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Fasciola hepatica is a trematode infecting ruminants worldwide, occasionally reported in a wide range of animal species, including humans. According to the WHO, fasciolosis is recognized as a re-emerging neglected tropical disease, responsible for endemic and epidemic outbreaks in humans. Although the main hosts of the parasite are represented by cattle, sheep and goats, wildlife may be involved in its circulation. Here we firstly report F. hepatica in a wild boar from Italy (southern area) and characterize it both morphologically and molecularly. The nad1 gene analysis of specimens analyzed, revealed a high genetic similarity with those of humans from Iran and Peru, as well as a close phylogenetic relationship to those in ruminants from Brazil, Ecuador and Egypt. Considering the increase in the wild boar populations in urban and peri-urban areas, a potential role of this ungulate in the circulation of this zoonotic trematode is suggested., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

In light of rapidly spreading triclabendazole resistance alternative fasciocidal drugs are urgently needed. Following up on promising results obtained with artemether in Fasciola hepatica infected sheep, we here report the efficacy and safety of artesunate in sheep with a natural F. hepatica infection. Artesunate was administered intravenously and intramuscularly, adverse events were monitored and drug efficacy was elucidated by means of faecal egg and worm burden reductions. A single 40 mg/kg intravenous dose of artesunate induced an egg count reduction of 68.9% and a worm burden reduction of 77.4%. Intramuscular artesunate at 40 mg/kg reduced faecal egg count and worm burden by 97.6% and 91.9%, respectively; whereas at 60 mg/kg it caused 93.2% and 87.1% reduction in faecal egg count and worm burden, respectively. Three sheep died 24-72 h post-treatment with a double dose of 40 mg/kg intramuscular artesunate, showing lethargy, sialorrhoea, reduced rumination and tremors. Egg and worm burden reductions of 93.3% and 83.9%, respectively, were calculated in the three surviving sheep. In conclusion, the interesting fasciocidal properties of artesunate in sheep warrant further investigations with an emphasis on toxicity studies., (Copyright 2009 Elsevier Ltd. All rights reserved.)

Background: Gastrointestinal nematode (GIN) infection is one of the main constraints to sheep production both in temperate and tropical countries. Economic losses caused by GIN are related to decreased production, treatment costs and even animal death. The present paper was aimed at assessing the anthelmintic efficacy (based on faecal egg count reduction) of moxidectin and ivermectin both administered per os at dose rate of 0.2 mg/Kg body weight and the benefit of anthelmintic treatments on milk production in a commercial dairy sheep farms in central Italy whose animals were naturally infected by GIN., Results: The treatment with moxidectin was highly effective (> 98%) from day 7 until day 75, and effective (90-98%) until day 105. The treatment with ivermectin was highly effective (> 98%) from day 7 until day 14, effective (90-98%) at day 28 and moderately effective (80-89%) on day 45. The milk productions in the treated groups were significantly higher than those of the control group., Conclusion: In conclusion, the results of the present study demonstrated that moxidectin and ivermectin administered per os according to the manufacturer's instructions were both effective and safe anthelmintics in sheep. The total milk production was higher in the treated groups than the control group. Overall, animals treated with moxidectin had a milk production 40.8% higher than control group; whereas animals treated with ivermectin had a milk production 32.2% higher than control group.

The aim of the present paper was to assess the benefit of targeted selective treatments (TSTs) on milk production and faecal worm egg count, in four commercial dairy sheep farms in southern Italy where animals were exposed to natural infection with gastrointestinal (GI) strongyles. On each farm, three similar groups of 20 animals each were formed and randomly assigned to the following treatments: strategic prophylactic treatment (SPT) group, targeted selective treatment group, and control (C) group. The TST schemes used were different between farms with regard to the indicators for treatment (FAMACHA or faecal egg count (FEC) or milk production) and the timing of treatment (periparturient or postparturient). Regarding milk production, on the four study farms the mean daily milk production in the SPT ewes was higher than those of the controls on Farms 1, 2, 3 and 4 by 13.6%, 30.4%, 37.0% and 5.5%, respectively. In the case of Farms 2 and 3 these differences were significant (P<0.05). Average milk production in the TST ewes was higher than that in the control group, by 7.2% in Farm 1; 10.1% in Farm 2; 13.0% in Farm 3; and 1.4% in Farm 4; however, none of these differences were statistically significant. Regarding the effects on FEC, FAMACHA-based TST (Farms 1 and 2) had no positive effects on GI strongyle egg counts. In the FEC-based TST group on Farm 3 both the TST and SPT treatments reduced FEC. In the milk production-based TST group (Farm 4), no conclusion could be drawn regarding strategic efficacy since there was also a decrease in GI strongyle FEC in the control group. The findings of the present study demonstrate that TST can be considered as a potentially useful anthelmintic strategy in southern Italy, but the timing of treatments seems to be more crucial for the strategic and production efficacy than the indicators used for those treatments. From a practical point of view using milk production as a TST indicator appeared to be the most user-friendly approach for farmers; however, further studies will be needed to confirm its validity and acceptability.

