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Comparison of the fecal microbiota of domestic commercial meat, laboratory, companion, and shelter rabbits (Oryctolagus cuniculi)
Jennifer Kylie1,
J. Scott Weese1 &
Patricia V. Turner1
Rabbits are cecotrophic, hindgut-fermenters that rely heavily on their gastrointestinal microbiota for optimal digestion of plant-based diets. Dysbiosis, caused by disruption of the gastrointestinal microbiota, is known to predispose rabbits to rabbit enteritis complex (REC), a major cause of morbidity and mortality. The objectives of this study were to describe the fecal microbiota of domestic rabbits from a variety of settings (commercial meat, companion, laboratory, and shelter) and to identify how factors such as age, season, and routine antimicrobial use affect the fecal microbiota composition.
A total of 86 pooled commercial meat, 54 companion, 14 pooled laboratory, and 14 shelter rabbit fecal samples were evaluated using 16S rRNA gene sequencing of the V4 region. In all sample types, the predominant bacterial phylum was Firmicutes. Other commonly identified phyla (composing ≥ 1% of the total microbiota composition) were Verrucomicrobia, Proteobacteria, and Bacteroidetes. Significant differences in composition were noted between commercial, companion, laboratory, and shelter rabbit samples for proportions of Verrucomicrobia (P < 0.01), Proteobacteria (P < 0.01), and Lentisphaerae (P = 0.01) within the total microbiota. Within the commercial meat rabbit samples, significant differences between the microbiota composition of growers (n = 42) and does (n = 44) were limited to one unclassified Firmicutes (P = 0.03) and no differences were identified at the phylum level. Significant differences were present between fecal samples taken from rabbits during the summer (n = 44) compared to the winter (n = 42), with Firmicutes (P = 0.04), Verrucomicrobia (P = 0.03), Proteobacteria (P = 0.02), Deinococcus-Thermus (P = 0.04), Armatimonadates (P = 0.003), and Actinobacteria (P = 0.03) forming significantly different proportions of the microbiota. The only significant difference in composition between those farms that routinely reported antimicrobial use and those that did not was in one unclassified Bacteroidetes (P < 0.05) and no differences were identified at the phylum level.
Rabbit husbandry and diet, in addition to season, significantly influence the fecal microbiota composition of domestic rabbits, while age of the rabbit post-weaning has minimal impact.
Rabbits are herbivorous, monogastric, hindgut-fermenting mammals that rely on cecotrophy to ensure maximum nutrient absorption from their diet. Management of this unique gastrointestinal (GI) physiology can be challenging, and, as a result, enteric disease is common in domestic rabbits. One of the most difficult aspects of managing rabbit digestion is maintaining the normal commensal gastrointestinal microbiota. In rabbits, disruption of the normal microbiota, termed dysbiosis, is commonly implicated as a cause of enteritis, with ensuing diarrhea, subsequent dehydration, inadequate nutrition, and, potentially, death as a result.
The etiopathogenesis of enteritis in rabbits is complex and generally multifactorial. Enteric pathogens such as Escherichia coli, Clostridium spiriforme, Lawsonia intracellularis, rotavirus, and coronavirus are commonly associated with diarrhea outbreaks in rabbits, as are other agents such as Clostridium piliforme, Salmonella spp., parvovirus, and astrovirus [1,2,3,4,5,6]. However, much is still unknown about initiating factors as infection with these organisms is not synonymous with disease and subclinical infection may occur with no overt clinical signs [7]. Most cases of enteritis in rabbits are caused by a combination of factors, including feeding of a low fiber diet, debility and overall health status, management-related stress, and age, along with the presence of one or more potentially pathogenic organisms [7,8,9].
