With respect to the epidemiology of leishmaniasis, it is crucial to take into account the ecoclim... more With respect to the epidemiology of leishmaniasis, it is crucial to take into account the ecoclimatic and environmental characteristics that influence the distribution patterns of the vector sand fly species. It is also important to consider the possible impact of on-going climate changes on the emergence of this disease. In order to map the potential distribution of Phlebotomus perniciosus, the main vector species of canine leishmaniasis in north-eastern Italy, geographical information systems tools, ecological niche models (ENM) and remotely sensed environmental data were applied for a retrospective analysis of an entomological survey conducted in north-eastern Italy over 12 years. Sand fly trapping was conducted from 2001 to 2012 in 175 sites in the provinces of Veneto, Friuli-Venezia Giulia and Trentino-Alto Adige. We developed a predictive model of potential distribution of P. perniciosus using the maximum entropy algorithm software, based on seasonal normalized difference vege...
Vector borne infections are now again a priority worldwide. Emergence or re-emergence of these di... more Vector borne infections are now again a priority worldwide. Emergence or re-emergence of these diseases are usually associated with climatic change, but also other factor are involved, such as demographic, socio-economic and accidental factors. The results of 3 years epidemiolocal investigations about ruminants babesiosis and canine leishmaniosis in north-eastern Italy are presented. These two case studies show that these infections are a potential threat to Public Health. Outcomes of the research confirm the importance of a proper surveillance of vector-borne infections also in north-eastern Italy.
Host-parasite relationships have been frequently investigated in mountain dwelling ungulates, tho... more Host-parasite relationships have been frequently investigated in mountain dwelling ungulates, though mostly focusing on gastrointestinal nematodes. On the contrary, very few studies were conducted on broncopulmonary nematodes, which may result in severe parenchymal lesions and act as predisposing factor for multifactorial pneumonia. The epidemiological and ecological features of lungworms infecting an Alpine ibex population in the Eastern Alps, Italy, were non-invasively investigated by means of a modified Baermann technique with an original quantitative methodology. Out of a total of 269 samples collected monthly from July to November 2013 and from July to October 2014, 212 (78.8%) were positive for Muellerius and 26 (9.7%) for Protostrongylus, whereas Neostrongylus and Cystocaulus were less prevalent (4.1% and 0.7%, respectively). None of the investigated samples tested positive for dictyocaulids. The genus Muellerius showed the highest larval output intensity (134.2L1/g), followe...
Host-parasite relationships have been frequently investigated in mountain dwelling ungulates, tho... more Host-parasite relationships have been frequently investigated in mountain dwelling ungulates, though mostly focusing on gastrointestinal nematodes. On the contrary, very few studies were conducted on broncopulmonary nematodes, which may result in severe parenchymal lesions and act as predisposing factor for multifactorial pneumonia. The epidemiological and ecological features of lungworms infecting an Alpine ibex population in the Eastern Alps, Italy, were non-invasively investigated by means of a modified Baermann technique with an original quantitative methodology. Out of a total of 269 samples collected monthly from July to November 2013 and from July to October 2014, 212 (78.8%) were positive for Muellerius and 26 (9.7%) for Protostrongylus, whereas Neostrongylus and Cystocaulus were less prevalent (4.1% and 0.7%, respectively). None of the investigated samples tested positive for dictyocaulids. The genus Muellerius showed the highest larval output intensity (134.2L1/g), followed by Protostrongylus with 33.8L1/g. A contrasting age-related pattern of Muellerius and Protostrongylus was revealed, with the former significantly more prevalent and abundant in adult animals, while the latter in kids. Due to the limited accessibility of the study area during winter and spring, it was difficult to describe clear seasonal trends in larval output, although Muellerius showed a minimum in the late summer and a rise in the autumn. The newly developed diagnostic method showed a fair repeatability, thus representing an interesting tool to investigate the ecology of lungworms in protected species, such as the A. ibex. Based on results, ibex in the Marmolada massif seem to have an ecologically stable relationship with their lungworm community.
