title: Longitudinal study on the detection of canine Leishmania infections by conjunctival swab analysis and correlation with entomological parameters.
abstract: A longitudinal study was carried out on kennelled stray dogs in a canine leishmaniasis (CanL) endemic area, to evaluate early and late diagnostic performance of a non-invasive conjunctival swab (CS) nested (n)-PCR analysis for Leishmania detection in 2 cohorts of dogs, respectively. (A) Sixty-five IFAT- and CS n-PCR-negative dogs exposed to, and followed up once or twice a month during a full sand fly season (July-November 2008). In parallel, a sand fly survey was performed on site using standard sticky traps set twice a month, for a cumulative surface of 63 m(2). (B) Seventeen IFAT- and CS n-PCR-negative dogs found positive in July 2008 at the peripheral blood buffy-coat (BC) n-PCR. These dogs were examined again by BC n-PCR in September and November 2008, and before the subsequent transmission season (May 2009) along with CS n-PCR and IFAT. None of the cohort (A) dogs converted to positive CS n-PCR during the transmission season. Although approximately 2500 phlebotomine specimens were collected with peaks of 100-147 specimens/m(2) sticky trap, the cumulative density of the only proven CanL vector in the area (Phlebotomus perniciosus) was found to be very low (0.5/m(2)). All cohort (B) dogs remained substantially seronegative; BC n-PCR showed an intermittent positive trend during the period surveyed, resulting in 82% conversions to negative by the end of the study, in contrast with 71% conversions to positive at the CS n-PCR analysis. In conclusion, while CS n-PCR was not found effective for the early detection of Leishmania contacts in dogs exposed to a low pressure of vectorial transmission, this assay showed to slowly convert to positive in a high rate of dogs, in the absence of seroconversion. CS n-PCR technique can be a suitable marker for assessing Leishmania exposure in dogs as a non-invasive alternative to current serological and molecular tools.
authors.affiliation: Unit of Vector-borne Diseases & International Health, MIPI Department, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Gramiccia
authors.forename: Trentina
authors.initials: T
authors.lastname: Di Muccio
authors.forename: Eleonora
authors.initials: E
authors.lastname: Fiorentino
authors.forename: Aldo
authors.initials: A
authors.lastname: Scalone
authors.forename: Gioia
authors.initials: G
authors.lastname: Bongiorno
authors.forename: Silvia
authors.initials: S
authors.lastname: Cappiello
authors.forename: Rosa
authors.initials: R
authors.lastname: Paparcone
authors.forename: Valentina
authors.initials: V
authors.lastname: Foglia Manzillo
authors.forename: Michele
authors.initials: M
authors.lastname: Maroli
authors.forename: Luigi
authors.initials: L
authors.lastname: Gradoni
authors.forename: Gaetano
authors.initials: G
authors.lastname: Oliva
mesh_terms: D000818:Animals; D015331:Cohort Studies; D003228:Conjunctiva / Q000469:parasitology*; D004283:Dog Diseases / Q000175:diagnosis* / Q000469:parasitology; D004285:Dogs; D005260:Female; D007896:Leishmaniasis / Q000175:diagnosis / Q000662:veterinary*; D008297:Male; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011576:Psychodidae; D012621:Seasons