Sheep In Italy Research Articles

Molecular phylogenetic analysis of Echinococcus granulosus sensu lato infecting sheep in Italy.

Cystic Echinococcosis (CE), caused by the larval form of Echinococcus granulosus sensu lato, is a neglected zoonosis still threatening public health worldwide. In Italy different epidemiological scenarios were reported depending on the geographical area and associated socio-economic activities. Although in northern Italy the occurrence of E. granulosus is considered sporadic, in the southern regions and, particularly in Sardinia, CE prevalence reaches high levels. We analysed CE cysts collected from infected sheep from various areas of mainland Italy and the Sardinia island, with the main objective to investigate intergenotypic and intragenotypic variations at national level. CE cysts were collected from slaughtered sheep following post mortem inspection at local abattoirs. Total genomic DNA was extracted and amplification and sequencing of the partial mitochondrial genes nad5 and cox1 were performed. A Bayesian phylogenetic tree was estimated on a nad5 dataset (n=260) composed of E. granulosus samples from this study (n=126) and all the nad5 haplotypes available in GenBank (n=134). In addition, haplotype network, diversity and neutrality analysis were performed on nad5 and cox1 sequences of Italian origin obtained in this study. E. granulosus sensu stricto (s.s.) was found to be the only Echinococcus species infecting sheep in Italy, mainly represented by G1 genotype (76%) and, to a lower extent, by G3 genotype (24%). Phylogenetic analyses revealed 40 nad5 and 33 cox 1 haplotypes, and the presence of two founder haplotypes, belonging to G1 and G3 genotype, showing 100% similarity with DNA sequences from different geographic regions. The lack of geographical segregation, high haplotype and low nucleotide diversity coupled with significant negative values of Tajima's D and Fu's Fs observed in this study indicated high genetic variation and demographic expansion of E. granulosus s.s. in Italy.

Gastrointestinal Parasite Infections and Environmental Sustainability of the Ovine Sector: Eimeria spp. Infections and Nitrogen and Phosphorus Excretions in Dairy Sheep in Italy.

In sheep farming, gastrointestinal parasites can be responsible for significant reductions in animal health and production. Nitrogen (N) and phosphorus (P) fecal excretions are the main determining factors for N2O emissions from manure management and may pose other environmental problems, such as the acidification and eutrophication of natural habitats. By using the Mini-FLOTAC technique on fecal samples from sheep of different ages and physiological status from 19 dairy sheep farms in Tuscany (central Italy), gastrointestinal parasite infections were evaluated. The animal N and P fecal contents were also assessed, with the aim of evaluating possible relationships between the identified parasites and the environmental sustainability of the examined farms. The obtained results showed that Eimeria spp. (86.36%) and gastrointestinal strongyle (54.55%) infections are prevalent in the examined farms. Moreover, significantly higher (p ≤ 0.05) P and Eimeria oocyst/gram-of-feces (OPG) values were found in fecal samples from animals < 1 year of age, and a significant (p ≤ 0.05) positive correlation resulted between N content and Eimeria OPG in fecal samples from animals in the first month of lactation. The findings from this study suggest for the first time that Eimeria spp. infections may have an impact on the environmental sustainability of sheep farming.

Polymorphism of prion protein gene (PRNP) in Nigerian sheep

ABSTRACT Polymorphism of the prion protein gene (PRNP) gene determines an animal’s susceptibility to scrapie. Three polymorphisms at codons 136, 154, and 171 have been linked to classical scrapie susceptibility, although many variants of PRNP have been reported. However, no study has investigated scrapie susceptibility in Nigerian sheep from the drier agro-climate zones. In this study, we aimed to identify PRNP polymorphism in nucleotide sequences of 126 Nigerian sheep by comparing them with public available studies on scrapie-affected sheep. Further, we deployed Polyphen-2, PROVEAN, and AMYCO analyses to determine the structure changes produced by the non-synonymous SNPs. Nineteen (19) SNPs were found in Nigerian sheep with 14 being non-synonymous. Interestingly, one novel SNP (T718C) was identified. There was a significant difference (P < 0.05) in the allele frequencies of PRNP codon 154 between sheep in Italy and Nigeria. Based on the prediction by Polyphen-2, R154H was probably damaging while H171Q was benign. Contrarily, all SNPs were neutral via PROVEAN analysis while two haplotypes (HYKK and HDKK) had similar amyloid propensity of PRNP with resistance haplotype in Nigerian sheep. Our study provides valuable information that could be possibly adopted in programs targeted at breeding for scrapie resistance in sheep from tropical regions.

