Effects of enzymatic hydrolysis and ultrasounds pretreatments on corn cob and vine trimming shoots for biogas production.

Due to their lignocellulosic nature, corn cob and vine trimming shoots (VTS) could be valorized by anaerobic digestion for biogas production. To enhance the digestibility of substrates, pretreatments of lignocellulosic materials are recommended. The effect of enzymatic hydrolysis, ultrasounds pretreatments (US) and the combination of both was assayed in lignocellulosic composition, methane, and biogas yields. The pretreatments leaded to a reduction in lignin and an increase in neutral detergent soluble compounds making corn cob and VTS more amendable for biogas conversion. The US were negative for biogas production from both substrates and in particular strongly detrimental for VTS. On the opposite side, the enzymatic hydrolysis was certainly beneficial increasing 59.8% and 14.6% the methane production from VTS and corn cob, respectively. The prior application of US did not potentiate (or not sufficiently) the improvement in the methane production reflected by the enzymatic hydrolysis pretreatment of VTS and corn cob.

Data management systems for the bovine viral diarrhoea eradication programme in Switzerland.

A programme to eradicate bovine viral diarrhoea was launched in Switzerland in 2008 with the aim of eradicating the causal virus. During the first year of the programme, the entire population of 1.6 million cattle were tested for the presence of the virus; in the following three years an additional 1.8 million calves were tested. The complexity of information generated during the eradication programme, together with a tight schedule, made computerised data management a necessity. To organise, coordinate and supervise the programme, extensions were made to the computerised information system ISVet, of the Swiss Veterinary Service, which provides automated documents for both the Veterinary Service and private veterinarians. Specific data are accessible by user groups via the BVD-Web platform, ISVet and the Swiss animal movement database. The functionalities of the structure and the reports needed to control the progress of the programme are described in detail. The authors also discuss the major advantages, disadvantages and pitfalls when planning an eradication programme using a national centralised database over a distributed computer network.

Risk assessment for the design of a risk-based surveillance programme for fish farms in Switzerland (in accordance with Council Directive 2006/88/EC of the European Union).

Swiss aquaculture farms were assessed according to their risk of acquiring or spreading viral haemorrhagic septicaemia (VHS) and infectious haematopoietic necrosis (IHN). Risk factors for the introduction and spread of VHS and IHN were defined and assessed using published data and expert opinions. Among the 357 aquaculture farms identified in Switzerland, 49.3% were categorised as high risk, 49.0% as medium risk and 1.7% as low risk. According to the new Directive 2006/88/EC for aquaculture of the European Union, the frequency of farm inspections must be derived from their risk levels. A sensitivity analysis showed that water supply and fish movements were highly influential on the output of the risk assessment regarding the introduction of VHS and IHN. Fish movements were also highly influential on the risk assessment output regarding the spread of these diseases.

Toxoplasma gondii in Switzerland: a serosurvey based on meat juice analysis of slaughtered pigs, wild boar, sheep and cattle.

Toxoplasmosis is one of the most important zoonotic diseases worldwide and is caused by the protozoan Toxoplasma gondii. Besides vertical infection during pregnancy, humans can get infected post-natally either by peroral uptake of sporulated Toxoplasma oocysts or by ingestion of tissue cysts upon consumption of raw or undercooked meat. The aim of this study was to approximate the risk of human infection via meat consumption by estimating the seroprevalence of T. gondii in slaughtered animals in Switzerland and to compare data with prevalences assessed 10 years ago. The study included pigs, cattle, sheep and wild boar of different age groups and housing conditions whenever possible and applicable. A P-30-ELISA was used to detect T. gondii-specific antibodies and to determine seroprevalences in meat juice of slaughtered animals. A total of 270 domestic pigs (120 adults, 50 finishing, 100 free-ranging animals), 150 wild boars, 250 sheep (150 adults, 100 lambs) and 406 cattle (47 calves, 129 heifers, 100 bulls, 130 adult cows) were tested. Seropositivity increased with the age of the assessed animals. Independent of the age-group, the overall seroprevalence was lowest in wild boars (6.7%), followed by pigs (23.3%), cattle (45.6%) and sheep (61.6%), respectively. Conventional fattening pigs and free-ranging pigs surprisingly had comparable seroprevalences (14.0% and 13.0%, respectively). Unlike in other European countries, where generally a decrease in the number of seropositive animals had been observed, we found that the prevalence of seropositive animals, when compared with that of 10 years ago, had increased for most species/age groups. Conclusively, the results demonstrated a high seroprevalence of T. gondii in animals slaughtered for meat production and revealed that increasing age of the animals is a more important risk factor than housing conditions in Switzerland.

