Cryptosporidium infections in sheep farms from Italy.

Cryptosporidiosis is recognized as being a significant cause of gastrointestinal illness due to its wide range of vertebrate hosts, including humans. Infection with Cryptosporidium spp. is especially common in young domestic ruminants (calves, lambs and goat kids) and has been associated with economic losses worldwide. In contrast to cattle, to date, detailed studies on Cryptosporidium infections in sheep from Europe are still limited; thus, their importance as reservoirs of Cryptosporidium species with implications on animal and public health still needs to be clarified. This study evaluates the prevalence and zoonotic potential of Cryptosporidium spp. in sheep farms in Italy. A total of 915 individual faecal samples divided into three different animal categories were collected from 61 sheep farms. Each sample was examined by microscopy of faecal smears stained by modified Ziehl-Neelsen and by biomolecular techniques. Cryptosporidium oocysts were detected in 10.1% of the animals examined and in 34.4% of the farms. The prevalence of Cryptosporidium spp. was significantly higher (χ2 = 51.854; P < 0.001) in diarrhoeic samples than in pasty or normal faeces. Genotype analyses showed the presence of two Cryptosporidium species: C. parvum and C. ubiquitum. Subtyping analysis of C. parvum isolates revealed the presence of subtypes IIa15G2R1 and IIdA20G1 and of subtype XIIa for C. ubiquitum. These findings have public health implications since both Cryptosporidium species identified are considered zoonotic, and C. parvum is the second-most common Cryptosporidium species infecting humans. Our data reveal that lambs, especially those excreting diarrhoeic faeces, may be important reservoirs of Cryptosporidium. We also highlight the need to establish adequate control and monitoring programmes for the control of this infection in sheep farms primarily through coprological monitoring.

Molecular characterisation and risk factor analysis of Cryptosporidium spp. in calves from Italy.

To provide up-to-date information on the occurrence of Cryptosporidium in pre-weaned calves from Sardinia (Italy), the species implicated and their zoonotic potential, 147 faecal samples from 22 cattle herds were microscopically examined for Cryptosporidium oocysts; positive isolates were molecularly characterised. A questionnaire was developed to identify risk factors for Cryptosporidium infection. Overall, the percentage of positive calves and farms was 38.8 and 68.2%, respectively. The SSU rRNA-based PCR identified two Cryptosporidium species, Cryptosporidium parvum (95.8%) and C. bovis (4.2%). Sequence analyses of the glycoprotein (gp60) gene revealed that all C. parvum isolates belonged to the subtype family IIa (IIaA15G2R1 and IIaA16G3R1), with the exception of three isolates that belonged to the subtype family IId (IIdA20G1b and IIdA20). Mixed logistic regression results indicated that calves aged 15-21 days were more likely to be Cryptosporidium-positive. The risk of being positive was also significantly higher in herds from Central Sardinia and in farms using non-slatted flooring. In addition, the application of disinfectants and milk replacers was significantly associated with higher Cryptosporidium prevalence. In contrast, the risk of being positive was significantly reduced in halofuginone-treated calves. Our results reveal that a significant percentage of suckling calves are carriers of zoonotic subtypes of C. parvum. Thus, both healthy and diarrhoeic calves younger than 1 month may represent a risk for the transmission of cryptosporidiosis in humans and animals.

Seroepidemiological and biomolecular survey on Toxoplasma gondii infection on organic pig farms.

Pigs are an important reservoir of Toxoplasma gondii, and pork meat is considered one of the main sources of human infection. The present survey assesses the prevalence of T. gondii in organically reared pigs in Sardinia found on the island using various diagnostic methods. Firstly, a seroepidemiological survey was carried out on 63 farms, examining individual blood samples from 414 adult pigs. Secondly, a biomolecular investigation carried out on another 62 farms, examining the brain samples from 107 adult pigs. The seroprevalence detected for T. gondii at the farm level was 85.7% (54 out of the 63 pig farms examined). However, the prevalence rate in the investigated pigs was 51.7% (214 positives out of 414 examined). A total of 54.8% (34/62) of the monitored pig farms were found to be positive by PCR. The presence of T. gondii DNA in individual brain samples was detected in 47.7% (51/107) of the examined animals. The high prevalence of T. gondii found is alarming and suggests that pigs reared on organic farms should be considered as a significant source of infection for humans in the areas investigated.