This study was carried out to compare different diagnostic techniques to reveal the presence of p... more This study was carried out to compare different diagnostic techniques to reveal the presence of piroplasms in asymptomatic cattle kept at pasture. Nineteen blood samples were collected from animals of two different areas of Emilia Romagna Region of Italy and processed for microscopic observation, PCR, serological test (IFAT) for Babesia bovis and Babesia bigemina antibodies and in vitro cultivation. The cultures were performed on both bovine and ovine erythrocytes. Seventeen blood smears (89%) were positive for piroplasms, while PCR was positive on 18 samples (95%). DNA sequencing of 18S rRNA identified the piroplasms as Theileria spp. In vitro cultures were successful for 6 samples (32%) cultured on bovine blood and subsequent identified these as Babesia major by PCR. On IFAT analyses of 16 samples, 36.8% resulted positive for B. bovis and 31.6% positive for B. bigemina. These results show, in the same animals, the co-infection with Babesia spp. and Theileria spp.; the detection of B. major was possible only using the in vitro cultures.
In the last decade an increase of the number of red foxes in anthropized habitats across European... more In the last decade an increase of the number of red foxes in anthropized habitats across European countries, including Italy, has been observed. This pones implications in terms of disease transmission between wildlife and domestic animals; in fact, there are evidences of the role of foxes as reservoirs and amplifiers of a broad spectrum of parasites infecting pets. The present study evaluated the prevalence of Eucoleus boehmi, an emerging extra-intestinal nematodes of the Capillariinae subfamily, in red foxes. The nasal passages and sinuses of 179 red foxes culled from several areas of northern and central Italy were inspected and the mucosal surfaces were scrapped and examined for adult nematodes and eggs, microscopically and genetically identified. Overall 55 foxes (30.7%) were found to be infected with E. boehmi, i.e. 27 on inspection of the nasal passages and sinuses and 28 on mucosal flush and scraping. The occurrence of E. boehmi was significantly (p < 0.05) correlated to the sampling location, the age and gender of the animals examined; the higher rates of prevalence were observed in animals culled in Piedmont (43.3%) and in female (60.6%) and adult (38.1%) subjects. A total of 184 adult parasites were recovered, with a mean intensity of infection of 3.34, and a more frequent localization of E. boehmi in the nasal passages rather than in the sinuses. A significant (p < 0.05) relationship was found between the parasite burden and body condition and age of the animals; the intensity of infection was significantly higher in juveniles (mean: 6.3 specimens) and in animals showing poor fox body condition (mean: 7.8 specimens). These results show that E. boehmi is highly prevalent in the red fox populations of certain areas of Italy. Epidemiological implications are discussed, with a special focus on the role that this wild canid may have in the increasing transmission of nasal eucoleosis to domestic dogs.
Babesia EU1, a potentially important emerging zoonotic pathogen, already detected in ticks and wi... more Babesia EU1, a potentially important emerging zoonotic pathogen, already detected in ticks and wild ruminants of different European Countries, was found in three pools of Ixodes ricinus nymphs in three different sites located in a single District of north-eastern Italy. Totally 356 ticks (60 pools) were collected from the environment during a surveillance activity in the year 2006. Babesia EU1 estimated individual tick prevalence in the area is 0.85%. The finding that also in northern Italy the tick population is carrying Babesia EU1 suggests a wide geographical spreading of this zoonotic pathogen in Europe.
Leishmania species can cause diseases with a wide spectrum of clinical manifestations. Different ... more Leishmania species can cause diseases with a wide spectrum of clinical manifestations. Different types of diseases are caused by the same species group, whereas different species may cause the same type of disease. In this study we report the preliminary results of a multilocus molecular and phylogenetic analysis of Leishmania isolated from Sudanese patients. Samples were obtained from the eastern part of the country where the disease is endemic and from the Institute of Endemic Diseases, University of Khartoum. PCR and direct sequencing were used to analyse the polymorphisms in gp63 and minicircle kDNA genes of Leishmania strains from patients showing different clinical manifestations. The interspecific difference in the gp63 sequences ranged from 1.8% to 2.7%. A higher interspecific difference (0.8%-7.7%) was recorded for the kDNA sequences. The intraspecific nucleotidic variation ranged from 0.0% to 0.8% for both genes. phylogenetic analysis of the Leishmania species sequenced in...
