A prevalence survey and risk analysis of filariosis in dogs from the Mt. Vesuvius area of southern Italy.

A dog microfilariae prevalence and risk factor survey was conducted in 51 contiguous municipalities of the Mt. Vesuvius area (Campania region, southern Italy) in order to add data to the limited epidemiological information available regarding filarial worms in this zone. Between May 1999 and June 2000, blood samples were collected from 351 asymptomatic dogs. Blood samples were examined using a modified Knott's technique and histochemical staining in order to count and identify microfilariae. The results were subjected to statistical analysis and choroplethic municipal maps (MMs) were drawn by a geographical information system (GIS) software. Microfilariae were detected in 63 of the 351 dogs surveyed, constituting a total filarial prevalence of 17.9%. In particular, 56 dogs (15.9%) showed only microfilariae of Dipetalonema reconditum; three dogs (0.8%) only microfilariae of Dirofilaria repens; two dogs (0.6%) microfilariae of both D. reconditum and D. repens and two dogs (0.6%) microfilariae of both Dirofilaria immitis and D. repens. High D. reconditum prevalence was associated with hunting practice, masculine gender and older dogs. There was also a tendency to find high prevalence in dogs sampled in the afternoon.In conclusion, the presence of microfilariae of D. reconditum in 92% of microfilaraemic dogs indicates that this filarial worm was the predominant filarial species in dogs in the Mt. Vesuvius area.In addition, the general trends of the MMs showed that D. immitis and D. repens were present only in a few municipalities, whereas D. reconditum was widely and homogeneously spread throughout the entire study area.     

A cross-sectional coprological survey of liver flukes in cattle and sheep from an area of the southern Italian Apennines

A cross-sectional coprological survey of liver flukes (Fasciola hepatica and Dicrocoelium dendriticum) was conducted on 81 bovine farms and 197 ovine farms with animals pasturing in an area (3971 km(2)) of the southern Italian Apennines. The farms were selected to be uniformly distributed throughout the study area using geographical information system (GIS) software. Between June 1999 and March 2000, faecal samples were collected from 975 cattle and 3940 sheep and examined using a modified McMaster technique. The results were subjected to statistical analysis and point distribution maps (PDMs) were drawn by GIS.Cattle of 9 of the 81 (11.1%) farms were positive for F. hepatica and of 43 (53.1%) for D. dendriticum. Sheep of 8 of the 197 (4.1%) farms were positive for F. hepatica and of 133 (67.5%) for D. dendriticum. Co-infection was found in cattle of 2 (2.5%) farms, and in sheep of 8 (4.1%) farms.The findings of the present survey show that D. dendriticum was the predominant liver fluke found in cattle and sheep with respect to egg count numbers for both farms and animals. In addition, the general trends of the PDMs show that D. dendriticum was widely and homogeneously spread throughout the study area, whereas F. hepatica was present only in a few concentrated zones of the study area that had both positive bovine and positive ovine farms.     

Comparative pharmacokinetics of levamisole-oxyclozanide combination in sheep and goats following per os administration

Since there is no registered anthelmintic drug available for use in goats, extra-label use of drugs is a common practice in most countries. The aim of the present study was to compare the pharmacokinetic disposition of levamisole (LVM)-oxyclozanide (OXZ) combination in sheep and goats following per os administration. Goats (n = 8) and sheep (n = 8) 12- to 16-months-old were used for this study. The animals received tablet formulation of LVM and OXZ combination orally at a dose of 7.5 mg/kg and 15 mg/kg body weight, respectively. Blood samples were collected by jugular vein at different times between 5 min and 120 h after drug administrations. The plasma concentrations of LVM and OXZ were analyzed by HPLC following liquid-liquid phase extraction procedures. The plasma concentrations and systemic availabilities of both LVM and OXZ in goats were lower and the plasma persistence of LVM was shorter compared with those observed in sheep. Terminal half-lives (t_1/2λz) of both molecules are shorter in goats compared with those in sheep. Goats treated with LVM-OXZ combination at the recommended dose for sheep may result in a reduced efficacy, because of under-dosing, which may increase the risk of drug resistance in parasites. Increased or repeated dose could be a strategy to provide higher plasma concentration and thus to improve the efficacy against the target parasites in goats compared with sheep. However, some adverse reactions may occur since LVM has relatively very narrow therapeutic index due to its nicotine-like structure and effect.     

