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Dr. Luca Villa
Università degli Studi di Milano, Milan, Italy

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0 Epidemiology
0 Parasitic diseases
0 ruminants
0 Neosporosis
0 Apicomplexa

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Journal article
Published: 01 April 2021 in Microorganisms
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Toxoplasma gondii and Neospora caninum (Apicomplexa, Sarcocystidae) are protozoan parasites infecting a wide range of intermediate hosts worldwide, including birds. Raptors acquire the infections through the ingestion of both infected preys and oocysts in the environment suggesting they might be used as indicators of the spread of these pathogens. Here, we report an epidemiological survey with the aim of determining the prevalence of T. gondii and N. caninum infections in wild birds of prey, hospitalized in two Wildlife Recovery Centres (WRCs) in Northern Italy. Genomic DNA extracted from brain tissue samples was submitted to Real Time PCR targeting T. gondii B1 and N. caninum Nc5 genes. T. gondii genotyping was then performed by multilocus sequence typing (MLST) analysis, targeting three polymorphic genes (GRA6, BTUB, and altSAG2). T. gondii DNA was found in 35 (62.5%) out of 56 examined samples; concerning genotyping, it was possible to amplify at least one gene for 26 animals, and obtained sequences belonged to Type II. N. caninum DNA was only detected in two (3.6%) common kestrels (Falco tinnunculus), adding a new species to the list of suitable intermediate hosts for this pathogen. Data obtained in the present study thus confirmed the spread of both T. gondii and N. caninum in wild bird of prey, endorsing the role of WRCs in the epidemiological surveillance of wildlife.

ACS Style

Alessia Gazzonis; Luca Villa; Emanuele Lubian; Sara Ressegotti; Guido Grilli; Stefano Raimondi; Sergio Zanzani; Maria Manfredi. Molecular Survey on Toxoplasmagondii and Neosporacaninum Infection in Wild Birds of Prey Admitted to Recovery Centers in Northern Italy. Microorganisms 2021, 9, 736 .

AMA Style

Alessia Gazzonis, Luca Villa, Emanuele Lubian, Sara Ressegotti, Guido Grilli, Stefano Raimondi, Sergio Zanzani, Maria Manfredi. Molecular Survey on Toxoplasmagondii and Neosporacaninum Infection in Wild Birds of Prey Admitted to Recovery Centers in Northern Italy. Microorganisms. 2021; 9 (4):736.

Chicago/Turabian Style

Alessia Gazzonis; Luca Villa; Emanuele Lubian; Sara Ressegotti; Guido Grilli; Stefano Raimondi; Sergio Zanzani; Maria Manfredi. 2021. "Molecular Survey on Toxoplasmagondii and Neosporacaninum Infection in Wild Birds of Prey Admitted to Recovery Centers in Northern Italy." Microorganisms 9, no. 4: 736.

Protozoology original paper
Published: 16 March 2021 in Parasitology Research
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Besnoitiosis is an emerging parasitic disease of equids. Italy is one of the few European countries where the circulation of Besnoitia spp. antibodies was demonstrated. In this study, a case of clinical besnoitiosis in two donkeys in northern Italy is reported. The two animals were clinically examined. Serum and blood samples were analyzed for the detection of Besnoitia spp. antibodies and for hematology, biochemistry, and enzyme activity, respectively. ITS-1 PCR and sequencing were carried out on DNA extracted from skin biopsies. Clinical examination revealed numerous scleral pearls in eyes of both animals; alopecia and hyperkeratosis with skin nodules in the region of the neck, hind leg, and on the pinnae were detected. No cysts were evidenced by endoscopy in respiratory and genital tracts. Both animals resulted seropositive to Besnoitia spp. antibodies by Western Blot. Hematology evidenced light anemia, leukocytosis with eosinophilia, and lymphocytosis; biochemistry and enzyme activity revealed hypoalbuminemia with decreased albumin/globulin ratio and elevated alkaline phosphatase values. Parasitic DNA extracted from skin biopsies of both donkeys demonstrated a homology of 100% with Besnoitia spp. This first clinical case of besnoitiosis in two donkeys in Italy both confirms the circulation of Besnoitia spp. in Italian equids and demonstrates that the distribution area of equine besnoitiosis in Europe could be wider than expected. Further studies are needed to infer its relevance, in relation to seroprevalence and clinical disease, and to identify the species of Besnoitia infecting donkeys. Besnoitiosis may be a neglected disease of donkeys in Europe: an early and accurate diagnosis is fundamental to implement adequate control measures to prevent a “silent” spread of Besnoitia spp. infection in equids populations.

ACS Style

Luca Villa; Alessia Libera Gazzonis; Carlos Diezma-Diaz; Chiara Perlotti; Sergio Aurelio Zanzani; Francesco Ferrucci; Gema Álvarez-García; Maria Teresa Manfredi. Besnoitiosis in donkeys: an emerging parasitic disease of equids in Italy. Parasitology Research 2021, 120, 1811 -1819.

AMA Style

Luca Villa, Alessia Libera Gazzonis, Carlos Diezma-Diaz, Chiara Perlotti, Sergio Aurelio Zanzani, Francesco Ferrucci, Gema Álvarez-García, Maria Teresa Manfredi. Besnoitiosis in donkeys: an emerging parasitic disease of equids in Italy. Parasitology Research. 2021; 120 (5):1811-1819.

