Identification and further characterization of Enterobacteriaceae and Cronobacter spp. in a milk powder and infant processing plant

Due to technical reasons, milk powder and powdered infant formulae (PIF) are not sterile products. In order to achieve the requirements set by the Swiss and European regulations for microbiologic criteria extensive epidemiological studies are needed on each individual plant level. In this way contamination routes can be identified and appropriate measurements taken. A legally reglemented pasteurization process eliminates Enterobacteriaceae. Therefore recontamination must now be focussed on. PIF contaminated with Cronobacter spp. can lead to severe infections in neonates such a sepsis, meningitis or necrotizing enterocolitis. The reported prevalence of commercially available PIF appears to be gradually decreasing from estimates of 14% in 1988 to 2.0-2.5%, where it now seems to have become stabilized. In order to make a reasonable estimate concerning the prevalence of Cronobacter spp. on an individual plant level, 950 samples (raw material, finished products, environmental samples) were analysed. The high prevalence of 16% comes from the intentional sampling of critical raw material and environment samples. The PFGE analysis, however, did not reveal any correlation between raw material and environmental samples which would indicate a possible contamination via finished products. 470 PIF Enterobacteriaceae isolates were identified through biochemical tests as well as by rpoB sequencing. E. cloacae (35%), Pantoea spp. (11%) and K. pneumoniae (8%) were the most prevalent genus and species. In order to reveal possible contamination routes, a subtyping was conducted. The species E. cloacae, which can be found in the same niches as Cronobacter spp., could be used as a significant hygienic indicator organism. To complete the epidemiological picture, 363 milk based samples were analysed (raw milk, milk concentrate, milk powder). Raw milk contains Enterobacteriaceae but no Cronobacter spp. were detected. However, 12/172 samples of milk powder contained Cronobacter spp. due to recontamination (during the packaging process and/or further processing steps). In order to increase the sensitivity and specificity of today’s available analysis for the detection of Cronobacter spp. methodological improvements had to be undertaken. The currently used enrichment media (mLST, EE) contain components of too selective nature which can therefore lead to false negative results. The new “Cronobacter Screening Broth” (CSB) contains sucrose and 5-bromo-4-chloro-3-indolyl-α-D-Glucopyranoside which now leads to a sensitivity of 100% and a negative predicting value of 100% as well. The change in colour of the broth indicates a presumptively positive result whereby only these samples need to be streaked onto chromogenic agar. The visual intermediate result leads to a reduction in costs and working time. In order to increase specificity as well as the commercial pressure of fast product release, a PCR-based system where positive and negative results are clearly available in short time is recommended. Several real-time PCR based systems for detection of Cronobacter spp. have become commercially available. Two systems (one open platform (Biotecon Diagnostics, Potsdam, Germany) and one dedicated system (BioControl, Bellvue, USA)) generated neither false positive nor false negative results. Both systems were able to detect 9 target and 13 non-target strains. The dedicated system has the advantage of shorter hands-on and analysis time. In addition, contaminations due to handling faults are reduced. The existing rpoB based Cronobacter species PCR was upgraded for the recently described species C. condimenti which can now be detected with high reliability. Additional epidemiological data is needed in order to monitor the microbiological situation in industrial plants constantly as well as consequently. Based on information on individual plant level it is possible to implement adequate measurements such as HEPA filters, exact time for adding heat labile ingredients, personal and material flow, air management and cleaning (type, time). Scientific support is needed concerning adequate analytical methods, formation of biofilm, desiccation data, types of enrichment media, sample size as well as additional epidemiological data. Our recent study concerning genetic diversity showed that different Cronobacter isolates from one sample can contain different PFGE fingerprints. This observation suggests that analysis of one isolate per sample may not be sufficient for trace back studies. The analysis of at least five colony forming units per sample is suggested. This example shows that through a close collaboration between industrial companies and scientific institutes, knowledge can be actively turned into practice. – This helps prevent pre-term babies and newborns from falling ill to Cronobacter spp

Design Makes a Difference

There is more behind a pretty product than appearance: consumer safety, ease of use, production efficiency, and competitive marketing are essential ingredients in today\u27s industrial design process