Serological and molecular detection of Leishmania infantum in cats of Northern Sardinia, Italy.

Leishmania infections are endemic in dogs in central and southern regions of Italy while feline leishmaniosis has only been sporadically described in these areas and few studies have been reported on serological and molecular investigation of infections in domestic cats. The purpose of this study is to assess the presence of Leishmania infantum infection in cats from Sardinia, a typical endemic area of leishmaniasis in Italy. Ninety cats attended at the Veterinary Teaching Hospital of the University of Sassari, Italy, were tested for L. infantum using the indirect immunofluorescent antibody (IFA) test and PCR. Serological (10%) and molecular (5.5%) detection in this population was low, with antibody titres ranging from 1/40 to 1/80. Only one cat was simultaneously positive by IFAT and PCR. This study shows the presence of L. infantum infection in cats living in Sardinia, with a prevalence that falls within the ranges described in other endemic areas and confirms the need of further research to better characterize the epidemiologic role of cats in the transmission of this infection.

First record of Gongylonema nepalensis in domestic and wild ruminants in Europe.

The gullet worm, Gongylonema pulchrum, is cosmopolitan in distribution, infecting a variety of mammals including domestic and wild ruminants. Gongylonema nepalensis recently collected from the esophageal epithelium of water buffaloes in Nepal was separated from G. pulchrum based on its distinctly shorter left spicule relative to body length and unique nucleotide sequences of the ribosomal RNA gene (rDNA) and the mitochondrial cytochrome c oxidase subunit 1 gene (cox-1). During meat inspections at four abattoirs on Sardinia Island, Italy, 25 Gongylonema worms were collected from one each individual of cattle (n=8), sheep (n=7), goats (n=4), and mouflon (Ovis aries musimon; n=6), and characterized morphologically and genetically. Intriguingly, all of the collected worms from these ruminants were G. nepalensis, exhibiting comparable body lengths to G. pulchrum in cattle from other regions but with significantly shorter left spicules like G. nepalensis (less than 20.9% of the entire body length in contrast to 21.8-65.6%, the reported proportion of G. pulchrum). Furthermore, the rDNA nucleotide sequences of these worms from different ruminant species on Sardinia Island were almost identical to each other and to Nepalese G. nepalensis isolates. With the exception of one worm from a sheep (displaying a single nucleotide substitution), the 369-bp cox-1 nucleotide sequences of all the Sardinian G. nepalensis isolates from the different host sources were absolutely identical, forming a clade with Nepalese G. nepalensis isolates and not G. pulchrum isolates. The present study reveals that G. nepalensis is not a local parasite in the Indian subcontinent (Nepal), but instead has a certain geographical distribution in Europe and takes several ruminant species as a definitive host.

Endoparasites detected in faecal samples from dogs and cats referred for routine clinical visit in Sardinia, Italy.

This study aimed to update data on the prevalence of intestinal and lung parasitic infections in owned dogs and cats in Sardinia, Italy. Examinations on faecal samples from 619 dogs and 343 cats routinely referred to the Veterinary Teaching Hospital of the University of Sassari were performed between the years of 2011 and 2015. Individual faecal samples were analysed using the Wisconsin technique for copro-microscopic examination and the Baermann technique for the presence of lungworm larvae. Endoparasites were found in 34.9% and 43.4% of examined dogs and cats, respectively. Helminthic infections (21.2% in dogs and 32.6% in cats) occurred more frequently than protozoan infections (17.9% in dogs and 17.8% in cats). In both dogs and cats, the most common parasites were ascarids (12.1% and 15.7%), Cystoisospora spp. (10.2% and 10.8%), Giardia duodenalis (9.4% and 8.5%), and hookworms (7.9% and 5.5%). Evidence of bronchopulmonary nematode infections were found in 0.8% of examined dogs and in 15.8% of examined cats. Age was identified as a risk factor, with animals younger than 6months more frequently infected than older animals, while no significant association was observed for gender. This study demonstrated that endoparasites in owned dogs and cats of Sardinia have considerably high prevalence. Veterinary practitioners and pet owners should be more aware of these infections and should adopt more effective and standardized control practices.

