Field observations and experiences gained from the implementation of control measures against lumpy skin disease in South-East Europe between 2015 and 2017.

The first epidemics of lumpy skin disease (LSD) reported in Europe in 2015 severely affected the cattle farming sector in several Balkan countries. After the first incursion into Greece in 2015, the disease quickly spread across the Balkan region with over 7000 outbreaks reported by the end of 2016. Thanks to a coordinated regional control and eradication policy, the spread of the disease was halted by the end of 2017. Regional large-scale vaccination campaign with effective homologous vaccines and high vaccination coverage revealed to be essential for the successful control the disease, supported by other measures such as early detection of outbreaks, total or partial stamping out and restrictions on cattle movements. The aim of this paper is to discuss the field observations, challenges and lessons learnt while dealing with the first LSD epidemics in Europe. The cross-border collaboration by the veterinary authorities of all affected countries, coordinated by the European Commission and the technical support provided by many other international organizations played a fundamental role in stopping the spread of a disease that otherwise could have expanded further to the European territory causing a large damage to the whole European cattle farming industry. The experience obtained during the control of LSD epidemics indicates that in the future LSD spread can be effectively halted, provided that appropriate surveillance plans and vigilance remains in place in the areas at risk of re-incursion, especially those bordering endemic countries.

Occurrence and molecular typing of Giardia isolates in pet rabbits, chinchillas, guinea pigs and ferrets collected in Europe during 2006-2012.

A total of 1180 faecal samples (528 from rabbits, 531 from chinchillas and 121 from guinea pigs) collected during 2006-2012 by veterinarians in Germany and in other European countries were submitted to a diagnostic laboratory for Giardia testing by means of coproantigen ELISA. Of these samples, 40 rabbits (7.6 per cent), 326 chinchillas (61.4 per cent) and five guinea pigs (4.1 per cent ) were found to be positive. To gain insights into the genetic identity of Giardia in small mammals, ELISA-positive samples from 23 chinchillas, five ferrets, a rabbit, and a Desmarest's hutia were investigated by PCR and sequencing of fragments of the small subunit ribosomal DNA (ssu), the triose phosphate isomerase (tpi) and the ß-giardin (bg) genes. At the ssu locus, assemblage B was identified in 28 of 30 isolates, whereas assemblage A and D were each detected in one sample. The majority of isolates from chinchillas and those from ferrets had Giardia duodenalis sequences identical to sub-assemblages AI or BIV, based on either a single locus (tpi or bg) or multiple loci (tpi and bg). As sub-assemblages AI or BIV are associated with human infection, these results indicate that small mammals can act as reservoirs of cysts potentially infectious to humans.

Polymerase chain reaction - restriction fragment length polymorphism analysis for the differentiation of Trichinella nativa and Trichinella britovi.

Recently, Trichinella nativa was identified in foxes in Germany and Poland, indicating that the geographical distribution of T. nativa is not restricted to areas north of the isotherm -4°C in January. In the European Union, legislation requires that a regular monitoring of the occurrence of Trichinella spp. in indicator animals such as foxes or raccoon dogs be carried out. The Trichinella isolates must also be identified on a species level. The multiplex PCR recommended by the Community Reference Laboratory for Trichinella allows species identification, yet the differentiation of T. nativa and Trichinella britovi, a widespread Trichinella species in the temperate regions of Europe, is unstable. We therefore describe an easy and reliable method for the differentiation of the two species, which can be utilised to monitor a potential spread of T. nativa in Central Europe.

Molecular typing of Giardia duodenalis isolates from German travellers.

Giardia duodenalis isolates from German travellers returning from tropical areas were characterised by PCR amplification and sequencing of fragments of the beta-giardin (bg), glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) genes. Assignment of isolates to specific G. duodenalis assemblages was found to differ according to the marker used. Indeed, at the bg locus, assemblages A and B were identified, with a higher prevalence of the former over the latter, whereas at the tpi and gdh loci, most samples were classified as assemblage B. In agreement with previous studies, sequence analysis showed that assemblage B isolates have a higher genetic polymorphism than assemblage A isolates, and novel variants were described. The degree of polymorphism was shown in a graphical representation of the polymorphic sites generating a novel sequence, the heterogeneous positions common to assemblages A and B (double peaks), that may represent mixed assemblage infection and the heterogeneous positions detected at random sites. Notably, assemblage D, which is considered to be adapted to dogs, was found at the gdh locus in two samples originating from southern Asia, as novel genotypes. By comparing the geographical origin of the infected cases and the number of German travellers visiting the areas considered, India and west Africa appeared to be the areas associated to the highest risk of acquiring Giardia infection. The analysis of the geographical distribution of the genotypes did not suggest any particular geographical clustering pattern, but it may be useful to evaluate these results with a higher number of isolates. Most of the samples typed at the three markers could not be assigned unequivocally to either assemblage A or B, and this was confirmed also by a real-time PCR assay, using a set of assemblage-specific primers. The results of this study reinforce the notion that genetic exchanges and allelic sequence heterogeneity represent major obstacles towards understanding the epidemiology of giardiasis and that exposure to Giardia parasites in endemic areas often results in mixed infections in returning travellers.

