Monitoring and detection of new endemic foci of canine leishmaniosis in northern continental Italy: An update from a study involving five regions (2018-2019).

Canine leishmaniosis (CanL) is an emerging zoonosis caused by Leishmania infantum and transmitted in southern Europe by phlebotomine sand flies of the subgenus Phlebotomus (Larroussius). Endemic foci of CanL have been recorded in northern continental Italy since early 1990s and attributed to the northward expansion of vector populations due to climatic changes in association with travelling/relocated infected dogs from the southern Mediterranean littoral. In this study, further spread of endemic Leishmania foci was monitored during 2018-2019 in five regions (Aosta Valley, Piedmont, Lombardy, Veneto and Friuli-Venezia Giulia), with focus to territories where investigations were not performed, or they have been inconclusive. Clinical cases of CanL identified by local veterinary practitioners and confirmed by reference diagnosis centers were regarded as autochthonous if their origin from, or travel to, areas endemic for CanL were excluded in the previous ≥2 years. Around these index cases, i) serosurveys for L infantum were carried out where indicated, ii) sampling from potential autochthonous cases in healthy or clinically-suspected resident dogs was intensified by collaborating veterinary practitioners, and iii) suitable sites were investigated for the presence of competent phlebotomine vectors. Fifty-seven municipalities whose enzootic status of CanL was unreported before 2018, were identified as endemic. The stability of 27 foci recorded over the past decade, was also confirmed. Competent phlebotomine vectors, mainly Phlebotomus perniciosus, were collected for the first time in 23 municipalities. The newly recorded endemic municipalities appear to be distributed over a west-to-east decreasing gradient: 30 in Piedmont, 21 in Lombardy, 4 in Veneto and 2 in Friuli-Venezia Giulia. As regards Veneto, it should be noted that a relatively restricted territory was investigated as several municipalities of the region had already been surveyed and detected as endemic for CanL in the past. Cold climate conditions of the easternmost region of Friuli-Venezia Giulia bordering non-endemic territories of Slovenia, are probably less favorable to L infantum transmission.

Effectiveness of the spot-on combination of moxidectin and imidacloprid (Advocate®) in the treatment of ocular thelaziosis by Thelazia callipaeda in naturally infected cats.

BACKGROUND: The present study evaluated the therapeutic effectiveness of moxidectin 1.0% (w/v) and imidacloprid 10% (w/v) (Advocate® spot-on solution for cats, Bayer Animal Health) against natural infections with the eyeworm Thelazia callipaeda in cats. This study was conducted as a GCP, negative-controlled, blinded and randomised field study in privately owned cats living in an area in southern Italy where T. callipaeda is enzootic. METHODS: The study was carried out in 30 cats (19 females and 11 males, aged from 8 months to 5 years, weighing 1.2-5.2 kg) of different breeds, naturally infected by T. callipaeda. At study inclusion (Day 0), animals were physically examined and the infection level was assessed by examination of both eyes for clinical score and live adult T. callipaeda count. Each cat was weighed and randomly assigned to one of the treatment groups (G1: Advocate, G2: untreated control). Clinical assessments and T. callipaeda adult counts were performed on Day 14. At the study completion visit on Day 28, clinical assessments and counts of T. callipaeda adults and larvae were performed. All cats were daily observed by their owners and general health conditions were recorded during the entire period of the study. RESULTS: The primary effectiveness variable was the percentage of animals in G1 group (Advocate) showing a complete elimination (parasitological cure) of adult eye worms at Day 14 and Day 28 . The effectiveness of the treatment in the G1 group was 93.3 and 100% at Day 14 and Day 28 , respectively, when compared to group G2. Total worm count reduction from both eyes for Advocate was 96.3% on Day 14 and 100% on Day 28. Clinical data were confirmed by the examination of conjunctival pouch flushing. An overall reduction in the number of cats with lacrimation and conjunctivitis was observed following treatment despite the fact that in a few cats treated with Advocate clinical signs persisted due to the chronic nature of the disease. CONCLUSIONS: Based on the results of the present trial, a single dose of Advocate was found to be safe and highly effective in the treatment of natural T. callipaeda infection in cats.

Prevention of feline leishmaniosis with an imidacloprid 10%/flumethrin 4.5% polymer matrix collar.