Since it was first developed in South Africa, the FAMACHA system has been successfully tested in different geographic area where Haemonchus contortus is the major gastrointestinal (GI) strongyle species of sheep. The present study, the first on the use of the FAMACHA system in Italy, was aimed at obtaining a complete haematological profile in sheep with GI strongyle infection, in order to evaluate the validity of the FAMACHA system for detecting the severity of anaemia in sheep from southern Italy. Between April and September 2006, 793 blood and faecal samples were collected from 137 adult sheep of different local breeds of the Campania region (southern Italy). Each study animal was monthly subjected to blood collection, GI strongyle fecal egg counts (FEC) and a complete clinical examination, with particular attention to the colour of ocular mucous membranes. The FAMACHA score values (from 1 to 5) were compared to Hematocrit (Hct) and Haemoglobin (Hb) values. Considering the FAMACHA score number "3" as borderline value for anaemia, the eye score values for anaemic animals were considered in the categories of 4 and 5 (Group 4-5) or of 3, 4 and 5 (Group 3-4-5), respectively. Anaemia due to GI strongyle infection was considered when the Hct and/or Hb values were under 27% and 9g/dl, respectively. The correlation between FAMACHA score and both Hb and Hct was significant (P<0.001) and negative; however, the low correlation coefficients indicated a weak negative association between FAMACHA and either Hb and Hct parameters. The sensitivity, specificity, predictive negative value (NPV) and predictive positive value (PPV) of the FAMACHA system were calculated. The highest values of sensitivity were 66% and 60% in the Group 3-4-5 for Hct and Hb, respectively. The highest values of specificity were 98% in the Group 4-5, for Hb and Hct, respectively. In the Group 3-4-5, the specificity was 60% and 62% for Hb and Hct, respectively. The NPV was 92% (Hct) and 84% (Hb) in the Group 3-4-5, while it was 87% (Hct) and 78% (Hb) in the Group 4-5. The PPV, for the others categories considered (Hb, Hct, Group 4-5 and 3-4-5), was always below 54%. In conclusion, in the present study, the FAMACHA system showed a low sensitivity in detecting anemic sheep; these findings indicate that in southern Italy, under our experimental conditions, FAMACHA may represent an additional element that can integrate clinical examinations. It is important that such evaluations are performed in each region before the system is implemented, as there could be local issues that affect the accuracy of the system.

The present paper was aimed at assessing the benefit of strategic anthelmintic treatments on milk production in four commercial dairy sheep farms in Southern Italy whose animals were naturally infected by gastrointestinal (GI) strongyles and Dicrocoelium dendriticum. The scheme was based on two treatments timed in relationship to parturition, i.e. the first with moxidectin in the periparturient period and the second with netobimin at the mid/end of lactation. On each farm, two similar groups (20 animals each) were formed, one untreated control group and one group treated according to the above-mentioned scheme. Faecal egg counts (FEC) were performed on each study animal at the start of the trial and then monthly until the end of the study; in addition, milk production was recorded for each animal fortnightly in each farm for the lactation period. The results showed a significant increase in milk yield across all four farms that ranged from 19% to 44% improvement in milk yield. The benefit in milk yield in addition to considerably reduced egg output of the treated animals provide clear evidence that the two main aims of prophylactic parasite control, i.e. to maintain or improve animal performance and to reduce pasture contamination, can be achieved using strategic anthelmintic treatments.