A preliminary microbiological assessment of process hygiene of traditional outdoor camel slaughter in Sahrawi refugee camps.

The aim of this study was to investigate the hygiene performance of a camel (Camelus dromedarius) slaughtering process as carried out with the traditional method in the Sahrawi refugee camps located in southwestern Algeria. The camel slaughtering process in this region differs significantly from that carried out in commercial abattoirs. Slaughtering is performed outdoors in desert areas, and dehiding of the carcass is approached via the dorsoventral route rather than the classic ventrodorsal route. Samples were taken from 10 camel carcasses from three different areas: the hide, the carcass meat immediately after dehiding, and the meat after final cutting. Enterobacteriaceae counts (EC) were enumerated employing conventional laboratory techniques. Carcass meat samples resulted in EC below the detection limit more frequently if the hide samples from the same carcass had also EC counts below the detection limit. Because of the low number of trials, the calculation of statistical significance of the results was not possible. Further experimental research is needed in order to validate the results presented in this study. The comparison of the microbiological hygiene performance between dorsal dehiding and traditional ventral dehiding of slaughtered animals could serve to validate the hypothesis of the potential positive impact of the dorsal dehiding method in carcass meat hygiene.

Changing dietary habits in a changing world: emerging drivers for the transmission of foodborne parasitic zoonoses.

Changing eating habits, population growth and movements, global trade of foodstuff, changes in food production systems, climate change, increased awareness and better diagnostic tools are some of the main drivers affecting the emergence or re-emergence of many foodborne parasitic diseases in recent years. In particular, the increasing demand for exotic and raw food is one of the reasons why reports of foodborne infections, and especially waterborne parasitosis, have increased in the last years. Moreover increasing global demand for protein of animal origin has led to certain farming practices (e.g. aquaculture) increasing in emerging or developing countries, where health monitoring may not be sufficiently implemented. Therefore, high quality epidemiological data are needed which together with biological, economic, social and cultural variables should be taken into account when setting control programs for these increasingly popular production systems in emerging economies. This review focuses on the dietary, social, economic and environmental changes that may cause an increase in human exposure to foodborne parasites. Some examples illustrating these new epidemiological dynamics of transmission foodborne parasitic disease are presented.

Evaluation of a Western Blot and ELISA for the detection of anti-Trichinella-IgG in pig sera.

Human trichinellosis is a foodborne disease caused by ingestion of infective Trichinella muscle larvae via pork or meat of other food animals which are susceptible to this zoonotic parasite. There are new approaches for a risk-oriented meat inspection for Trichinella in pigs which are accompanied by monitoring programmes on herd level to control freedom from this parasite. For this purpose, testing schemes utilizing serological tests with a high sensitivity and specificity are required. This study aimed at the evaluation of an ELISA and a Western Blot (WB) for the detection of anti-Trichinella-IgG in terms of sensitivity and specificity taking results of artificial digestion as gold standard. For this purpose, 144 field sera from pigs confirmed as Trichinella-free as well as 159 sera from pigs experimentally infected with T. spiralis (123), T. britovi (19) or T. pseudospiralis (17) were examined by ELISA (excretory-secretory antigen) and WB (crude worm extract). Sera from pigs experimentally infected with four other nematode species were included to investigate the cross-reactivity of the antigen used in the WB. For all Trichinella-positive pig sera, band pattern profiles were identified in the WB and results were analysed in relation to ELISA OD% values. Testing of pig sera revealed a sensitivity of 96.8% for the ELISA and 98.1% for the WB whereas the methods showed a specificity of 97.9 and 100%, respectively. WB analysis of Trichinella-positive pig sera revealed five specific band patterns of 43, 47, 61, 66, and 102 kDa of which the 43 kDa protein was identified as the predominant antigen. The frequency of the band pattern profile was irrespective of the dose and the period of infection as well as the Trichinella species investigated. In conclusion, monitoring in swine farms for Trichinella antibodies should be based on screening pig sera by means of ELISA followed by confirmatory testing through WB analysis.

Experimental ELISA for diagnosis of ovine dicrocoeliosis and application in a field survey.

Dicrocoeliosis is a parasitic disease caused by trematodes belonging to the genus Dicrocoelium. It usually produces mild symptoms, and for this reason, dicrocoeliosis often remains undetected. Its diagnosis is mostly based on postmortem examination of the liver or on coprological assays for in vivo diagnosis. However, the latter method has scant sensitivity and because of the long pre-patency of Dicrocoelium spp. only permits late diagnosis. In the present study, an enzyme-linked immunosorbent assay (ELISA) was developed based on excretory/secretory antigen of Dicrocoelium dendriticum. The ELISA detected antibodies (IgG) in experimental infected sheep starting from day 30 post-infection (d.p.i.), whereas coprological samples were positive from 58 d.p.i. The ELISA was used in a field study in order to assess dicrocoeliosis seroprevalence in ovine flocks from the province of Trento (northeastern Italy), and this was the first sero-epidemiological study of ovine dicrocoeliosis in northern Italy. Altogether, 842 sheep sera were tested. In accordance with previous surveys carried out in other regions of Italy, a high prevalence of 80% to 100% was found.