Chicago/Turabian Style

Luca Villa; Alessia Libera Gazzonis; Carlos Diezma-Diaz; Chiara Perlotti; Sergio Aurelio Zanzani; Francesco Ferrucci; Gema Álvarez-García; Maria Teresa Manfredi. 2021. "Besnoitiosis in donkeys: an emerging parasitic disease of equids in Italy." Parasitology Research 120, no. 5: 1811-1819.

Journal article
Published: 15 March 2021 in Parasites & Vectors
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Background Besnoitia besnoiti is an Apicomplexan protozoa causative of bovine besnoitiosis, a chronic and debilitating disease of cattle, with a variety of pathological findings that could alter some laboratory parameters. A study was conducted in a bovine besnoitiosis endemically infected dairy herd located in Italy characterized by high intra-herd seroprevalence and cattle with clinical signs of the disease. In the study, alterations in laboratory parameters, i.e. hematological and biochemical parameters, enzyme activities and serum cortisol levels, in Besnoitia besnoiti naturally infected cows were investigated in depth. Methods Laboratory parameters in 107 cows, of which 61 were seronegative and 46 were seropositive to B. besnoiti, including 27 with clinical signs of bovine besnoitiosis, were compared. Generalized linear models were used to evaluate the effect of Besnoitia infection on the considered laboratory parameters. Results Hematological analyses revealed that B. besnoiti infection determined a significant alteration to the leukocyte differential, with a higher percentage of granulocytes and a lower percentage of lymphocytes in seropositive and clinically affected animals (Mann–Whitney U-test, P = 0.022); erythrocyte and platelet counts did not show any difference between the considered groups of cows. Biochemistry tests evidenced that the parasite infection influenced serum protein values in seropositive cows and glutamate dehydrogenase values in clinically affected animals. No or only slight differences were revealed for all of the other biochemical and enzyme activity parameters in B. besnoiti-infected animals. In addition, despite the lack of statistical significance, seropositive and clinically affected cows evidenced higher concentrations of serum cortisol values compared to seronegative animals. Conclusions Although physiological, pathological and farm-related factors could have influenced the results in investigated animals, further studies involving more animals from different farms would be advisable to infer the role of B. besnoiti on these alterations, since laboratory parameters could help veterinarians in the diagnosis of bovine besnoitiosis in cattle.

ACS Style

Luca Villa; Alessia Libera Gazzonis; Sergio Aurelio Zanzani; Silvia Mazzola; Alessia Giordano; Maria Teresa Manfredi. Exploring alterations in hematological and biochemical parameters, enzyme activities and serum cortisol in Besnoitia besnoiti naturally infected dairy cattle. Parasites & Vectors 2021, 14, 1 -13.

AMA Style

Luca Villa, Alessia Libera Gazzonis, Sergio Aurelio Zanzani, Silvia Mazzola, Alessia Giordano, Maria Teresa Manfredi. Exploring alterations in hematological and biochemical parameters, enzyme activities and serum cortisol in Besnoitia besnoiti naturally infected dairy cattle. Parasites & Vectors. 2021; 14 (1):1-13.

Chicago/Turabian Style

Luca Villa; Alessia Libera Gazzonis; Sergio Aurelio Zanzani; Silvia Mazzola; Alessia Giordano; Maria Teresa Manfredi. 2021. "Exploring alterations in hematological and biochemical parameters, enzyme activities and serum cortisol in Besnoitia besnoiti naturally infected dairy cattle." Parasites & Vectors 14, no. 1: 1-13.

Journal article
Published: 13 January 2021 in Parasites & Vectors
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Background Neospora caninum, a coccidian protozoan, represents an important cause of bovine abortion. Available N. caninum strains show considerable variation in vitro and in vivo, including different virulence in cattle. To which extent sexual recombination, which is possible in the intestines of domestic dogs and closely related carnivores as definitive hosts, contributes to this variation is not clear yet. Methods Aborted bovine foetuses were collected between 2015 and early 2019 from Italian Holstein Friesian dairy herds suffering from reproductive problems. A total of 198 samples were collected from 165 intensive farms located in Lombardy, northern Italy. N. caninum samples were subjected to multilocus-microsatellite genotyping using ten previously established microsatellite markers. In addition to our own data, those from a recent study providing data on five markers from other northern Italian regions were included and analysed. Results Of the 55 samples finally subjected to genotyping, 35 were typed at all or 9 out of 10 loci and their individual multilocus-microsatellite genotype (MLMG) determined. Linear regression revealed a statistically significant association between the spatial distance of the sampling sites with the genetic distance of N. caninum MLMGs (P < 0.001). Including data from this and a previous North Italian study into eBURST analysis revealed that several of N. caninum MLMGs from northern Italy separate into four groups; most of the samples from Lombardy clustered in one of these groups. Principle component analysis revealed similar clusters and confirmed MLMG groups identified by eBURST. Variations observed between MLMGs were not equally distributed over all loci, but predominantly observed in MS7, MS6A, or MS10. Conclusions Our findings confirm the concept of local N. caninum subpopulations. The geographic distance of sampling was associated with the genetic distance as determined by microsatellite typing. Results suggest that multi-parental recombination in N. caninum is a rare event, but does not exclude uniparental mating. More comprehensive studies on microsatellites in N. caninum and related species like Toxoplasma gondii should be undertaken, not only to improve genotyping capabilities, but also to understand possible functions of these regions in the genomes of these parasites.

ACS Style

Luca Villa; Pavlo Maksimov; Christine Luttermann; Mareen Tuschy; Alessia L. Gazzonis; Sergio A. Zanzani; Michele Mortarino; Franz J. Conraths; Maria Teresa Manfredi; Gereon Schares. Spatial distance between sites of sampling associated with genetic variation among Neospora caninum in aborted bovine foetuses from northern Italy. Parasites & Vectors 2021, 14, 1 -14.

