An integrated approach to control Cystic Echinococcosis in southern Italy.

Cystic echinococcosis (CE) is a severe zoonosis, caused by the larval stage of the tapeworm Echinococcus granulosus. This helminth infection is of increasing public health and socio-economic concern due to the considerable morbidity rates that cause economic losses both in the public health sector and in the livestock industry. Control programmes against E. granulosus are considered long-term actions which require an integrated approach and high expenditure of time and financial resources. Since 2010, an integrated approach to control CE has been implemented in a highly endemic area of continental southern Italy (Campania region). Innovative procedures and tools have been developed and exploited during the control programme based on the following strategies: i) active and passive surveillance in livestock (using geospatial tools for georeferencing), ii) diagnosis in dogs (using the FLOTAC techniques and molecular analysis), iii) targeted treatment of farm dogs (using purpose-built confinement cages), iv) early diagnosis in livestock (by ultrasonography), v) surveillance in humans (through hospital discharge records analysis), vi) monitoring the food chain (analysing raw vegetables), vii) outreach activities to the general public (through dissemination material, e.g. brochures, gadgets, videos, virtual reality). Over eight years, the integrated approach and the new strategies developed have resulted in a noteworthy reduction of the parasite infection rates in livestock (e.g. up to 30 % in sheep). The results obtained so far highlight that using a one health multidisciplinary and multi-institution effort is of pivotal importance in preparing CE control programmes at regional level and could be extended to other endemic Mediterranean areas.

Analysis of potential risk factors of caprine coccidiosis.

Caprine coccidiosis represents a major concern in goat industry worldwide causing high economic losses, mainly due to poor weight gains, treatment costs and even death of kids. Therefore, analyses on risk factors involved in the establishment and outcome of this disease are important to improve infection status and herd management. In Portugal, approximately 400,000 goats are reared every year. The majority of them are allocated in the southern part of the country. In the current study, a representative number of animals (382 goats) and herds (37 herds) from Alentejo were monitored. Faecal samples from five young and five adult goats of each farm were collected rectally and examined individually by using the modified McMaster technique for oocyst counts (oocyst per gram of faeces - OPG). Additionally, a questionnaire on the husbandry conditions and anti-parasitic treatments was performed. Overall, a prevalence of Eimeria infections of 99% and 100% was detected on animal and herd level, respectively. Eimeria arloingi and E. ninakohlyakimovae as the two most pathogenic caprine species also constituted the most prevalent ones (34.91% and 29.90%, respectively), both in the young and the adult goat groups. As expected, OPG vales were significantly higher (p = 0.031) in young than in adult goats. Furthermore, risk factor analyses revealed young age, higher total number of animals on the farm and outdoor kidding as strongly associated with a higher risk of caprine coccidiosis. Overall, the high prevalence of pathogenic Eimeria species on Portuguese goat farms in combination with the occurrence of related risk factors strengthen the importance of coccidiosis control in caprine industry.

Initial assessment of the economic burden of major parasitic helminth infections to the ruminant livestock industry in Europe.

We report a European wide assessment of the economic burden of gastrointestinal nematodes, Fasciola hepatica (common liver fluke) and Dictyocaulus viviparus (bovine lungworm) infections to the ruminant livestock industry. The economic impact of these parasitic helminth infections was estimated by a deterministic spreadsheet model as a function of the proportion of the ruminant population exposed to grazing, the infection frequency and intensity, the effect of the infection on animal productivity and mortality and anthelmintic treatment costs. In addition, we estimated the costs of anthelmintic resistant nematode infections and collected information on public research budgets addressing helminth infections in ruminant livestock. The epidemiologic and economic input data were collected from international databases and via expert opinion of the Working Group members of the European Co-operation in Science and Technology (COST) action COMbatting Anthelmintic Resistance in ruminants (COMBAR). In order to reflect the effects of uncertainty in the input data, low and high cost estimates were obtained by varying uncertain input data arbitrarily in both directions by 20 %. The combined annual cost [low estimate-high estimate] of the three helminth infections in 18 participating countries was estimated at € 1.8 billion [€ 1.0-2.7 billion]. Eighty-one percent of this cost was due to lost production and 19 % was attributed to treatment costs. The cost of gastrointestinal nematode infections with resistance against macrocyclic lactones was estimated to be € 38 million [€ 11-87 million] annually. The annual estimated costs of helminth infections per sector were € 941 million [€ 488 - 1442 million] in dairy cattle, € 423 million [€ 205-663 million] in beef cattle, € 151million [€ 90-213 million] in dairy sheep, € 206 million [€ 132-248 million] in meat sheep and € 86 million [€ 67-107 million] in dairy goats. Important data gaps were present in all phases of the calculations which lead to large uncertainties around the estimates. Accessibility of more granular animal population datasets at EU level, deeper knowledge of the effects of infection on production, levels of infection and livestock grazing exposure across Europe would make the largest contribution to improved burden assessments. The known current public investment in research on helminth control was 0.15 % of the estimated annual costs for the considered parasitic diseases. Our data suggest that the costs of enzootic helminth infections which usually occur at high prevalence annually in ruminants, are similar or higher than reported costs of epizootic diseases. Our data can support decision making in research and policy to mitigate the negative impacts of helminth infections and anthelmintic resistance in Europe, and provide a baseline against which to measure future changes.