Local and regional ecological morphology of dung beetle assemblages across four biogeographic regions

Aim Niche partitioning within species assemblages is thought to influence species packing and/or total niche space occupied. The evolution of dung beetles (Scarabaeinae) is likely to have been strongly influenced by inter-specific competition, leading to niche partitioning. We consider whether local-scale processes leave a signature in regional patterns of functional diversity in dung beetle assemblages, and investigate the correlation between total exploited ecomorphological space and density of species packing with increased species richness. We test whether ecomorphological space occupied by local assemblages reflects that of their regional species pool, and the extent to which ecomorphological space is convergent or divergent within functional groups across regional pools. Location Neotropics, Africa, Australia and Madagascar. Methods Dung beetle assemblages were collected in a standardized manner from four biogeographic regions. Ecomorphological similarity among the assemblages was assessed by multivariate analysis of 19 linear measurements for 300 species and three functional nesting types (roller, tunneller or dweller), firstly on a local level within the Neotropics and Afrotropics, and then between the regional species pools. Results Key body measurements, in particular the hind tibia, separated rollers and tunnellers into largely non-overlapping entities along the first three axes of the shape analysis. Three Neotropical assemblages, which vary widely in species numbers, each harboured a similar amount of morphometric variation, resulting in increasingly dense species packing with greater species richness. Similar findings were obtained in two South African assemblages. Assemblages in the four biogeographic regions showed largely similar distributions of ecomorphological variation, including the separation of rollers and tunnellers, despite their distant phylogenetic relationships. Ecomorphological similarity among regions was particularly high in tunnellers, whilst the rollers exhibited greater regional differentiation. Main conclusions Local assemblages evidently represent the full diversity of functional groups available in the regional pool, even in species-poor assemblages. There is a strong trend towards convergence in morphology separating tunnellers and rollers in phylogenetically independent lineages. The ecomorphological similarity of regional assemblages suggests that morphological convergence is the result of common selective forces active within the assemblages themselves. This lends support to the widely hypothesized effect of inter-specific interactions and niche partitioning in determining assemblage composition and lineage evolution in the Scarabaeinae. © 2011 Blackwell Publishing Ltd

Observations écologiques sur le Néotrague de Bates ( Neotragus Batesi de Winton, 1903, artiodactyle, ruminant, bovide) du nord-est du Gabon

Bates’s Pygmy Antelope Neotragus batesi was studied for 15 consecutive months in North East Gabon. Fourteen animals were individually marked and six of them (5 9 9 and 1 3) monitored by radio-tracking. During the same period, 128 specimens were col lected outside the study area, to examine the stages of growth of their jugal teeth, tooth wear and reproductive condition. Four age-categories can be recognized among young animals, and five among adults. In the mature rain forest Neotragus batesi is commonly found in tree falls and along riversides. It thrives in secondary growth vegetation and native plantations where it reaches its highest population densities. Both its morphology and habits are well adapted to locomotion through dense undergrowth. Bates’s Pigmy Antelope is active by day as well as by night, with peaks in the morning and the evening. It rests in the middle of the day, and in the beginning and middle of the night. The home ranges are small (1 to 3 ha for females ; 2 to 4 ha for males). This area is not used evenly throughout the year ; the core-area of the range changes in keeping with the seasons. Fur thermore, every home range includes areas which are used only by day and others which are visited only by night, vegetation struc ture differing accordingly. The more regularly tended native plantations are those which produce the most food for Neotragus batesi, but periodical clearing of the undergrowth also disturbs the animals. Bates’s Pygmy Antelope has therefore to make use of both well managed native plantations and neglected ones or secondary growth to ensure its food supply and shelter. About 200 species of plants are eaten by Neotragus batesi in N.F. Gabon. Usually this pigmy antelope makes use of a given area when its plant food is most readily available, thus quickly adapting to new situations. Some of the most actively sought plant species, however, may definitely be scarce. On the whole this animal tends to browse upon the most nutritious parts of the plants it selects. Bates’s Pigmy Antelope is a solitary animal ; 76 % of the individuals encountered were alone. More than one female can share a single home range, whereas males never do so. Interindividual communication is mostly achieved through short groans (between a young animal and its mother, or between adult males and females), and through olfactory marking by the secretion of males’ anteorbital glands. Females do not display any territorial behaviour, whereas males might do so. Males are apparently polygamous, their large home ranges overlapping those of several females. Females give birth throughout the year, with peaks at the end of each of the two rainy seasons. In this way the young animals are weaned at a time when fresh grass is readily available. Mortality rate among males apparently increases at the M2 stage, when the young leave their mothers’ ranges, and also among the oldest adults. Among females mortality seems to be the highest amongst subadults, at the onset of the reproductive period. Females, as usual, outlive males. Competition with sympatric ungulates is avoided by diffe rences in size, a protracted activity rythm, and a strictly folivorous diet

