Echinococcus multilocularis genetic diversity in Swiss domestic pigs assessed by EmsB microsatellite analyzes.

Assessing the genetic diversity of the parasite Echinococcus multilocularis provides key information about the temporal and spatial strain flow in a given area. Previous studies indicated that a historical endemic area conventionally presents a relatively high genetic diversity, whereas peripheral or newly endemic areas exhibit a more restricted variability of the parasite. The Swiss plateau region is part of the European historically endemic area, and the genetic diversity has already been investigated by assessing either human metacestode isolates or adult worms from foxes. To date, there have been no studies covering the whole geographical area affected by the parasite. The aim of the present study was to make use of the domestic pig to investigate the genetic diversity of E. multilocularis in relation to spatial distribution. A total of 55 E. multilocularis-induced hepatic lesions from slaughtered pigs from Switzerland were studied using EmsB microsatellite analyzes, and findings were compared to already published data (originating from human, primate, foxes, and rodent samples). A total of 12 EmsB profiles were described among the domestic pigs, some of them presenting a clear spatial organization in the Swiss plateau, with three of the main profiles geographically separated. One of the 12 EmsB profiles has been newly identified for Switzerland in this study, while the other 11 profiles had been previously described in other Swiss E. multilocularis isolates from other hosts. Overall, a total of 18 EmsB profiles have so far been described within the Swiss endemic area. Six profiles appeared only among human, primate, rodent, and fox samples. Based on a richness and diversity accumulation analysis, the sampling efficiency for the whole studied area has now been improved considerably by compilation of 178 E. multilocularis specimens obtained from four different intermediate and one definitive host species in Switzerland.

Combined cross-sectional and case-control study on Echinococcus multilocularis infection in pigs in Switzerland.

The canid tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE) in humans and other intermediate hosts. Depending on the permissiveness of the intermediate host, the larval form of E. multilocularis (metacestode) may be either fertile, e.g. in rodents, and thus supporting the life cycle of the parasite, or infertile, e.g. in pigs, and thus interrupting the life cycle. Pigs have been shown to act as aberrant hosts for the metacestode and consequently develop liver lesions but represent a dead-end for the parasite. Routine liver inspection at slaughter provided the basis for a large-scale surveillance study on E. multilocularis infection in pigs. The aim of this combined cross-sectional and case-control study was to estimate the minimal prevalence of E. multilocularis in pigs in Switzerland, to find factors associated with infection, and to assess potential regional clusters of infection. During the 12-month-study period, approximately 85% of all pigs slaughtered in Switzerland were assessed. In total, 450 pig livers with macroscopic lesions suggestive of E. multilocularis infection were analysed. Of those, 200 samples were positive by E. multilocularis-PCR. Thus, the overall minimal prevalence detected by molecular means was 0.009% in all slaughter pigs (200 of 2'143'996), 0.008% in finishing pigs (177 of 2'123'542), and 0.11% in breeding pigs (22 of 20'454). Histology revealed the unique presence of a laminated layer in 105 cases, and an additional germinal layer detected in a single case. Protoscoleces could not be observed in any of the lesions. Factors positively associated with infection were "foxes seen in the pig shed", "foxes on premises", "presence of other animals in the shed", "absence of a hygiene barrier", "outdoor feeding", "feeding grass", "lack of rodent control", "not having own dogs on the farm" and "infrequent deworming of sows". Infection was present in all regions sampled and was representative of the important pig rearing areas of Switzerland, without evidence of any obvious geographical cluster. Conclusively, our study provided further evidence of widespread environmental contamination with E. multilocularis eggs in Switzerland. Furthermore, the absence of protoscoleces in any of the lesions supported the concept that pigs act only as a dead-end host and thus do not contribute to the life cycle of the parasite. Factors associated with E. multilocularis infection were in-line with parasite biology, and many can be addressed by increasing hygiene and management standards.

Combined cross-sectional and case-control study on Echinococcus multilocularis infection in pigs in Switzerland.