AMA Style

Luca Villa, Pavlo Maksimov, Christine Luttermann, Mareen Tuschy, Alessia L. Gazzonis, Sergio A. Zanzani, Michele Mortarino, Franz J. Conraths, Maria Teresa Manfredi, Gereon Schares. Spatial distance between sites of sampling associated with genetic variation among Neospora caninum in aborted bovine foetuses from northern Italy. Parasites & Vectors. 2021; 14 (1):1-14.

Chicago/Turabian Style

Luca Villa; Pavlo Maksimov; Christine Luttermann; Mareen Tuschy; Alessia L. Gazzonis; Sergio A. Zanzani; Michele Mortarino; Franz J. Conraths; Maria Teresa Manfredi; Gereon Schares. 2021. "Spatial distance between sites of sampling associated with genetic variation among Neospora caninum in aborted bovine foetuses from northern Italy." Parasites & Vectors 14, no. 1: 1-14.

Journal article
Published: 22 October 2020 in Parasites & Vectors
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Background Leishmania infantum is a vector-borne pathogen endemic in countries in the Mediterranean basin, including Italy. Dogs act as the primary reservoir for this parasite, but other animal species may also be infected. Low-to-moderate seroprevalence levels of infection have been reported in apparent healthy equine populations in southern Europe, reinforcing the importance of exploring those species, including horses, that act as a food source for vectors and may thus participate in the epizoological scenario of canine leishmaniosis (CanL) and zoonotic visceral leishmaniosis (ZVL). Since little is known regarding the exposure to L. infantum in horses in Italy, we assessed the seroprevalence in healthy equine populations from different CanL endemic areas. Methods The survey was conducted on 660 apparently healthy horses distributed throughout central and northern regions of Italy between 2016 and 2019. Blood samples were collected and the presence of anti-Leishmania antibodies (IgG) was investigated by the immunofluorescence antibody test. Information on the location and altitude of the stables, along with the horses’ breed, age, sex, and reproductive status was obtained by filling in a questionnaire. This was then used for statistical analysis by generalized linear models to explore risk factors associated with seroreactivity to L. infantum. Results An average seroprevalence of 13.9% was detected for L. infantum in the equine populations investigated, with statistically significant associations between seroprevalence, geographical variables (northern vs central Italy, origin and altitude) and individual factors (i.e. age and breed morphotype). Conclusions Our results highlight that horses are frequently exposed to L. infantum. Further prevalence surveys in horses, also using direct methods (e.g. PCR), are warranted to clarify the role of these hosts in the epidemiology of Leishmania in Italy.

ACS Style

Alessia Libera Gazzonis; Filippo Bertero; Iolanda Moretta; Giulia Morganti; Michele Mortarino; Luca Villa; Sergio Aurelio Zanzani; Benedetto Morandi; Riccardo Rinnovati; Fabrizio Vitale; Maria Teresa Manfredi; Luis Cardoso; Fabrizia Veronesi. Detecting antibodies to Leishmania infantum in horses from areas with different epizooticity levels of canine leishmaniosis and a retrospective revision of Italian data. Parasites & Vectors 2020, 13, 1 -9.

AMA Style

Alessia Libera Gazzonis, Filippo Bertero, Iolanda Moretta, Giulia Morganti, Michele Mortarino, Luca Villa, Sergio Aurelio Zanzani, Benedetto Morandi, Riccardo Rinnovati, Fabrizio Vitale, Maria Teresa Manfredi, Luis Cardoso, Fabrizia Veronesi. Detecting antibodies to Leishmania infantum in horses from areas with different epizooticity levels of canine leishmaniosis and a retrospective revision of Italian data. Parasites & Vectors. 2020; 13 (1):1-9.

Chicago/Turabian Style

Alessia Libera Gazzonis; Filippo Bertero; Iolanda Moretta; Giulia Morganti; Michele Mortarino; Luca Villa; Sergio Aurelio Zanzani; Benedetto Morandi; Riccardo Rinnovati; Fabrizio Vitale; Maria Teresa Manfredi; Luis Cardoso; Fabrizia Veronesi. 2020. "Detecting antibodies to Leishmania infantum in horses from areas with different epizooticity levels of canine leishmaniosis and a retrospective revision of Italian data." Parasites & Vectors 13, no. 1: 1-9.

Short communication
Published: 18 January 2020 in Parasitology International
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The consumption of ovine and caprine meat is considered one of the major transmission routes for Toxoplasma gondii infection in humans. The present study aimed at obtaining epidemiological and molecular data on T. gondii infection in small ruminants slaughtered or commercialized in Italy. Meat juices from 227 sheep and 51 goats were analyzed with a commercial ELISA and antibodies were detected in 28.6% sheep and 27.5% goats. A significant difference was highlighted between adult sheep and the other considered categories (young sheep, young and adult goats) concerning the detection of antibodies (94.1%; p-value = .008). Muscles of positives samples were submitted to molecular analysis, and T. gondii DNA was detected in 15 sheep and three goats; sequencing of B1 gene showed that all belonged to Type II. The present study confirmed small ruminants' meat as a possible source of T. gondii infection for consumers eating raw or undercooked meat, particularly in those countries where the consumption of sheep and goats' meat products is a traditional gastronomic habit.

ACS Style

Alessia Libera Gazzonis; Sergio Aurelio Zanzani; Luca Villa; Maria Teresa Manfredi. Toxoplasma gondii infection in meat-producing small ruminants: Meat juice serology and genotyping. Parasitology International 2020, 76, 102060 .