Because of its critical importance in rabbit health, several studies have focused on understanding the composition of the rabbit enteric microbiota. Historically, researchers used culture-based techniques; however, the development of culture-independent techniques has permitted much greater in-depth analysis [10, 11]. Culture-based studies concluded that Bacteroidetes was the predominant bacterial phylum within the rabbit gastrointestinal tract, regardless of age [12]. With culture-independent sequencing Bacteroidetes are still reported as predominant in the intestinal tract of very young rabbits, but the predominant phylum in post-weaned and adult rabbits is Firmicutes, with Bacteroidetes accounting for only a small fraction [13,14,15,16]. Other predominant phyla routinely identified using culture-independent sequencing include Verrucomicrobia and Proteobacteria [14, 16].
Previous studies examining the fecal microbiota of rabbits have used small numbers of laboratory rabbits kept in well-controlled environments exposed to minimal environmental or husbandry variations, making broad applicability to other domestic rabbit conditions less likely. Factors such as diet, husbandry, and seasonal effects have been demonstrated to have effects on the fecal microbiota in both humans and other animal species [17,18,19], but have not been explored in rabbits. The objectives of this study were to characterize the microbiota of domestic rabbits kept in a variety of environments and to identify factors contributing to enteric microbiota variations using a culture-independent, high-throughput sequencing method. We hypothesized that significant fecal microbiota differences would be seen between commercial meat rabbits and domestic rabbits kept in other settings, largely because of differences in husbandry, including dietary composition, routine use of antimicrobials, and environmental management. Additionally, we hypothesized that these differences would be least significant between commercial meat rabbits and rabbits housed in a shelter environment (i.e., a facility where previously-owned animals have been surrendered for re-homing) due to the likely inconsistencies in the environment of both of these types of rabbits and diet (in the case of shelter rabbits), while differences would be most significant between commercial meat rabbits and both companion and laboratory rabbits due to more consistent environmental conditions, diet, care, and reduced infectious disease within these latter two groups of rabbits.
A total of 168 rabbit fecal samples from various sources were included in the final analysis: 86 pooled commercial meat, 54 companion, 14 pooled laboratory, and 14 shelter animals. Twenty-three samples (14 commercial meat, 8 companion, and 1 shelter) were not included in the final analysis either due to an inability to produce bands following DNA extraction, amplification, and purification, an insufficient quantity of DNA within the sample (< 12.5 nM) at the time of normalization prior to sequencing or an insufficient number of sequences (< 5000) present following the completion of all quality control filters. A total of 10,897,154 V4 16S rRNA gene sequences passed all quality control filters. Sequence numbers per sample ranged from 6359 to 325,029, with a mean of 64,864, median of 53,721, and a standard deviation of 46,279.
The domestic rabbit fecal microbiota
The analysis was based on 168 samples, with a subsample of 4577 sequences per sample for normalization. A total of 31 bacterial phyla were identified; however, only five were present at a relative abundance of ≥ 1%. Of these five phyla, Firmicutes composed 66.4% of the total microbiota, Verrucomicrobia composed 14.1%, Proteobacteria composed 9.5%, and Bacteroidetes composed 1.54% while 6.9% of the sequences could not be identified at the phylum level (see Additional file 1: Table S1 for further information).
Comparison of the rabbit fecal microbiota based on animal source
There were several statistically significant differences in relative abundance of bacteria within all taxonomic levels based on animal source. These differences remained consistent regardless of whether the specific source from which the sample came was included as a random effect in the analysis; therefore, all relative abundance analyses that are reported below exclude specific source as a random effect. Significant differences were identified between commercial rabbits, and companion and laboratory rabbits in relative abundances of Proteobacteria (P < 0.01), between commercial rabbits, companion rabbits, and laboratory and shelter rabbits in relative abundances of Verrucomicrobia (P < 0.01), and between commercial rabbits, and companion and shelter rabbits in relative abundances of Lentisphaera (P < 0.01) when the p-value was corrected for false discovery rates (Fig. 1). These differences were also reflected at all levels of taxonomic classification (see Additional file 2: Table S2).