Triclabendazole is the current drug of choice against Fasciola spp. infections in livestock, but resistance has become a major problem. In this study, we assessed the efficacy and safety of artemether, a derivative of artemisinin, in sheep with a low natural Fasciola hepatica infection. Artemether was administered orally or intramuscularly; sheep were monitored for 8 h posttreatment and then once daily for adverse events, and drug efficacy was estimated by fecal egg count reductions and worm burden reductions. Single 40- and 80-mg/kg oral doses of artemether showed no effect on F. hepatica egg and worm burden. Treatment with a single 160-mg/kg intramuscular dose of artemether significantly reduced the egg burden (64.9%) and worm burden (91.3%). At half this dose, a worm burden reduction of 65.3% was obtained, which was still statistically significant (P < 0.05). The lowest intramuscular dose of artemether investigated (40 mg/kg) yielded no effect on egg counts and worm burden. There were no adverse events due to artemether; however, two abortions were observed 7 days posttreatment. In conclusion, artemether shows interesting fasciocidal properties in sheep, but embryotoxicity is of concern. Further studies are warranted to assess the potential of additional artemisinin derivatives and other peroxidic compounds for the treatment of Fasciola spp. infections in different ruminants.

The aim of the present paper was to assess benefit of strategic anthelmintic treatments on milk production in six commercial dairy sheep farms, located in southern Italy, whose animals were naturally infected with gastrointestinal strongyles. On each farm, two similar groups were formed, one untreated control group and one treated group. In all the treated groups, the strategic anthelmintic schemes were based on: (i) only one treatment with moxidectin in the periparturient period (February, Farm No. 6), or; (ii) two treatments, i.e. the first with moxidectin performed in the periparturient period (February, Farms Nos. 1, 2, 3 and 4) or in the postparturient period (April, Farm No. 5), and the second with netobimin at the mid/end of lactation (June, Farms Nos. 1, 2, 3, 4 and 5). Faecal egg count reduction (FECR) tests were performed on each farm in order to asses the anthelmintic efficacy of the drugs used. In addition, milk yield measurements for each animal fortnightly in each farm for the lactation period were performed. In terms of FECR, both moxidectin and netobimin were effective in all the 6 studied farms. Regarding milk production, overall in the 6 study farms the mean daily milk productions of the treated groups were higher than those of the control group. However, there were important differences between the 6 farms, i.e. the increase of milk production in the treated groups versus the control groups was as follows: +18.9% (Farm 1), +30.4% (Farm 2), +4.0% (Farm 3), +37.0% (Farm 4), +5.5% (Farm 5) and +40.8% (Farm 6). The results of the study showed that the economic efficacy of an anthelmintic treatment is not a cause-effect issue, but is a multifactorial issue which depends upon the quali-quantitative parasitological status of the animals, the pathogenesis of the species of parasites, the virulence of the strains of parasites, the local epidemiology, the timing of treatment, the breed of animal in terms of genetics and production types, nutrient supply.

A cross-sectional survey was carried out on 2004-2005 in the Campania region, southern Italy, to study the multivariate geographical distribution of four different sheep helminths, i.e. Fasciola hepatica (liver fluke), Calicophoron (Paramphistomum) daubneyi (rumen fluke), Dicrocoelium dendriticum (lancet fluke), and the gastrointestinal strongyle Haemonchus contortus. A series of multivariate Bayesian hierarchical models based on square root transformation of faecal egg counts were performed. The results were consistent with theoretical knowledge of the biology and epidemiology of the four studied helminths. In particular, the impact of common intermediate hosts (F. hepatica and C. daubneyi share the same intermediate host species) was quantified and evidence of previously unknown ecological components was given. D. dendriticum was correlated to F. hepatica and H. contortus was found not to be spatially associated with the previously mentioned helminths.

A clinical longitudinal field trial was conducted on a dairy sheep farm in southern Italy to assess the effectiveness of a novel anthelmintic treatment strategic scheme against Fasciola hepatica. The scheme utilizes a dual anthelmintic treatment (DAT), i.e., the use of either one of two different anthelmintics on the flock, albendazole sulphoxide (SO) at 1-month intervals and rafoxanide at 2-month intervals, administered to the lactating and non-lactating animals, respectively. The DAT strategic scheme lasted 3 years. In Year 1 and Year 2, shotgun monthly DATs for 5 consecutive months (July, August, September, October, and November) were performed on the flock. In Year 3 there was only one monthly DAT, in July. Overall, the DAT scheme reduced the prevalence of F. hepatica infection by 94.4% (from an average prevalence of 71.1% during the pre-DAT period to an average prevalence of 4.0% during Year 3), and the eggs/gram of faeces (EPG) from 29.3 to 1.3. In conclusion, the DAT strategic scheme reported in the present study successfully reduced both the prevalence and EPGs of F. hepatica to a level at which there were no longer any clinical symptoms of the disease. This scheme did not influence the albendazole SO efficacy against GI nematodes and might be used for the treatment of fasciolosis in dairy sheep farms.