AMA Style

Alessia Libera Gazzonis, Sergio Aurelio Zanzani, Luca Villa, Maria Teresa Manfredi. Toxoplasma gondii infection in meat-producing small ruminants: Meat juice serology and genotyping. Parasitology International. 2020; 76 ():102060.

Chicago/Turabian Style

Alessia Libera Gazzonis; Sergio Aurelio Zanzani; Luca Villa; Maria Teresa Manfredi. 2020. "Toxoplasma gondii infection in meat-producing small ruminants: Meat juice serology and genotyping." Parasitology International 76, no. : 102060.

Short communication
Published: 07 November 2019 in Parasitology International
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A form of generalized demodectic mange in two dairy cows infected with Besnoitia besnoiti is described. The herd was endemically infected with bovine besnoitiosis; an overall seroprevalence of B. besnoiti antibodies of 23.5%, that increased up to 43.5% considering only cows, was reported. Two out of the cows seropositive to B. besnoiti, at clinical examination presented skin nodules, widespread all over the body, and in particular in anterior regions. Skin biopsies from the region of the neck were collected and the nodules were microscopically examined through compression method. B. besnoiti tissue cysts were not revealed but a semi-solid yellowish content was evidenced with the presence of several mites, morphologically identified as Demodex bovis. Histological examination of skin biopsies evidenced slight acanthosis and hyperkeratosis of the epidermis and superficial dermatitis with oedema and macrophagic and eosinophilic infiltration. Cystic formations located in the deep dermis were lined by metaplastic squamous epithelium and severe cellular infiltration. A treatment with eprinomectin was attempted and clinical improvement of both cows was observed, particularly at the fifteenth day after treatment, with nodules reduced in size and mites in there degenerated. This is the first report of the co-infection of D. bovis infestation and bovine besnoitiosis in cattle. Furthermore, it was demonstrated that D. bovis circulates in the Italian cattle population, but subclinical forms could be underdiagnosed.

ACS Style

Luca Villa; Alessia Libera Gazzonis; Chiara Perlotti; Sergio Zanzani; Giuseppe Sironi; Maria Teresa Manfredi. First report of Demodex bovis infestation in bovine besnoitiosis co-infected dairy cattle in Italy. Parasitology International 2019, 75, 102021 .

AMA Style

Luca Villa, Alessia Libera Gazzonis, Chiara Perlotti, Sergio Zanzani, Giuseppe Sironi, Maria Teresa Manfredi. First report of Demodex bovis infestation in bovine besnoitiosis co-infected dairy cattle in Italy. Parasitology International. 2019; 75 ():102021.

Chicago/Turabian Style

Luca Villa; Alessia Libera Gazzonis; Chiara Perlotti; Sergio Zanzani; Giuseppe Sironi; Maria Teresa Manfredi. 2019. "First report of Demodex bovis infestation in bovine besnoitiosis co-infected dairy cattle in Italy." Parasitology International 75, no. : 102021.

Journal article
Published: 04 November 2019 in Animals
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The objectives of this study were: (i) To investigate possible geographical or environmental factors influencing the infections by Toxoplasma gondii and Neospora caninum in sheep and goats in northern Italy; (ii) to identify areas at risk of infection to set up preventive measures. Forty-three sheep and goat farms were included. Their locations were plotted and associated with T. gondii and N. caninum seroprevalence, then the distribution of farms’ prevalence was evaluated by spatial analysis. Significant clusters for both low and high prevalence were obtained, and a generalized linear model with ordinal logistic regression was implemented to verify if spatial clustering could be due to climate factors (temperature, rainfall, and their interaction). Clusters of high (80.0%) and low prevalence (28.12%) resulted for T. gondii seroprevalence in sheep farms. No significant clusters resulted for goat farms. Clusters of high (38.68%) and low prevalence (21.23%) resulted for N. caninum seroprevalence in sheep farms. One high-prevalence cluster (15.62%) resulted for goat farms. For goats, spatial analysis and analysis on climatic data showed the absence of environmental significant risk factors associated with T. gondii or N. caninum infection. On the contrary, for sheep, annual temperature, rainfall, and their association affected the risk of T. gondii and N. caninum infection. Particularly, high temperatures and abundant rainfalls were related to T. gondii seroprevalence, while low temperatures and scarce rainfalls were related to N. caninum seroprevalence.

ACS Style

Alessia Gazzonis; Luca Villa; Mariateresa Manfredi; Sergio Zanzani. Spatial Analysis of Infections by Toxoplasma gondii and Neospora caninum (Protozoa: Apicomplexa) in Small Ruminants in Northern Italy. Animals 2019, 9, 916 .

AMA Style

Alessia Gazzonis, Luca Villa, Mariateresa Manfredi, Sergio Zanzani. Spatial Analysis of Infections by Toxoplasma gondii and Neospora caninum (Protozoa: Apicomplexa) in Small Ruminants in Northern Italy. Animals. 2019; 9 (11):916.

Chicago/Turabian Style

Alessia Gazzonis; Luca Villa; Mariateresa Manfredi; Sergio Zanzani. 2019. "Spatial Analysis of Infections by Toxoplasma gondii and Neospora caninum (Protozoa: Apicomplexa) in Small Ruminants in Northern Italy." Animals 9, no. 11: 916.