Median relative abundances of predominant bacterial phyla of the rabbit fecal microbiota separated by animal source. Significant differences (p ≤ 0.05) are present between sources for Proteobacteria, Verrucomicrobia, and Lentisphaera (included in 'Other' and composed < 1% of the total composition)
Commercial meat and companion rabbits had distinctly different fecal microbiota community structures, while those of laboratory and shelter rabbits tended to overlap with those from companion rabbits (Fig. 2). The dendrogram of community structure (Yue and Clayton index of dissimilarity) is presented in Fig. 3.
Principle co-ordinates analysis (PCoA) of the rabbit fecal microbiota based on the Jaccard Index. a blue circles = commercial meat rabbits, red circles = companion rabbits, yellow circles = laboratory rabbits, green circles = shelter rabbits; b) blue circles = does, red circles = fryers; c) blue circles = winter, red circles = summer; d) blue circles = routine antimicrobial use, red circles = no routine use of antimicrobials. Clustering of markers within a plot indicates similarity in microbiota community membership. Significant differences are present between rabbit sources and between seasons, but not between ages and antimicrobial use statuses
Dendrogram of the rabbit fecal microbiota community structure based on the Yue and Clayton Index of Dissimilarity. Blue = commercial rabbits, red = companion rabbits, yellow = laboratory rabbits, green = shelter rabbits. Clustering of lines indicates similarity in microbiota community structure. Significant differences are present between commercial meat rabbits and all other sources. The top cluster delineated by the vertical black line indicates clustering of samples collected from commercial meat rabbits during the winter months while the clusters indicated by the lower vertical black lines indicate clustering of samples collected from commercial meat rabbits during the summer months
In both cases, clustering of ≥ 10 samples can be seen within large numbers of commercial and pet samples, with the laboratory and shelter rabbit fecal samples scattered intermittently between, and primarily amongst, the companion samples. Using the Jaccard tree, significant differences were noted between all possible group pairings using a parsimony test (P < 0.02 for all comparisons), something that was also noted with unweighted UniFrac tests (P ≤ 0.04 for all comparisons), indicating that the fecal microbiota community membership differed significantly between all sources. Using the parsimony test with the Yue and Clayton tree, significant differences were identified between commercial meat and laboratory samples (P = 0.03), commercial meat and companion animal samples (P < 0.01), and commercial meat and shelter samples (P < 0.01). These differences were also detected using unweighted UniFrac (all P < 0.01), indicating that fecal microbiota community structure was significantly different between commercial meat rabbits and rabbits from all other sources, but the other sources did not differ amongst one another. Thirty-seven OTUs were identified to be differentially abundant via LEfSe (P ≤ 0.05) with LDA scores ≥ 3.0 (Table 1).
Table 1 OTUs identified as significantly differentially abundant (p ≤ 0.05) from all other rabbit sources with LDA scores ≥ 3.0
Comparison of the rabbit fecal microbiota based on animal age
Forty-four samples, each consistently of pooled feces from 3 does per commercial meat farm, and 42 samples composed of pooled feces from 3 growers per commercial meat farm, were compared, with a subsample of 4577 sequences per sample. No significant differences were observed in relative abundance between the two age groups at the bacterial phylum, order, family, and genus levels (all adjusted P > 0.05). At the class level, there was a significant difference between does and growers for an unclassified Firmicutes, where the relative abundance in does was 5.8% and in growers was 7.7% (P = 0.03).
Community structure differences are visualised in the PCoA (Fig. 2b). There is significant overlap between the community structures of the two different age groups with no obvious grouping based on age. A significant difference between the fecal microbiota community structure of does and fryers was only noted using the Yue and Clayton tree with the parsimony test (P = 0.04); however, no more than five samples were noted to be clustered by age within commercial meat samples in the Yue and Clayton tree analysis (Fig. 3). Six OTUs were identified to be significantly different between ages (P ≤ 0.05) with LEfSe LDA scores ≥ 3.0 (Fig. 4a).