A survey on Apicomplexa protozoa in sheep slaughtered for human consumption

Infections with the Apicomplexa Toxoplasma gondii, Neospora caninum, and Sarcocystis spp. are common causes of reproductive disorders in sheep. However, few epidemiological studies regarding co-infections with these three protozoa are reported in sheep in Italy. For this reason, this study aims to evaluate possible co-infections with T. gondii, N. caninum, and Sarcocystis spp. in sheep slaughtered for human consumption. From April to July 2019, individual blood, brain, heart, and diaphragm samples were collected from 138 sheep after slaughtering. The presence of IgG anti-Toxoplasma in serum samples was evaluated through ELISA. DNA of the three protozoa was investigated using specific PCRs. Co-infection with T. gondii, N. caninum and Sarcocystis spp. was found in 66.7% of the examined sheep. Antibodies against T. gondii were found in the 36.2% of serum samples. The presence of T. gondii DNA was detected in the 67.4%, 77.5%, and 21.7% of the brain, heart, and diaphragm samples, respectively. Neospora caninum DNA was found in 72.5% of the examined brain samples. Sarcocystis spp. DNA was detected in 92% and 52.2% of the heart and diaphragm samples, respectively. Sequence analysis of the Sarcocystis spp. revealed the sole presence of Sarcocystis tenella. The present study demonstrates that sheep have a high risk of infection with the three Apicomplexa investigated, suggesting the need to adopt adequate measures to prevent the spread of these parasitic infections considering their clinical and economic impact on ovine production. Furthermore, the possible role sheep play in the zoonotic transmission of toxoplasmosis to humans was highlighted.

Hepatitis E virus in sheep in Italy.

Hepatitis E virus (HEV) is the leading cause of human enterically transmitted viral hepatitis occurring around the world both as outbreaks and as sporadic cases. The accumulating literature indicates that domestic pigs and wild boars are the main reservoirs of genotype 3 and genotype 4 for human infections in industrialized countries. However, the recent identification of HEV from various animal species poses additional potential concerns for HEV zoonotic infection. In this study, the role of sheep as potential host of hepatitis E virus (HEV) was investigated. By screening 192 sheep from seven farms located in Abruzzo Region (Southern Italy), HEV-specific antibodies were detected in the sera of 41 animals (21.3%) whilst the RNA of HEV, genotype 3, was detected in 20 faecal (10.4%) and three serum samples (1.6%). Upon sequence analyses of a partial ORF2 gene region of eight HEV positive samples, the sheep sequences all grouped together within HEV genotype 3 subtype c, being most closely related to HEV strains identified in goat and wild boar from Abruzzo. This is the first study that demonstrates, serologically and molecularly, the presence of HEV in sheep population in a European country.

The feasibility of using passive injectable transponders in field operations for individual identification of sheep in Italy

This trial evaluates the feasibility of using passive injectable transponders (PITs) in field operations by testing identification procedures on 185 one-year-old Biellese ewes reared under nomadic farming conditions. Commercial PITs of 3.85 ± 0.05 mm × 31.2 mm were used at two application sites, the armpit and the retro-auricular region. The two application sites were compared taking into account the ease of injection, animal reaction, injection duration, inflammatory response, PIT readability at up to 12 months post-injection and PIT recovery at the slaughterhouse. The injection site influenced the reaction of the animal and the ease and duration of the injection, but it did not affect the palpation and the reading findings during the rearing period. The injection site also influenced the PIT retrieval at the slaughterhouse depending on the operator who recovered the PIT. The readability values observed at the end of the observation period for both injection sites (83% in the retro-auricular region and 79% in the armpit region) were too low to be suitable for long-term animal identification. In addition, the failed retrieval of PITs at slaughter, determined the destruction of 40 (22%) carcases to avoid any risks in the food chain. These findings suggest that the limited use of PITs is indicated when other methods of electronic identification cannot be employed, while a wider application of the device tested in the present study is not recommended in farming practices.

Origins of Wohlfahrtia magnifica in Italy based on the identification of mitochondrial cytochrome b gene haplotypes.

To identify the geographical origins of larvae of Wohlfahrtia magnifica (Diptera: Sarcophagidae) causing myiasis of sheep in Italy, comparative DNA sequence analysis of the mitochondrial cytochrome b gene was performed, based on gene fragments amplified by PCR from genomic DNA isolated from individual specimens. DNA extractions of 19 larvae from Lazio, Molise, Puglia, and Sicilia generated 17 readable sequences homologous to 2 haplotypes, either CB_magn01 or CB_magn02; DNA extracts from 4 adult flies from Calabria (reared from larvae) produced 4 readable sequences belonging to the haplotype CB_magn01. The two haplotypes found represent both the East and West phylogenetic lineages of W. magnifica, which is consistent with the species' arrival from central/southeast Europe (East lineage) and/or from southwest Europe/northwest Africa (West lineage). This is the first report of the sympatric occurrence of the two lineages, which could have resulted from natural or human-assisted dispersal. Polymorphic nuclear loci will have to be characterized in order to explain the origins and lack of mitochondrial haplotype diversity of this pest in Italy, where it poses increasing veterinary problems.