A County Grand Jury Indictment Manual

Because of the recent change in law, as a result of passage of the "Crime Victims Justice Reform Act," Proposition 115, (Cal. Const. Art. I, sect. 24) which created a number of specific effects on the Administration of Criminal Justice which included the application of legal effect in the matter of Hawkins v. Superior Court, (1978) 22 Cal.3d 584 requiring a "Post Indictment Preliminary Hearing," it is anticipated that more counties will return to the use of the grand jury for its indictment process. In order to insure proper review of criminal matters by this citizen review panel, and to provide fair administration of justice, there is a need for a comprehensive, detailed, impartial education tool to teach members of a county grand jury the obligations placed upon them by the law. A manual is an efficient method to fill this need. This Thesis is the beginnings of such a manual. This Thesis serves as the focus for curriculum development for a training manual to be used by members of California county grand juries. The manual is intended as an introduction to the duties and obligations of grand jurors in the indictment process. It gives a general overview of the prosecutor's responsibilities and an explanation of the independence that the grand jury exercises in deciding whether or not to indict. It is recommended that a survey be completed, which narrows the topics perceived to be of importance by the people involved in the criminal justice system that have a direct relationship with county grand juries. The results of the survey, after a thorough analysis and interpretation of the data, will lend itself to the final topics to be discussed in the manual. It should be noted that this manual is not intended to replace advice nor instruction from the superior court, nor the prosecutor who would be advising the grand jury during a time of inquiry on whether to indict

Modifications of the rainforest frugivore community are associated with reduced seed removal at the community level

International audienceTropical rainforests worldwide are under increasing pressure from human activities, which are altering key ecosystem processes such as plant-animal interactions. However, while the direct impact of anthropogenic disturbance on animal communities has been well studied, the consequences of such defaunation for mutualistic interactions such as seed dispersal remains chiefly understood at the plant species level. We asked whether communities of endozoochorous tree species had altered seed removal in forests affected by hunting and logging and if this could be related to modifications of the frugivore community. At two contrasting forest sites in French Guiana, Nouragues (protected) and Montagne de Kaw (hunted and partly logged), we focused on four families of animal-dispersed trees (Sapotaceae, Myristicaceae, Burseraceae and Fabaceae) which represent 88 % of all endozoochorous trees which were fruiting at the time and location of the study. We assessed the abundance of the seed dispersers and predators of these four focal families by conducting diurnal distance sampling along line transects. Densities of several key seed dispersers such as large-bodied primates were greatly reduced at Montagne de Kaw, where the specialist frugivore Ateles paniscus is probably extinct. In parallel, we estimated seed removal rates from fruit and seed counts conducted in one-square-meter quadrats placed on the ground beneath fruiting trees. Seed removal rates dropped from 77 % at Nouragues to 47 % at Montagne de Kaw, confirming that the loss of frugivores associated with human disturbance impacts seed removal at the community level. In contrast to Sapotaceae, whose seeds are dispersed by mammals only, weaker declines in seed removal for Burseraceae and Myristicaceae suggest that some compensation may occur for these bird- and mammal-dispersed families, possibly because of the high abundance of toucans at the disturbed site. The defaunation process currently occurring across many tropical forests could dramatically reduce the diversity of entire communities of animal-dispersed trees through seed removal limitation