Long-term Electrocardiography Recording with Holter Monitoring in 15 Donkeys

The heart rates (HR) and rhythms of 15 healthy donkeys of various ages, both sexes, and various breeds were analyzed throughout 24-hour Holter monitoring. The animals were evaluated at rest in their daily environment without the presence of investigators causing stress, using a three-channel digital Holter recorder. Analysis revealed a maximal HR range from 62.5 to 93.7 beats/min (mean, 72.50 ± 7.51 beats/min), whereas the minimal HR ranged from 29.7 to 42.2 beats/min (mean, 34.90 ± 4.22 beats/min). The daily mean HR was 47.55 ± 3.70 beats/min. The daytime mean HR was 49.39 ± 2.77 beats/min, whereas the night time rate was 46.22 ± 4.38 beats/min. Sinus bradycardia and sinus tachycardia were detected in 7 and 10 of 15 studied donkeys, respectively. Cardiac dysrhythmias due to high vagal tone such as sinoatrial heart block and second-degree atrioventricular heart block were occasionally recorded in 1 and 3 donkeys, respectively. A higher mean HR and fewer cardiac dysrhythmias were observed in donkeys than in horses and ponies at rest. These findings could be justified by differences in the autonomic nervous system tone.     

Use of remote sensing and geographical information systems to identify environmental features that influence the distribution of paramphistomosis in sheep from the southern Italian Apennines

A geographic information system (GIS) was constructed using remote sensing (RS) and landscape feature data together with Calicophoron daubneyi positive survey records from 197 georeferenced ovine farms with animals pasturing in a 3971 km(2) area of the southern Italian Apennines. The objective was to study the spatial distribution of this rumen fluke, identify environmental features that influence its distribution, and develop a preliminary risk assessment model. The GIS for the study area was constructed utilizing the following environmental variables: normalized difference vegetation index (NDVI), land cover, elevation, slope, aspect, and total length of rivers. These variables were then calculated for "buffer zones" consisting of the areas included in a circle of 3 km diameter centered on 197 farms. The environmental data obtained from GIS and RS and from data taken by the veterinarians on the field (stocking rate and presence of streams, springs and brooks on pasture) were analyzed by univariate (Spearman and ANOVA) and multivariate (discriminant) statistical analyses using the farm coprological status (positive/negative) as the dependent variable. Sheep on 32 of the 197 (16.2%) farms, were positive for C. daubneyi, with an average intensity of 52 epg. A multivariate stepwise discriminant analysis model was developed that included moors and heathland, sclerophyllous and coniferous forest vegetation, autumn-winter NDVI and presence of streams, springs and brooks on pasture. The variables entered in the model were also correlated with C. daubneyi positive farms in the univariate tests and are consistent with the environmental requirements of C. daubneyi and its snail intermediate host.     

The influence of flotation solution, sample dilution and the choice of McMaster slide area (volume) on the reliability of the McMaster technique in estimating the faecal egg counts of gastrointestinal strongyles and Dicrocoelium dendriticum in sheep

The present study was aimed to evaluate the influence of flotation solution, sample dilution, and the choice of McMaster slide area (volume) on the reliability of the McMaster technique in estimating the faecal egg counts of gastrointestinal (GI) strongyles and Dicrocoelium dendriticum in a composite sample of faeces from naturally infected sheep. Fourteen flotation solutions having densities between 1.200 and 1.450, and six sample dilutions, 1:10, 1:15, 1:20, 1:30, 1:40 and 1:50 were used. Each of the six dilutions was divided into 70 aliquots in order to have five replicates of each of the 14 flotation solutions at each of the six dilutions. For each McMaster slide, the GI strongyle and D. dendriticum egg counts were performed under one grid (McM 0.15 ml), two grids (McM 0.3 ml), one chamber (McM 0.5 ml), and both chambers (McM 1.0 ml). Mean eggs per gram (EPG) of faeces of GI strongyles and D. dendriticum were calculated and statistical analyses were performed on the resulting data. The type of flotation solution used significantly influenced the EPG in the GI strongyles and in the D. dendriticum egg counts. All the sucrose-based solutions at density between 1.200 and 1.350 floated more GI strongyle eggs than the others. With respect to D. dendriticum, only six solutions were capable of floating eggs and the potassium iodomercurate solution (density 1.440) floated more eggs than the others. The reliability of the McMaster technique regarding sample dilution was high for both GI strongyle and D. dendriticum EPG at 1:10 and 1:15, and then progressively decreased with increasing dilution. The reliability of the McMaster technique regarding the choice of the McMaster slide area (volume) was high for both GI strongyle and D. dendriticum EPG at the McMaster slide area (volume) of 1.0 ml, i.e. the total area of the McMaster slide. The EPG counts resulting from choosing any of the other three McMaster slide areas (volumes), i.e. McM 0.15 ml, McM 0.3 ml, or McM 0.5 ml, produced unreliable over-estimates. The findings of the present study show that the highest reliability of the McMaster technique for estimating GI strongyle and D. dendriticum egg counts in faeces from pastured sheep is obtained when using flotation solutions based on sucrose for GI strongyles, and potassium iodomercurate for D. dendriticum, dilutions which do not exceed 1:15, and the McMaster slide area (volume) of 1.0 ml.     