OTUs noted to be significantly different between a) ages and b) seasons with LEfSe scores > 3.0
Comparison of the rabbit fecal microbiota based on season
A comparison was conducted of 44 pooled fecal samples collected from meat rabbits during winter months and 42 pooled fecal samples collected during summer months, with a subsample of 4577 sequences per sample. Significant differences at all taxonomic levels of classification were identified (Table 2).
Table 2 Relative abundance and FDR p-values for significantly different (P ≤ 0.05) samples from summer (n = 44) and winter (n = 42)
The community structure differences are presented in the PCoA in Fig. 2c. While there is significant visible overlap between the groups, there is a distinct subpopulation of samples from the summer group that is separate from the overlying samples. Significant differences were identified between summer and winter fecal microbiota community membership and structure using a parsimony test (P < 0.01), and unweighted UniFrac tests (P < 0.01) with both the Jaccard tree and the Yue and Clayton tree (P < 0.01 for both tests). One distinct cluster comprised of ≥ 10 commercial meat rabbits samples collected during the winter months was present, as was one distinct cluster of ≥ 10 samples collected during the summer months within the Yue and Clayton Tree calculated based on animal source (Fig. 3). Twenty-three OTUs were identified to be significantly different between season (P ≤ 0.05) with LDA scores ≥ 3.0 (Fig. 4b).
Comparison of the rabbit fecal microbiota based on routine antimicrobial use
Routine use of antimicrobials in feed or water was reported in 17 of 23 (73.9%) farms, while 3 farms did not respond and one farm had closed at the time of follow-up questioning. Farms for which antimicrobial use could not be confirmed were excluded from the analysis. Antimicrobials reported to be used routinely included, in order of descending frequency of use, salinomycin sodium 6%, virginiamycin, tylosin, chlortetracycline, bacitracine methylene disalicylate, sulfamethazine, sulfadimethoxine, and one anti-coccidial agent that was not further specified. Twelve of the 17 (70.6%) farms reporting routine antimicrobial use reported using ≥ 2 antimicrobials in combination, making statistical analysis of the data beyond the general category of "routine antimicrobial use" not feasible. Antimicrobials were not reported to be routinely used in any of the laboratory rabbits sampled, and antimicrobial use could not be confirmed for the companion or shelter rabbit samples; thus, results from these groups were also excluded from this analysis.
Seventy-five pooled commercial meat rabbit fecal samples were compared on the basis of routine antimicrobial use following sequence filtering and the removal of non-responders, 58/75 (77.3%) with routine antimicrobial use, with a subsample of 4577 sequences per sample. The only significant difference observed at all levels of taxonomic classification, aside from the phylum level where no significant differences were observed, was in one Bacteroidetes unclassified at the class level (P < 0.01 at all taxonomic levels), which was significantly higher in samples from farms where no routine antimicrobial use occurred (0.85, 0.75, 0.75, 0.57% at class, order, family, and genus levels, respectively) compared to when routine use did occur (0.29, 0.31, 0.30, 0.32%, respectively).
No significant differences in community membership or structure were identified using Jaccard or Yue and Clayton trees with both the parsimony and unweighted UniFrac tests (p ≥ 0.29 for all tests); samples from farms that did not report routine use of antimicrobials can be seen as well-distributed amongst samples from farms that did in the PCoA in Fig. 2d. Thirteen OTUs were identified to be significantly different between use status (P ≤ 0.05) with LDA scores ≥ 2.0; single OTUs with an LDA score ≥ 3.0 that were more frequently identified with routine antimicrobial use were from each of the following genera: Ignatzschineria, Erysipelothrix, and Wohlfahrtiimonas. One OTU with an LDA score ≥ 3 was significantly more prevalent when antimicrobials were not being routinely used was present from each of the following genera: Acetitomaculum, Clostridium cluster III, and Parasporobacterium, in addition to one unclassified Ruminococcaceae, one unclassified Desulfovibrionaceae, and one unclassified Bacteroidetes.