Protozoology original paper
Published: 28 October 2019 in Parasitology Research
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Bovine besnoitiosis (Besnoitia besnoiti) is an emerging parasitic disease of cattle in Europe. This study reports a case of bovine besnoitiosis in a dairy farm housing 217 cattle in Italy. A serological screening was performed on the whole herd using the recommended approach of ELISA and confirmatory Western Blot. Seropositive animals were clinically examined to reveal symptoms and lesions of besnoitiosis. Risk factors and the effects of the parasite infection on reproductive and productive performances were evaluated. Histopathology and molecular analyses on tissues from a slaughtered cow affected by the chronic phase of the disease were carried out. An overall seroprevalence of 23.5%, which increased up to 43.5% considering only cows, was recorded. Clinical examination of 33 of the seropositive cows evidenced the presence of tissue cysts in at least one of the typical localizations (sclera, vulva, or skin) in 25 animals. Statistical analysis did not evidence any significative impact of the parasite infection on herd efficiency; however, a decrease of productive parameters was recorded in cows showing cutaneous cysts. Concerning the chronically affected cow, histopathology revealed B. besnoiti tissue cysts in the skin of the neck, rump, hind legs, eyelid and vulva, in the muzzle, in mucosal membranes of the upper respiratory tract, and in the lungs. Parasite DNA was detected also in masseter muscles, tonsils, mediastinal lymph nodes, liver, cardiac muscle, aorta wall, ovaries, uterus, and vulva. Bovine besnoitiosis continues to spread in the Italian cattle population. Breeders and veterinarians should be aware of this parasitic disease, and control programs should be developed based on surveillance through a diagnostic procedure including both clinical examination and laboratory tests.

ACS Style

Luca Villa; Alessia Libera Gazzonis; Sergio Zanzani; Chiara Perlotti; Giuseppe Sironi; Maria Teresa Manfredi. Bovine besnoitiosis in an endemically infected dairy cattle herd in Italy: serological and clinical observations, risk factors, and effects on reproductive and productive performances. Parasitology Research 2019, 118, 3459 -3468.

AMA Style

Luca Villa, Alessia Libera Gazzonis, Sergio Zanzani, Chiara Perlotti, Giuseppe Sironi, Maria Teresa Manfredi. Bovine besnoitiosis in an endemically infected dairy cattle herd in Italy: serological and clinical observations, risk factors, and effects on reproductive and productive performances. Parasitology Research. 2019; 118 (12):3459-3468.

Chicago/Turabian Style

Luca Villa; Alessia Libera Gazzonis; Sergio Zanzani; Chiara Perlotti; Giuseppe Sironi; Maria Teresa Manfredi. 2019. "Bovine besnoitiosis in an endemically infected dairy cattle herd in Italy: serological and clinical observations, risk factors, and effects on reproductive and productive performances." Parasitology Research 118, no. 12: 3459-3468.

Journal article
Published: 01 October 2019 in Preventive Veterinary Medicine
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The zoonotic protozoa Toxoplasma gondii is one of the major abortive pathogens in small ruminants. Nevertheless, data on T. gondii infection in goats during lactation and on the presence of T. gondii in goat milk are lacking. A longitudinal study was planned in a T. gondii naturally infected dairy goat farm with the aim of (i) evaluating the variation of anti-T. gondii antibodies in blood and milk during the lactation; (ii) identifying the optimal phase during lactation for T. gondii monitoring; (iii) detecting the presence of T. gondii DNA in the milk. From March to July 2017, 30 goats in a farm were fortnightly visited seven times and sampled for blood and, when in lactation, for milk. Individual data regarding age, reproductive disorders, and the day of lactation were recorded. For the detection of anti-T. gondii antibodies in blood and milk a commercial ELISA kit was used. Milk samples (n = 63) of selected nine seropositive animals were also molecularly analysed to amplify a sequence within the ITS1 region of T. gondii. The seroprevalence of T. gondii infection was 63.3% (19/30); a high agreement was obtained between serum and milk results (Spearman's coefficient = 0.793 and Kendall's tau = 0.624), particularly between the 15th and the 60th day of lactation. In the statistical analysis, performed with generalized linear mixed models (GLMMs), the variable "phase of lactation" was strongly associated to ELISA values obtained in both serum and milk (p-value = 0.0001, F = 5.197, and p-value = 0.016, F = 2.755, respectively). Finally, molecular analyses revealed the presence of parasitic DNA in 20.6% (13/63) of milk samples, with a discontinuous parasite excretion; statistical analyses did not reveal any association among the parasite excretion and the considered variables. Milk could be considered as a valid alternative to blood for monitoring T. gondii infection in goat herds. Moreover, the detection of T. gondii DNA in milk enhanced the possibility for raw goat's milk consumption to be considered as a risk to public health.

ACS Style

Alessia Libera Gazzonis; Sergio Aurelio Zanzani; Luca Villa; Maria Teresa Manfredi. Toxoplasma gondii in naturally infected goats: Monitoring of specific IgG levels in serum and milk during lactation and parasitic DNA detection in milk. Preventive Veterinary Medicine 2019, 170, 104738 .

AMA Style

Alessia Libera Gazzonis, Sergio Aurelio Zanzani, Luca Villa, Maria Teresa Manfredi. Toxoplasma gondii in naturally infected goats: Monitoring of specific IgG levels in serum and milk during lactation and parasitic DNA detection in milk. Preventive Veterinary Medicine. 2019; 170 ():104738.

Chicago/Turabian Style

Alessia Libera Gazzonis; Sergio Aurelio Zanzani; Luca Villa; Maria Teresa Manfredi. 2019. "Toxoplasma gondii in naturally infected goats: Monitoring of specific IgG levels in serum and milk during lactation and parasitic DNA detection in milk." Preventive Veterinary Medicine 170, no. : 104738.