Intestinal parasites are common in dogs worldwide, and their importance has recently increased fo... more Intestinal parasites are common in dogs worldwide, and their importance has recently increased for a renewed awareness on the public health relevance that some of them have. In this study, the prevalence of helminths and protozoa was evaluated by microscopy in 318 canine faecal samples collected from eight rescue shelters in the North-eastern Italy; 285 of them were also submitted to the molecular characterization of Giardia duodenalis and Cryptosporidium spp. isolates. An analysis was performed to evaluate the prevalence rates in relation to canine individual data, shelter provenance and anthelmintic treatments. Overall, 52.5 % (167/318) of faecal samples were positive for at least one parasite. Trichuris vulpis showed the highest overall prevalence rate (29.2 %), followed by G. duodenalis (15.1 %), Toxocara canis (9.7 %), ancylostomatids (8.2 %) and Cystoisospora (5.7 %). The prevalence of G. duodenalis, evaluated by real-time PCR, was 57.9 % (165/285), and 79 isolates were characterized by nested PCR on the β-giardin gene. The assemblages found were mainly the host-specific genotypes C and D, while only one assemblage was identified as the human-specific genotype B1. Isolates of Cryptosporidium spp., recorded in 3/285 (1.1 %) stool samples, were Cryptosporidium parvum based on the characterization of the Cryptosporidium oocyst wall protein (COWP) gene. Although the results describe a relatively limited risk of dog-originating zoonoses, there is the need to improve the quality of shelter practices towards better health managements for safe pet-adoption campaigns and a minimization of the environmental faecal pollution with canine intestinal parasites.
Aelurostrongylus abstrusus is a metastrongyloid nematode infesting the respiratory system of dome... more Aelurostrongylus abstrusus is a metastrongyloid nematode infesting the respiratory system of domestic cats worldwide. Troglostrongylus brevior and Troglostrongylus subcrenatus, two lungworms thought to infest wild felids, have been found recently in domestic cats from Spain and Italy. These unexpected findings have raised doubts about the assumed past and present occurrence of Troglostrongylus spp., especially T. brevior, in domestic hosts and suggest that there may have been missed detection or misdiagnosis. The present retrospective study evaluated the presence of lungworms in cats from Italy with a diagnosis of respiratory parasitism or with compatible lung lesions from 2002 to 2013. Sixty-eight samples of DNA and larvae from cats with a diagnosis of aelurostrongylosis, and 53 formalin-fixed paraffin-embedded lung samples from cats confirmed as lungworm infested or with compatible lesions, were investigated using two DNA-based assays specific for A. abstrusus or T. brevior. All DNA and larval samples were positive for A. abstrusus and one was additionally positive for T. brevior. Most paraffin-embedded lung tissues were positive only for A. abstrusus, but two samples tested positive for both lungworms and one for T. brevior only. This study supports the major role of A. abstrusus in causing feline respiratory parasitism in endemic areas of Italy.
In Italy, canine piroplasmosis is believed to be widespread, but few data are available on its pr... more In Italy, canine piroplasmosis is believed to be widespread, but few data are available on its presence in most areas. In 2005 and 2006, vertebrate and invertebrate hosts were investigated in Central and Northern Regions of the Country. Microscopy on blood smears, molecular tools and serological tests were applied to 420 blood samples collected from dogs, in order to evaluate the presence of these protozoa and to identify possible risk factors. Moreover, ticks were analyzed by molecular techniques. Microscopy identified as positive 2.8% of the animals, all from Central Italy, and PCR detected 'piroplasm' DNA in 6.0%. Serology evidenced a mean prevalence of 34.0% with a decreasing trend from Central to Northern areas. The 507 collected ticks were identified as belonging to 8 species, mostly represented by Rhipicephalus sanguineus (n=376) and Ixodes ricinus (n=58). Molecular analyses evidenced the presence of babesial parasites (Babesia canis canis, B. canis vogeli, B. microti-like) in 25 (4.9%) of them; in Rh. sanguineus there was also demonstration of the vertical transmission of B. canis canis. Statistical analysis identified 'kennel' as risk factor for Babesia infection. Our findings evidenced that different species of piroplasms potentially infectious for dogs are circulating in Italy, and that epidemiological aspects of these infections are more complex than expected. Vector importance of both Rh. sanguineus and I. ricinus is hypothesized, but further investigation is needed.
