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Abstract
The main purpose of this study was to detect B. melitensis in amniotic fluids of the pregnant ewes by ELISA and real time PCR, the investigated animals were divided into two groups, group A (n=36) have not abortion history and group B (n= 32) have a previous abortion history. The results of serum samples showed a positive reaction to ELISA for B. melitensis in both groups (A&B) were 19.45%, 37.5%, respectively, while in fetal fluid samples of qPCR result was 30.56% in group A and 56.25% in group B. In conclusion, the real time PCR technique can be successfully used for early diagnosis of B. melitensis by amniocentesis in both known previously aborted ewes as well as the unknown once.
Keywords: amniocentesis, Brucella melitensis, ewe, real time PCR
INTRODUCTION
Brucellosis is a contagious disease affecting many organs such as placenta, fetal fluids and testis (Narnaware et al. 2017; Saxena et al. 2018), causing reproductive losses in mammal's due to high rates of morbidity and lifetime sterility and consequently causes economic losses (Khan and Zahoor, 2018). Brucella melitensis (B. melitensis) is the main cause of brucellosis in the sheep (Noakes et al. 2018). It is a gram negative coccobacillus, an intracellular organism (Corbel, 2006). Brucella genus members are closely correlated, and some microbiologists have suggested reclassifying this genus into a single species B. melitensis (Bi, 2008; Akpan, & Udo, 2017). It is endemic in Central America, Africa and many Mediterranean countries, causing abortion in late pregnancy, stillbirth or weak lambs (Noakes et al. 2018). Amniocentesis is a diagnostic test follow out during pregnancy (Hussein, 2019). It is the first invasive tool carried out in fetal medicine for prenatal diagnosis and therapy (Alwan and Friedman, 2019). It is a necessary appliance in obstetrics (Connolly and Eddleman, 2016).
An amniocentesis technique for sample collection was used in the 2nd third of gestation with minimal risk of the fetus and uterus using a sheep model and tested to make it a routine procedure in veterinary medicine (Mota et al. 2002; Prestes et al. 2001). It was commonly used in human medicine to identify fetal viability, early determination of fetal sex, chromosomal anomalies, biochemical defects (Bravo et al. 1995; Sadłecki et al. 2014), and to confirm the suspected...