Effect of different temperatures on survival and development of Aelurostrongylus abstrusus (Railliet, 1898) larvae.

Aim of the study was to get in-depth information on Aelurostrongylus abstrusus first-stage larvae (L1s) survival at different temperatures and to assess the capability of these larvae to develop into the third infective stage (L3s). Faeces of a naturally infected cat were split into two aliquots: the first was divided in subsamples assigned to four groups (F1-F4); from the second aliquot, L1s were extracted by Baermann technique, suspended in water and divided into four groups (W1-W4). Groups were stored at different temperatures (F1/W1 -20 ± 1°C; F2/W2 +4 ± 1°C; F3/W3 +14 ± 1°C; and F4/W4 +28 ± 1°C) and L1s vitality assessed every seven days. The capability of L1s stored in water to develop into L3s in snails was evaluated at the beginning and every 21 days. The L1s of W2 and F2 groups remained viable for a longer period (231 and 56 days, respectively) compared to those of other groups. The capability of L1s to moult into L3s in snails showed a decreasing trend; the group W2's L1s maintained the capability to moult into L3s for the longest time (day 189) compared to the other groups. The time of survival of A. abstrusus L1s is influenced by temperature. However, the species seems to be more resistant to temperature variations than other feline lungworms, and this may explain its wider distribution across Europe.

Cross-sectional survey on Tritrichomonas foetus infection in Italian cats.

The feline genotype of Tritrichomonas foetus is a widespread cause of large-bowel diarrhoea in cats. The aim of this study was to determine the prevalence of the T. foetus infection in cat populations across Italy. Fresh, individual faecal samples were collected from 267 cats, kept in different environments (i.e., private households, breeding structures, municipal catteries and colonies) in three different sites across Italy. The faecal samples were tested by PCR to detect T. foetus. Moreover, the same samples were subjected to a concentration-flotation technique and a commercial direct fluorescent-antibody (DFA) test to detect additional enteric parasites, including Giardia duodenalis. The overall prevalence of T. foetus infection was 5.2%. All the infected cats showed diarrhoea at the time of sampling: 9 out of 14 positive cats were co-infected with G. duodenalis, 1 with Toxocara cati and 3 with Dipylidium caninum. The risk factor analysis showed that not only the breed, but also co-infections with G. duodenalis and Dipylidium caninum were significantly associated with the presence of T. foetus. This study confirms the presence of T. foetus in cats living in Italy, suggesting that this protozoan parasite should always be included in the differential diagnosis of patients referred with large-bowel disease symptoms, especially if they were purebred animals, or affected by other enteric protozoa, such as G. duodenalis.

The European wildcats (Felis silvestris silvestris) as reservoir hosts of Troglostrongylus brevior (Strongylida: Crenosomatidae) lungworms.

The increasing reports of Troglostrongylus brevior lungworm in domestic cats from Italy and Spain raised questions on its factual distribution and on the role wildcats play as reservoirs of these parasites. Carcasses of 21 wildcats were collected in natural parks of southern Italy (i.e., Catania, Sicily n=5 and Matera, Basilicata n=16) and biometrically and genetically identified as Felis silvestris silvestris, but two as hybrids. Troglostrongylus brevior and Eucoleus aerophilus lungworms were found in 15 (71.4%) and 7 (33.3%) individuals, respectively, being five (23.8%) co-infected by the two species. Both lungworms showed an aggregated distribution in the host population, assessed by k-index (i.e., 0.69 for T. brevior and 0.42 for E. aerophilus). Although no statistical significant difference was recorded among age, gender and geographical location of wildcats, a larger rate of infection by T. brevior was assessed in yearlings (85.7%) than adults (64.3%). This is the first epidemiological study reporting T. brevior infection in the European wildcat and discusses the potential threat this may represent for the conservation of this endangered species of felids. In addition, given the large frequency of lungworm infection herein recorded, the role of wildcats as reservoir hosts of these parasites to domestic cats is discussed.