Acute visceral cysticercosis by Taenia hydatigena in lambs and treatment with praziquantel.

An acute outbreak of Taenia hydatigena cysticercosis, causing mortality in 5 of 21 (23.8%) female lambs, is reported. Gross post-mortem examinations and histology showed Cysticercus tenuicollis as the cause of death. Biochemical parameters in infected lambs confirmed severe hepatitis. Praziquantel, given once at 15 mg/kg body weight (bw), was administered and a dramatic improvement in the clinical condition and biochemical parameters was observed up to 30 days following treatment.

Anthelmintics efficacy against intestinal strongyles in horses of Sardinia, Italy.

Intestinal strongyles (IS) are the most important parasites of equids, due to their high prevalence worldwide, pathogenicity and the spread of drug-resistant populations. Despite the large number of horses bred in Sardinia Island, Italy, no data are available on the efficacy of anthelmintic compounds in the control of horse strongylosis. Therefore the aim of the present study was to evaluate the efficacy of five commercial anthelmintic formulations containing fenbendazole (FBZ), pyrantel (PYR), moxidectin (MOX) and two ivermectin formulations (IVM1 and IVM2) against IS in Sardinia by performing a fecal egg count reduction test (FECRT) and investigating the egg reappearance period (ERP) after treatment. In total, 74 horses from 7 farms were examined. Coprocultures performed for individual fecal samples collected at the day of the treatment revealed that cyathostomins were the predominant parasitic species (98.6%). The FECR for all horses belonging to the treatment groups after two weeks was ≥ 95% with a 95% C.I. > 90%. The expected ERP did not decrease in any of the treatment group as FECR values < 90% were found at D60 for FBZ, at D90 for PYR and IVM1, at D150 for IVM2. All horses treated with MOX showed FECRT > 90% for the entire duration of the trial until D150. The results of the present survey indicate that drug-resistant cyathostomin populations are not present in the examined horse population, contrariwise to what observed in other Italian and European regions. The reasons and implications of these results are discussed.

Epidemiological updates and economic losses due to Taenia hydatigena in sheep from Sardinia, Italy.

The aim of this study was to investigate the epidemiology and transmission of Taenia hydatigena in sheep and dogs from Sardinia and the economic estimation of losses due to this metacestodosis in lambs. A total of 7781 Sarda breed lambs were examined at abattoirs for the detection of Cysticercus tenuicollis or necrotic-haemorrhagic tracks of their migration. Morphological and molecular identification of parasites was carried out. Individual faecal samples from 300 dogs were examined for copromicroscopic investigations and coproELISA assay. An overall prevalence of 14.6% for T. hydatigena cysticercosis was found in the examined lambs. In total, 10,807 parasitary tracks were found, with an abundance of 1.39 and an average intensity of 9.52. The molecular analysis of the isolates showed an overall pairwise nucleotide divergence for the CO1 and ND1 was of 0-3.1 and 0-3.3%, respectively. Low intra- and interspecific variation was recorded for C. tenuicollis isolates used in this study which suggested the absence of differentiation. Microscopic examination of dog faeces showed a total prevalence of 31.3% for endoparasites in the examined samples (94/300). Taeniid eggs were found in 8.3% of the dogs. The results of the monoclonal antibody ATH4 ELISA test showed a prevalence of 11% (33/300) for T. hydatigena coproantigens. The total economic costs related to cysticercosis amounted to almost € 330,000. The prevalence of C. tenuicollis in 14.6% of 30-40-day-old lambs highlights the high parasitic pressure by T. hydatigena in the territory of Sardinia, Italy.

Simultaneous infection by four feline lungworm species and implications for the diagnosis.

