Diagnosis of diseases depends on histology using formaldehyde-fixed paraffin embedded tissue .The polymerase chain reaction (PCR)helps the pathologists to confirm the diagnosis of many disease including mycotic infections.Aborted placentas from 40 ewes were formalin fixed, paraffin embedded and sectioned. Tissue sections were stained with haematoxylin and eosine, and periodic acid Schiff (PAS) for histopathology examination. Fresh tissue parts were cultivated ,subjected to DNA extraction for the purpose of real-time PCR amplification.Histopathological examination of placental tissue revealed extensive necrotizing placentitis with the presence of spherule of CoccidioidesPosadasiiin the necrotic area. Only eight samples gave positive growth for coccidioidesPosadasii out of forty  aborted placental samples.DNA amplification by real-time PCR revealed positive amplification for the eight samples which confirmed the detection of coccidiodesposadasii.

Brucellosis is one of the common bacterial zoonosis in the worldwide caused by organisms  belong to the  genus  Brucella, However, animal  Brucellosis is  a serious problem worldwide and is endemic  globally and  disease of  sexually matured animals and commonly transmitted to other animals by direct or indirect contact with infected animals or discharges such as: aborted fetuses, placental membranes or fluids. Fifty samples from internal organ of aborted fetus and the same number of aborted placenta were collected from aborted ewes positive to Rose Bengal test. For isolation and identification of Brucella from placenta and stomach contents , the standard procedures (Alton et al., 1988) were followed. The isolates were further confirmed by molecular techniques. The results showed that Br.melitensis was isolated from all animals that expressed Rose Bengal and serum agglutination test positive .The agglutination with monospecific A and M antisera were performed and these test revealed that the bacterial isolates expressed three Biovar as following ,Biovar 1(M,50%),Biovar 2 (A,26%) and Biovar 3 (0,24%). The number of this Biovar were varied according to collect area of the samples, high percentage (44%) was seen in the ALmanitherh ,,followed by AL Shabeka (30%) and AL Kuzweenah (26%)     The results of PCR assay explained that 22 bacterial isolates showed a single amplified DNA product (44%) ,two bands at the level of 273 and 680 bp were seen in 8 isolates (16% ) and four bands at 273,680,750 and 850 bp were reported in 20 isolates(40%).

The aims of the present study is  to isolate the fungal species associated with ewe abortion and to examine the histopathological  changes in the placenta associated with fungal isolates .In addition to study the genetic information  of TNF alpha gene, intensity and score of TNF-alpha  protein in  aborted and normal placenta by PCR and IHC respectivly. Frozen placental tissue and paraffin embedded tissue   belong to aborted and normal groups were used for assessment of TNF alpha gene by studying the   genetic information of TNF-alpha   gene   in aborted and normal   placental sample by PCR .immmunohistochemical study was carried out to determine TNF protein in  the placental sample. The same frozen placenta tissue samples  were also used for DNA extraction to detect TNF alpha gene in the two groups.    A fragment of (238 bp ) of the TNF alpha gene was amplified by PCR using primers ,the sequence of sense and antisense primer for TNF  alpha  gene was: GAA TAC CTG GAC TAT GCC GA,CC TCA CTT CCC TAC ATC CCT (bp 238) Gene Bank:   X56756.    The study showed that 10 fungal species were isolated from aborted  placenta with high percentage of these isolates was recorded in March ,.The fungal isolates were variable according to region of sample collecting ,high percentage in the  AL-Manathera(36%),followed by Al-Kuzweenia (34%) and Al-Shabaka(30%).The fungal isolates include Coccidioides immitis (16%). Asperigllus.fumigitus (14%), Trichophyton, Rhodetella rubra,  and Aspergillus.ochraus ,(10%) for each one . Candida  kruzei,(8%). Aspergillus.flavus and Geotrichum ,6%) for each one . Prototheca..zopfi, Saccharomyces.cerevisia, Blast.capitatuss, Cryptococcus uniguttultus and Candida zeylanoides,(4%) for each one .           This study revealed the presence of ~238bp band in addition to the ~300 ,400 and 600  bp desired fragment in PCR products that  related to aborted samples compared to normal one .,this may be  due to polymorphism within the TNF gene itself.       We  found a significant elevated TNF alpha  protein   in inflammatory cell (macrophage) of aborted sheep placenta (increasing of intensity and score), while no changes could be observed for pro-inflammatory  molecules in the control sample (normal delivery)      The current study expressed   correlation between polymorphisms in TNF- alpha gene ,immunohistochimistry and the fungal isolated species.        The histopathological examination showed acute suppurative  placentitis, necrosis of chorion villi, congestion of  blood vessels with fibrin deposition in the  villus stroma and inter villus space, in addition to calcification in the villus stroma and  sloughing and desquamation of cytotrophoblast .numerous  yeast  and spheriols were detected in the placental tissue  by periodic acid Schiff reaction(PAS).