Absence of Viable Toxoplasma gondii in Artisanal Raw-Milk Ewe Cheese Derived from Naturally Infected Animals

The presence of viable Toxoplasma gondii was investigated in artisanal cheeses made from milk of naturally infected ewes. Ewe milk was analyzed beforehand for the presence and vitality of T. gondii by loop-mediated isothermal amplification (LAMP) and reverse-transcriptase PCR (RT-PCR), respectively. Cheeses were prepared from raw milk following a traditional cheesemaking process. The cheese obtained from T. gondii-positive milk was analyzed by LAMP to detect Toxoplasma DNA-positive samples. RT-PCR was then carried out to assess the viability of the parasites in T. gondii-positive milk samples and fresh cheese, after 5 and 15 days of ripening. Physical-chemical parameters of cheeses were also investigated. All cheese samples derived from T. gondii-positive milk were positive according to LAMP, at both 5 and 15 days of ripening, while none of the samples were positive according to RT-PCR. Thus, while the presence of the parasite was demonstrated by the detection of specific DNA, the absence of detectable T. gondii RNA supports the hypothesis that changes in the chemical and physical characteristics occurring during the cheesemaking process and ripening period, could be sufficient to inactivate viable T. gondii in milk, minimizing the risk of human infection through consumption of raw sheep milk cheese

Prevalence and risk factors associated with cat parasites in Italy: a multicenter study

Background: Parasites that infect cats include protozoa, helminths and arthropods, many of which are transmissible to humans. Effective control relies on a good knowledge of parasite distribution and the risk factors for infection. The present study was aimed at evaluating the prevalence of major feline parasites in Italy and the risk factors associated with their occurrence. Methods: Over a 12-month study period, feces, hair and ectoparasites from naturally infected cats from feral colonies, shelters and private households were analyzed at 13 study centers across Italy. Samples from these cats (n = 987) were analyzed at all centers using the same diagnostic methods. Prevalence values and risk factors were evaluated statistically for the identification of predictors of risk. Results: The overall prevalence of gastro-intestinal and broncho-pulmonary (BP) nematodes was 35.9% (354/987). Toxocara cati was the most prevalent species (253/987; 25.6%), followed by Ancylostomatidae (98/987; 9.9%). Among BP nematodes, Aelurostrongylus abstrusus was the most common (76/987; 7.7%). Approximately 35.7% (352/987) of the study population was infested by ectoparasites, of which the most common were fleas (29.4%, 290/987), followed by ear mites Otodectes cynotis (9.8%, 97/987). Predictors of risk for parasite infection included age, a predominantly or exclusively outdoor lifestyle, geographic area and lack of antiparasitic treatment. Conclusions: Both ecto- and endoparasites are still common in cats throughout Italy, many of them being of zoonotic concern and vectors of pathogens to humans. Given the presence of parasites throughout the entire study period, year-round treatment should be considered. Furthermore, data confirm the need to protect the human–animal bond using proper endo- and ectoparasiticides to reduce the risk of human infection, in application of the One-Health concept

Eucoleosi nasale clinicamente manifesta in cani provenienti dal Centro Italia

In the past few years the respiratory nematode Eucoleus boehmi has been increasingly reported in dogs from Europe, including Italy. Nonetheless, knowledge of the clinical importance of this parasitosis in canine practice is still poor. The present work describes 20 clinical cases of canine nasal eucoleosis (CNE) from Central Italy in 2012-2013, to provide new insights into the clinical features of the infection and to promote an appropriate awareness among veterinarians. Twenty dogs, scored positive for E. boehmi eggs at copromicroscopical examination, were selected and history details and clinical data were collected. Most of the infected animals (70%) consisted on hunting dogs which lived in rural or suburban areas of central Italy. All parasitised dogs showed different clinical pictures, being nasal discharge (16/20) the most frequent sign, followed by repeated sneering (11/20) and cough (7/20). Other respiratory signs, i.e. epistaxis, and impairment of scenting ability were observed. These results confirm that E. boehmi has a clinical importance on first airway diseases and demonstrate that veterinarians should be vigilant on its occurrence

Ascaridia galli: a report of erratic migration

This paper describes a case of an unusual recovery of adult Ascaridia galli in hen’s egg. Several data are available on this occurrence but it appears to be the first case described in Italy. The worm was identified as an adult female, 6.8 cm in length, with three trilobed lips, cervical narrow alae, oesophagus club-shaped without posterior bulb, vulva near the middle of body, with gravid uteri containing a large number of eggs. The presence of Ascaridia galli in hen’s eggs cannot be considered as hazard for public health but may be cause of a potential consumer complaint. Moreover it is a sign of presence of ascaridiosis, parasitosis that still produces economic losses in modern poultry production system

Detection of Toxoplasma gondii in faeces of privately owned cats using two PCR assays targeting the B1 gene and the 529-bp repetitive element