Species diversity and spatial distribution of ixodid ticks on small ruminants in Greece.

A cross sectional field study on the distribution of Ixodidae ticks was carried out over two consecutive tick seasons (2012 and 2013) in small ruminants (309 farms) located in mainland and five islands of Greece. A total of 2108 ticks (1199 females, 908 males, and 1 nymph) were collected from sheep and goats. Two species of Rhipicephalus, two of Ixodes, five of Hyalomma, three of Haemaphysalis, and one of Dermacentor were detected. The species with the widest distribution were Rhipicephalus sanguineus s.l. (64.8 %), R. bursa (25.9 %), and Dermacentor marginatus (4.1 %), whereas the least frequently collected species were Ixodes ricinus, I. gibbosus, Haemaphysalis parva, H. sulcata, H. punctata, Hyalomma marginatum, H. excavatum, H. dromedarii, H. rufipes, and H. impeltatum representing together less than 5.3 % of the collections. R. sanguineus s.l. was the predominant species, while D. marginatus and I. gibbosus were restricted to the northeastern Aegean Sea islands. This is the first survey on tick fauna from mainland and islands in Greece.

Evaluation of mean echogenicity of tendons and ligaments of the metacarpal region in neonatal foals: A preliminary study.

The aims of this research were to evaluate mean echogenicity (ME) of the deep and superficial digital flexor tendons (DDFT and SDFT), the interosseous muscle (IM), and the accessory ligament of the deep digital flexor tendon (ALDDFT) of the metacarpal region in neonatal foals, and determine the effect of sex, side and body weight on this quantitative ultrasonographic evaluation. Thirteen orthopedically sound neonatal foals were examined. Four areas of study (1A, 1B, 2A, 2B) were identified. Transverse scans of the DDFT, SDFT, IM and ALDDFT were obtained, recorded, and analyzed. The most echogenic structures were the ALDDFT and DDFT, while the SDFT was significantly less echogenic than all other structures (P<0.05). No influence of sex, forelimb, or body weight was observed. The echogenicity of the tenodesmic structures of foals partially overlapped that reported in the metacarpal region in adult horses, except for IM.

miR-30e* is an independent subtype-specific prognostic marker in breast cancer.

BACKGROUND: Breast cancer clinical outcome is affected by tumor molecular features, and the identification of subtype-specific prognostic biomarkers is relevant for breast cancer translational research. Gene expression signatures proved to be able to complement prognostic information provided by classical clinico-pathological features. Recently, microRNAs (miRNAs) have been causally linked to tumorigenesis and cancer progression and have been associated with patient outcome, also in breast cancer. METHODS: MicroRNAs associated with the development of distant metastasis were identified in a cohort of 92 ESR1+/ERBB2- lymph node-negative breast cancers from patients not receiving adjuvant treatment. Results were confirmed and further investigated in a total of 1246 miRNA and gene expression profiles of the Molecular Taxonomy of Breast Cancer International Consortium data set. Moderated t-test, univariable and multivariable Cox regression models were used for statistical analyses. RESULTS: miR-30e* was identified as independent protective prognostic factor in lymph node-negative untreated patients with ESR1+/ERBB2- tumours and retained a significant association with a good prognosis in treated patients with the same tumor subtype as well as in the ERBB2+ subtype, but not in ESR1-/ERBB2- tumours. CONCLUSIONS: We highlighted a relevant and subtype-specific role in breast cancer for miR-30e* and demonstrated that adding miRNA markers to gene signatures and clinico-pathological features can help for a better prognostication.

Cross-sectional area and mean echogenicity of shoulder and elbow tendons in adult German Shepherd dogs.