Besides Aelurostrongylus abstrusus, other parasites belonging to the superfamily Metastrongyloidea, namely Oslerus rostratus, Troglostrongylus brevior and to the family Trichuridae, i.e. Eucoleus aerophilus (syn. Capillaria aerophila), have also been reported as agents of respiratory infection in domestic cats. A case of simultaneous infection by four feline lungworm species in Sardinia is herein described. An adult female cat (Felis silvestris catus), road-killed in the southeast part of Sardinia (municipality of Villacidro, province of Cagliari), Italy, was referred to the Laboratory of Parasitology of the Veterinary Teaching Hospital in Sassari. At necropsy, the lungs were examined and dissected under a stereomicroscope for the presence of parasites, and first-stage larvae (L1) of broncho-pulmonary nematodes were searched for in a faecal sample using the Baermann method. Parasites collected in the lungs were morphologically identified as A. abstrusus, E. aerophilus, and O. rostratus. In addition to the above species, L1s of Troglostrongylus spp. were detected at coproscopy but no adult specimen was found in the lungs. The morphological identification was confirmed by the molecular amplification and sequencing of cox1 mitochondrial gene, 18S and ITS2 ribosomal DNA. This finding stands as the first simultaneous infection by four feline lungworm species in the same animal, and as the first report of O. rostratus and E. aerophilus in Sardinia.

New insights on metastrongyloid lungworms infecting cats of Sardinia, Italy.

In addition to the well-known Aelurostrongylus abstrusus (Strongylida: Angiostrongylidae), Troglostrongylus brevior (Strongylida: Crenosomatidae) has recently been diagnosed as a causative agent of bronco-pulmonary infections of cats in Spain and Italy. However, information concerning the impact of this species of lungworm on feline population is limited to a few case reports. From July 2011 to May 2013 an epidemiological survey was carried out on Sardinia island (Italy), where 107 individual faecal samples were examined by Baermann technique, and first-stage larvae were identified based on their morphology and characterization of molecular markers. The 29.9% (32/107) of cats examined were infested by broncho-pulmonary nematodes and, although A. abstrusus was the most frequently detected (n=27; 25.2%), larvae of T. brevior were also found (n=7; 6.5%). In addition, two cats (1.9%) were co-infested by both species. Overall metastrongyloid infection was higher in female cats (n=22; 38.6%) than in males (n=10; 20%) (χ(2)=4.39; p=0.036). The mean age of positive animals was 21.1 (±29.8) months, being infected animals from 2 months to 10 years of age. Of the 32 animals that scored positive for lungworms only 6 (18.8%) displayed a respiratory condition associated with lungworm infestations. Biomolecular characterization confirmed the morphological diagnosis of A. abstrusus. Positive samples that were identified at genus level as Troglostrongylus spp. were molecularly characterized as T. brevior. This study represents the first epidemiological survey on metastrongyloid lungworms of domestic cats in Sardinia and the first report of T. brevior on this island.

Ultrasound as a monitoring tool for cystic echinococcosis in sheep.

An ultrasound-based survey for cystic echinococcosis (CE) in sheep was carried out in Sardinia in 2012. The study was done on three farms (A, B, C) which had been pre-selected for different CE prevalence levels (A: >80%, B: 50-80%, C: <50%). In total, 129 sheep were examined on the farms using portable ultrasound equipment (A: n=51, B: n=30, C: n=48). Within a period of 20 days after ultrasound examination, all sheep were slaughtered and underwent a parasitological post-mortem examination for cysts in the liver and lungs. With post-mortem as gold standard, ultrasonography gave a test sensitivity of 88.7% and a specificity of 75.9%, while the positive and negative predictive values were 81.8% and 84.6%, respectively. When only sheep with fertile cysts were considered, the sensitivity of the test increased to 100%. We conclude that the ultrasound examination of the liver in sheep - using state-of-the-art technology - is a sensitive and specific diagnostic tool, which is cost-effective, highly appropriate for field use and requires only moderate time (no shaving required). The method can also be applied to other livestock species and will be useful tool in epidemiological studies, monitoring schemes and vaccination/control trials.