Our study provides significant insight into the composition of fecal microbiota of domestic rabbits, as well as factors that may promote changes in its composition. The large number of animals sampled, combined with the variety of factors incorporated into the study, suggests that the results are applicable to rabbits in a variety of domestic settings. Consistent with previous culture-independent sequencing studies, such as those conducted by Eshar and Weese [14] and Zhu et al. [16], the predominant phylum identified in the rabbit fecal microbiota, regardless of source, age, season or reported antimicrobial use was Firmicutes. Within this phylum, the Clostridia were the most abundant class identified, of which the Ruminococcaceae and the Lachnospiraceae were the most abundant families present. While the prevalence of Firmicutes in our study is lower than that reported by Eshar and Weese (66% vs. 82%, respectively), other studies report levels of Firmicutes varying between 61 and 82% with significant individual variation, suggesting that the relative abundance of Firmicutes in the rabbit enteric microbiota falls somewhere within this range [16, 20]. In addition to being identified as the predominant phylum present in the rabbit gut, Firmicutes has been described as the most abundant phylum in the gastrointestinal microbiota of most healthy adult mammals, including humans [21,22,23,24], emphasizing its likely importance as a commensal in GI health.
Other predominant phyla identified include Verrucomicrobia, Proteobacteria, and Bacteroidetes. The relative levels identified for these phyla are consistent with previous reports, although proportions of Verrucomicrobia appear to vary considerably with the method of analysis [14, 16, 20, 25]. While Verrucomicrobia is still a relatively newly-defined group of organisms, Akkermansia (within this phylum) has been suggested to have a key role in hydrolysis of diverse ingested polysaccharides, contributing to more complete digestion of dietary cellulose as well as methane metabolism [26,27,28]. Thus, its presence as an important constituent of the rabbit fecal microbiota is unsurprising. The most well-known species in this genus is Akkermansia muciniphila, a mucin-degrading bacterium that has been demonstrated to be beneficial in both human and animal health [29, 30]. For example, increases in the proportion of fecal Akkermansia mucinophila have been associated with a healthier metabolic status in both humans and mice [29, 30]. While the impact of Akkermansia muciniphila on rabbit health is still unknown, it is known that the ability to breakdown mucin is especially important during cecotrophy for optimal nutrient extraction [8]. In vitro studies have also identified a positive correlation between the proportion of Verrucomicrobiaceae and concentrations of proprionate and acetate, volatile fatty acids that are found in high concentrations within rabbit ceca following the breakdown of fiber-rich carbohydrates, and significant energy sources [20, 31]. Akkermansia mucinophila levels have also been inversely correlated with several inflammatory markers in mice [32], indicating that a decrease in Akkermansia mucinophila levels may result in a more pro-inflammatory gut environment. A decreased proportion of Verrucomicrobia within the fecal microbiota of commercial meat rabbits overall, and particularly during the summer months, as found in this study, suggests less optimal gut health and nutrient extraction in this group of rabbits, as well as a pro-inflammatory state. This most likely reflects the lack of hay in the diet of commercial meat rabbits, compared to rabbits from other sources, as well as metabolic distress potentially due to heat stress, discussed in further detail below.
While small proportions of Proteobacteria are routinely observed within the enteric microbiota of all mammals, it has been suggested that increased relative abundance of Proteobacteria should be considered as a diagnostic indicator of underlying dysbiosis, predisposing individuals to enteric disease or indicating the presence of disease [33]. Relative increases in Proteobacteria with or without a concurrent reduction in Firmicutes have been observed in several species in cases of metabolic gastrointestinal disorders, such as genetically-related and diet-induced obesity [34, 35], as well as in chronic intestinal inflammation, such as inflammatory bowel disease and ulcerative colitis [36, 37]. It is hypothesized that the resultant bacterial population shift caused by increasing relative proportions of Proteobacteria stimulates the host gastrointestinal immune system, resulting in a pro-inflammatory response [38]. Therefore, the increased relative proportion of Proteobacteria observed within commercial meat rabbits overall, and specifically during the summer, suggests a shift away from the normal gut microbiota in these animals, increasing their risk for disease development. This idea is supported by the higher proportion of potentially pathogenic bacteria associated with REC identified in commercial meat rabbit fecal samples overall as well as in the summer at the OTU level. In both cases, elevations in Escherichia/Shigella accounted for significant differences in fecal microbiota composition when compared to feces from rabbits from other sources.
