Termination of pregnancy in a border region between Switzerland and Italy (2008-2015).

In Switzerland, voluntary termination of pregnancy (VTP) can be performed in all public and private hospitals with an obstetrics/gynaecology department. For various reasons, many Italian women use the Swiss healthcare system, in particular in Canton Ticino, a border region adjacent to Italy in the southern part of Switzerland, when they want to have a VTP. In this study, we aimed to illustrate trends in the VTPs in the Canton Ticino between 2008 and 2015 and demonstrate differences between the Swiss women resident in Switzerland (SSR), foreign women resident in Switzerland (FSR) and foreign women resident abroad (FAR), focusing in particular on the Italian women as during this period there were legal changes in Italy. The number of VTPs was constant on a national level (10,924 in 2008, 10,255 in 2015); in contrast, since 2012 the number has progressively decreased (41%) in Ticino, mainly because of the significant reduction in VTPs in women resident in Italy (decrease of 75.7%). In addition, we wanted to evaluate the impact of the pre-VTP counselling at a family planning centre (FPC) on the VTP decision. The high number of pre-VTP consultations suggests that this service is appreciated and helpful. We observed an encouraging trend in changing the decision to have a VTP after the consultation at the FPC, where 12% of the pregnant women decided to continue the pregnancy. Because of its location, the Canton Ticino is an example how availability of certain drugs, methods and laws can influence the cross-border flow of the patients.

Phenotypic and genotypic identification of streptococci and related bacteria isolated from bovine intramammary infections.

BACKGROUND: Streptococcus spp. and other Gram-positive, catalase-negative cocci (PNC) form a large group of microorganisms which can be found in the milk of cows with intramammary infection. The most frequently observed PNC mastitis pathogens (major pathogens) are Streptococcus uberis, Strep. dysgalactiae, and Strep. agalactiae. The remaining PNC include a few minor pathogens and a large nonpathogenic group. Improved methods are needed for the accurate identification and differentiation of PNC. A total of 151 PNC were collected from cows with intramammary infection and conclusively identified by 16S rRNA sequencing as reference method. Nine phenotypic microbiological tests (alpha-hemolysis, CAMP reaction, esculin hydrolysis, growth on kanamycin esculin azide agar and on sodium chloride agar, inulin fermentation, hippurate hydrolysis, leucine aminopeptidase and pyrrolidonyl peptidase activity), multiplex PCR for the three major pathogens (target genes for Strep. uberis, Strep. dysgalactiae and Strep. agalactiae: pauA, 16S rRNA, and sklA3, respectively), and mass spectroscopy using the matrix-assisted laser desorption ionization-time of flight (MALDI-TOF MS) were evaluated for the diagnosis and discrimination of the three clinically most relevant PNC. RESULTS: The probability that a strain of Strep. uberis, Strep. dysgalactiae and Strep. agalactiae was correctly identified by combining the results of the 9 phenotypic tests was 92%, 90%, and 100%, respectively. Applying the multiplex PCR, all strains of the three major pathogens were correctly identified and no false positive results occurred. Correct identification was observed for all strains of Strep. uberis and Strep. agalactiae using MALDI-TOF MS. In the case of Strep. dysgalactiae, some variability was observed at the subspecies level, but all strains were allocated to one single cluster. CONCLUSIONS: The results of the present study show that reliable identification of the clinically most relevant PNC (Strep. uberis, Strep. agalactiae and Strep. dysgalactiae) can be obtained by use of a combination of colony morphology, hemolysis type and catalase reaction, and a multiplex PCR with specific primers restricted to these 3 pathogens. The MALDI-TOF MS is a fast method that shows promising results, although identification of Strep. dysgalactiae at the subspecies level is not yet satisfactory.

Meteorological factors and risk of community-acquired Legionnaires' disease in Switzerland: an epidemiological study.