New insights onto cardiopulmonary nematodes of dogs in Sardinia, Italy.

Dog heartworms Angiostrongylus vasorum and Dirofilaria immitis cause severe parasitological diseases; the importance of these parasitosis is growing due to their health impact on animals, the possible zoonotic implications and the recent spreading across several European countries and previously non-endemic areas. The aim of this study is to update the epidemiological scenario of cardiopulmonary nematodes A. vasorum and D. immitis in dogs of Sardinia island and to perform a morphological identification of larvae by the use of the Baermann and Knott techniques respectively and the molecular characterization of the mitochondrial cytochrome c oxidase subunit I (cox1) and the second ribosomal transcribed spacer region (ITS-2) of larvae L1 of A. vasorum. In the present study, 3.4% (5/146) of dogs resulted positive at Baermann technique for A. vasorum while 8.9% (61/684) to D. immitis. If on one side A. vasorum can be considered an emerging parasite in Sardinia, the parasitic pressure and the risk of infection for D. immitis in the island seems to be increased compared with the recent past.

Cercopithifilaria spp. in dogs in Sardinia Island (Italy).

A survey on Cercopithifilaria spp. was carried out on owned and kennelled dogs in Sardinia, Italy. A total of 180 dogs were sampled and tested by microscopic detection or PCR of dermal microfilariae in skin snip sediments. The overall prevalence for Cercopithifilaria spp. at both microscopy and molecular tests was 9.4 % (17/180), while 8.3 % (15/180) of dogs scored positive at microscopic detection of sediments only. Of the 225 microfilariae measured, 212 were identified as Cercopithifilaria bainae and the remaining as Cercopithifilaria sp. II. All samples were molecularly processed for specific amplification of cytochrome oxidase subunit 1 (cox1) and ribosomal 12S gene fragments. The Basic Local Alignment Search Tool analysis of the cox1 and 12S sequences here obtained showed a high nucleotide similarity (99 and 100 %, respectively) with those of C. bainae available in GenBank. In particular, cox1 haplotype I (HI; n=14), haplotype HXVIII (n=2), and a new haplotype, named HXIX (n=1), differing for a single polymorphism from HI, were detected. This study reports data on the occurrence, distribution, and genetic makeup of C. bainae and Cercopithifilaria sp. II infesting dogs in Sardinia, suggesting that these filarioids are spread in areas where Rhipicephalus sanguineus sensu lato ticks occur.

Morphological and molecular characterization of bovine coenurosis in Sardinia, Italy.

Coenurosis is a central nervous system disease of wild and domestic ruminants caused by Coenurus cerebralis, a bladder worm stage of Taenia multiceps). Even in Sardinia island, this metacestode seems to be widespread in sheep (Scala et al. Vet Parasitol 143(3-4):294-298, 2007) where coenurosis is an important health problem (Varcasia et al. Parasitol Res 99(5):622-626, 2006) the last and unique report of coenurosis in cattle was in 1990 (Cubeddu et al. 1990). In the present paper, a case of bovine coenurosis in Sardinia was described 22 years after the first report with a morphological a biomolecular characterization. A 2-year-old Limousine bull was euthanized in the Bolotana (NU) municipality (Central Sardinia). The remote anamnesis achieved from the farmer reporting that the bull showed neurological symptoms from 1 year of age previously classified as nutritional problems by the farm's veterinary. The breeder also says that the bull have by self-produced the skull fracture by hitting a gaff in the farm. The skull was opened and the brain removed and carefully examined showing two coenurus cysts containing clear fluid with numerous scoleces both in the right hemisphere. Morphological features of the cysts and mt-DNA sequencing confirm that the parasites were T. multiceps Coenuri.

Epidemiology, chronobiology and taxonomic updates of Rhinoestrus spp. infestation in horses of Sardinia Isle, Western Mediterranean (Italy).

