Seroepidemiologcal investigation of visceral leishmaniasis in stray and owned dogs in alborz province, central Iran using direct agglutination test

The aim of present study was to determine the seroprevalence of canine visceral leishmaniasis [CVL] among stray and owned dogs in Kouhsar district of Alborz Province, central Iran. The study was performed from March 2011 to July 2011 using Direct Agglutination Test [DAT]. Three hundred and thirty seven dogs including 257 stary and 80 owned dogs were selected by random sampling. The agreement between serological data and sex, age, life style of dogs and clinical signs were assessed by Chi-square. DAT showed that from 337 serum samples collected from owned and stray dogs, 12sera [3.6%] were positive. The seroprevalance was 10% [8/80] among owned dogs and 1.6% [4/257] among stray dogs. A significant difference in seroplevalance was seen between owned and stray dogs [P = 0.01]. The highest seroprevalence rate [14%] was observed among the ownership dogs of 5 years old and above. Statistical analysis revealed significant relation between seroprelvalence and age [P= 0.02]. There was no statistically significant relation between male [6.3%] and female [2.2%] seroprevalence [P= 0.085]. This survey indicates the importance and necessity of serologic screening of visceral leishmaniasis in human and dogs in Kouhsar district

Seroprevalence of canine visceral leishmaniasis in asymptomatic dogs in Iran

Clinically infected dogs have been identified as the main reservoir hosts of visceral leishmaniasis [VL] caused by Leishmania infantum. Recently asymptomatic infected dogs were regarded to be as important as clinically ill dogs. This study was aimed to determine the seroprevalence of L. infantum infection among asymptomatic dogs in the southwest and central parts of Iran and to investigate possible risk factors associated with this infection. 548 serum samples were collected from dogs in three Iranian provinces and subjected to direct agglutination test [DAT] in dilutions of 1:80 to 1:20480. Fifty three [9.67%] of the dogs had detectable anti-L. infantum antibodies at dilutions of >/= 1:80. Living status of the dogs [household or free roaming] was a potential risk factor for the infection; seroprevalence was significantly higher in free roaming dogs [P<0.001]. Dogs of more than 2-year-old had a significantly higher infection rate in comparison with younger dogs [P<0.001]. No significant statistical differences were seen between seroprevalences of the male and female dogs. The results of this study show relatively high sero-prevalence of L. infantum infection in evaluated regions and higher seroprevalence in old and free roaming dogs, which shows the importance of environmental contamination and access of the dogs to the other reservoir hosts

Epidemiological aspects of visceral leishmaniasis in Baft district, Kerman province, southeast of Iran

Visceral leishmaniasis [kala-azar] is an endemic disease in some areas of Iran. A cross-sectional study was conducted for sero-epidemiological survey of visceral leishmaniasis [VL] in Baft district from Kerman Province, southeast of Iran. Blood samples were collected from children up to 12 years old and 10% of adult population from Baft villages with a multi-stage randomized cluster sampling. In addition, blood samples were collected from 30 domestic dogs from the same areas. All the collected blood samples were tested by direct agglutination test [DAT] for the detection of anti-Leishmania antibodies in both human and dog using the cut-off value of >/= 1:3200 and >/= 1:320, respectively. Parasitological, molecular, and pathological were performed on infected dogs. Chi-square and Fisher exact tests were used to compare sero-prevalence values. From 1476 collected human serum samples, 23 [1.55%] showed anti-Leishmania antibodies at titers of 1:800 and 1:1600 whereas 14 [0.95%] showed anti-Leishmania infantum antibodies at titers of >/= 1:3200. No statistically significant difference was found between male [1.18%] and female [0.69%] sero-prevalence [P=0.330]. Children of 5-8 years showed the highest sero-prevalence rate [3.22%]. Seven out of 30 domestic dogs [23%] showed anti-Leishmania antibodies at titers >/= 1:320. Leishmania infantum was identified in five infected dogs by nested -PCR assay. It seems that visceral leishmaniasis is being endemic in southern villages of Baft district, southeast of Iran