BACKGROUND: Leishmaniosis caused by Leishmania infantum is one of the most important vector-borne diseases affecting animals and humans worldwide. Dogs are considered main reservoirs of the zoonotic forms, though in the last years the role of cats as reservoirs has been increasingly investigated. Feline leishmaniosis (FeL) occurs in endemic areas and no specific preventive measures have been investigated so far. In this study the efficacy of a 10% imidacloprid/4.5% flumethrin polymer matrix collar, licensed for tick and flea prevention, has been assessed against FeL in a longitudinal study on 204 privately owned cats from the Aeolian islands (Sicily), an area highly endemic for the disease. From March to May 2015 [Study Day 0 (SD 0)], cats negative for FeL were collared (G1, n = 104) or left untreated (G2, n = 100). Diagnosis consisted of serology and qPCR on blood and conjunctival swabs, which were collected at baseline (SD 0) and at the end of the study (SD 360). Interim clinical examinations were performed on SD 210 (when collars were replaced in G1) and SD 270. RESULTS: Of the 159 cats which completed the study, 5 in G1 and 20 in G2 were positive for L. infantum infection, in at least one of the diagnostic tests leading to a yearly crude incidence of 6.3% and 25.0% in G1 and G2, respectively (P = 0.0026). This translates into an efficacy of the collar of 75.0% in preventing feline Leishmania infection. The collar was generally well tolerated with no systemic adverse reactions and few local skin reactions were observed in the application area in four out of 104 treated cats (3.8%). CONCLUSIONS: The 10% imidacloprid/4.5% flumethrin collar significantly reduced the risk of L. infantum infection in cats. To our knowledge, this is the first study in which a preventative strategy against feline Leishmania infection is assessed under natural conditions. These findings close a gap in veterinary medicine, in that they confirm this collar as a tool in reducing the risk of Leishmania infection in cats. Such a preventative tool could contribute to the reduction of the risk of the disease in animal and in human populations when included in integrated leishmaniosis control programmes.

Efficacy of moxidectin 2.5% and imidacloprid 10% in the treatment of ocular thelaziosis by Thelazia callipaeda in naturally infected dogs.

Thelazia callipaeda (Spirurida, Thelaziidae) has been documented as agent of ocular infection in domestic animals (dogs and cats), wildlife (e.g., foxes, hares, rabbits), and humans. In the last two decades, this parasitosis has been increasingly reported in several European countries. Both adult and larval stages of the eyeworm are responsible for symptoms ranging from mild (e.g., lacrimation, ocular discharge, epiphora) to severe (e.g., conjunctivitis, keratitis, and corneal opacity or ulcers). The present study evaluated the clinical efficacy and safety of imidacloprid 10% and moxidectin 2.5% spot on (Advocate(®), Bayer Animal Health) in comparison to milbemycin oxime/praziquantel tablets (Milbemax(®), Novartis-Animal Health), as positive control, in the treatment of canine thelaziosis in naturally infected dogs and, a third group was used as an untreated control. Forty-seven dogs (27 females and 20 males) harbouring at least one live adult worm of T. callipaeda in one eye were enrolled from an endemic area of southern Italy. Each dog was then weighed and assigned in accordance with a random treatment allocation plan to one of the treatment groups (G1: imidacloprid 10% and moxidectin 2.5% spot on, G2: Untreated control and G3: milbemycin oxime/praziquantel tablets). On Day (D) 7, 14, 28 and 35 dogs were physically examined and the infection level was assessed by examination of both eyes, including conjunctival pouch and third eyelid for live adult T. callipaeda count and clinical scores. Dogs in G1 were treated on D0 and D28, whereas those in G3 on D0 and D7. Efficacy in G1 was 100% at each day post treatment (p<0.01). For the G3 group efficacy was 57.39% on D7 (p<0.05), 92.79% on D14 and 100% on D28 and D35 (p<0.01). The application of the spot on formulation moxidectin 2.5% and imidacloprid 10% was highly effective in the treatment of canine thelaziosis caused by T. callipaeda. Advocate(®) spot on can be recommended for the control of T. callipaeda infection, considering that this formulation is currently licensed in Europe for the treatment of a wide range of parasites affecting dogs.

Field Evaluation of Two Different Treatment Approaches and Their Ability to Control Fleas and Prevent Canine Leishmaniosis in a Highly Endemic Area.

This study investigated the efficacy of two collars for the treatment and prevention of flea infestations. Additionally the effect of these collars on the incidence of Leishmania infantum infection as compared with a group of vaccinated dogs was evaluated. A total of 224 young dogs from private animal shelters were enrolled in April/May into four groups: G1, 55 dogs treated with 10% imidacloprid + 4.5% flumethrin collar (Seresto, Bayer Animal Health); G2, 60 dogs treated with 4% deltamethrin collar (Scalibor protector band, MSD Animal Health); G3, 54 dogs vaccinated with CaniLeish (Virbac Animal Health); and G4, 55 dogs left non-treated as controls. Dogs were followed up at days 120 (September), 210 (December), and 360 (April-May). At those time points, clinical assessments, ectoparasite counts and blood, bone marrow and skin samples, to detect the presence of L. infantum, were performed. The efficacy of Seresto in protecting dogs from flea infestation was 100% (P < 0.01) on day 120 and 210, while animals treated with Scalibor showed a prevalence of the infestation ranging from 23.3% to 33.3% on day 120 and 210, respectively. At the end of the study, the incidence of L. infantum infection in collared dogs-based on animals being positive in any of the tests-was 5.5% in Seresto-treated dogs and 20% in Scalibor-treated dogs, resulting in overall efficacy of prevention of 88.3% for Seresto and 61.8% for Scalibor. No statistical difference was detected in L. infantum positive dogs for bone marrow PCR and/or cytology at day 360 between the CaniLeish (15.4%) and non-treated control dogs (10.0%). Both collars proved to be effective (P < 0.01) in preventing L. infantum infection throughout one transmission season, whereas no significant difference was recorded in the frequency of active infections between dogs vaccinated with CaniLeish and control dogs, emphasizing the importance of using repellent/insecticide actives as a priority measure for protection against canine leishmaniosis.