From January to December 2008, 265 horses slaughtered in Sardinia (Italy) were examined for the presence of Rhinoestrus spp. (Diptera: Oestridae) through the examination of the nasal cavities and pharynges. Larvae were detected in 49% of the horses, with a mean intensity of infestation of 16.09 and abundance of 7.95. A total of 2108 larvae were collected, 66% of which were classified in first instar (L1), 22% in second instar (L2) and 12% in third instar (L3). The most frequent localization of larvae was the ethmoid, while the less one the larynx. According to the dynamics of Rhinoestrus larval stages, three periods in the chronobiology can be considered, the diapause (September-February) characterized by an absolute prevalence of first larval stage; the active phase of the endogenous phase (February-September) with an increase in the percentages of L2 and L3, and the exit phase (May-September), pointed by a further increase of L1. Morphological examination of L3 larvae revealed the presence of the Rhinoestrus purpureus features in 8% of the examined larvae, of 8% of the Rhinoestrus usbekistanicus features, while in 84% of the larvae were evidenced intermediate features. Contrastingly biomolecular analysis of the COI gene of the larvae evidenced uniformity at genetic level, confirming the presence of a unique species in the Mediterranean area. The results of the present paper, reveal the wide diffusion of rhinoestrosis among Sardinian horses, and suggest the need for applying appropriate control measures. Chemotherapy should be very useful if administered during the diapause period, for reducing the presence of L1 stages and interrupting thus the life cycle of this myiasis.

Molecular characterization of subcutaneous and muscular coenurosis of goats in United Arab Emirates.

Coenurosis is a parasitic disease caused by the larval stage of Taenia multiceps, which affects various ruminants species, particularly sheep and goats, and occasionally humans. In this note, non-cerebral coenurosis in goats with the goal to compare morphological and biomolecular characteristics is focused in order to determine if the non-cerebral specimens should be considered a new specie or a strain of T. multiceps. Three hundred goats slaughtered with macroscopic lesions due to the presence of metacestodes were examined in an abattoir in Dubai, United Arab Emirates in order to evaluate the presence of extra-cerebral coenuri. Forty-eight coenuri were found under the skin, between fasciae of the skeletal muscles, diaphragm and in the abdominal cavities. Morphologically, the examined non-cerebral coenuri recovered showed the same features reported by other authors for Coenurus cerebralis, but their location outside the central nervous system suggests that they may be a different strain or genetic variants of T. multiceps. Nine caprine coenuri were then processed for sequencing of mitochondrial partial COI (396 bp) and ND1 (471 bp) genes indicating that they had a pairwise distance of 1.0-1.3% and 2.4-4.1% compared with parasite' COI sequences from Italy (Tm1-Tm3 strains) and Erzurum strains of ovine origin, respectively; whereas it had 0.6-1.3% and 0.4-1.1% pairwise distance for ND1. Phylogenetic trees of their ND1 and COI sequences using the maximum composite likelihood method with MEGA showed that these nine parasites recovered in Emirates abattoir could be grouped into a new strain. In conclusion, morphological and molecular characterization of caprine coenurosis from Dubai, United Arab Emirates, shows that the specimens are significantly different from other strains or genotypes in COI sequence, which suggests that they should belong to different genotypes or strains of T. multiceps.

Cystic echinococcosis in Sardinia: farmers' knowledge and dog infection in sheep farms.