Protozoology original paper
Published: 15 August 2019 in Parasitology Research
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Blastocystis is a ubiquitous protozoan with a wide range of hosts. In humans, its presence has been associated with gastrointestinal disorders, although its role as a pathogen still needs to be elucidated. Until now, 17 Blastocystis subtypes (STs) have been identified, with ST1–ST4 the most commonly found in humans. Among domestic animals, the same STs reported in humans have been detected in dogs. An epidemiological survey on dog kennels was carried out to evaluate the prevalence of Blastocystis and the STs involved. Overall, 99 faecal samples were collected from the rescue shelters. Blastocystis detection was performed through conventional barcoding PCR targeting the 1800-bp SSU-rDNA, followed by sequencing and phylogenetic analysis. Blastocystis DNA was found in 21 faecal samples (21.2%), and all samples were successfully sequenced and identified as ST3 in a unique monophyletic group. The presence of Blastocystis was reported for the first time in dogs from Italy, with the identification of ST3, the subtype most commonly found in humans.

ACS Style

Alessia Libera Gazzonis; Marianna Marangi; Sergio Aurelio Zanzani; Luca Villa; Annunziata Giangaspero; Maria Teresa Manfredi. Molecular epidemiology of Blastocystis sp. in dogs housed in Italian rescue shelters. Parasitology Research 2019, 118, 3011 -3017.

AMA Style

Alessia Libera Gazzonis, Marianna Marangi, Sergio Aurelio Zanzani, Luca Villa, Annunziata Giangaspero, Maria Teresa Manfredi. Molecular epidemiology of Blastocystis sp. in dogs housed in Italian rescue shelters. Parasitology Research. 2019; 118 (10):3011-3017.

Chicago/Turabian Style

Alessia Libera Gazzonis; Marianna Marangi; Sergio Aurelio Zanzani; Luca Villa; Annunziata Giangaspero; Maria Teresa Manfredi. 2019. "Molecular epidemiology of Blastocystis sp. in dogs housed in Italian rescue shelters." Parasitology Research 118, no. 10: 3011-3017.

Protozoology original paper
Published: 20 February 2019 in Parasitology Research
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A sample of the diaphragm was collected from each of 100 wild boars legally hunted in the Val Grande National Park in north-western Italy and examined for the presence of Sarcocystis infection by histological and molecular methods. In histological sections, thick-walled sarcocysts consistent with those of Sarcocystis miescheriana were detected in 32 wild boars. Genomic DNA extracted from diaphragm samples was initially subjected to PCR amplification of the internal transcribed spacer 1 (ITS1) region, and 97 wild boars were found to harbour a Sarcocystis infection at this screening. Selected DNA samples were then subjected to PCR amplification and sequencing of the ITS1 region and the 18S and 28S ribosomal RNA (rRNA) genes of the nuclear ribosomal DNA unit, while all positive samples were subjected to PCR amplification of the mitochondrial cytochrome c oxidase subunit I (cox1) gene. S. miescheriana was identified in 97 wild boars (97%), while the zoonotic Sarcocystis suihominis was identified in one wild boar (1%), which also harboured S. miescheriana. Intra-specific sequence variation was found in all four DNA regions of S. miescheriana examined and in the 18S rRNA gene and ITS1 region of S. suihominis. The partial cox1 gene was amplified and sequenced from 72 isolates of S. miescheriana, yielding 43 haplotypes with pairwise sequence identities of 97.6–99.9%. These haplotypes were 79.1–79.8% identical with the cox1 sequence of S. suihominis. Phylogeny based on cox1 sequences placed S. miescheriana and S. suihominis as sister species within a clade comprising mainly Sarcocystis spp. of ruminants with felids as known or presumed definitive hosts. The same was true for the phylogeny based on 18S rRNA gene sequences.

ACS Style

Alessia Libera Gazzonis; Bjørn Gjerde; Luca Villa; Stefano Minazzi; Sergio Aurelio Zanzani; Pietro Riccaboni; Giuseppe Sironi; Maria Teresa Manfredi. Prevalence and molecular characterisation of Sarcocystis miescheriana and Sarcocystis suihominis in wild boars (Sus scrofa) in Italy. Parasitology Research 2019, 118, 1271 -1287.

AMA Style

Alessia Libera Gazzonis, Bjørn Gjerde, Luca Villa, Stefano Minazzi, Sergio Aurelio Zanzani, Pietro Riccaboni, Giuseppe Sironi, Maria Teresa Manfredi. Prevalence and molecular characterisation of Sarcocystis miescheriana and Sarcocystis suihominis in wild boars (Sus scrofa) in Italy. Parasitology Research. 2019; 118 (4):1271-1287.

Chicago/Turabian Style

Alessia Libera Gazzonis; Bjørn Gjerde; Luca Villa; Stefano Minazzi; Sergio Aurelio Zanzani; Pietro Riccaboni; Giuseppe Sironi; Maria Teresa Manfredi. 2019. "Prevalence and molecular characterisation of Sarcocystis miescheriana and Sarcocystis suihominis in wild boars (Sus scrofa) in Italy." Parasitology Research 118, no. 4: 1271-1287.