Mitochondrial DNA analysis of medieval sheep (Ovis aries) in central Italy reveals the predominance of haplogroup B already in the Middle Ages.

We retrieved 34 medieval ovicaprine remains, from three archaeological sites of central Italy dating to about 1000 years old, and analyzed them using mitochondrial DNA. We compared the reconstructed haplogroups with modern sheep samples from Europe and the Middle East and sequences from the literature. In modern sheep, haplogroup HA is present in countries with access to the Mediterranean and close to the domestication center, whereas it is very rare or absent in the rest of Europe. The haplogroup HB was predominant in ancient samples (90%), whereas haplogroup HA was found at 10%. Ancient haplogroups match the present distribution in modern sheep in Italy, indicating that the current proportion of HA/HB was already established in the Middle Ages and is not the result of subsequent events such as selective breeding practices.

Theileria sp. OT3 and other tick-borne pathogens in sheep and ticks in Italy: Molecular characterization and phylogeny

PCR Reverse Line Blot (RLB) hybridization and sequencing were used to determine the dynamics of infection with tick-borne pathogens in one hundred apparently healthy sheep in Italy. Blood samples were tested once prior to the onset of the grazing season (June 2010) and once after the end of the grazing season (August 2010). Ticks collected from sheep and from the vegetation were also tested by PCR/RLB.Before grazing, 56% of the sheep harbored several tick-borne pathogens: Anaplasma ovis was the most prevalent (41%), followed by A. ovis co-infected with Theileria sp. OT3 (14%). After grazing, 87% of sheep were positive for A. ovis alone (41%), co-infected with Theileria sp. OT3 (8%) or co-infected with Babesia motasi (5%). Other sheep were infected with Anaplasma phagocytophilum alone (20%), co-infected with B. motasi (7%) or with Theileria sp. OT3 (5%) (p<0.001). After grazing, sheep were significantly more infected with tick-borne pathogens than before grazing. Ticks collected were all Haemaphysalis punctata (n-89) and 36% were positive for A. ovis, Ehrlichia ovina and A. ovis combined with A. phagocytophilum.Phylogenetic analysis including isolates from countries in the Mediterranean Basin show circulation of the same variants of Theileria sp. OT3, whereas two different geographical origins for the isolates of A. ovis and A. phagocytophilum were identified. This is the first report from Italy of Theileria sp. OT3 in sheep, whereas the detection of Ehrlichia ovina in ticks is worth noting, and the presence of A. phagocytophilum in sheep and in ticks poses a potential public health risk.

Molecular epidemiology of bluetongue virus serotype 1 circulating in Italy and its connection with northern Africa

Western BTV-1 emerged in the Mediterranean basin in 2006 and it has since been isolated in southern and northern European countries. Six BTV-1 strains isolated from infected sheep in Italy between 2006 and 2013 and a BTV-1 strain isolated from an infected sheep in Tunisia in 2011 were fully sequenced. The seven strains were shown to be nearly identical in each gene segment. The Seg-2 sequences of the BTV-1 strains group according to the year of isolation reflecting the time of BTV incursions in Italy. Combined results suggest that BTV-1 strains isolated in Sardinia, Sicily and mainland Italy in 2012 and 2013 have a direct northern African origin. The Italian strains originated from a strain closely related to a BTV-1 strain isolated in Tunisia in 2011. Better surveillance programs with northern and sub-Saharan African countries should be implemented making the control of spread of BTV easier and effective.

Evaluation of targeted selective treatments in sheep in Italy: Effects on faecal worm egg count and milk production in four case studies

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.

The effect of different legume‐based swards on intake and performance of grazing ruminants under Mediterranean and cool temperate conditions

AbstractIntake and performance of sheep or cattle grazing legume‐based swards were assessed over 2 years at four locations in Europe with different climatic conditions: Sardinia (Italy), southern France, northern Germany and south‐west England (UK). Legume species were sown in mixtures with locally appropriate companion grass species. Standard legume species commonly used at the location (Medicago polymorpha in Italy, Medicago sativa in France, and Trifolium repens in Germany and UK) were compared with two alternative legume species characterized by different agronomic or nutritional characteristics. They were: Trifolium subterraneum and Hedysarum coronarium in Italy; Trifolium incarnatum and Onobrychis sativa in France; Trifolium pratense and Lotus corniculatus in Germany; and Trifolium ambiguum and L. corniculatus in the UK. Lactating milk sheep in Italy, non‐lactating ewes in southern France, growing cattle in Germany, and ewes and lambs in the UK were used in three replicates per treatment. Intake and performance of sheep and cattle in various treatments varied with location, year and period within year. In Germany, intake and performance by cattle were not affected by treatment. In contrast, at the other locations, sheep grazing standard or alternative legume species known to contain condensed tannins (sulla, sainfoin and birdsfoot trefoil) had higher proportions of legume and crude protein (CP) concentrations in the diet and higher CP intakes than those grazing the other alternative legume species. Sheep performance paralleled these results. It is concluded that there is potential for a greater use of alternative legume species, at least for sheep‐grazing systems, in both the Mediterranean and cool temperate zones of Europe.