With respect to the epidemiology of leishmaniasis, it is crucial to take into account the ecoclim... more With respect to the epidemiology of leishmaniasis, it is crucial to take into account the ecoclimatic and environmental characteristics that influence the distribution patterns of the vector sand fly species. It is also important to consider the possible impact of on-going climate changes on the emergence of this disease. In order to map the potential distribution of Phlebotomus perniciosus, the main vector species of canine leishmaniasis in north-eastern Italy, geographical information systems tools, ecological niche models (ENM) and remotely sensed environmental data were applied for a retrospective analysis of an entomological survey conducted in north-eastern Italy over 12 years. Sand fly trapping was conducted from 2001 to 2012 in 175 sites in the provinces of Veneto, Friuli-Venezia Giulia and Trentino-Alto Adige. We developed a predictive model of potential distribution of P. perniciosus using the maximum entropy algorithm software, based on seasonal normalized difference vege...
Vector borne infections are now again a priority worldwide. Emergence or re-emergence of these di... more Vector borne infections are now again a priority worldwide. Emergence or re-emergence of these diseases are usually associated with climatic change, but also other factor are involved, such as demographic, socio-economic and accidental factors. The results of 3 years epidemiolocal investigations about ruminants babesiosis and canine leishmaniosis in north-eastern Italy are presented. These two case studies show that these infections are a potential threat to Public Health. Outcomes of the research confirm the importance of a proper surveillance of vector-borne infections also in north-eastern Italy.
Host-parasite relationships have been frequently investigated in mountain dwelling ungulates, tho... more Host-parasite relationships have been frequently investigated in mountain dwelling ungulates, though mostly focusing on gastrointestinal nematodes. On the contrary, very few studies were conducted on broncopulmonary nematodes, which may result in severe parenchymal lesions and act as predisposing factor for multifactorial pneumonia. The epidemiological and ecological features of lungworms infecting an Alpine ibex population in the Eastern Alps, Italy, were non-invasively investigated by means of a modified Baermann technique with an original quantitative methodology. Out of a total of 269 samples collected monthly from July to November 2013 and from July to October 2014, 212 (78.8%) were positive for Muellerius and 26 (9.7%) for Protostrongylus, whereas Neostrongylus and Cystocaulus were less prevalent (4.1% and 0.7%, respectively). None of the investigated samples tested positive for dictyocaulids. The genus Muellerius showed the highest larval output intensity (134.2L1/g), followe...
Host-parasite relationships have been frequently investigated in mountain dwelling ungulates, tho... more Host-parasite relationships have been frequently investigated in mountain dwelling ungulates, though mostly focusing on gastrointestinal nematodes. On the contrary, very few studies were conducted on broncopulmonary nematodes, which may result in severe parenchymal lesions and act as predisposing factor for multifactorial pneumonia. The epidemiological and ecological features of lungworms infecting an Alpine ibex population in the Eastern Alps, Italy, were non-invasively investigated by means of a modified Baermann technique with an original quantitative methodology. Out of a total of 269 samples collected monthly from July to November 2013 and from July to October 2014, 212 (78.8%) were positive for Muellerius and 26 (9.7%) for Protostrongylus, whereas Neostrongylus and Cystocaulus were less prevalent (4.1% and 0.7%, respectively). None of the investigated samples tested positive for dictyocaulids. The genus Muellerius showed the highest larval output intensity (134.2L1/g), followed by Protostrongylus with 33.8L1/g. A contrasting age-related pattern of Muellerius and Protostrongylus was revealed, with the former significantly more prevalent and abundant in adult animals, while the latter in kids. Due to the limited accessibility of the study area during winter and spring, it was difficult to describe clear seasonal trends in larval output, although Muellerius showed a minimum in the late summer and a rise in the autumn. The newly developed diagnostic method showed a fair repeatability, thus representing an interesting tool to investigate the ecology of lungworms in protected species, such as the A. ibex. Based on results, ibex in the Marmolada massif seem to have an ecologically stable relationship with their lungworm community.
