Leishmania infantum infection serosurveillance in stray dogs inhabiting the Madrid community: 2007-2018.

BACKGROUND: Leishmaniosis is an endemic zoonotic disease in the Mediterranean basin caused by Leishmania infantum and transmitted by phlebotomine sandflies. While in dogs disease may be severe, leishmaniosis is also a public health concern as was shown in the largest outbreak of human leishmaniosis (HL) in Europe in 2009 occurring in the Madrid region. The aim of the present study was to assess the applicability of the Leishmaniosis Surveillance Program (LeishSP) established in Madrid in 1996 by examining trends in L. infantum seroprevalence and associated epidemiological risk factors based on data for the 2007-2018 period. METHODS: The study population consisted of 3225 stray dogs from 17 animal shelters collaborating with the LeishSP. Seroprevalences were recorded twice annually (April and November) from 2007 to 2018. In each yearly period, a minimum of 100 dogs were tested to detect dogs infected before and after the sandfly risk season in Madrid area. Each dog was subjected to the same protocol of blood sample collection and clinical examination to collect epidemiological data and clinical signs. Anti-Leishmania-specific IgG was determined by IFAT cut-off ≥ 1:100. RESULTS: Overall seroprevalence was 6.1% (198 positive dogs). Epidemiological data indicate a significantly higher seroprevalence in dogs > 4 years old, purebred dogs (Pit Bull and related breeds), and medium to large size dogs. There were no seroprevalence differences according to sex and/or season (April and November). In addition, no significant differences were observed according to whether dogs lived inside or outside the HL outbreak area. Remarkably, of 198 dogs testing positive for L. infantum, 64.6% had no clinical signs, indicating a high proportion of clinically healthy infected dogs that could be a potential source of infection. CONCLUSIONS: Results indicate a stable seroprevalence of L. infantum infection after 2006 in stray dogs in Madrid but with a recent slightly increasing trend. These observations support the need to continue with the LeishSP implemented by sanitary authorities of the Madrid Community as an early warning strategy for human and animal leishmaniosis and to enable continued assessment of the epidemiological role of dogs with subclinical infection in this important zoonotic disease.

Epidemiological role of dogs since the human leishmaniosis outbreak in Madrid.

BACKGROUND: Canine leishmaniosis (CanL) has been in the spotlight since the 2009 outbreak of human leishmaniosis in Madrid. In the framework of the Leishmaniosis Surveillance Programme set up in Madrid, this study examines Leishmania-specific seroprevalences in stray dogs for the outbreak area and rest of the Madrid region over the period spanning from the outbreak to the present (2009-2016). These data are of interest because stray dogs could be sentinels for disease surveillance in endemic areas. Since 2011, we have also been monitoring owned dogs in the outbreak area. METHODS: Over the study period, Leishmania infantum seroprevalence was determined in 2,123 stray dogs from the outbreak and non-outbreak areas. A serological study was also performed for owned dogs in the outbreak area: high-risk dogs such as hunting or farm dogs (n = 1,722) and pets (n = 1372). All dogs were examined and blood was collected. The variables recorded for each animal were: breed, age, sex, and clinical history indicating if the animal was healthy or clinically suspected of having any disease, and if they showed a clinical picture compatible with CanL. RESULTS: Seroprevalences of L. infantum in stray dogs were similar in the two areas examined: 4.7% (20 out of 346) in the outbreak area and 5.4% (96 out of 1,777) in the remaining Madrid region (χ 2 = 0.080, P = 0.777). A significant association was found between seroprevalence and age (z = -6.319; P < 0.001). Seroprevalence in owned dogs in the outbreak area was 2.1% in high-risk dogs (37 out of 1,722) and 1.2% in pets (17 out of 1,372) (χ 2 = 3.561, P = 0.0591). CONCLUSIONS: Both stray and owned dogs do not seem to play an important role in maintaining the transmission cycle of L. infantum in the Madrid outbreak area. The stable seroprevalence of infection observed in sentinel dogs suggests the good clinical management and prevention of CanL by local practitioners in owned dogs.

Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test.

An outbreak of human leishmaniasis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.