OBJECTIVES: The aim of this study was to identify meteorological factors that could be associated with an increased risk of community-acquired Legionnaires' disease (LD) in two Swiss regions. DESIGN: Retrospective epidemiological study using discriminant analysis and multivariable Poisson regression. SETTING: We analysed legionellosis cases notified between January 2003 and December 2007 and we looked for a possible relationship between incidence rate and meteorological factors. PARTICIPANTS: Community-acquired LD cases in two Swiss regions, the Canton Ticino and the Basle region, with climatically different conditions were investigated. PRIMARY OUTCOME MEASURES: Vapour pressure, temperature, relative humidity, wind, precipitation and radiation recorded in weather stations of the two Swiss regions during the period January 2003 and December 2007. RESULTS: Discriminant analysis showed that the two regions are characterised by different meteorological conditions. A multiple Poisson regression analysis identified region, temperature and vapour pressure during the month of infection as significant risk factors for legionellosis. The risk of developing LD was 129.5% (or 136.4% when considering vapour pressure instead of temperature in the model) higher in the Canton Ticino as compared to the Basle region. There was an increased relative risk of LD by 11.4% (95% CI 7.70% to 15.30%) for each 1 hPa rise of vapour pressure or by 6.7% (95% CI 4.22% to 9.22%) for 1°C increase of temperature. CONCLUSIONS: In this study, higher water vapour pressure and heat were associated with a higher risk of community-acquired LD in two regions of Switzerland.

Detection limits of Legionella pneumophila in environmental samples after co-culture with Acanthamoeba polyphaga.

BACKGROUND: The efficiency of recovery and the detection limit of Legionella after co-culture with Acanthamoeba polyphaga are not known and so far no investigations have been carried out to determine the efficiency of the recovery of Legionella spp. by co-culture and compare it with that of conventional culturing methods. This study aimed to assess the detection limits of co-culture compared to culture for Legionella pneumophila in compost and air samples. Compost and air samples were spiked with known concentrations of L. pneumophila. Direct culturing and co-culture with amoebae were used in parallel to isolate L. pneumophila and recovery standard curves for both methods were produced for each sample. RESULTS: The co-culture proved to be more sensitive than the reference method, detecting 10²-10³ L. pneumophila cells in 1 g of spiked compost or 1 m³ of spiked air, as compared to 105-106 cells in 1 g of spiked compost and 1 m³ of spiked air. CONCLUSIONS: Co-culture with amoebae is a useful, sensitive and reliable technique to enrich L. pneumophila in environmental samples that contain only low amounts of bacterial cells.

Putative risk factors for infections with Toggenburg Orbivirus in goat herds in Southern Switzerland (Canton of Ticino).

Toggenburg Orbivirus (TOV), only detected in goats, has been described as a member of the Bluetongue virus (BTV) serogroup. The transmission pathway of the virus seems different from other Bluetongue viruses (BTVs). The objective of this study was to explore risk factors, especially the influence of alpine pasture and the presence of other livestock species, for the presence of TOV infected goats on farms. Between February 2008 and September 2009, blood samples were collected and analyzed for TOV and hereupon a total of 60 goat farm owners (37 TOV-positive and 23 TOV-negative holdings) were interviewed. Additionally, goatlings were tested for TOV by rRT-PCR prior and after alpine pasture in 2009. These goatlings were positive for TOV only after the alpine pasture. The final logistic regression model included: "exposure to goats from other farms" (OR=10.12, p=0.007), "exposure of the goats to red deer" (OR=4.79, p=0.04) and "exposure to sheep from other farms" (OR=0.05, p=0.002). These variables do not implicitly include direct contact, and the findings are only vaguely indicative for a contact-driven transmission. Furthermore, it is likely that they are only associated with, and thus indicative for, an unknown risk factor associated with alpine pasture not measured in the study. The results of this screening study do not indicate iatrogenic transmission pathways as a main transmission mode and stimulate the formulation of hypotheses on the origin, the transmission pathway and other host species for TOV. Especially, the involvement of an insect vector in transmission on alpine pasture and the relevance of vertical transmission are to be clarified.

Rapid identification of Legionella spp. by MALDI-TOF MS based protein mass fingerprinting.