This study was carried out to compare different diagnostic techniques to reveal the presence of p... more This study was carried out to compare different diagnostic techniques to reveal the presence of piroplasms in asymptomatic cattle kept at pasture. Nineteen blood samples were collected from animals of two different areas of Emilia Romagna Region of Italy and processed for microscopic observation, PCR, serological test (IFAT) for Babesia bovis and Babesia bigemina antibodies and in vitro cultivation. The cultures were performed on both bovine and ovine erythrocytes. Seventeen blood smears (89%) were positive for piroplasms, while PCR was positive on 18 samples (95%). DNA sequencing of 18S rRNA identified the piroplasms as Theileria spp. In vitro cultures were successful for 6 samples (32%) cultured on bovine blood and subsequent identified these as Babesia major by PCR. On IFAT analyses of 16 samples, 36.8% resulted positive for B. bovis and 31.6% positive for B. bigemina. These results show, in the same animals, the co-infection with Babesia spp. and Theileria spp.; the detection of B. major was possible only using the in vitro cultures.
In the last decade an increase of the number of red foxes in anthropized habitats across European... more In the last decade an increase of the number of red foxes in anthropized habitats across European countries, including Italy, has been observed. This pones implications in terms of disease transmission between wildlife and domestic animals; in fact, there are evidences of the role of foxes as reservoirs and amplifiers of a broad spectrum of parasites infecting pets. The present study evaluated the prevalence of Eucoleus boehmi, an emerging extra-intestinal nematodes of the Capillariinae subfamily, in red foxes. The nasal passages and sinuses of 179 red foxes culled from several areas of northern and central Italy were inspected and the mucosal surfaces were scrapped and examined for adult nematodes and eggs, microscopically and genetically identified. Overall 55 foxes (30.7%) were found to be infected with E. boehmi, i.e. 27 on inspection of the nasal passages and sinuses and 28 on mucosal flush and scraping. The occurrence of E. boehmi was significantly (p < 0.05) correlated to the sampling location, the age and gender of the animals examined; the higher rates of prevalence were observed in animals culled in Piedmont (43.3%) and in female (60.6%) and adult (38.1%) subjects. A total of 184 adult parasites were recovered, with a mean intensity of infection of 3.34, and a more frequent localization of E. boehmi in the nasal passages rather than in the sinuses. A significant (p < 0.05) relationship was found between the parasite burden and body condition and age of the animals; the intensity of infection was significantly higher in juveniles (mean: 6.3 specimens) and in animals showing poor fox body condition (mean: 7.8 specimens). These results show that E. boehmi is highly prevalent in the red fox populations of certain areas of Italy. Epidemiological implications are discussed, with a special focus on the role that this wild canid may have in the increasing transmission of nasal eucoleosis to domestic dogs.
Babesia EU1, a potentially important emerging zoonotic pathogen, already detected in ticks and wi... more Babesia EU1, a potentially important emerging zoonotic pathogen, already detected in ticks and wild ruminants of different European Countries, was found in three pools of Ixodes ricinus nymphs in three different sites located in a single District of north-eastern Italy. Totally 356 ticks (60 pools) were collected from the environment during a surveillance activity in the year 2006. Babesia EU1 estimated individual tick prevalence in the area is 0.85%. The finding that also in northern Italy the tick population is carrying Babesia EU1 suggests a wide geographical spreading of this zoonotic pathogen in Europe.
Leishmania species can cause diseases with a wide spectrum of clinical manifestations. Different ... more Leishmania species can cause diseases with a wide spectrum of clinical manifestations. Different types of diseases are caused by the same species group, whereas different species may cause the same type of disease. In this study we report the preliminary results of a multilocus molecular and phylogenetic analysis of Leishmania isolated from Sudanese patients. Samples were obtained from the eastern part of the country where the disease is endemic and from the Institute of Endemic Diseases, University of Khartoum. PCR and direct sequencing were used to analyse the polymorphisms in gp63 and minicircle kDNA genes of Leishmania strains from patients showing different clinical manifestations. The interspecific difference in the gp63 sequences ranged from 1.8% to 2.7%. A higher interspecific difference (0.8%-7.7%) was recorded for the kDNA sequences. The intraspecific nucleotidic variation ranged from 0.0% to 0.8% for both genes. phylogenetic analysis of the Leishmania species sequenced in...