Cystic Echinococcosis (CE) is one of the most widespread parasitic diseases in Sardinia, the second largest Mediterranean island where almost 3,558,000 milk sheep were raised extensively. The aim of this survey was to evaluate the level of farmers' knowledge on CE transmission, focusing on the role of human to facilitate the persistence of this zoonosis in Sardinia after 14 years after the last campaign against CE. The other goal of the survey is to update on presence of Echinococcus granulosus in its definitive hosts through three ELISA coproantigen tests. An interview was carried out with 172 farmers. The questionnaire was designed to include possible factors associated with the transmission of Echinococcosis: ownership and number of dogs, the use of anthelmintic drugs against dog cestode, frequency of anthelmintic treatment in dogs, home slaughtering and offal disposal. Individual faecal samples were retrieved from 300 dogs, and after a preliminary macroscopic examination to discover adult worms and/or proglottids, was submitted to copromicroscopic examination. Coproantigens were then extracted according to the protocol described by Allan et al. (1992), and subsequently stored at -20°C until use. Faecal soluble antigens from E. granulosus were detected using three different ELISA coproantigen assays: (a) the commercially produced Chekit Echinotest (Bommeli, Bern, CH) based on polyclonal antibodies against adult excretory/secretory (E/S) antigens; (b) a sandwich ELISA that uses rabbit polyclonal antibodies against adult E/S antigens and biotinylated monoclonal antibody EmA9 produced against adult Echinococcus multilocularis somatic extract (Malgor et al., 1997); and (c) a sandwich assay that uses monoclonal antibody EgC3 produced by immunization with adult E. granulosus E/S products (Casaravilla et al., 2005). Questionnaire results reveal that on all farms home-slaughtering was done, and offal was used as dog meal raw (17%) or after boiling (37%), discarded in the trash (23%), or buried superficially (15%). Most farmers (69%) declared to deworm their dogs, but only 10% used cestodicidal drugs. The coprological survey of 300 farm dogs using sedimentation, flotation and three different coproantigen (CA) ELISAs resulted in a faecal prevalence of 8.3% for taeniid eggs, while the CA tests gave prevalences of 3% (Chekit Echinotest, Bommeli), 6% (EmA9 sandwich ELISA) and 10% (EgC3 sandwich ELISA). Our results show that this is not only an educational problem, but also an economic one, stressing the need that future control plans should follow an integrative approach including veterinary and medical services, farmers, breeders' associations and the Government.

Preliminary field trial of a vaccine against coenurosis caused by Taenia multiceps.

Taenia multiceps is a taeniid cestode that in its adult stage lives in the small intestine of dogs and other canids. In the intermediate hosts, the larval stage of T. multiceps causes coenurosis, a common disease in the CNS of ruminants, which typically leads to the death of the infected animals. Recent research into new methods for control of coenurosis and other taeniid cestode infections such as hydatidosis has identified vaccination as a potentially valuable new tool. In order to test the applicability of vaccination as an approach for control of T. multiceps infection in sheep, a field trial was carried out against natural infection in Sardinian farms (Italy) with recombinant proteins of T. multiceps. The recombinant proteins with Quil A as adjuvant were injected subcutaneously, the first administered to lambs at 10-12 weeks of age and a booster dose given after 2-4 weeks. A total of 632 sheep were selected, belonging to the "replacement quota" of six different farms, of which 424 were used as controls (unvaccinated) and 208 were vaccinated. After a period of more than 40 months from the beginning of the field trial, 33 episodes of cerebral coenurosis occurred in the monitored farms, including 32 cases in control sheep and l case in a vaccinated animal. Statistical analysis revealed a significant reduction in the number of coenurosis cases in the vaccinated animals (chi(2)=14.08, P<0.001). This is the first successful field test of a practical vaccine against T. multiceps and, considering the high degree of effectiveness achieved, could be a prelude to routine application in field situations of particular risk, such as Sardinia.

Cystic echinococcosis in equids in Italy.

Between March 2003 and February 2007, the livers and the lungs of 2,231 horses from various Italian regions were examined for cystic echinococcosis presence at the time of slaughter. Hydatid cysts were found in six horses, namely four from Sardinia, one from Sicily, and one from Tuscany. The location, number, morphology, and fertility of the cysts found were determined. DNA was extracted from the germinal layers and protoscoleces of the fertile cysts and polymerase chain reactions (PCR) were performed in order to strain type DNA isolates for reduced nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1), cytochrome c oxidase subunit 1 (CO1) and 12S partial genes. The PCR products were then purified and sequenced in forward and reverse. Hydatid materials obtained from positive animals were identified as Echinococcus granulosus s.s. (old G1, sheep strain) and Echinococcus equinus (old G4, horse strain) for ND1, CO1, and 12S partial genes. This allowed us to record the presence of the E. equinus in Italy for the first time with molecular tools and also to report new data on the epidemiological situation of this parasite in Italy.