Lungworms of the genus Troglostrongylus (Strongylida: Crenosomatidae): neglected parasites for domestic cats.

Feline lungworms belonging to the Troglostrongylus genus have been neglected for a long time. However, recent reports of Troglostrongylus brevior and Troglostrongylus subcrenatus in domestic cats have stimulated the interest of the scientific community on these lungworms. Troglostrongylus spp. have an indirect life cycle, which overlaps that of the better known Aelurostrongylus abstrusus. Nonetheless, adult worms of both genera have a distinct morphology and localization within the respiratory system of definitive hosts and, potentially, a different epidemiology and pathogenicity. As copromicroscopy has a low specificity for metastrongyloids due to the similarities of first-stage larvae, specific morphometrical keys and/or PCR diagnostic tools are advocated. Accordingly, more clinical studies and necropsy data are needed to elucidate the impact of Troglostrongylus spp. on the health of domestic cats. This article reviews current information on Troglostrongylus spp. as well as data on their occurrence in Europe and reports specific key morphological characters for the identification of adults and larvae, which is important to refine their diagnosis and for a better understanding of the feline lungworm infections.

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.

Rhipicephalus sanguineus (Ixodida, Ixodidae) as intermediate host of a canine neglected filarial species with dermal microfilariae.

The life cycles of filarioids of dogs presenting dermal microfilariae have been little studied. Following the recent retrieval of dermal microfilariae identified as Cercopithifilaria sp. in a dog from Sicily (Italy), this study was designed to assess the role of the brown dog tick Rhipicephalus sanguineus as an intermediate host of this filarial species. An experimental tick infestation was performed on an infected dog using 300 nymphs of R. sanguineus. Engorged nymphs were collected and examined by both microscopic dissection and molecular analysis at five time points (i.e., the same day of tick detachment and 10, 20, 30 and 50 days post-detachment) to detect the presence and developmental stage of filariae in the ticks. A total of 270 engorged nymphs were collected from the dog and developing filarioid larvae detected in 10 (5%) out of 200 ticks dissected. Infective third-stage larvae were observed in 4 (2%) of the all dissected ticks, 30 days post-detachment. Twelve (6.6%) out of 181 samples molecularly tested were positive for Cercopithifilaria sp. This study demonstrates that nymphs of R. sanguineus feeding on a dog naturally infected by Cercopithifilaria sp. can ingest microfilariae, which develop up to the third infective stage thus suggesting that this tick species might act as an intermediate host of this little known canine filarioid.

Risk for the introduction of exotic ticks and pathogens into Italy through the illegal importation of tortoises, Testudo graeca.

In April 2008, 585 tortoises illegally imported into Italy from North Africa were examined for the presence of ticks. Of these, 221 tortoises (37.8%) were infested with a mean intensity of 3.9 +/- 3.1 ticks (range 1-17 ticks). A total of 798 ticks (672 males, 125 females and one nymph) were collected and identified as Hyalomma aegyptium (L.) (Acari: Ixodidae). The overall male : female ratio observed was 5 : 1. The prevalence and mean intensity [+/-standard deviation (SD)] of ticks were higher among male (67.4%, 4.0 +/- 3.2) than female (55.6%, 3.8 +/- 3.1) tortoises, although this difference was not significant. By contrast, the prevalence and mean intensity of ticks were significantly higher on tortoises weighing >100 g (61.5%, 4.0 +/- 3.2) compared with tortoises weighing <100 g (12.1%, 2.1 +/- 1.1). Of the infested tortoises, 89.8% had ticks on their hind limbs, 21.0% on forelimbs, 18.6% on the tail and pre-anal area, and 4.8% on the head; thus the hind limbs were evidently the preferred attachment site. The present report highlights the need to develop surveillance systems to prevent the introduction and spreading of exotic ticks and tick-borne pathogens in Italy and other European countries.

Influence of gastrointestinal trichostrongylidosis on ram fertility.

