2009; Forghani 2010; Intisar et al. difficult to distinguish from those caused by other respiratory viruses. is an RNA virus of the family that causes mild respiratory disease DL-Carnitine hydrochloride of ruminants when it is the only pathogen (Underwood et al. 2015). In combination with other pathogenic agents, BPI3V is one of the most significant respiratory pathogens in ruminants (Stevenson and Hore 1970; Lehmkuhl et al. 1985; Bechmann 1997; Alkan et al. 2000). This virus can cause pneumonia on its own but it is more generally a part of the etiological complex of enzootic pneumonia (Dungworth 1993). The confirmative diagnosis of PI3V infection is made by virus isolation in cell culture, polymerase chain reaction (PCR), and immunohistochemical (IHC) examinations of the lower respiratory tract (Alkan et al. 2000; Grubor et al. 2004; Gafer et al. 2009; Forghani 2010; Intisar et al. 2010). The only histological examination of sections stained with hematoxylin and eosin (H&E) has been reported to be insufficient for the diagnosis, because pathological changes observed in lungs infected with are similar to each other (Caswell and Williams 2007). There are no detailed studies that employed the direct immunofluorescence antibody technique to investigate the role of BPI3V in sheep pneumonia cases in Algeria. The objective of the present study was to evaluate the prevalence and characterize the gross and histopathological pneumonic lesions and to determine the presence of BPI3V using direct fluorescent antibody technique (DFAT) DL-Carnitine hydrochloride on frozen pneumonic lung sections of sheep from Batna municipal slaughterhouse (Eastern Algeria). Materials and methods Study area This study was carried out at Batna municipal slaughterhouse (Eastern Algeria). Batna region is semi-arid which is characterized by cold and humid winters and hot and dry summers. This agro-pastoral region has a potential of 56.052 cattle with 35.445 dairy cows and 1.137.361 sheep with 638.423 ewes (DSA 2019). Samples collection The samples were represented by lungs of sheep that were slaughtered at Batna municipal slaughterhouse during the period from December 2018 to December 2019. A total of 13084 lungs were examined for research of lesions. The studied animals were young, aged between 6 to 18 months, originating from farms of Batna region and its surrounding areas. The samples taken from the affected lungs were divided into two parts: one part was fixed in 10% buffered formalin and the other part was stored at C 20 C until analysis. Microscopic examination Tissue samples taken from pneumonic lesions fixed in 10% buffered formalin were dehydrated in graded ethanol and incorporated in paraffin before sectioning. Sections of 4 m in thickness were stained Rabbit Polyclonal to USP30 with H&E and examined using a light microscope according to the protocol of Luna (Luna 1968). Direct fluorescent antibody technique examination Direct fluorescent antibody technique was employed for the detection of BPI3V using monoclonal mouse anti-PI3 FITC conjugate (Cat No: BIO 030, BioxJemelle, Belgium). The immunofluorescence staining was performed according to the manufacturers protocol. Lung tissues stored at ? 20 C were cut to obtain sections of 4 m thickness that were placed on silanized slides (SuperFrost? Plus Gold) and air-dried. At that time, DL-Carnitine hydrochloride the sections were fixed in ethanol for 15 min at room temperature. After washing in phosphate-buffered saline (PBS), the sections were treated with the conjugate diluted in a 1:20 ratio with PBS-Blue Evans at room temperature for 1 h in a dark place. DL-Carnitine hydrochloride After the second washing in PBS, the slides were mounted with glycerol. Finally, the sections obtained were examined under a fluorescent microscope. The DFAT was only performed in tissue with interstitial pneumonia, fibrinous bronchopneumonia, and acute bronchopneumonia, because BPI3V is one of the respiratory germs incriminated in the appearance and development of these pneumonias. Statistical analysis Data.