The aim of this study was to describe the cross-sectional area and mean echogenicity of the main tendons of the shoulder and elbow joints in adult German Shepherd dogs and to determine the effects of sex, weight, and age on these parameters. No previous publications in the veterinary literature have reported information regarding quantitative ultrasonographic tendon measurements in dogs. Thirty German Shepherd dogs were examined: 13 males and 17 females. The cross-sectional area was significantly higher in males than in females (p <0.05) for the distal tendon of the triceps brachii muscle and the tendons of the flexor carpi ulnaris and common digital extensor muscles. The influence of sex on mean echogenicity was not significant. According to age, mean echogenicity was higher in older dogs, while the cross-sectional areas were similar in the two groups. Cross-sectional area and mean echogenicity of the tendons showed a direct increase with an increase in body weight. The data gained from this study can help support the clinician to discriminate between normal and pathological conditions.

Geographical information systems as a tool in the control of heartworm infections in dogs and cats.

Geospatial tools (e.g., geographical information systems, remote sensing, global positioning systems, and virtual globes) are very useful for the simultaneous visualization of health data with environmental data, which holds promise to understand environmental-health linkages and to generate new hypotheses to be tested in future research. Current epidemiological studies clearly show that the distribution patterns of vector-borne infections are changing; for example, in Europe, heartworm infection and subcutaneous dirofilariosis are spreading throughout areas that previously had little to no incidence of heartworm. In view of the changes of the distribution patterns of Dirofilaria immitis and Dirofilaria repens, geospatial tools are now more useful for mapping (including territorial sampling), monitoring, ecological analysis, risk assessment, forecasting (including the choose of the timing of treatment), early warning, and surveillance of both heartworm and subcutaneous dirofilariosis. All these issues have control of these infections as the ultimate goal.

Intestinal parasites in immigrants in the city of Naples (southern Italy).

The present study was aimed at carrying out a cross-sectional copromicroscopic survey of helminths and intestinal protozoa in immigrants in Naples (southern Italy). Between October 2008 and November 2009, a total of 514 immigrants were tested comparing the FLOTAC dual technique and the ethyl acetate concentration technique. Combined results of the two techniques served as a diagnostic 'gold' standard and revealed an overall prevalence of parasitic infections of 61.9% (318/514). The ethyl acetate concentration technique detected a low number of positive results (49.0%) and this was confirmed for each helminth/protozoa species detected. Among helminths, Trichuris trichiura (3.9%), hookworms (3.7%) and Ascaris lumbricoides (1.4%) were the most prevalent. Strongyloides stercoralis (0.4%), Enterobius vermicularis (0.4%), Schistosoma mansoni (1.0%), Hymenolepis nana (1.6%) and Taenia spp. (0.2%) were also found, as well as zoonotic helminths, as Trichostrongylus spp. (0.8%) and Dicrocoelium dendriticum (0.8%). As regard to pathogenic protozoa, Blastocystis hominis was the most commonly detected (52.7%), followed by Entamoeba histolytica/Entamoeba dispar/Entamoeba moshkovskii (11.9%) and Giardia duodenalis (4.5%). Several issues concerning diagnosis, epidemiology and public health impact of parasitic infections in immigrants are offered for discussion. In conclusion, the present paper pointed out the need of better diagnosis and cure of the immigrant population in order to improve access to health care of this neglected and marginalised population group, for its own protection and care.

Calibration and diagnostic accuracy of simple flotation, McMaster and FLOTAC for parasite egg counts in sheep.

The present study was aimed at carrying out a calibration and a comparison of diagnostic accuracy of three faecal egg counts (FEC) techniques, simple flotation, McMaster and FLOTAC, in order to find the best flotation solution (FS) for Dicrocoelium dendriticum, Moniezia expansa and gastrointestinal (GI) strongyle eggs, and to evaluate the influence of faecal preservation methods combined with FS on egg counts. Simple flotation failed to give satisfactory results with any samples. Overall, FLOTAC resulted in similar or higher eggs per gram of faeces (EPG) and lower coefficient of variation (CV) than McMaster. The "gold standard" for D. dendriticum was obtained with FLOTAC when using FS7 (EPG=219, CV=3.9%) and FS8 (EPG=226, CV=5.2%) on fresh faeces. The "gold standard" for M. expansa was obtained with FLOTAC, using FS3 (EPG=122, CV=4.1%) on fresh faeces. The "gold standard" for GI strongyles was obtained with FLOTAC when using FS5 (EPG=320, CV=4%) and FS2 (EPG=298, CV=5%). As regard to faecal preservation methods, formalin 5% and 10% or freezing showed performance similar to fresh faeces for eggs of D. dendriticum and M. expansa. However, these methods of preservation were not as successful with GI strongyle eggs. Vacuum packing with storage at +4°C permitted storage of GI strongyle eggs for up to 21 days prior to counting. Where accurate egg counts are required in ovine samples the optimum method of counting is the use of FLOTAC. In addition, we suggest the use of two solutions that are easy and cheap to purchase and prepare, saturated sodium chloride (FS2) for nematoda and cestoda eggs and saturated zinc sulphate (FS7) for trematoda eggs and nematoda larvae.