Journal article
Published: 13 October 2018 in Comparative Immunology, Microbiology and Infectious Diseases
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Toxoplasma gondii is a zoonotic parasite infecting a wide range of intermediate hosts, including birds. Nevertheless, scant information on the spread of infection in wild bird populations is available to date. With the aim of updating information on T. gondii infection in birds of prey and possible risk factors associated with the infection, a serosurvey was planned on both wild and captive raptors. An overall of 93 raptors from Northern Italy were tested for the presence of anti-T. gondii antibodies with a commercial modified agglutination test (MAT). A T. gondii prevalence of 10.7% was recorded; the highest seroprevalence was observed within the Family Strigidae (12.5%). Only wild animals tested positive; any statistical difference among species, taxonomic family, age, origin, use, migratory behavior and diet composition was not highlighted. Toxoplasmosis in birds of prey, sentinel species for the environmental spread of T. gondii, should always be considered within sanitary programs devoted to avian species protection.

ACS Style

Alessia Libera Gazzonis; Sergio Zanzani; Azzurra Santoro; Fabrizia Veronesi; Emanuela Olivieri; Luca Villa; Emanuele Lubian; Silvia Lovati; Fulvio Bottura; Sara Epis; Maria Teresa Manfredi. Toxoplasma gondii infection in raptors from Italy: Seroepidemiology and risk factors analysis. Comparative Immunology, Microbiology and Infectious Diseases 2018, 60, 42 -45.

AMA Style

Alessia Libera Gazzonis, Sergio Zanzani, Azzurra Santoro, Fabrizia Veronesi, Emanuela Olivieri, Luca Villa, Emanuele Lubian, Silvia Lovati, Fulvio Bottura, Sara Epis, Maria Teresa Manfredi. Toxoplasma gondii infection in raptors from Italy: Seroepidemiology and risk factors analysis. Comparative Immunology, Microbiology and Infectious Diseases. 2018; 60 ():42-45.

Chicago/Turabian Style

Alessia Libera Gazzonis; Sergio Zanzani; Azzurra Santoro; Fabrizia Veronesi; Emanuela Olivieri; Luca Villa; Emanuele Lubian; Silvia Lovati; Fulvio Bottura; Sara Epis; Maria Teresa Manfredi. 2018. "Toxoplasma gondii infection in raptors from Italy: Seroepidemiology and risk factors analysis." Comparative Immunology, Microbiology and Infectious Diseases 60, no. : 42-45.

Journal article
Published: 01 October 2018 in Parasitology International
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Among Apicomplexa protozoa infecting equids, Besnoitia spp., Toxoplasma gondii and Neospora spp. represent important issues from a sanitary and zootechnical viewpoint. However, only scarce epidemiological data are available on the spread of the infections in horses and donkeys in Europe. Therefore, a serosurvey was planned to estimate the prevalence of these Sarcocystidae species in Italian equids. Serum samples from 268 horses and 18 donkeys raised in Italy were collected and serologically analyzed to detect anti-Besnoitia spp., anti-T. gondii and anti-Neospora spp. antibodies: an approach based on an initial screening by in-house ELISA followed by a confirmatory WB was used. Two horses (0.7%) and four donkeys (22.2%), showed antibodies anti-Besnoitia spp. Ten horses (3.7%) resulted positive to T. gondii and one of these (0.4%) was seropositive also to Neospora spp. This is the first detection of anti-Besnoitia spp. specific antibodies in Italian horses and donkeys. The study confirmed the circulation of Besnoitia spp. among equids in Europe. Low prevalence of T. gondii and Neospora spp. in horses raised in Italy was reported. Nevertheless, it is noteworthy to consider that consumption of horse meat could represent a source for human toxoplasmosis.

ACS Style

Luca Villa; Alessia Libera Gazzonis; Gema Álvarez-García; Carlos Diezma Díaz; Sergio Aurelio Zanzani; Maria Teresa Manfredi. First detection of anti- Besnoitia spp. specific antibodies in horses and donkeys in Italy. Parasitology International 2018, 67, 640 -643.

AMA Style

Luca Villa, Alessia Libera Gazzonis, Gema Álvarez-García, Carlos Diezma Díaz, Sergio Aurelio Zanzani, Maria Teresa Manfredi. First detection of anti- Besnoitia spp. specific antibodies in horses and donkeys in Italy. Parasitology International. 2018; 67 (5):640-643.

Chicago/Turabian Style

Luca Villa; Alessia Libera Gazzonis; Gema Álvarez-García; Carlos Diezma Díaz; Sergio Aurelio Zanzani; Maria Teresa Manfredi. 2018. "First detection of anti- Besnoitia spp. specific antibodies in horses and donkeys in Italy." Parasitology International 67, no. 5: 640-643.

Journal article
Published: 01 October 2018 in Journal of Parasitology
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Alessia Libera Gazzonis; Sergio Aurelio Zanzani; Katia Stradiotto; Emanuela Olivieri; Luca Villa; Maria Teresa Manfredi. Toxoplasma gondii Antibodies in Bulk Tank Milk Samples of Caprine Dairy Herds. Journal of Parasitology 2018, 104, 560 -565.

AMA Style

Alessia Libera Gazzonis, Sergio Aurelio Zanzani, Katia Stradiotto, Emanuela Olivieri, Luca Villa, Maria Teresa Manfredi. Toxoplasma gondii Antibodies in Bulk Tank Milk Samples of Caprine Dairy Herds. Journal of Parasitology. 2018; 104 (5):560-565.

Chicago/Turabian Style

Alessia Libera Gazzonis; Sergio Aurelio Zanzani; Katia Stradiotto; Emanuela Olivieri; Luca Villa; Maria Teresa Manfredi. 2018. "Toxoplasma gondii Antibodies in Bulk Tank Milk Samples of Caprine Dairy Herds." Journal of Parasitology 104, no. 5: 560-565.