A study on the influence of gastrointestinal trichostrongyles on ram fertility was performed. Two groups of semen donor rams (Sarda breed) were utilized; one was experimentally infected with Haemonchus contortus, Trichostrongylus colubriformis and Teladorsagia circumcincta and the other was used as a control group. In all animals, coprological, haematological and reproductive parameters were studied. The results suggest that the parasites had a limited effect with some changes in phosphorus, cholesterol and chlorine levels. In our experience the parasitic burden produced no significant change on ram fertility.

Two species of Philometra (Nematoda: Philometridae) from sparid fishes (porgies) off Sicily, Italy, including Philometra obladae sp. n. from the body cavity of Oblada melanura (Sparidae).

Two species of Philometra Costa, 1845 (Nematoda: Philometridae), were, for the first time, recorded from fishes of the family Sparidae (porgies) from the Tyrrhenian Sea off Sicily, Italy: Philometra obladae sp. n. from the body cavity of the saddled seabream Oblada melanura (Linnaeus) and Philometra filiformis (Stossich, Boll Soc Adriat Sci Nat 17:121-136, 1896) from the gonads of the common pandora Pagellus erythrinus (Linnaeus). The new species (a single gravid female available) is characterised mainly by a large body (335 mm long, 3.4 mm wide), minute cephalic papillae (14 in number) arranged in two circles, a relatively short (1.29 mm) oesophagus with a distinct anterior inflation, a rounded caudal end without any projections and the length of larvae (549-600 mum) from the uterus. A key to species of Philometra with females located in the body cavity of marine and brackish-water fishes is given. Some new data on the female morphology of a little-known gonad-infecting species P. filiformis are provided; new observations revealed, for the first time in this species, the presence of lateral cephalic papillae and the absence of previously reported black intestinal corpuscles in the larvae.

Use of the FLOTAC technique for the diagnosis of Aelurostrongylus abstrusus in the cat.

Aelurostrongylus abstrusus is a lungworm of the domestic cats distributed worldwide. Its presence, however, is often underestimated due to limitation of common coprological diagnostic techniques. The present study reports, for the first time, the use of the FLOTAC technique for A. abstrusus diagnosis in feline faecal samples with results compared to the following three standard copromicroscopic techniques: the Baermann technique, the McMaster technique and the Wisconsin technique. The comparison study showed that the FLOTAC technique produced mean larvae per gram of faeces significantly (P<0.05) greater than that produced by the other more widely used diagnostic tools. Thus, the FLOTAC technique can be utilised for quantifying lungworm larva burdens in feline faecal samples because of its high sensitivity, as already shown for various parasites of veterinary and human importance.

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.

Efficacy of oxfendazole and fenbendazole against tortoise (Testudo hermanni) oxyurids.

Thirty-six tortoises (Testudo hermanni) with naturally acquired oxyurids infections were used to assess the anthelmintic efficacy of oxfendazole (Dolthene; Merial) and fenbendazole (Panacur; Hoechst Roussel Vet). Animals were randomly assigned to three groups (A, B, and C) based on sex and weight. Animals in group A (seven males and six females) were orally treated with oxfendazole at dose rate of 66 mg/kg, group B animals (nine males and eight females) were orally treated with fenbendazole at dose rate of 100 mg/kg, and group C animals (three males and three females) were not treated and served as controls. All animals were individually stabled in plexiglas boxes under controlled conditions of temperature, humidity, and light beginning 7 days pretreatment and continuing for the duration of the trial. Individual tortoises feces were examined daily by the McMaster technique and drugs efficacy was assessed by the fecal eggs count reduction (FECR) test. Both drugs showed 100% of FECR. However, oxfendazole reached this level 12 days after treatment, whereas 31 days after treatment were necessary to obtain the same stable result with fenbendazole. The two drugs were well tolerated by all the animals and no adverse reactions were observed after treatment.

[Biomorphology of gastrointestinal nematodes of small ruminants]. / Biomorfologia dei nematodi gastrointestinali dei piccoli ruminanti.