Distribution of Cryptosporidium parvum subtypes in calves in Germany.

Cryptosporidium DNA was extracted from 134 faecal specimens from pre-weaned calves from different German Federal States (age range, 3-15 days old), which tested positive for oocysts by microscopic analysis. The 18S rDNA gene and the oocyst wall protein gene (COWP) were used as targets for PCR and RFLP techniques. Cryptosporidium species were identified by using SspI, MboII and RsaI endonucleases for the digestion of 18S rDNA and COWP amplified fragments, respectively. In all samples, restriction patterns corresponding to Cryptosporidium parvum were identified, which is in agreement with abundant literature data indicating C. parvum as the most common species in pre-weaned calves. In order to estimate the genetic heterogeneity among C. parvum calf isolates, 53 samples chosen to represent different German Federal States were successfully subtyped by sequence analysis of the highly polymorphic 60-kDa glycoprotein gene. All isolates belonged to the allele IIa (with seven subtypes), with the exception of one isolate that belonged to the allele IId. Moreover, three novel subtypes of the allele family IIa have been found. This study confirms the utility of genotyping and subtyping tools in characterizing the transmission of Cryptosporidium spp. This is the first molecular epidemiological report about subtyping of Cryptosporidium bovine isolates in Germany.

Frequency and complications of chronic iridocyclitis in ANA-positive pauciarticular juvenile chronic arthritis.

Chronic iridocyclitis (CI) is among the most important extra-articular manifestations of juvenile chronic arthritis (JCA) and is most often observed in young girls with pauciarticular JCA and circulating antinuclear antibodies (ANAs). The frequency of CI found in 39 children with ANA-positive pauciarticular JCA was 56%. None of the patients developed ocular complications during follow-up. We think that this favourable outcome was primarily due to the early diagnosis and to our choice of using atropine as a mydriatic agent and to prolong atropine, after having spaced its administration further apart, up to one year or longer after the remission of ocular manifestations.

[Temporary myopia and subjective symptoms in video display terminal operators]. / Miopizzazione temporanea e sintomatologia soggettiva in operatori al videoterminale.

The study covered a group of female VDT operators assigned to data entry and data acquisition. Subjective VDT-related symptoms of asthenopia were assessed by means of a questionnaire. All operators were examined by an ophthalmologist. Visual acuity was measured using vision tables with optotypes. In order to achieve an objective assessment of VDT-induced visual fatigue, refractive power was measured at the beginning and at the end of the shift, using an infra-red autorefractometer. Changes in refractive power were then related to VDT work and asthenopia symptoms. Visual acuity defects were observed in 63.5% of the operators; in 36% of the cases the subjects were either unaware of the defects or the defects were not adequately corrected. 62.5% of the operators complained of subjective asthenopia symptoms. Asthenopia was not related to the number of hours at the VDT. The results suggest that ametropic subjects are likely to be more susceptible to visual fatigue than emmetropic subjects, since there was a tendency for the prevalence of asthenopia to increase in the former group. A significant decrease in refractive power (temporary myopization) was observed in 20% of the operators at the end of the shift at the VDT: all these subjects also complained of asthenopia, compared to 50% of the workers without end-of-shift myopization. The difference between the groups was highly significant (p less than 0.01); contrarywise, none of the subjects without asthenopia developed myopization. On the basis of these results, it can be concluded that end-of-shift myopization, as measured by an automatic refractometer, may be used as a reliable objective measurement of VDT-related visual fatigue.

Visual fatigue in video display terminal operators: objective measure and relation to environmental conditions.

The lighting conditions, luminance, contrast, and design of the workplace were studied in video display terminal (VDT) work stations operated by a group of female VDT data-acquisition clerks. VDT-induced symptoms were assessed by means of subject answers to a questionnaire. To measure VDT-induced ocular fatigue objectively, refraction power was determined before and at the end of workshift by an infrared autorefractometer. Job-induced refraction changes were then related to visual complaints and conditions in the workplace. The results confirmed that VDT data-acquisition work can lead to temporary myopia (myopization) in a remarkable percentage of operators; a significant correlation between eye discomfort, ocular asthenopia, and myopization was also found. Illumination levels, luminance, and contrast seem to be of paramount importance regarding visual symptoms: neither asthenopia nor myopization was observed when all of these conditions were adequate. If the ergonomic design of the workplace and the viewing distance are adequate, there are also usually fewer musculoskeletal symptoms. Our results suggest that changes in the ocular refraction status before and at the end of the work-shift, as determined by an automatic refractometer, provide a good objective index of VDT-induced "ocular fatigue", which in our study proved to be significantly related to workplace conditions.