Molecular update on cystic echinococcosis in cattle and water buffaloes of southern Italy

Cystic echinococcosis (CE)--caused by the larval stage (hydatid cyst) of the cestode Echinococcus granulosus--is one of the most widespread zoonoses of veterinary and medical importance. Molecular techniques have allowed the identification of 10 different genotypes (G1-G10) of the parasite. The present paper is an update regarding the E. granulosus genotypes infecting water buffaloes and cattle bred in the Campania region of southern Italy. The molecular study was performed on 30 hydatid cysts (11 from water buffaloes and 19 from cattle). Two different mitochondrial DNA genes, namely the cytochrome c oxidase subunits 1 and the 12S ribosomal DNA (12S rDNA) were used as genetic markers. Three different genotypes of E. granulosus were unequivocally identified, i.e. the G1 (common sheep), G2 (Tasmanian sheep) and G3 (buffalo) genotypes, as well as some G1 and G2 variants. It should be noted that the present study demonstrated for the first time: (i) the presence of the G2 genotype in water buffaloes from a Mediterranean area; and (ii) the fact that the analysed portion of the 12S rDNA gene can not discriminate between the G2 and G3 genotypes of E. granulosus. The finding of the G1, G2 and G3 genotypes in large ruminants from southern Italy is of epidemiological relevance and immediate public health importance because of their recognized infectivity in humans.     

Neospora caninum in pastured cattle: determination of climatic, environmental, farm management and individual animal risk factors using remote sensing and geographical information systems

A cross-sectional serological survey was conducted on cattle pasturing in an area of the southern Italian Apennines to evaluate the seroprevalence to Neospora caninum, and to investigate the climatic, environmental, farm management, and individual animal factors that influence the distribution of this protozoan. Blood samples were collected from 864 pastured cattle raised on 81 farms. Serum samples were tested for antibodies to N. caninum using an ELISA assay (HerdCheck), IDEXX). A geographical information system (GIS) for the study area was constructed using the following remote sensing (RS) and landscape feature data: autumn-winter, spring and summer normalized difference vegetation index (NDVI), land cover, elevation, slope, aspect, mean rainfall and minimum, mean, and maximum temperature in spring, summer, autumn and winter. Data on each of these features were then extracted for "buffer zones" consisting of the area included in a circle of 3 km diameter centered on the 81 geo-referenced centroids of the main cattle pastures. Climatic and environmental data obtained from RS and GIS and individual animal characteristics and farm management data obtained from a questionnaire were analyzed in relation to N. caninum seropositivity and antibody titres both by linear and logistic regression models. Out of the 81 farms sampled, 63 (77.8%) had at least one tested animal positive for N. caninum. Out of the 864 bovine sera samples, 266 (30.8%) were found to have antibodies to N. caninum. The results of the logistic regression model show that significantly high seroprevalence to N. caninum was found in heifers/steers and adults, in cattle raised on farms having a large number of dogs, and in cattle raised in buffer zones having high minimum temperatures in the spring, and a narrow extension of summer NDVI. In addition, positive linear correlations were found between N. caninum antibody titres and the number of dogs on farm, and the minimum temperature in spring. All the above determined risk factors for N. caninum seroprevalence indicate that horizontal infection resulting from the ingestion of oocysts shed by dogs is the most probable route of N. caninum infection in pastured cattle of the southern Italian Apennines.     

Epidemiology of bovine tick-borne diseases in southern Italy

This investigation was carried out in an area covering part of three southern Italian regions: Campania, Basilicata and Apulia. Eighty-one farms were involved using the formula suggested by Thrusfield; they were equally distributed over the area which was subdivided into 81 geo-referenced sub-areas. In May and June 1999 from a total of 506 cattle, older than 18 months, blood-samples were taken and ticks were collected and identified. Serum samples were tested for antibodies of Bahesia bigemina, Babesia bovis and Anaplasma marginale with an ELISA technique. Eight farms (9.8%) out of the 81 examined were positive for B. bigemina only, 3 (3.7%) for A. marginale only, and 70 (86.4%) for both. None of the animals of any farm was found to be positive for B. bovis. Out of the 506 sera tested, 117 (23.1 %) were positive for B. bigemina only, 58 (11.5%) forA. marginale only and 250 (49.4%) for both species; 81 (16.0%) were negative for all of them. Ticks were collected on animals on 62 (76.5%) out of the 81 farms. Adult ticks (1 410) were collected and identified; the highest number belonged to the Rhipicephalus bursa species (65.5%), followed by Rhipicephalus turanicus (8.6) and Haemaphysalis punctata (8.4). The results showed that B. bigemina, A. marginale and their potential vectors are common in the area examined and indicated that there is a risk for animals imported from tick-borne disease-free areas.