Toxoplasma gondii infection is a worldwide parasitic zoonosis with a high-health risk for humans. The key epidemiological role played by felids is related to oocyst shedding. The present study compared two amplification protocols for the molecular diagnosis of Toxoplasma infection in owned cats. A total of 78 owned cats referred to an Italian university-teaching hospital and exposed to various T. gondii-associated risk factors were sampled for blood and faeces. Faecal specimens were processed by flotation and tested using 2 copro-PCRs targeting the widely used B1 gene and the 529-bp repetitive element (RE). The sera were tested by the indirect immunofluorescent antibody test (IFAT) for the detection of immunoglobulins against T. gondii. Sixteen faeces (20.52%) tested positive for T. gondii DNA; 12 samples were positive only at B1-PCR, two at 529-bp RE-PCR and two at both genetic targets (overall agreement = 82.11%). The amplicons obtained were sequenced, and the Basic Local Alignment Search Tool analysis showed a high homology with the T. gondii strains available in reference databases. Two stool samples were microscopically positive for T. gondii-like oocysts and also tested positive by both B1 and 529-bp RE-PCRs. Thirty-three (42.3%) sera tested positive for antibodies; of which, seven were found to have T. gondii DNA-positive results using the B1 genetic target (overall agreement = 57.77%). The amplification sets targeting B1 and 529-bp RE showed substantially different yields. Further research is needed to better understand the significance and the sensitivities of using these multi-copy-targeted molecular methods from cat faeces before being used for routine diagnosis

Metastrongyloid infection by Aelurostrongylus abstrusus, Troglostrongylus brevior and Angiostrongylus chabaudi in a domestic cat

The "cat lungworm", Aelurostrongylus abstrusus, affects cats worldwide. Recently, other cardio-pulmonary parasites, e.g. Troglostrongylus brevior and Angiostrongylus chabaudi, have been isolated from cats either for the first time or a long time after they were first described. This paper describes the first known mixed infection by A. abstrusus, T. brevior and A. chabaudi in a domestic cat. Biological and epidemiological implications of old and "new" metastrongyloid-caused infections in cats are discussed

Parasitological Survey in European Brown Hare (Lepus europaeus Pallas, 1778) Breeding Facilities in Southern Italy

Parasites are considered important regulating factors of hosts’ population dynamics, not only in free-ranging wildlife, but also in captive bred animals. To date, only few studies have been carried out to assess the parasitic communities of the European brown hare in Southern Italy, and only one focused on animals in captivity. The aim of the present survey was to assess the composition of the endoparasite community in game hares bred for restocking purposes. For this purpose, 215 fecal pools collected in eight different breeding facilities were examined by qualitative and quantitative coprological techniques. Parasites characterized by a direct life cycle, including six species of coccidia from the genus Eimeria and the nematode Trichostrongylus retorataeformis, proved to be the most prevalent parasites. Further helminthic infestations by Passalurus ambiguous, Strongylosides papillosus, Cittotenia spp. and Dicrocoelium dendriticum were also detected, but with an overall prevalence lower than 20%. The present study contributes to increasing knowledge on the health status of a poorly investigated species, and is useful for optimizing breeding efforts in captivity

Sequence variation in the B1 gene among Toxoplasma gondii isolates from swine and cats in Italy

The evaluation of the genetic variations of Toxoplasma gondii among isolates of a wide variety of animal hosts can provide significant information for better understanding the epidemiology and population structure of the parasite in different geographical areas. The aim of this study was to provide information on T. gondii genetic diversity in host species living in central Italy, which could act as a potential source of human infection. Seventyseven feline faecal samples, and 36 and 20 diaphragm pillar tissue samples from pigs and wild boars were collected in Umbria (central Italy). The samples were tested by a nested-PCR protocol amplifying an informative region within the B1 gene, a multi-copy genetic target, showing a good rate of variability. Thirty-six specimens (27.07%) belonging to 10 pigs, 13 wild boars and 13 cats, tested positive to the B1 nested-PCR screening. Of these, 23 good quality sequences (8 from wild boars, 5 from pigs, and 10 from cats) were analyzed. A comparison of the B1 DNA sequences showed that a single homogeneous nucleotide substitution (C/T) was present at position 31 in the isolates from pigs and wild boars compared with the sampled cats and other hosts (including humans) available in GenBankr (TM). The present results suggest the existence of a T. gondii genetic diversity for swine host species, based on a SNP (C/T) of the B1 gene. Further studies are needed to draw more solid conclusions on the discriminatory power of the B1 target by collecting more swine samples from much broader geographical areas

Morishitium polonicum as a Cause of Severe Respiratory Disease in Eurasian Blackbirds (Turdus merula) in Central Italy

Two Eurasian Blackbirds (Turdus merula) from central Italy were found with severe cyclocoelid trematodosis associated with airsacculitis. The birds were submitted with severe respiratory distress; one died shortly after hospitalization, while the second bird was euthanized. At necropsy, a massive presence of cyclocoelid flukes was observed in the coelomic cavity and air sacs of both birds. The air sacs were diffusely opaque, thickened, and covered by scant fibrinous exudate mixed with numerous parasites. Histologically, the air sacs showed diffuse and severe oedema with fibrinous exudate. Diffuse mononucleated and heterophilic infiltration mixed with multiple granulomas contained degenerated trematodes. Morishitium polonicum was identified using morphologic keys and molecular analysis of extracted DNA. Infections caused by M. polonicum are poorly documented in blackbirds and the findings in these birds support the pathogenic role of this trematode as a potential cause of death in blackbirds in Italy. Extended epidemiologic surveys are required to properly assess the potential importance of M. polonicum as a life-threatening pathogen in Blackbird populations