[Establishment and integration of a visceral leishmaniasis surveillance system in the primary health care system and its evaluation, Meshkin-Shahr District, Iran]

Visceral leishmaniasis is a systemic parasitic disease with a high fatality rate in under-5-year-old children. The disease is endemic in some parts of Iran, particularly in the north-west region. In 2001 a visceral leishmaniasis [VL] surveillance system was established for children aged>/=12 years in the primary health system in Meshkin-Shahr District, Ardebil Province, situated in the north-west of Islamic Republic of Iran. All cases with clinical signs and symptoms of VL and confirmed positive by the direct agglutination test [DAT] were referred for physical examination and treatment. The mean annual incidence of VL decreased significantly from 1.88 per 1000 children before [1985-2000], to 0.77 per 1000 child population after [2001-07], the intervention. In the control area with no surveillance, it increased from 0.11 to 0.23 per 1000. Early detection of VL using serological tests and timely treatment of cases can decrease the mortality and morbidity rates of VL in endemic areas

Disseminated leishmaniasis caused by Leishmania tropica in a puppy from Karaj, central Iran

A 5-month old puppy with muco-cutaneous lesions in the chin, around lips and eyes was examined physically and microscopically for leishmaniasis. Muco-cutaneous lesions containing a large number of amastigotes of Leishmania spp. were observed. Amastigotes were also detected in liver and spleen of the puppy. The animal was positive with Dipstick rK39 kit and high level of anti-Leishmania antibodies was detected by direct agglutination test [DAT]. DNA, Using PCR-RFLP technique extracted from cultured Leishmania promastigotes and L. tropica was identified. This is the first report of concurrent mucosal and visceral involvement of L. tropica in a puppy from Iran

Molecular and seroepidemiological survey of visceral leishmaniasis among humans and domestic dogs in Mazandaran province, north of Iran

New cases of visceral leishmaniasis [VL] have been reported recently in some parts of Mazandaran Province, north of Iran where the first human case of VL was reported in 1949. This study aimed to determine the present status of Leishmania infantum infection among humans and domestic dogs using serological and molecular methods in central parts of Mazandaran Province. In this cross-sectional study, blood samples were randomly collected from 402 humans and fortynine domestic dogs throughout 2009 and 2010 in the central part of Mazandaran Province including Semeskadeh and Kiakola districts where recent cases of human visceral leishmaniasis had been reported there. All the collected samples were tested by direct agglutination test [DAT] for the detection of anti- Leishmania infantum antibodies as well as convenience PCR assay on whole blood samples for detection of leishmanial infection and identification of Leishmania species. None of 402 collected human [402] and dog [49] blood samples showed anti Leishmania infantum antibodies at titers 1:3200 and 1:320 as cut-off values of DAT, respectively but only 2 of domestic dogs [4.1%] were found PCR-positive corresponding to L .infantum. This study confirms the circulation of L. infantum at least among domestic dogs and highlights the sporadic pattern of VL in the studied areas. Further investigations regarding to sand flies fauna and wild canines as reservoir hosts of the disease, are recommended

Neospora caninum and leishmania infantum co-Infection in domestic dogs [canis familiaris] in Meshkin-Shahr district, northwestern Iran

Mediterranean visceral leishmaniasis [MVL] is an infectious disease that affects both human and animals. Domestic dogs [Canis familiaris] are principal reservoir hosts of MVL caused by Leishmania infantum. Dogs are definitive hosts for Neospora caninum and a risk factor for infecting intermediate hosts. The immunosuppression caused by visceral leishmaniasis [VL] can promote the occurrence of co-infections with other agents such as neosporosis. This study aimed to determine the frequency of co-infection of the both protozoan parasites in the endemic areas of VL from Meshkin-Shahr District, north-west of Iran. Altogether, 171 serum samples were collected from domestic dogs of Meshkin- Shahr District by multistage cluster sampling from October 2008 to August 2009. The collected serum samples were tested for the detection of simultaneous infection of L. infantum and N. caninum using direct agglutination test [DAT] and indirect ELISA, respectively. Of the 171 domestic dogs, 27 [15.8%] and 52 [30.4%] were showed antibodies against L. infantum and N. caninum, respectively. Simultaneous infections of N. caninum and L. infantum was found in 16 [9.4%] of the dogs. In VL-positive and VL-negative dogs, N. caninum infection was found in 59.3% and 25.0%, respectively. A statistically significant difference was found between VL-positive and VL-negative dogs with N. caninum infection [P= 0.001]. These findings indicate that Meshkin-Shahr District in northwestern Iran is an active focus of canine visceral leishmaniasis [CVL]. Neospora caninum and L. infantum co-infection is prevalent in the area and infection by L. infantum seems to enhance susceptibility to N. caninum infection in domestic dogs