Covariate selection in multivariate spatial analysis of ovine parasitic infection.

Gastrointestinal (GI) strongyle and fluke infections remain one of the main constraints on health and productivity in sheep dairy production. A cross-sectional survey was conducted in 2004-2005 on ovine farms in the Campania region of southern Italy in order to evaluate the prevalence of Haemonchus contortus, Fasciola hepatica, Dicrocoelium dendriticum and Calicophoron daubneyi from among other parasitic infections. In the present work, we focused on the role of the ecological characteristics of the pasture environment while accounting for the underlying long range geographical risk pattern. Bayesian multivariate spatial statistical analysis was used. A systematic grid (10 km×10 km) sampling approach was used. Laboratory procedures were based on the FLOTAC technique to detect and count eggs of helminths. A Geographical Information System (GIS) was constructed by using environmental data layers. Data on each of these layers were then extracted for pasturing areas that were previously digitalized aerial images of the ovine farms. Bayesian multivariate statistical analyses, including improper multivariate conditional autoregressive models, were used to select covariates on a multivariate spatially structured risk surface. Out of the 121 tested farms, 109 were positive for H. contortus, 81 for D. dendriticum, 17 for C. daubneyi and 15 for F. hepatica. The statistical analysis highlighted a north-south long range spatially structured pattern. This geographical pattern is treated here as a confounder, because the main interest was in the causal role of ecological covariates at the level of each pasturing area. A high percentage of pasture and impermeable soil were strong predictors of F. hepatica risk and a high percentage of wood was a strong predictor of C. daubneyi. A high percentage of wood, rocks and arable soil with sparse trees explained the spatial distribution of D. dendriticum. Sparse vegetation, river, mixed soil and permeable soil explained the spatial distribution of the H. contortus. Bayesian multivariate spatial analysis of parasitic infections with covariates from remote sensing at a very small geographical level allowed us to identify relevant risk predictors. All the covariates selected are consistent with the life cycles of the helminths investigated. This research showed the utility of appropriate GIS-driven surveillance systems. Moreover, spatial features can be used to tailor sampling design where the sampling fraction can be a function of remote sensing covariables.

FLOTAC: an improved method for diagnosis of lungworm infections in sheep.

The FLOTAC techniques involve the spinning of faecal samples onto the surface of counting chambers to permit enumeration of parasitic elements (eggs, larvae, oocysts and cysts) to an accuracy of one parasitic element per gram of faeces. In the present study it is demonstrated that FLOTAC provides a rapid and very sensitive method for counting of lungworm larvae of sheep. The optimum flotation solution for lungworm larvae is zinc sulphate and mercury II iodide (s.g. 1.45) although zinc sulphate (s.g. 1.20 or 1.35) on its own also gave good results. Samples preserved in 5% formalin gave the highest counts but fresh, frozen and samples in 10% formalin also gave higher counts than McMaster and simple flotation. Larval counts of 307 field samples gave up to 1.27x more positives samples than use of Baermann funnels and up to 4.18x more larvae per sample. As FLOTAC is faster than Baermannisation of samples it offers a better method of counting larvae in ruminant faecal samples.

Long-term physical training allows high exercise performance in moderate chronic obstructive pulmonary disease (COPD).

Chronic obstructive pulmonary disease (COPD) is a major health problem, because of its prevalence, morbidity and mortality. As a result of symptoms such as cough and dyspnea patients with COPD suffer from exercise limitation and reduced health related quality of life. The present paper reports the case of a 67-year old ex-smoker patient with COPD, who had exercised regularly since when he was young, and maintained a better exercise capacity than healthy people of the same age, despite a forced expiratory volume in 1 second of the 60% of the predicted normal value. This case suggests that regular physical exercise in COPD patients may prevent the loss of exercise capacity despite significant airway obstruction.