Journal article
Published: 11 May 2018 in Parasitology Research
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Wild boar is a source of human infections with zoonotic pathogens, including food-borne parasites. With the aim of a characterization of the human exposure risk, a survey on wild boars intended for human consumption was planned, selecting three pathogens, Toxoplasma gondii, Alaria alata, and Trichinella spp., as markers of meat infection. Diaphragm muscle samples from 100 wild boars hunted in Piedmont region (Northern Italy) in two hunting seasons (2015-2016) were collected. Concerning T. gondii, a combined approach of antibody detection and molecular techniques with genotyping was performed. For the detection of A. alata and Trichinella spp., the larva migration technique and the magnetic stirrer method were employed, respectively; in addition, molecular confirmation of the morphological identification of the recovered specimen was performed. Anti-T. gondii antibodies were found in meat juice samples (43.3%) and T. gondii DNA (type II) was detected in three animals (7.1%) out of 42 seropositive examined. In none of the sampled wild boars (0%), Trichinella spp. larvae were found, whereas one animal (1%) scored positive to A. alata mesocercariae. The molecular diagnosis proved the morphological identification of the trematode. This is the first finding of A. alata in Italian wild boar population. The present study confirmed the role of wild boars as a source of parasitic zoonotic diseases and thus the risk derived for humans posed by the consumption of game meat. Considering the zoonotic implications, the results underline the importance of monitoring and surveillance of zoonotic parasites in Italian wild boar populations.

ACS Style

Alessia Libera Gazzonis; Luca Villa; Katharina Riehn; Ahmad Hamedy; Stefano Minazzi; Emanuela Olivieri; Sergio Aurelio Zanzani; Maria Teresa Manfredi. Occurrence of selected zoonotic food-borne parasites and first molecular identification of Alaria alata in wild boars (Sus scrofa) in Italy. Parasitology Research 2018, 117, 2207 -2215.

AMA Style

Alessia Libera Gazzonis, Luca Villa, Katharina Riehn, Ahmad Hamedy, Stefano Minazzi, Emanuela Olivieri, Sergio Aurelio Zanzani, Maria Teresa Manfredi. Occurrence of selected zoonotic food-borne parasites and first molecular identification of Alaria alata in wild boars (Sus scrofa) in Italy. Parasitology Research. 2018; 117 (7):2207-2215.

Chicago/Turabian Style

Alessia Libera Gazzonis; Luca Villa; Katharina Riehn; Ahmad Hamedy; Stefano Minazzi; Emanuela Olivieri; Sergio Aurelio Zanzani; Maria Teresa Manfredi. 2018. "Occurrence of selected zoonotic food-borne parasites and first molecular identification of Alaria alata in wild boars (Sus scrofa) in Italy." Parasitology Research 117, no. 7: 2207-2215.

Original paper
Published: 03 January 2018 in Parasitology Research
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Toxoplasma gondii is a widespread protozoan affecting animals and humans. One of the major routes of human infection is through the consumption of raw or under-cooked meat, particularly of certain animal species, including pigs. Although T. gondii represents an important public health issue, its control at slaughter is not mandatory. Consequently, available information on T. gondii infection in domestic animals destined for human consumption is scarce. Thus, an epidemiologic survey was designed to update information on T. gondii infection in pigs from intensive production. Fattening pigs and sows from conventional farms were sampled. Sera were tested with a commercial ELISA for anti-T. gondii antibodies, whereas molecular analysis by 529 bp repetitive element PCR and B1 real-time PCR with subsequent genotyping was performed on heart samples. Statistical analysis was carried out to detect farm management features and sanitary procedures enhancing the risk of infection. At the farm level, 63.6% (7/11) of farms housing sows and 6.7% (1/15) housing fattening pigs scored positive, with individual prevalences of 8.6% (13/151) in sows and 0.5% (1/219) in fattening pigs. T. gondii DNA was detected in eight sows and one fattening pig, and sequencing revealed the presence of all three genotypes (types I, II, and III). Furthermore, the decrease in the biosecurity level enhanced the risk of infection within a farm. The present survey outcomes confirm the spread of T. gondii infection in pig farms in an area of intensive swine production. The application of stricter sanitary procedures may contribute to a further reduction of infection.

ACS Style

Alessia Libera Gazzonis; Marianna Marangi; Luca Villa; Maria Elena Ragona; Emanuela Olivieri; Sergio Aurelio Zanzani; Annunziata Giangaspero; Maria Teresa Manfredi. Toxoplasma gondii infection and biosecurity levels in fattening pigs and sows: serological and molecular epidemiology in the intensive pig industry (Lombardy, Northern Italy). Parasitology Research 2018, 117, 539 -546.

AMA Style

Alessia Libera Gazzonis, Marianna Marangi, Luca Villa, Maria Elena Ragona, Emanuela Olivieri, Sergio Aurelio Zanzani, Annunziata Giangaspero, Maria Teresa Manfredi. Toxoplasma gondii infection and biosecurity levels in fattening pigs and sows: serological and molecular epidemiology in the intensive pig industry (Lombardy, Northern Italy). Parasitology Research. 2018; 117 (2):539-546.

Chicago/Turabian Style

Alessia Libera Gazzonis; Marianna Marangi; Luca Villa; Maria Elena Ragona; Emanuela Olivieri; Sergio Aurelio Zanzani; Annunziata Giangaspero; Maria Teresa Manfredi. 2018. "Toxoplasma gondii infection and biosecurity levels in fattening pigs and sows: serological and molecular epidemiology in the intensive pig industry (Lombardy, Northern Italy)." Parasitology Research 117, no. 2: 539-546.