Rapid measurement of the yield stress of anaerobically-digested solid waste using slump tests.

The anaerobic digestion of solid waste is usually performed using dry or semi-dry technology. Incoming waste and fermenting digestate are pasty media and thus, at the industrial scale, their suitability for pumping and mixing is a prerequisite at the industrial scale. However, their rheology has been poorly characterised in the literature because there is no suitable experimental system for analysing heterogeneous media composed of coarse particles. We have developed a practical rheometrical test, a "slump test", for the analysis of actual digested solid waste. It makes it possible to estimate yield stress from the final slump height. From the slump behavior, we conclude that digestates behave as visco-elastic materials. The yield stress of different digested waste was measured between 200 and 800Pa. We show that the media containing smaller particles or with higher moisture content are characterised by smaller yield stresses. This study thus demonstrates the impact of the origin of the digestate on the yield stress.

Prevalence and genotypes of Toxoplasma gondii in feline faeces (oocysts) and meat from sheep, cattle and pigs in Switzerland.

The protozoan parasite Toxoplasma gondii infects almost all warm blooded animal species including humans, and is one of the most prevalent zoonotic parasites worldwide. Post-natal infection in humans is acquired through oral uptake of sporulated T. gondii oocysts or by ingestion of parasite tissue cysts upon consumption of raw or undercooked meat. This study was undertaken to determine the prevalence of oocyst-shedding by cats and to assess the level of infection with T. gondii in meat-producing animals in Switzerland via detection of genomic DNA (gDNA) in muscle samples. In total, 252 cats (44 stray cats, 171 pet cats, 37 cats with gastrointestinal disorders) were analysed coproscopically, and subsequently species-specific identification of T. gondii oocysts was achieved by Polymerase Chain Reaction (PCR). Furthermore, diaphragm samples of 270 domestic pigs (120 adults, 50 finishing, and 100 free-range animals), 150 wild boar, 250 sheep (150 adults and 100 lambs) and 406 cattle (47 calves, 129 heifers, 100 bulls, and 130 adult cows) were investigated by T. gondii-specific real-time PCR. For the first time in Switzerland, PCR-positive samples were subsequently genotyped using nine PCR-restriction fragment length polymorphism (PCR-RFLP) loci (SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) for analysis. Only one of the cats shed T. gondii oocysts, corresponding to a T. gondii prevalence of 0.4% (95% CI: 0.0-2.2%). In meat-producing animals, gDNA prevalence was lowest in wild boar (0.7%; 95% CI: 0.0-3.7%), followed by sheep (2.0%; 95% CI: 0.1-4.6%) and pigs (2.2%; 95% CI: 0.8-4.8%). The highest prevalence was found in cattle (4.7%; 95% CI: 2.8-7.2%), mainly due to the high prevalence of 29.8% in young calves. With regard to housing conditions, conventional fattening pigs and free-range pigs surprisingly exhibited the same prevalence (2.0%; 95% CI: 0.2-7.0%). Genotyping of oocysts shed by the cat showed T. gondii with clonal Type II alleles and the Apico I allele. T. gondii with clonal Type II alleles were also predominantly observed in sheep, while T. gondii with mixed or atypical allele combinations were very rare in sheep. In pigs and cattle however, genotyping of T. gondii was often incomplete. These findings suggested that cattle in Switzerland might be infected with Toxoplasma of the clonal Types I or III, atypical T. gondii or more than one clonal Type.

Long-term estrogen exposure of whitefish Coregonus lavaretus induces intersex but not Lake Thun-typical gonad malformations.

