tests were proved to be non-informative to diagnose tuberculosis on infected farms. Complement fixing and haemagglutinating antibodies were found to be active in tuberculin anergic animals. A higher antigenicity of Ukrainian RIEVM TB antigen complex as compared to Siberian RVI one was revealed by complement fixation test as well as by indirect haemagglutination test using VIEV polysaccharide antigen; the detection rate was 68 (7.0%), 28 (2.9%) and 299 (30.6%) respectively. The correlation between seropositivity and immunoreactivity was not established. Animals, positive in complement fixation and indirect haemagglutination tests, did not react to tuberculin. Nineteen out of twenty tuberculin reactive animals showed post mortem lesions, consistent with their seropositivity during post-mortem inspection; moreover, the postmortem lesions of animals, positive in complement fixation test using Siberian RVI antigen, were consistent in all cases. The results obtained suggest a high performance of allergy test and serological test combination and a promising potential of their complex use for tuberculosis diagnosis in cattle.

Development of new antigens for serologic diagnosis of animal trypanosomosis and their use for epizootic monitoring

The purpose of the research is developing a method for obtaining erythrocyte antigens containing and not containing Trypanosoma equiperdum and T. evansi DNA, which can later be used in serological reactions to differentiate these types of Trypanosoma.Materials and methods. The studies were conducted in the Protozoology Laboratory and the Vyshnevolotsk Branch of the Federal State Budget Scientific Institution “Federal Scientific Centre VIEV RAS”, as well as livestock farms of the Russian Federation and other countries using clinical, microscopic, hematological, parasitological, biomolecular and serological methods.Results and discussion. Studies carried out for the first time have shown that it is possible to use erythrocyte antigens containing the T. equiperdum and T. evansi DNA obtained after 3-fold administration to mice and rabbits of a mixture of trypanosomal antigen with addition of 1.0 ml of an adjuvant (aluminum hydroxide), and bleeding of animals at 25 to 30 days. The formed precipitate was used as an antigen for serological tests. Experiments have shown that blood for preparation of positive serum can be taken when antibodies are in titers of 1:20 in the Prolonged Complement Fixation Test, and at least 1:400 in the Indirect Hemagglutination Test and ELISA, and for negative serum when horse blood serum reacts negatively with antigens of T. equiperdum and T. evansi in the Prolonged Complement Fixation Test, Indirect Hemagglutination Test and ELISA. The test systems of the Prolonged Complement Fixation Test, Indirect Hemagglutination Test and ELISA prepared by us with antigens containing and not containing T. equiperdum and T. evansi DNA resulted in creating a universal test system (Indirect Hemagglutination Test) for differentiating T. equiperdum from T. evansi.

THE STABILITY OF WET T1 CONTAGIOUS BOVINE PLEUROPNEUMONIA VACCINE

THE stability of wet T1 contagious bovine plenropneumonia (CBPP) Vaccine stored at 4°C was studied. Two large batches of the vaccines were stored at 4°C for up to 5 months. From the third month of storage, the vaccines were used to vac­cinate CBPP-free animals. The immune levels of the animals were monitored by complement fixation test (CET) and challenge trial. The results indicated that all the vaccinated animals produced reasonable an­tibody titres even after 5 months storage. Also, all the animals vaccinated with vaccines stored between 3 to 5 months resisted infection by contact while ail the incubated and control animals came down with the disease. The results indicated that the vaccine could actually be stored for up to 5 months at 4°C before use.