No significant differences were observed between does and growers in this study. This is consistent with previous reports suggesting that the rabbit fecal microbial community rapidly reaches a steady state at, or just after, weaning [20]. This stabilization has been observed in several other species, including mice [39] and pigs [40]. Additionally, no significant differences were observed between samples from farms in which antimicrobials were routinely used compared to farms where they were not. In 2007, Abecia et al. examined the effects of bacitracin and tiamulin on the cecal microbiota of lactating rabbit does. The study demonstrated that significant microbiota compositional changes were highly dependent on the specific antimicrobial used, with bacitracin having minimal effects on the cecal microbiota composition and tiamulin causing significant changes [41]. In the current study, a variety of antimicrobials with different pharmacologic mechanisms were reported in use alone or in combination, including those with anticoccidial activity and those used to treat respiratory disease. Effects of specific agents on the gut microbiota may have been masked by the concurrent use of other antimicrobial agents; additional studies into the effects of specific antimicrobial agents on the microbiota composition will help to differentiate those antimicrobials likely to have a significant effect versus those that have minimal effect.
Several factors that may contribute to gut microbiota composition are thought to differ significantly between domestic rabbit sources – in particular, the diet. Once animals are weaned, commercial meat rabbits are almost exclusively fed extruded pellet diets relatively low in fiber (15–20%) and high in carbohydrates to encourage rapid growth prior to slaughter [42]. This contrasts with recommended feeding practices for companion rabbits, which suggest a high fiber hay-based diet (e.g., timothy hay) that is supplemented (< 5%) by lower fiber pellets to ensure that all nutritional requirements are met while optimizing gastrointestinal function [9, 43]. Companion rabbit owners also frequently supplement this recommended dietary regimen with other fiber-rich fruits and vegetables. While the diet of laboratory rabbits is also primarily that of high carbohydrate pellets, the fiber content of many of the commercially available laboratory diets tends to be closer to 25%. Laboratory rabbits are also frequently given hay and vegetables as enrichment, although generally at more restricted levels than companion rabbits, while the dietary history of domestic rabbits in shelters is largely unknown. There is also significant variation in how long these animals may have been in a shelter receiving a balanced diet. Dietary fiber source, content, and digestibility are critical components of the rabbit diet, in which changes are directly and significantly correlated with enteritis and mortality in growing meat rabbits [44, 45]. It was beyond the scope of this study to evaluate specific changes in dietary fiber source, content, and type in relation to the rabbit fecal microbiota; however, this likely contributes to the majority of differences noted in microbiota between commercial meat and other sources of rabbits and is an area of further investigation.
Studies in other species examining the influence of season on the gut microbiota have identified decreases in the relative abundance of Verrucomicrobia during the summer as observed in the current study, but not relative increases in Bacteroidetes and Proteobacteria as were seen in this study [19]. The seasonal changes in the microbiota of Arctic ground squirrels observed in the Stevenson et al. 2014 study were attributed to diet availability and variation between seasons. In the current study, seasonality was only examined in commercial meat rabbits, in which the diet remains relatively consistent with fixed content proportions year-round (although changes in plant source and feed quality may vary from batch to batch). Thus, it is unlikely that the seasonal changes in relative abundance of different phyla were related to diet. Rabbits are much more tolerant of colder temperatures and low humidity and they are especially prone to heat stress [9, 46]. In general, Canadian rabbit barns do not have environmental controls for temperature or relative humidity and rabbits are exposed to hot and humid ambient conditions in July and August. The relative reduction in levels of the two beneficial phyla (Firmicutes and Verrucomicrobia), and the relative increase in less beneficial phyla (Proteobacteria) could relate to seasonal-related climate changes and directly impact rabbit health, susceptibility to enteritis, and possibly feed conversion efficiency. Additional studies are needed to explore these possibilities. Studies of potential seasonal differences in gut microbiota of rabbits from other sources are also needed.