A set of reference strains representing 38 different Legionella species were submitted to Whole Cell Mass Spectrometry (WCMS) with MALDI-TOF. The dendrogram computed from strain mass spectral patterns obtained by WCMS was compared to the phylogenetic tree obtained from macrophage infectivity potentiator (mip) sequences. The trees inferred from these two methods revealed significant homologies. Using 453 Legionella isolates previously characterized by genotyping, it was possible to create species-specific SuperSpectra, using appropriate sets of spectral masses, allowing unambiguous differentiation and identification of the most frequently isolated Legionella species. These SuperSpectra were tested for their suitability to identify Legionella strains isolated from water samples, cooling towers, potting soils and patient specimens deposited at the Swiss National Reference Centre for Legionella and previously identified by molecular methods such as mip gene sequencing. 99.1% of the tested strains isolated from the environment could be correctly identified by comparison with the new SuperSpectra. The identification of Legionella spp. by MALDI-TOF MS is rapid, easy to perform and has the advantage of being time- and cost-saving, in comparison to sequence-based identification.

Real-time PCR investigation of potential vectors, reservoirs, and shedding patterns of feline hemotropic mycoplasmas.

Three hemotropic mycoplasmas have been identified in pet cats: Mycoplasma haemofelis, "Candidatus Mycoplasma haemominutum," and "Candidatus Mycoplasma turicensis." The way in which these agents are transmitted is largely unknown. Thus, this study aimed to investigate fleas, ticks, and rodents as well as saliva and feces from infected cats for the presence of hemotropic mycoplasmas, to gain insight into potential transmission routes for these agents. DNA was extracted from arthropods and from rodent blood or tissue samples from Switzerland and from salivary and fecal swabs from two experimentally infected and six naturally infected cats. All samples were analyzed with real-time PCR, and some positive samples were confirmed by sequencing. Feline hemotropic mycoplasmas were detected in cat fleas and in a few Ixodes sp. and Rhipicephalus sp. ticks collected from animals but not in ticks collected from vegetation or from rodent samples, although the latter were frequently Mycoplasma coccoides PCR positive. When shedding patterns of feline hemotropic mycoplasmas were investigated, "Ca. Mycoplasma turicensis" DNA was detected in saliva and feces at the early but not at the late phase of infection. M. haemofelis and "Ca. Mycoplasma haemominutum" DNA was not amplified from saliva and feces of naturally infected cats, despite high hemotropic mycoplasma blood loads. Our results suggest that besides an ostensibly indirect transmission by fleas, direct transmission through saliva and feces at the early phase of infection could play a role in the epizootiology of feline hemotropic mycoplasmas. Neither the investigated tick nor the rodent population seems to represent a major reservoir for feline hemotropic mycoplasmas in Switzerland.

Use of mapping and statistical modelling for the prediction of bluetongue occurrence in Switzerland based on vector biology.

Due to the spread of bluetongue (BT) in Europe in the last decade, a sentinel surveillance programme was initiated for Switzerland in 2003, consisting of serological sampling of sentinel cattle tested for BT virus antibodies, as well as entomological trapping of Culicoides midges from June until October. The aim of this study was to create a 'suitability map' of Switzerland, indicating areas of potential disease occurrence based on the biological parameters of Obsoletus Complex habitat. Data on Culicoides catches from insect traps together with various environmental parameters were recorded and analysed. A multiple regression analysis was performed to determine correlation between the environmental conditions and vector abundance. Meteorological data were collected from 50 geo-referenced weather stations across Switzerland and maps of temperature, precipitation and altitude were created. A range of values of temperature, precipitation and altitude influencing vector biology were obtained from the literature. The final combined map highlighted areas in Switzerland which are most suitable for vector presence, hence implying a higher probability of disease occurrence given the presence of susceptible animals. The results confirmed the need for an early warning system for the surveillance of BT disease and its vectors in Switzerland.

Diversity of the population of Tick-borne encephalitis virus infecting Ixodes ricinus ticks in an endemic area of central Switzerland (Canton Bern).