A high prevalence of gonad morphological variations has been observed in whitefish Coregonus lavaretus from Lake Thun (Switzerland). To clarify the role of endocrine disruption as a possible cause of the gonad alterations, whitefish were reared in a long-term laboratory experiment under exposure to 17 beta-estradiol (E2). Fish were fed from first-feeding until 3 yr of age at a daily rate of 0 (control), 0.5 or 50 microg E2 kg(-1) fish. E2 exposure resulted in a time- and concentration-dependent increase of prevalence and intensity of intersex gonads, i.e. gonads that macroscopically appeared as either testis or ovary but microscopically contained both male and female germ cells. Four types of intersex could be distinguished: Types 1 and 2 were composed of mainly male tissue, with Type 1 containing single oocytes and Type 2 displaying an ovary-like lamellar structure of the tissue. In Type 3, an increased percentage of the tissue was occupied by female germ cells, while in Type 4, the majority of the gonad tissue consisted of female germ cells. Chronic E2 exposure additionally resulted in a concentration-dependent shift of the sex ratio towards females, a reduced condition factor, retarded gonad growth together with delayed maturation of germ cells, and elevated levels of hepatic vitellogenin mRNA. However, Lake Thun-typical alterations of gonad morphology were not induced by chronic E2 exposure. The results provide evidence that estrogen-active compounds unlikely play a role in the etiology of gonad malformations in Lake Thun whitefish.

How normal is abnormal? Discrimination between deformations and natural variation in gonad morphology of European whitefish Coregonus lavaretus.

The gonad morphology of whitefish Coregonus lavaretus collected in Lake Thun, Switzerland, and two neighbouring lakes was assessed in order to differentiate between 'normal' and 'abnormal' character states of gonad morphology, which had been previously described in C. lavaretus from Lake Thun (constrictions, asymmetries, aplasia, compartmentations, fusions and hermaphroditism). In total, 4668 fish were collected and analysed using two complementary sampling schemes: (1) monthly samples of catches by the commercial fishermen and (2) samples of ripe spawners of all known 33 spawning sites of the three lakes. Considerable variation in gonad morphology in C. lavaretus populations of all lakes was found. Notably, all deviation types were observed in fish of all three lakes. Asymmetries and constrictions were frequent in all three lakes and showed systematic differences in frequency between the two sampling strategies. This indicates that asymmetries and constrictions represent to a large extent natural variation in gonad morphology of C. lavaretus and are also prone to considerable measurement error. In contrast, aplasia, fusions, compartmentations and hermaphroditism occurred predominantly in one C. lavaretus form of Lake Thun and in particular in populations spawning at great depths. This suggests that these deviation types are probably reliable indicators for gonad deformations and supports the interpretation that Lake Thun harbours a unique case of deformed gonads in C. lavaretus of yet unknown origin.

Frequent and unexplained gonadal abnormalities in whitefish (central alpine Coregonus sp.) from an alpine oligotrophic lake in Switzerland.

The occurrence of a large number of whitefish with morphologically altered gonads in Lake Thun (Switzerland) has been reported by commercial fishermen since the year 2000. Macroscopic and microscopic examination of gonads revealed that 35% (281 out of 808) of whitefish from Lake Thun were affected. Frequency of gonadal abnormalities varied significantly with sex, age and ecotype of whitefish. Apparent females (26% [106/408]) showed significantly less gonadal alterations than apparent males (40% [151/225]). Fish with deformed gonads were observed in all age classes from 1 to 6 yr, but were most frequent in the year classes 3+ to 5+. The gonadal alterations showed different frequencies among the 3 ecotypes of whitefish living in the lake: 1 ecotype was less affected (26%) than the other 2 forms (41% and 32%, respectively). The gonadal alterations included adhesions/fusions to the peritoneal wall and the lateral trunk musculature (overall: 5%; in females: 5%; in males: 5%), asymmetry (4%; 6%; 4%), atrophy (4%; 6%; 1%), compartmentations (11%; 4%; 18%), constrictions (3%; 1%; 7%) and hermaphroditism (1.1% of sampled fish, and 10 additional specimens collected by fishermen during the filleting process). In some cases, more than one alteration was observed in the same fish. Hermaphroditism manifested as 3 different types: (1) the lobular type, in which a consecutive sequence of testicular and ovarian lobes are found on the same gonad strand; (2) discrete gonads within one fish, with one gonad strand being ovarian tissue and the other being testicular tissue; and (3) the mosaic type, in which oocytes lie in tissue with normal testicular morphology, or spermatids or sperm are present in ovarian tissue. Several features of the gonadal alterations described in this study of whitefish from Lake Thun have also been reported for whitefish from other areas. However, the Lake Thun situation appears to be unique because of the high prevalence of fish affected and the broad variation of abnormal features. The causes of the frequent appearance of these gonadal alterations in whitefish from Lake Thun remain unclear.