A potential limitation of this study is whether the fecal microbiota is reflective of the gastrointestinal microbiota of rabbits in general. Michelland et al. [47] demonstrated no significant differences in bacterial community diversity between soft and hard feces or cecal content in rabbits. In addition, Schoster et al. [48] demonstrated that while significant differences existed between the microbiota of the stomach, duodenum, ileum, large colon, and feces in healthy horses, another hindgut-fermenting species, the microbiota profile remained stable from the cecum to the feces. These studies suggest that, at least in the case of hindgut-fermenting species, the fecal microbiota is representative of the distal gastrointestinal tract. For rabbits, in which the cecal microbiota is of primary consideration for overall animal health, the fecal microbiota provides an accurate and non-invasive method for studying their gastrointestinal health.
This study exclusively characterized the bacterial fecal microbiota of rabbits. It is well known that other prokaryotic and eukaryotic microorganisms, such as viruses, bacteriophages, protozoa, archaea and fungi, are essential components of the gastrointestinal microbiota, and can play significant roles in human and animal health [23, 49]. As pathogenic viruses, in addition to pathogenic bacteria, are significant contributors to rabbit dysbiosis and REC, a greater understanding of the rabbit gut virome would provide further insight into understanding rabbit health.
Lastly, it is unknown whether the organisms identified using next generation sequencing methods are functionally or metabolically active. Future computational studies are needed to specifically examine the metabolomics of the bacteria identified.
This study characterized the composition of the domestic rabbit fecal microbiota, as well as potential factors influencing its composition, such as rabbit source, age, and season, and the routine use of antimicrobials. By gaining an increased understanding of the rabbit enteric microbiota and factors that influence its composition, rabbit health and welfare can be better managed.
Animals and study approval
Commercial meat rabbit producers were contacted through Ontario Rabbit, a rabbit producer group, and asked to voluntarily participate in the study. Feces from domestic rabbits from 27 commercial farms (representing approximately 25% of large rabbit farms in Ontario, Canada) were collected for this study. Rabbit feces were sampled based on age (does and growers, aka fryers) and fecal samples were collected during summer and winter months. Feces from an additional 62 clinically healthy adult companion rabbits of varying ages and breeds were included. Rabbit fecal samples from one commercial laboratory rabbit vendor and seven laboratory research facilities in Ontario were also included, as were rabbit feces from four shelters in southwestern Ontario. All known history for samples collected is listed in Table 3. In all cases, participation in the study was voluntary, with companion, laboratory, and shelter organizations contacted directly by the researchers of the study.
Table 3 History of samples collected by source
Fecal sample collection
Hard feces of commercial meat rabbits (n = 100 samples) and laboratory rabbits (n = 14 samples) were collected from pans beneath rabbit home cages. In all cases, attempts were made to collect the freshest samples from the top layer of feces in the collecting pan (i.e., the most recently voided samples) or by placing a clean pan within 48 h prior to collection. Pooled fecal samples were collected from 3 separate cages per age group that were well-dispersed throughout each barn or facility and mixed thoroughly in sterile plastic bags. For commercial rabbits, one sample bag was submitted for each of two age groups, growers (aged 5–12 weeks) and does (reproductively active, adult, females) per farm. Samples from commercial rabbits were collected during summer (July-Aug) and winter (Feb-Mar) months, when possible. Producers were also asked to verbally report any antimicrobial use, both routine and sporadic, at the time of collection; this information was later confirmed by email or telephone follow-up. At research facilities, when possible, multiple samples (up to 3) were collected from the same facility if there were enough number of rabbits kept in the same room to allow for multiple collections (1 facility), rabbits were kept in multiple rooms or multiple sites throughout the facility (1 facility), significantly different age groups were present within the facility (1 facility), an opportunity was provided to collect samples in repeated years (1 facility), and an opportunity occurred to collect samples immediately following rabbit shipping/arrival to the facility and then following a period of acclimation (1 facility). Fecal samples from companion rabbits were collected from clinically normal rabbits whose owners attended a regional rabbit exposition (n = 55) and from healthy patients visiting the Ontario Veterinary College's Avian and Exotic Service for dental procedures (n = 7). Samples were chilled or placed on ice and transported for coding and processing to a central laboratory where 0.2 g of feces from each sample were isolated for DNA extraction while the remaining sample was frozen at -80 °C.