Integrated visceral leishmaniasis surveillance system in primary care for children in Meshkin-Shahr district, north-western Islamic Republic of Iran

In 2001 a visceral leishmaniasis [VL] surveillance system was set up for children aged

Seroepidemiological study of visceral leishmaniasis in Booyerahmad district, south-west Islamic Republic of Iran

Visceral leishmaniasis [VL] is endemic in parts of Islamic Republic of Iran. A cross-sectional seroprevalence study was carried out in children in Booyerahmad district in the south-west of the country. Serum samples were taken from 1628 children up to 10 years old from different areas in Booyerahmad in 2005-06. Anti-Leishmania antibody was detected in 50 out of 1628 children [3.1%] by direct agglutination test [antibody titre > 1:3200]. There was no significant difference in seropositivity between the sexes [2.8% males and 3.3% females]. The highest rate of infection [5.2%] was in the age group 10 years. Further studies are needed to explore the reservoirs and vectors of the disease in this region

[Comparison between serology and PCR methods for the diagnosis of visceral Leishmaniasis]

Canine visceral leishmaniasis [CVL] caused by Leishmania Infantum is endemic in most Mediterranean basin and its seroprevalence ranges from 10 to 37%. Diagnosis of Infection is very important especially in asymptomatic dogs for control of human leishmaniasis for control of human visceral leishmaniasis. This study was aimed to compare three methods for detection of canine visceral leishmaniasis. In this research process study, 71 dogs were selected from 4 endemic villages in Meshkin-Shahr district. Peripheral blood samples were tested by serologic [DAT and Dipstick rK39] and molecular [PCR] methods. Skin samples were tested by molecular [PCR] methods. Twelve samples of PCR products were sequenced that all of them were identified as Leishmania infantum and 2 nucleotide sequence data submitted to the GenBank database. From 71 dogs that were studied, 21.1% were symptomatic and others were asymptomatic[78.9%]. 17 dogs [23.9%] had >/= 1:320 titer of antibody by direct agglutination test [DAT]. Twenty two dogs[31%] were positive by Dipstick rK39 test, 21 dogs [29.6%] were positive by PCR on skin samples, 31 dogs [43.7%] were positive in blood PCR and 38 dogs [53.5%] were positive by skin/blood PCR. The highest correlation was between DAT and Dipstick test [76%].According to the results of this study, we can diagnose infection in symptomatic and asymptomatic dogs by DAT as a suitable method and PCR is suitable to follow parasite DNA in skin and other tissues of dogs

[Seroepidemjological survey of visceral leishmaniasis by direct agglutination test in bojnoord district, north Khorasan province in 2007]