An inactivated vaccine for the control of bluetongue virus serotype 16 infection in sheep in Italy

Because no suitable products are at the moment available to safely control the spread of BTV-16 in Europe, an inactivated vaccine was produced from the reference field isolate of bluetongue virus serotype 16. One group of six sheep was vaccinated subcutaneously with the inactivated vaccine twice, on days 0 and 28, whereas a second group of eight sheep was inoculated with saline solution and used as mock-vaccinated control animals. Seventy-eight days after the first vaccination, all sheep were inoculated subcutaneously with a suspension containing 10 6.3 TCID 50 of a virulent reference BTV-16 isolate. Apart from a transient inflammatory reaction at the injection site, no adverse effects were reported following vaccination. All vaccinated animals developed high titres (7.3–9.3 log 2(ED50%/50 μl)) of virus-specific neutralising antibodies and were resistant to challenge with BTV-16. Conversely, following challenge, control animals developed hyperthermia and long lasting high-titre viraemia.

Are goats naturally resistant to gastric Helicobacter infection?

Gastric Helicobacter species are widespread and have been reported in wild and domestic mammals of different dietary habits such as humans, dogs, cats, macaques, mice, cheetahs, ferrets, swine and cattle. All have been associated with gastric pathologies. Recently, gastric Helicobacter species were shown to be widespread in cattle and swine in Europe, and there is a report of Helicobacter pylori in sheep in Italy. However, there are no reports of Helicobacter infection in the goat, another important domestic animal of human consumption. The aim of our study was to assess whether Helicobacter abomasal infection was common in goats slaughtered for human consumption. Infection was detected through PCR analysis of DNA extracted from gastric biopsies, using genus- and species-specific primers. Bovine and porcine gastric samples were also analyzed as positive controls. None of the 70 goats were positive for Helicobacter spp.; however, Candidatus Helicobacter bovis and Candidatus Helicobacter suis were detected in 85% of the bovine and 45% of the porcine samples, respectively. We discuss the possibility that goats may exhibit natural resistance to abomasal infection by Helicobacter spp.

Quantitative and compositional variations of massese sheep milk by parity and stage of lactation

Effects of parity and length of lactation on physico-chemical characteristics of sheep milk were evaluated on milk samples from the first to the sixth lactation of 100 Massese sheep in Italy. Milk samples were collected twice a month and the following items were studied: quantity of milk, length of lactation, physico-chemical characteristics, amino acids, fatty acids and mineral content of the milk. Milk production showed a decreasing curvilinear trend with days of lactation, according to the formula y=2.1409−0.012 x+0.00002 x 2. Parity had a significant effect on total milk yield ( P⩽0.006), and on physico-chemical milk characteristics ( P⩽0.001), but had no effect on amino acid, fatty acid and mineral contents. Stage of lactation caused high variability in almost all analytical parameters expressed by linear or quadratic regression, with the exception of several amino acids which were independent of this factor.

Isolation of visna-maedi virus from the choroid plexus of an apparently healthy sheep in Italy

In this paper, the presence of visna-maedi in Italy, reported to exist since 1982 on clinical grounds, and the presence of anti-visna-maedi antibody is confirmed by the isolation of the viral agent from choroid plexus cells of an apparently healthy sheep. Gel diffusion test antigen, prepared from the explanted choroid plexus cells and from normal choroid plexus cells infected with the isolated agent, gave a positive reaction with one or two identity lines with reference antigens. The isolated agent gave both a characteristic cytopathic effect and a cytoplasmic fluorescence starting 24 hours after infection in normal choroid plexus cell cultures. Fluorescence was observed neither in control normal choroid plexus cells nor in infected normal choroid plexus cells incubated with a visna-maedi negative serum. By electron microscopy, budding forms arising from the cell membrane and extracellular particles of two distinct types were observed. One type was characterized by an electron-dense central core and a single membrane, and ranged from 80 to 110 nm, while the other had elongated bar-like cores and a double wall, and ranged from 100 to 140 nm. The characteristics observed for the isolated agent are identical to those reported by various authors for visna-maedi virus.