Isolates of the rumen fluke Calicophoron daubneyi (Digenea: Paramphistomidae) from various hosts and three locations in southern Italy were characterized genetically. The second internal transcribed spacer (ITS-2) of ribosomal DNA (rDNA) plus flanking 5.8S and 28S sequence (ITS-2+) was amplified from individual rumen flukes by PCR. PCR-linked restriction fragment length polymorphism (PCR-RFLP) analysis was performed using four different restriction endonucleases, and PCR products were sequenced. The PCR analyses from all the C. daubneyi specimens produced identical fragments, and the PCR-RFLP analyses did not show, with respect to any of the four restriction endonucleases, any differences between the C. daubneyi specimens. The sequence analyses of the ITS-2+ from each of the C. daubneyi specimens showed them all to be 428 bp, and composed of the entire ITS-2 sequence (282 bp) plus the two partial flanking conserved sequences, 5.8S (99 bp) and 28S (47 bp). No intra-specific variation was observed in the nucleotide composition of the ITS-2+ (homology=100%). There was, however, an observable inter-specific variation between the ITS-2+ of C. daubneyi and the ITS-2+ of both Calicophoron calicophorum (homology=97.2 %) and Calicophoronmicrobothrioides (homology=97.4 %), both previously deposited in the GenBank. The finding of the present study shows that, as has already demonstrated for other parasitic helminths, ITS-2 can serve as an effective genetic marker for the molecular identification of paramphistomes, and as a useful tool for developing molecular epidemiological techniques for the study of C. daubneyi transmission patterns and prevalence in definitive and intermediate hosts.

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.

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.

A cross-sectional survey of ticks was conducted on 197 ovine farms with animals pasturing in an area (3971 km2) of the southern Italian Apennines. The farms were selected to be uniformly distributed throughout the study area using Geographical Information System (GIS). Ticks were collected from 309 (31.4%) out of the 985 sheep sampled, belonging to 92 (46.7%) out of the 197 farms included in the study. The following tick species were found (farm prevalence): Dermacentor marginatus (37.6%), Haemaphysalis punctata (29.4%), H. sulcata (2.5%), H. parva (2.0%). H. inermis (0.5%), Ixodes gibbosus (2.0%), I. ricinus (0.5%), Rhipicephalus sanguineus group (1.0%), and R. bursa (0.5%). A point distribution map (PDM) was drawn by GIS in order to display the distribution of each tick genus in the study area. The general trends of the PDM show that Dermacentor marginatus and Haemaphysalis spp. were widely and homogeneously spread throughout the study area, whereas Rhipicephalus spp. and Ixodes spp. were present only in a few concentrated zones of the study area in accordance to their biological and ecological characteristics.

Two field trials were conducted in two farms (farms A and B) in southern Italy, to assess the efficacy of eprinomectin applied topically at the dose rate of 500 micro g/kg to sheep with naturally occurring infections of gastrointestinal nematodes (GIN). The nematode population determined by necropsy consisted of Teladorsagia circumcincta, Haemonchus contortus, Trichostrongylus vitrinus, T. capricola, Nematodirus sp., and Chabertia ovina in sheep from farm A, and of T. circumcincta, T. vitrinus, T. capricola, T. colubriformis, and C. ovina in sheep from farm B. In each farm, 42 female sheep were assigned to a eprinomectin treated group (E-group) and a control untreated group (C-group) of 21 animals each. On farm A, the percentage reductions in strongyle faecal egg counts from E-group compared to C-group were 99.1% on day 10; 97.4% on day 30; and 67.0% on day 60. On farm B, on the same days, they were 95.4, 84.9, and 69.4%, respectively. In the course of the two trials, eprinomectin was well tolerated by all the animals with no adverse reactions following the topical treatment.

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.