Presence and diffusion of Leishmaniasis and its vectors in the province of Rieti

Nel presente studio ci si è proposti di valutare la presenza e diffusione della leishmaniosi in una provincia dell’Italia centrale (Rieti) che, per caratteristiche ambientali (prevalentemente montuosa, clima freddo-secco) poco sembra prestarsi al ciclo della malattia. A questo scopo sono stati calcolati: i) sieroprevalenza grezza nella popolazione canina (2006-2013) e prevalenza media annuale ii) casi di leishmaniosi viscerale (LV) e cutanea (LC) (2000-2013). Catture di flebotomi sono state effettuate per due stagioni consecutive (2011-2012) per ogni sito sono stati registrati i dati climatici (temperatura, umidità etc.) ed altitudine. I flebotomi sono stati sottoposti a ricerca di Leishmania mediante PCR. La sieroprevalenza grezza per leishmania varia da 0 a 76,9% e la prevalenza media annuale non presenta un trend lineare. Sono stati registrati 6 casi di LV tutti in pazienti italiani tutti residenti in provincia di Rieti. I flebotomi sono stati rilevati in 5 dei 6 siti monitorati fino agli 800 m s.l.m., seppur con basse densità. Sono state identificate le seguenti specie: P. perniciosus (6,4 %), P. perfiliewi (1,8%), P. mascittii (0,1%) e S. minuta (91,7 %). E’ stata rilevata una correlazione statisticamente significativa (r=0,69, p<0,001) tra numero di flebotomi e temperatura giornaliera (Tmed°C) ed una correlazione negativa significativa (r= -0,51, p<0,05) con l’umidità relativa (Umed%). La ricerca di leishmania ha dato esito negativo in tutti i flebotomi analizzati. Questi rilievi suggeriscono l’endemia della leishmaniosi nella provincia di Rieti.The aim of this study is to describe the presence and diffusion of leishmaniasis in a province of Central Italy (Rieti) not optimal for the disease transmission for its environmental conditions (dry-cold weather, 70% highlands). We estimated i) Canine raw seroprevalence (2006-2013) and mean annual prevalence ii) cases of visceral and cutaneous leishmaniasis (VL, CL) (2000-2013). Phlebotomine sand flies were trapped during two seasons (2011-2012) and meteorological data and altitude were recorded for each site. The phlebotomines were submitted to Leishmania spp detection by PCR. The raw seroprevalence ranged from 0-76,9% and the mean annual prevalence does not show a linear trend. Six cases of VL were detected in italian patients who reside in the province. Phlebotomine sand flies were found in 5/6 sites until 800 m above s.l., with low density. Four species were identified: P. perniciosus (6,4 %), P. perfiliewi (1,8%), P. mascittii (0,1%) and S. minuta (91,7 %). The number of phlebotomines resulted positively correlated (r=0,69, p<0,001) with the mean daily temperature (Tmed°C) while a significant negative correlation (r= -0,51, p<0,05) was found with mean relative humidity (Umed%). Leishmania spp. was not detected. These findings suggest the endemic status of leishmaniasis in the province

Giardia duodenalis in colony stray cats from Italy

Giardia duodenalis is the most common intestinal protozoan in humans and animals worldwide, including eight morphologically identical assemblages, infecting pets, livestock, wildlife and human beings. Assemblages A and B are those with the higher zoonotic potential, and they have been detected in several mammals other than humans; the others (C to H) show a higher host specificity. Cats can harbour both the specific Assemblage F and the zoonotic ones A and B. Several studies have been carried out on G. duodenalis genotypes in cats; however, the role of this species in the epidemiology of giardiasis is still poorly understood. In this scenario, the present study carried out the detection and genetic characterization at sub-assemblage level of G. duodenalis from colony stray cats in central Italy. In the period 2018-2019, 133 cat faecal samples were analysed for the presence of G. duodenalis cysts by a direct immunofluorescence assay. Positive samples were subsequently subjected to molecular analyses for assemblage/sub-assemblage identification. Forty-seven samples (35.3%) were positive for G. duodenalis cysts by immunofluorescence. G. duodenalis DNA was amplified at SSU-rDNA locus from 39 isolates: 37 were positive for zoonotic Assemblage A and 2 showed a mixed infection (A + B). Positive results for the beta-giardin gene were achieved for 25 isolates. Sequence analysis revealed 16 isolates belonging to Sub-assemblage AII and 8 to Sub-assemblage AIII. One isolate resulted as ambiguous AI/AIII. Large sequence variability at the sub-assemblage level was detected, with several double peaks and mutations, making complex a proper isolate allocation. When compared with previous studies, the 35.3% prevalence of G. duodenalis in cats reported in the present article was surprisingly high. Moreover, all positive cats resulted to be infected with zoonotic assemblages/sub-assemblages, thus indicating stray cats as a possible source of human giardiasis and highlighting the sanitary relevance of cat colonies in the study area