Morphological organ alterations and infectious diseases in brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss exposed to polluted river water.

Poor water quality is discussed as a major factor causing a decline of brown trout populations in Swiss rivers. For our study we have chosen a river in the Swiss midlands, where the brown trout population has decreased dramatically during the last 10 yr and where feral fish have shown distinctive pathological alterations. The objective of our study was to investigate whether river water may be responsible for impaired fish health leading to an increased mortality in the river. In an active monitoring program, groups of brown and rainbow trout were exposed to polluted river water for 24 mo. Fish held in tap water served as a reference. Mortality, macroscopic and histopathologic changes, and infectious agents were investigated. Compared with the reference group, high mortality rates and severe pathological alterations of the inner organs were observed in fish held in river water. Especially gills, liver and kidney of these fish showed significantly higher changes than fish from tap water. These changes were dominated by degenerative and inflammatory reactions. Additionally, several infectious agents were diagnosed in fish exposed to river water. The most important findings were furunculosis and proliferative kidney disease. Brown trout seemed to be more sensitive than rainbow trout to environmental stress and infectious agents.

Effluent from a sewage treatment works causes changes in serum chemistry of brown trout (Salmo trutta L.).

To evaluate the impact of effluent from a sewage treatment works on fish health, serum chemistry variables were investigated in brown trout (Salmo trutta L.) held in cages (active monitoring) and wild brown trout (passive monitoring). Means of the measured serum parameters of the different treatment groups were close or within normal ranges. However, the results of the active monitoring demonstrated that the serum variables of reference trout held in tap water were clearly different from those of the river treatment groups. In the active monitoring, fish exposed to effluent from the sewage treatment works had significantly different blood urea nitrogen and bilirubin values than fish kept in river water. In the passive monitoring, total protein, blood urea nitrogen, and alkaline phosphatase were significantly different between the two groups. Of the numerous correlations between serum chemistry parameters and histological lesions, blood urea nitrogen and alkaline phosphatase were found to most strongly indicate gill and liver lesions, respectively. In the passive monitoring correlations between serum chemistry variables and histopathological lesions were restricted to bilirubin and liver lesions. This indicates that the application of serum chemistry variables as indicators of histological lesions in case of chronic exposure is questionable. A multivariate discriminant analysis was used to consider relationships between the single serum variables concurrently.

Increase of metallothionein-immunopositive chloride cells in the gills of brown trout and rainbow trout after exposure to sewage treatment plant effluents.

Metallothionein, a biomarker of exposure and toxicity of heavy metals, has been detected in the gills of brown trout (Salmo trutta fario L.) and rainbow trout (Oncorhynchus mykiss Richardson) by means of immunohistochemistry. A very prominent labelling of chloride cells was found after exposure to diluted sewage plant effluents. No significant increase was observed in either the number of labelled cells or their labelling intensity after exposure to water of a polluted river compared to fish kept in tap water. These results do not correlate with findings of a histopathological study, suggesting that the metal levels at the sewage treatment plant were too low to produce gross histopathology. A comparison between the species indicated that the rainbow trout showed a generally higher metallothionein expression than the brown trout.

Rat liver nucleoside diphosphosugar or diphosphoalcohol pyrophosphatases different from nucleotide pyrophosphatase or phosphodiesterase I: substrate specificities of Mg(2+)-and/or Mn(2+)-dependent hydrolases acting on ADP-ribose.