Tick-borne encephalitis virus (TBEV), a member of the genus Flavivirus, has a positive-strand RNA genome containing a single open reading frame flanked by non-coding regions (NCRs). Ixodes ricinus ticks (n = 307) were collected from vegetation in a natural TBEV focus in Belp, Switzerland. The presence and identity of the virus were determined by nested RT-PCR followed by sequencing of the 5'-terminal region that comprises the 5' NCR and the capsid-encoding region (C). The presence of the western European TBEV subtype (W-TBEV) genome was detected in 14.3 % of the ticks. Nucleotide sequence analysis revealed a high variability of 55.5 %. In particular, four DNA fragments (CS 'A', CS 'B', the folding-stem structure and the start codon) showed substantial heterogeneity, which has the potential of compromising replication, translation and packaging of the viral genome. This variability may reflect a viral strategy to select the fittest RNA molecule to produce a viral infection in the different vertebrate hosts that may be encountered by the ticks. It may also indicate a possible ancient introduction of TBEV to the Belp site. In addition, it may contribute to explaining the annual low incidence of tick-borne encephalitis in the natural focus of Belp, despite the high prevalence of TBEV genomes in ticks.

Presence of potentially pathogenic Babesia sp. for human in Ixodes ricinus in Switzerland.

We have designed and performed a new PCR method based on the 18S rRNA in order to individuate the presence and the identity of Babesia parasites. Out of 1159 Ixodes ricinus (Acari: Ixodidae) ticks collected in four areas of Switzerland, nine were found to contain Babesia DNA. Sequencing of the short amplicon obtained (411-452 bp) allowed the identification of three human pathogenic species: Babesia microti, B. divergens, for the first time in Switzerland, Babesia sp. EU1. We also report coinfections with B. sp. EU1-Borrelia burgdorferi sensu stricto and Babesia sp. EU1-B. afzelii.

Diversity within Borrelia burgdorferi sensu lato genospecies in Switzerland by recA gene sequence.

A total of 874 Ixodes ricinus ticks were collected in Switzerland to investigate the genetic diversity of the Borrelia population. We integrated to the RT-PCR method the DNA sequence analysis of a 162-bp fragment of the recA gene. Five genospecies were detected: Borrelia afzelii, Borrelia burgdorferi s.s., Borrelia garinii, Borrelia valaisiana, and Borrelia lusitaniae. A heterogeneous distribution was observed within the B. burgdorferi s.l. genospecies. The most prevalent and diverse genospecies found in Switzerland was Borrelia afzelii, which might suggest a rapid evolution of this genospecies.

Rhipicephalus ticks infected with Rickettsia and Coxiella in Southern Switzerland (Canton Ticino).

Ticks of the Rhipicephalus sanguineus species complex may be vector of various pathogens including Rickettsia conorii (the etiological agent of the Mediterranean spotted fever) and Coxiella burnetii (cause of the Query (Q) fever). R. sanguineus ticks have been imported in several parts of central and northern Europe, especially in environments such as kennels and houses providing the appropriate microclimatic conditions and the blood source necessary for their survival. Since 1940 these ticks have occasionally been recorded in Switzerland. In Ticino (the southern part of Switzerland), they have been reported since 1980 and their probable establishment in this area has been suggested in the '90s. By means of PCR and direct sequencing, we tested the identity of these ticks (using 12S rDNA gene) and the occurrence of Rickettsia spp. (using 16S rDNA, gltA and OmpA genes) as well as Coxiella sp. (using 16S rDNA). The results indicated that in Ticino, two different tick species coexist, i.e. R. sanguineus sensu stricto and Rhipicephalus turanicus. A few individuals of R. sanguineus sensu stricto are infected with Rickettsia massiliae/Bar29, which are strains of unknown pathogenicity. Coxiella sp., an endosymbiont of Rhipicephalus ticks, has also been identified in both tick species. Due to climatic changes towards global warming, imported tick species may therefore adapt to new area and might be considered as epidemiological markers for a number of infectious agents transmitted by them.