Lung parasites of the genus Metastrongylus Molin, 1861 (Nematoda: Metastrongilidae) in wild boar (Sus scrofa L., 1758) in Central-Italy: An eco-epidemiological study

The respiratory tracts of 57 wild boars (Sus scrofa L. 1758) hunted in central Italy during the 2011/2012 hunting season were examined to detect the presence of lung worms. Fifty-five out of 57 animals (96,5%) were positive. Five species of Metastrongylus were detected and their prevalence was as follows: Metastrongylus asymmetricus Noda, 1973 (91.2%), Metastrongylus confusus Jansen, 1964 and Metastrongylus salmi Gedoelst, 1923 (87.7%), Metastrongylus apri Gmelin, 1790 (80.7%), Metastrongylus pudendotectus Vostokov, 1905 (70.2%). In most cases multi-species infection was observed. The highest parasite load was found in young animals (<1 year old). The Metastrongylus genus sex ratio (M/F) had a range from 1:4.8 to 1:1.5 in favor of females. The Simpson and Shannon-Wiener indices showed a moderate uniformity in parasite community composition. The Fager index highlighted a high degree of affinity among all pairs of selected parasites. The whole parasite population showed an aggregate distribution. Our findings confirm that these parasites are widespread in the wild boar population. The establishment of outdoor domestic pig farming in the same area of the game preserve could pose the risk of infection to domestic animals. Further studies will be needed to understand the factors involved in the presence and prevalence of the intermediate host as well as the population dynamics of Metastrongylus spp

Seroprevalence and risk factors associated with exposure to Leishmania infantum in dogs, in an endemic Mediterranean region.

Canine leishmaniasis (CanL) is a zoonotic parasitic disease caused by Leishmania infantum in the Mediterranean area and transmitted by phlebotomine sand fly vectors. The domestic dog is the main reservoir host. The aim of this study was to assess the influence of different individual, environmental and spatial risk factors on the dog exposure to L. infantum and to estimate the seroprevalence among owned and kennel dogs, in the Lazio region (central Italy), where canine leishmaniasis is endemic. In the period 2010-2014, 13,292 sera from kennel and owned dogs were collected by official and private veterinarians. The presence of anti-Leishmania IgG was analysed by indirect fluorescent antibody test (IFAT), using a 1:80 titre cut-off. At the univariable analysis, CanL seropositivity was associated with sex, size, breed, coat length, living with other dogs and forest/semi-natural land cover. At the multivariable analysis, age, ownership and attitude were confirmed as risk factors, being more than 2 years old, owned, and hunting dogs at higher risk. Being a Maremma sheepdog was a protective factor. A true overall seroprevalence of 6.7% (95% CI: 6.2-7.2) was estimated in the whole population while 7.3% (95% CI: 6.8-7.8) was estimated in kennel dogs and 74.3% (95% CI: 70.8-77.6) in owned dogs. The role of kennels as a key component for CanL active and passive surveillance was also highlighted. This study confirmed the endemicity of CanL in the Lazio region and focused some factors that can influence the seropositivity of dogs in a Mediterranean region

Leishmanicidal activity of green tea leaves and pomegranate peel extracts on <i>L. infantum</i>

<p><i>Leishmania infantum</i> is responsible for the cutaneous and visceral form of this zoonotic disease, which is potentially lethal for humans and has dogs as natural reservoir. In the light of the antiparasitic properties displayed by several natural products, <i>L. infantum</i> promastigotes were exposed to green tea (<i>Camellia sinensis</i>) leaves extract (GTE) and pomegranate (<i>Punica granatum</i>) peel extract (PPE). Both extracts, characterized by NMR and HPLC analysis, inhibited parasite proliferation in a dose-dependent manner, as proved by IC₅₀ evaluation determined by MTT assay.Moreover, the reversibility assay showed that GTE and PPE have an aptotosis-mediated leishmanicidal effect, as evidenced by DNA degradation and confirmed by DNA fragmentation and real-time PCR analyses. Finally, for the first time morphological and ultrastructural alterations induced by a <i>P. granatum</i> extract on <i>Leishmania</i> were shown by the use of light, transmission and scanning electron microscopy.</p