Three rat liver nucleotides(5') diphosphosugar (NDP-sugar) or nucleoside(5') diphosphoalcohol pyrophosphatases are described: two were previously identified in experiments measuring Mg(2+)-dependent ADP-ribose pyrophosphatase activity (Miró et al. (1989) FEBS Lett. 244, 123-126), and the other is a new, Mn(2+)-dependent ADP-ribose pyrophosphatase. They are resolved by ion-exchange chromatography, and differ by their substrate and cation specificities, KM values for ADP-ribose, pH-activity profiles, molecular weights and isoelectric points. The enzymes were tested for activity towards: reducing (ADP-ribose, IDP-ribose) and non-reducing NDP-sugars (ADP-glucose, ADP-mannose, GDP-mannose, UDP-mannose, UDP-glucose, UDP-xylose, CDP-glucose), CDP-alcohols (CDP-glycerol, CDP-ethanolamine, CDP-choline), dinucleotides (diadenosine pyrophosphate, NADH, NAD+, FAD), nucleoside(5') mono- and diphosphates (AMP, CMP, GMP, ADP, CDP) and dTMP p-nitrophenyl ester. Since the enzymes have not been purified to homogeneity, more than three pyrophosphatases may be present, but the co-purification of activities, thermal co-inactivation, and inhibition experiments give support to: (i) and ADP-ribose pyrophosphatase highly specific for ADP(IDP)-ribose in the presence of Mg2+, but active also on non-reducing ADP-hexoses and dinucleotides (not on NAD+) when Mg2+ was replaced with Mn2+; (ii) a Mn(2+)-dependent pyrophosphatase active on ADP(IDP)-ribose, dinucleotides and CDP-alcohols; (iii) a rather unspecific pyrophosphatase that, with Mg2+, was active on AMP(IMP)-containing NDP-sugars and dinucleotides (not on NAD+), and with Mn2+, was also active on non-adenine NDP-sugars and CDP-alcohols. The enzymes differ from nucleotide pyrophosphatase/phosphodiesterase-I (NPPase/PDEaseI) by their substrate specificities and by their cytosolic location and solubility in the absence of detergents. Although NPPase/PDEaseI is much more active in rat liver, its known location in the non-cytoplasmic sides of plasma and endoplasmic reticulum membranes, together with the known cytoplasmic synthesis of NDP-sugars and CDP-alcohols, permit the speculation that the pyrophosphatases studied in this work may have a cellular role.

Rat liver mitochondrial ADP-ribose pyrophosphatase in the matrix space with low Km for free ADP-ribose.

A study involving markers of subcellular and submitochondrial fractions, gradient centrifugation, latency measurements and extraction with digitonin, demonstrates the association of a specific ADP-ribose pyrophosphatase with rat liver mitochondria and its localization in the matrix space. The enzyme hydrolyses ADP-ribose to AMP, with a Km of 2-3 microM. The results support the occurrence of a specific turnover pathway for free ADP-ribose and its relevance in mitochondria.

Location of dinucleoside triphosphatase in the matrix space of rat liver mitochondria.

The submitochondrial location of dinucleoside triphosphatase (EC 3.6.1.29), previously shown to be in part associated with mitochondria, has been studied in rat liver. The precipitability and latency of activity in organelle suspensions, and the profile of solubilization by digitonin, were like those of the matrix space marker glutamate dehydrogenase, and differed from those of other submitochondrial fractions. This, and the synthesis of diadenosine polyphosphates by mitochondrial aminoacyl-tRNA synthetases, suggest the occurrence of a pathway for the intramitochondrial turnover of diadenosine 5',5'''-P1,P3-triphosphate (Ap3A).

[Eisenmenger's syndrome and pregnancy. Apropos of 2 spontaneous deliveries, one of which was fatal]. / Syndrome d'Eisenmenger et grossesse. A propos de deux accouchements spontanés dont un mortel.

Two cases of spontaneous delivery at 27 weeks and 38 weeks are reported in two patients with Eisenmenger's syndrome. One case did not present any problems, while the other was followed by maternal death on the 6th day post-partum. The cause of death was similar to that of other cases reported in the literature. The pathophysiological explanation of the terminal clinical condition with refractory hypoxia remains unclear. In view of the maternal mortality of about 50 per cent, these patients should be prescribed effective contraception, free of thrombogenic and infectious risks. The therapeutic approach in patients who become pregnant is not well defined. However, a number of principles should be observed during delivery: oxygen therapy, rapid correction of haemorrhages, the timing and the modalities of anti-coagulation are controversial. The failure of the usual methods of resuscitation in the terminal stage raises the possibility of using fibrinolytics.