Dog visceral leishmaniasis is a significant public health problem that is endemic in tropical and sub tropical countries and is fatal in humans and dogs. dogs (397 males and 111 females mean age 3.24 from western and eastern parts of the Meshkin-Shahr were examined. A total of 508 dogs examined 119 dogs (23.4?%) had antibodies (titers of?≥1:320) against contamination (Moshfe et al. 2009; Cortes et al. 2012). Thus surveillance and control programs of reservoir hosts are essential (Silva et al. 2012; Tesh 1995). Serological methods are used widely for evaluation sero-prevalence of CVL (Kalayou et al. 2011). Direct agglutination test (DAT) is the first line Cerubidine (Daunorubicin HCl, Rubidomycin HCl) sero-diagnostic method in most of developing countries (Teran-Angel et al. 2007). This method has the advantages including simply high sensitivity specificity and reproducibility also easy to perform and it doesn’t need complicated equipments (Teran-Angel et al. 2007; Sousa et al. 2011). So Cerubidine (Daunorubicin HCl, Rubidomycin HCl) it is the most suitable method for using in field assays (Sundar Cerubidine (Daunorubicin HCl, Rubidomycin HCl) et al. 1998). The purpose of this study was to evaluate the sero-prevalence of visceral leishmaniasis in doggie population from in an endemic area of north west Iran. Materials and methods Study area The investigation was conducted in north west of Iran where HVL is usually endemic. Meshkin-Shahr district in the central northern part of the Ardabil province located at an altitude of 1490?m above sea level between longitudes 47°19′ and 48°17′ east and latitudes 38°57′ and 38°13′ north. Forty-two percent of their populations settle in urban areas Cerubidine (Daunorubicin HCl, Rubidomycin HCl) 58 live in rural areas and small population have nomadic living. The study area has moderate mountainous weather. In this area there are a lot of dogs with a friendly relationship with the human Rabbit Polyclonal to Bak. population which used as guard and herd dogs and also the stray doggie is found. In this region the amount of animal manure can be seen in yards and alleys which is considered as the resting place of dogs. Blood sampling A descriptive cross sectional study was carried out during 2011-2012. The Cerubidine (Daunorubicin HCl, Rubidomycin HCl) serum samples were collected from all domestic dogs in villages where human visceral leishmaniasis had been reported at least 5 cases during 3?years ago. After documented general information including age and gender from each doggie by interviewing dog owners whole blood sample collected from 508 ownership dogs into 10?ml polypropylene tubes. All the samples centrifuged at 800×for 5-10?min and then serum aliquots were stored at ?20?°C until examined. All of the serum samples were transferred leishmaniasis laboratory in the School of Public Health Tehran University of Medical Sciences and were tested by DAT. DAT test The DAT antigen used in this study was prepared in the protozoology unit of the School of Public Health Tehran University of Medical Sciences and stored at 4?°C until used. The principal phases of the procedure for preparing the DAT antigen were mass production of promastigotes of (Iranian strain) in RPMI1640 medium supplemented with 10?% fetal bovine serum trypsinization of the parasites staining with Coomassie Brilliant blue and fixing with 2?% formaldehyde (Mohebali et al. 2005; el Harith et al. 1989; Harith et al. 1986). The dog serum samples were tested by Cerubidine (Daunorubicin HCl, Rubidomycin HCl) DAT according to the methods described by (el Harith et al. 1989; Mohebali et al. 2011; Mohebali et al. 2006). For initial screening purposes two fold dilutions were prepared from 1:80 and 1:320 in doggie samples. Sera with titers 1:80 were diluted further to give maximum serum dilution of 1 1:20480. Unfavorable control antigen (antigen only) and known negative and positive controls were tested in each plate. In the present study we considered antibody titers?≥1:320 as cut off point for canine according to the previous studies (Bokaie et al. 1998; Mohebali et al. 2005; Edrissian et al. 1996). Data analysis Sero-prevalence values of anti-antibodies relative to gender age groups were statistically compared using the Chi squared (χ2) or Fisher’s exact tests. Analyses were carried out using SPSS software version 18 with a probability (antibody have been shown in 256 dogs (50.4?%). It has ranged from 1:80 to 1 1:20480 titers (Table?1). Table?1 Distribution of titers of anti antibodies in asymptomatic dogs in Meshkin-Shahr district northwest of Iran (2011-2012) Statistically significant was occurred between male and female (antibodies by glycerol-preserved (GP)-DAT antigen (Akhoundi et.