The canid tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE) in humans and other intermediate hosts. Depending on the permissiveness of the intermediate host, the larval form of E. multilocularis (metacestode) may be either fertile, e.g. in rodents, and thus supporting the life cycle of the parasite, or infertile, e.g. in pigs, and thus interrupting the life cycle. Pigs have been shown to act as aberrant hosts for the metacestode and consequently develop liver lesions but represent a dead-end for the parasite. Routine liver inspection at slaughter provided the basis for a large-scale surveillance study on E. multilocularis infection in pigs. The aim of this combined cross-sectional and case-control study was to estimate the minimal prevalence of E. multilocularis in pigs in Switzerland, to find factors associated with infection, and to assess potential regional clusters of infection. During the 12-month-study period, approximately 85% of all pigs slaughtered in Switzerland were assessed. In total, 450 pig livers with macroscopic lesions suggestive of E. multilocularis infection were analysed. Of those, 200 samples were positive by E. multilocularis-PCR. Thus, the overall minimal prevalence detected by molecular means was 0.009% in all slaughter pigs (200 of 2'143'996), 0.008% in finishing pigs (177 of 2'123'542), and 0.11% in breeding pigs (22 of 20'454). Histology revealed the unique presence of a laminated layer in 105 cases, and an additional germinal layer detected in a single case. Protoscoleces could not be observed in any of the lesions. Factors positively associated with infection were "foxes seen in the pig shed", "foxes on premises", "presence of other animals in the shed", "absence of a hygiene barrier", "outdoor feeding", "feeding grass", "lack of rodent control", "not having own dogs on the farm" and "infrequent deworming of sows". Infection was present in all regions sampled and was representative of the important pig rearing areas of Switzerland, without evidence of any obvious geographical cluster. Conclusively, our study provided further evidence of widespread environmental contamination with E. multilocularis eggs in Switzerland. Furthermore, the absence of protoscoleces in any of the lesions supported the concept that pigs act only as a dead-end host and thus do not contribute to the life cycle of the parasite. Factors associated with E. multilocularis infection were in-line with parasite biology, and many can be addressed by increasing hygiene and management standards.

A consensus rating method for small virus-retentive filters. I. Method development.

Virus filters are membrane-based devices that remove large viruses (e.g., retroviruses) and/or small viruses (e.g., parvoviruses) from products by a size exclusion mechanism. In 2002, the Parenteral Drug Association (PDA) organized the PDA Virus Filter Task Force to develop a common nomenclature and a standardized test method for classifying and identifying viral-retentive filters. One goal of the task force was to develop a test method for small virus-retentive filters. Because small virus-retentive filters present unique technical challenges, the test method development process was guided by laboratory studies to determine critical variables such as choice of bacteriophage challenge, choice of model protein, filtration operating parameters, target log10 reduction value, and filtration endpoint definition. Based on filtration, DLS, electrospray differential mobility analysis, and polymerase chain reaction studies, a final rating based on retention of bacteriophage PP7 was chosen by the PDA Virus Filter Task Force. The detailed final consensus filter method was published in the 2008 update of PDA Technical Report 41. Virus Filtration.

The temperature stability of mouse retroviruses depends on the cholesterol levels of viral lipid shell and cellular plasma membrane.

To delineate parameters contributing to the extracellular lifetime of retroviral vectors, we carried out stability tests of retroviruses derived from cell lines of different origin and kept under different cultivation conditions. Results show that amphotropic mouse retroviruses (MLV-A) derived from human and hamster cells exhibit 2- to 3-fold higher half-lives compared to retroviruses from mouse cells. Cultivation at 32 degrees C has been reported to yield high virus titers. However, the benefit of virus production in mouse cells at 32 degrees C is controversial. In our hands the cultivation temperature affected, hitherto not noticed, the half-life time of MLV-A. The 37/32 degrees C shift resulted in a 3-fold decrease of viral half-lifes compared to MLV-A released from mouse cells at 37 degrees C. Thus, MLV-A released at 37 degrees C is phenotypically different from MLV-A synthesized at 32 degrees C. Increased virus stability was inversely correlated with the level of cholesterol in the viral membrane. Finally, depletion of viral cholesterol in vitro resulted in intact virus with increased thermal stability. Thus, retrovirus lability depends on the host cell and parallels the cholesterol amount in the viral lipid shell.