Under the term gastrointestinal nematodes are included numerous parasites species of livestock belonging to the families Strongyloididae (Strongyloides), Strongylidae (Chabertia, Oesophagostomum) Trichostrongylidae (Trichostrongylus, Ostertagia, Teladorsagia, Cooperia, Marshallagia), Molineidae (Nematodirus), Ancylostomatidae (Bunostomum) and Trichuridae (Trichuris). This paper reviews the biomorphology aspects of these parasites as well as the controversy by the taxonomists in the classifications.

Agrometeorology and models for the parasite cycle forecast.

Insects are strongly influenced by meteorological variables in their natural environment. In agriculture, mathematical models have been developed to understand and forecast the cycle of pests based on climate data. By this manner, with the goal of reduce and rationalize plant chemical treatments, agrometeorological models have been realized to estimate the length and starting times of parasites phenological phases. In Sicily a new network of 95 GSM meteorological stations and a specific mathematical model for Aonidiella aurantii are used by Sicilian Agrometeorological Information System (SIAS) for the integrated pest management program of citrus orchards in the Island. As the plants parasites, vector borne diseases are influenced by climate in their appearance and abundance. In lights of the benefits that could derive from a model for the control of Leishmania vectors, SIAS experiences in modelling were used to develop a deductive model for Phlebotomus perniciosus which represents the major vector of human and canine leishmaniasis in Sicily.

Proposal of Health Information System (HIS) as tool for the epidemiological surveillance of leishmaniasis in urban areas.

In recent years leishmaniasis has become a serious Public Health problem in Italy. The coexistence of infected dogs, vectors and humans, makes urban areas suitable environment for the spread of this zoonotic disease. With the purpose to improve the current system for the control of leishmaniasis in urban areas, the authors report their experiences carrying out a Health Information System (HIS) for the surveillance of the disease in the city of Messina. The HIS project is built on a dataset containing geographical data on urban environment, census tracts at parish level and data of human and canine leishmaniasis cases reported in the last years. The HIS allows the spatial representation of the disease occurrence and can be used as a novel epidemiological tool to perform area-based surveillance.

[Animal echinococcosis in Italy: epidemiological update]. / Aggiornamenti epidemiologici sull'echinococcosi animale in Italia.

Cystic Echinococcosis (CE) is one of the most widespread parasitoses in the Mediterranean Region (MR). This is due to various factors, the most important being the close association between man, sheep and dogs in areas where open farming is practised. Although this disease has been known for several years and many studies have been carried out, nowadays in Italy there are no complete epidemiological data on its diffusion and distribution. The available data show that CE is mainly diffused in those districts where the sheep-dog cycle can be perpetuated, such as central and southern Italy, and the islands. Furthermore, no data are available on biomolecular characterisation of the strains of Echinococcus granulosus in Italy, apart form those in Sardinia, where the G1 (sheep-dog) and G7 (pig-dog) strains were recently isolated. One of the reasons why CE is a problem with no easy solution is undoubtedly the difficulty of making a certain diagnosis in the dog, the principal definitive host of E. granulosus.

Tissue damage and haematophagia due to praniza larvae (Isopoda: Gnathiidae) in some aquarium seawater teleosts.

Over a 5 yr period, severe infestations due to Gnathia vorax larvae affecting different teleosts have been registered at the Civic Aquarium of Messina, Italy. Here, the damage caused by these parasites is evaluated from a macroscopical and histopathological point of view. We also attempt to evaluate the amount of blood sucked up by a single parasite and make some observations about the haematophagia. The histopathological changes confirm mucosal electrolytic failure as the main problem, even though the mean value of blood sucked up by a single parasite, about 1.9 mg, suggests that haematophagia may play an important role in the pathogenesis of such infestation. In conclusion, it was found that the G. vorax larvae caused 2-fold damage that was responsible for the death of the fish. Little by little, the parasitic problem was successfully removed by washing the tank, using fresh water rather than costly and dangerous chemotherapeutans.