Intestinal parasites are common in dogs worldwide, and their importance has recently increased fo... more Intestinal parasites are common in dogs worldwide, and their importance has recently increased for a renewed awareness on the public health relevance that some of them have. In this study, the prevalence of helminths and protozoa was evaluated by microscopy in 318 canine faecal samples collected from eight rescue shelters in the North-eastern Italy; 285 of them were also submitted to the molecular characterization of Giardia duodenalis and Cryptosporidium spp. isolates. An analysis was performed to evaluate the prevalence rates in relation to canine individual data, shelter provenance and anthelmintic treatments. Overall, 52.5 % (167/318) of faecal samples were positive for at least one parasite. Trichuris vulpis showed the highest overall prevalence rate (29.2 %), followed by G. duodenalis (15.1 %), Toxocara canis (9.7 %), ancylostomatids (8.2 %) and Cystoisospora (5.7 %). The prevalence of G. duodenalis, evaluated by real-time PCR, was 57.9 % (165/285), and 79 isolates were characterized by nested PCR on the β-giardin gene. The assemblages found were mainly the host-specific genotypes C and D, while only one assemblage was identified as the human-specific genotype B1. Isolates of Cryptosporidium spp., recorded in 3/285 (1.1 %) stool samples, were Cryptosporidium parvum based on the characterization of the Cryptosporidium oocyst wall protein (COWP) gene. Although the results describe a relatively limited risk of dog-originating zoonoses, there is the need to improve the quality of shelter practices towards better health managements for safe pet-adoption campaigns and a minimization of the environmental faecal pollution with canine intestinal parasites.
Aelurostrongylus abstrusus is a metastrongyloid nematode infesting the respiratory system of dome... more Aelurostrongylus abstrusus is a metastrongyloid nematode infesting the respiratory system of domestic cats worldwide. Troglostrongylus brevior and Troglostrongylus subcrenatus, two lungworms thought to infest wild felids, have been found recently in domestic cats from Spain and Italy. These unexpected findings have raised doubts about the assumed past and present occurrence of Troglostrongylus spp., especially T. brevior, in domestic hosts and suggest that there may have been missed detection or misdiagnosis. The present retrospective study evaluated the presence of lungworms in cats from Italy with a diagnosis of respiratory parasitism or with compatible lung lesions from 2002 to 2013. Sixty-eight samples of DNA and larvae from cats with a diagnosis of aelurostrongylosis, and 53 formalin-fixed paraffin-embedded lung samples from cats confirmed as lungworm infested or with compatible lesions, were investigated using two DNA-based assays specific for A. abstrusus or T. brevior. All DNA and larval samples were positive for A. abstrusus and one was additionally positive for T. brevior. Most paraffin-embedded lung tissues were positive only for A. abstrusus, but two samples tested positive for both lungworms and one for T. brevior only. This study supports the major role of A. abstrusus in causing feline respiratory parasitism in endemic areas of Italy.
In Italy, canine piroplasmosis is believed to be widespread, but few data are available on its pr... more In Italy, canine piroplasmosis is believed to be widespread, but few data are available on its presence in most areas. In 2005 and 2006, vertebrate and invertebrate hosts were investigated in Central and Northern Regions of the Country. Microscopy on blood smears, molecular tools and serological tests were applied to 420 blood samples collected from dogs, in order to evaluate the presence of these protozoa and to identify possible risk factors. Moreover, ticks were analyzed by molecular techniques. Microscopy identified as positive 2.8% of the animals, all from Central Italy, and PCR detected 'piroplasm' DNA in 6.0%. Serology evidenced a mean prevalence of 34.0% with a decreasing trend from Central to Northern areas. The 507 collected ticks were identified as belonging to 8 species, mostly represented by Rhipicephalus sanguineus (n=376) and Ixodes ricinus (n=58). Molecular analyses evidenced the presence of babesial parasites (Babesia canis canis, B. canis vogeli, B. microti-like) in 25 (4.9%) of them; in Rh. sanguineus there was also demonstration of the vertical transmission of B. canis canis. Statistical analysis identified 'kennel' as risk factor for Babesia infection. Our findings evidenced that different species of piroplasms potentially infectious for dogs are circulating in Italy, and that epidemiological aspects of these infections are more complex than expected. Vector importance of both Rh. sanguineus and I. ricinus is hypothesized, but further investigation is needed.
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Papers by Rudi Cassini