This study aimed to determine of the seroprevalence of visceral leishmaniasis to design a prevention and control program in Bojnoord district. This was a descriptive and cross sectional study with multistage random sampling method. The study was carried out on children up to 12 years old and 10% of adult's population in 8 rural areas of Bojnoord district in 2007. In total, 1608 blood samples were collected to detect anti-Leishmania antibodies. All the samples were tested by direct agglutination test [DAT]. In order to determine Leishmania species, necropsy was performed on four suspected dogs in the studied areas. The species of Leishmania was determined by RAPD-PCR and PCR-RFLP methods using to amplify the ribosomal internal transcribed spacer 1 [ITS1]. Thirty and eight [2.36%] out of 1 608 collected blood samples had anti- Leishmania antibodies at titer 1:800 and nine [0.56%] were just positive at 1:3200 by DAT. There was no statistically significant difference between female and male seroprevalence [p>0/05]. The seoprevalence in children <=12 years old compared to individuals greater than 12 years old did not show any statistically significant [p>0/05]. Amastigotes were observed in all 4 necropsied dogs. The species of Leishmania isolated from 2 dogs, was determined as L.Infantum. Their ITS1 sequences were registered with Accession numbers EU810776 and EU810777 in NCBI. These findings showed that visceral leishmaniasis has been circulated with low endemicity in Bojnoord district. Therefore an appropriate monitoring system is needed for health services in this area

Seroepidemiological study on canine visceral leishmaniasis in Meshkin-Shahr district, Ardabil province, northwest of Iran during 2006-2007

This study aimed to determine the seroprevalence of canine visceral leishmaniasis in Meshkin-Shahr district as endemic areas of human visceral leishmaniasis [HVL] for presenting control program of HVL to health authorities. A Seroepidemiological study to determine seroprevalence of canine visceral leishmaniasis [CVL] among ownership dogs using direct agglutination tests [DAT] in 21 villages of Meshkin-Shahr district, Ardabil Province was carried out from June 2006 to August 2007. Three hundred and eighty four ownership dogs were selected by multi-stage cluster sampling. Chi-square and Fisher exact tests were used to compare seroprevalence values relative to gender, age and clinical signs. Of the 384 serum samples tested by DAT, 17.4% [95%C.I, 13.2%-20.8%] were positive [1:320 and higher]. No statistical significant difference was found between male [16.5%] and female [20.2%] seroprevalence [P=0.416]. The highest seroprevalence rate [64.2%] was observed among the ownership dogs of three years age and above. Only 25.4% of the seropositive dogs had clinical signs and symptoms. The most clinical signs among symptomatic dogs were cachexia [75%] and alopecia [36.5%]. The majority of seropositive dogs [74.6%] lived in endemic areas of Meshkin-Shahr district were asymptomatic. It seems that all symptomatic and asymptomatic infected dogs are the most important risk factors for human infection in VL endemic areas

Application of direct agglutination test [DAT] for the diagnosis and seroepidemiological studies of visceral leishmaniasis in Iran

Visceral leishmaniasis [VL] is one of the most important parasitic diseases which is endemic in different parts of Iran. Serological studies were conducted by direct agglutination test [DAT] on 12144 human serum samples, collected from four geographical zones of Iran. Sero prevalence, geographical distribution, clinical signs and symptoms for human visceral leishmaniasis based on DAT for the period of 2002 through 2005 were determined. From 516 kala-azar cases detected: 50.6% were from Meshkin-shahr and Moghan districts in Ardabil Province, northwest of Iran and 49.4% were detected from other areas of Iran. In physical examination of seropositive cases, which were detected by DAT with anti-leishmanial antibodies at titers of 1: 3200 to 1: 102400, almost 50% of suspected individuals showed the classical kala-azar signs and symptoms. Predominant signs and symptoms in 233 hospitalized patients with anti-Leishmania antibodies at 1:3200 and higher, were fever [88.0%] and splenomegaly [84.5%]. Statistically significant difference was found between males [58%] and females [42%] [P< 0.01]. Moreover, 93.6% of the VL patients were < 5 yr of age, and 6.4% were older than 5 yr that this difference was statistically significant [P< 0.01]. From 1383 serum samples collected from domestic dogs in the villages that are known as endemic foci of human leishmaniasis, 152 [11.0%] were positive by DAT [>/= 1:320]. Parasitological and serological examinations that were performed in 30 wild canines showed that 10% of these animals were infected by L. infantum. L. infantum Lon49 is the principal agent of the disease in human as well as animal reservoir hosts in different parts of Iran. For the first time in Iran, L. tropica isolated from both skin lesions in the face and bone marrow aspiration in a HIV+ man who co-infected with VL as well as in an infected dog from Ardabil Province