Is gastrointestinal strongyle faecal egg count influenced by hour of sample collection and worm burden in goats?

Gastrointestinal (GI) strongyle infections remain one of the main constraints to goat production worldwide. Although several indicators of GI strongyle infections have been proposed, faecal egg count (FEC) techniques are the most common approaches for the estimation of prevalence and intensity of these parasites. However, FEC may be subjected to a within-individual variation due to factors such as weather, season, random day-to-day variation and phase of the parasitic infection. A longitudinal study of GI strongyle FEC was conducted on 63 dairy goats to evaluate the effects of hour of sample collection on FEC and the relationship between FEC and worm burden. The goats were naturally infected with Teladorsagia circumcincta, Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum venulosum. Every 3 weeks for 14 months faeces were collected every 2 h for 24 h from three individually caged goats. The faeces were those passed by the goats during the 2 h preceding each collection. For each goat a sample from this material was analyzed using the Flotac double technique. To explore relationships between FECs and adult parasite counts, on the day following sampling, the three goats were euthanized and the nematodes present in the abomasa and intestines were recovered, identified and counted. The results of the general linear model did not show any significant effect of the hour of faecal sample collection on GI strongyle FEC, whereas a significant effect of the month of faecal sample collection was found. The results of the Pearson correlation showed a significant (P=0.000) positive relationship between FEC and worm burden (r=0.6), in particular regarding H. contortus (r=0.9). In conclusion, the present study showed that the hour of sample collection does not influence the GI strongyle FEC and that there is a good relationship between FEC and total GI strongyle worm burden in goats.

Hosts and habitats of Trichinella spiralis and Trichinella britovi in Europe.

Trichinella spiralis and Trichinella britovi are the two most common species of Trichinella circulating in Europe. Based on data provided to the International Trichinella Reference Centre over the past 20 years (data referring to 540 isolates of T. spiralis and 776 isolates of T. britovi), we describe the host species and habitat characteristics for these two pathogens in Europe. A Geographical Information System was constructed using administrative boundaries, a Corine Land Cover (CLC) map, and an elevation map. In most countries, T. britovi is more widespread (62.5-100% of the isolates) than T. spiralis (0.0-37.5%), although in Finland, Germany, Poland and Spain, T. spiralis is more prevalent (56.3-84.2% of the isolates). Trichinella britovi is more widespread than T. spiralis in sylvatic carnivores (89% versus 11%), whereas T. spiralis is prevalent in both wild boars (62% versus 38%) and domestic swine (82% versus 18%), as well as in rodents (75% versus 25%). Trichinella spiralis and T. britovi circulate in the same environments: 41.1% and 46.0%, respectively, in agricultural areas, and 45.5% and 46.6% in forested and semi-natural areas. Although both pathogens can be transmitted by domestic and sylvatic cycles, their epidemiology is strongly influenced by the higher adaptability of T. spiralis to swine and of T. britovi to carnivores. These results are important because they include information on the countries at risk for these pathogens, the role played by specific species as reservoirs, the role of the pathogens in domestic and sylvatic cycles, and the role of the habitat in their circulation. The results can also be used to identify the most suitable animal species for the monitoring of these pathogens in Europe.

Giardia and Cryptosporidium in canine faecal samples contaminating an urban area.

The aim of the present paper was to continue the study on the presence of parasitic elements in the canine faeces contaminating the urban environment of Naples (southern Italy), focussing on the protozoa Giardia and Cryptosporidium. The total number of sub-areas studied was 143, and the total number of canine faecal samples collected and examined was 415. Each faecal sample was tested for the presence of copro-antigens of Giardia and Cryptosporidium using two commercially available enzyme-linked immunosorbent assays. Giardia antigens were found in 19.6% (28/143) of the sub-areas and in 7.7% (32/415) of the canine faeces collected. Cryptosporidium antigens were found in 4.2% (6/143) of the sub-areas and in 1.7% (7/415) of the canine faeces collected. Co-infection was not found in any sample. The results of the logistic regression models did not show any association between the positivity to Giardia or Cryptosporidium and the independent demographic variables (human population density, male and female population density) taken into consideration. In conclusion, the findings of the present study revealed the presence of Giardia and Cryptosporidium in canine faecal samples from the urban environment of Naples; however, the zoonotic potential of these findings was not assessed due to the lack of information on species/genotypes detected.