DNA extraction and quality control
DNA extraction was performed using the E.N.Z.A. Stool DNA Kit (Omega Bio-Tek Inc., Doraville, Georgia, USA) following the manufacturer's protocol for pathogen detection. Quantity and quality of extracted nucleic acids were assessed by spectrophotometry (NanoDrop, Roche, Mississauga, ON, Canada).
16S rRNA gene amplification, purification, and sequencing
Using the protocol described by Caporaso et al. [50] and the primers recommended by Klindworth et al. [51], the V4 region of the 16S rRNA gene was amplified by PCR using the forward primer S-D-Bact-0564-a-S-15 (5'-AYTGGGYDTAAGNG-3′) and the reverse primer S-D-Bact-0785-b-A-18 (5'-TACNVGGGTATCTAATCC-3′), with an expected product size of 240 bp. These forward and reverse primers contained a region that overlapped the Illumina forward and reverse sequencing primers (TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG and GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG, respectively), allowing them to anneal to the primers containing the Illumina adaptors plus the 8 bp identifier indices (forward: AATGATACGGCGACCACCGAGATCTACAC-index-TCGTCGGCAGCGTC; reverse:CAGCAGAAGACGGCATACGAGAT-index-GTCTCGTGGGCTCGG). For each extracted sample, a 25uL reaction was performed using 12.5uL KAPA Ready Mix, 9.0uL sterile water, 0.5uL each forward and reverse primers (10pM/uL), and 2.5uL DNA template (5 ng/uL). The PCR conditions were as follows: 1) 3 min at 94 °C for denaturation, 2) 45 s at 94 °C for denaturation, 3) 60 s at 53 °C for denaturation, 4) 1.5 min at 72 °C for elongation, and 5) 10 min at 72 °C. Steps 2–4 were repeated for a total of 27 cycles. PCR products were stored at 4 °C until purification was completed. Electrophoresis of PCR products was conducted in 2% agarose gel to evaluate the products for the presence of bands of the appropriate length (~ 240 bp). Purification of PCR products was conducted using Agencourt AMPure XP (Beckman Coulter Inc., Mississauga, ON, Canada); 20uL of AMPure XP was mixed with the PCR product on a 96 well plate. Following 2 min at room temperature, the mixture was transferred to a magnetic plate, left for 2 min, and then the supernatant was discarded. Beads were then washed with 200uL freshly made 80% ethanol twice, air dried for 10 min, and separated from the magnet. The beads were then rinsed with 52.5uL of 10 mM Tris pH 8.5 buffer, placed back onto the magnetic plate, and 50uL of the supernatant of each sample was transferred to a new tube.
A second reaction was performed using 2.5uL purified product, 12.5uL KAPA Ready mix, 9.0uL sterile water, and 1.0uL each Illumina Forward Index Primer (I501-I508 or S513, S15-S18, S20-S22) (10pM/uL) and Illumina Reverse Index Primer (I701–712 or N716, N718-N729) for each sample. An additional PCR was conducted with the following conditions: 1) 3 min at 94 °C, 2) 45 s at 94 °C, 3) 60 s at 50 °C, 4) 1.5 min at 72 °C, and 5) 10 min at 72 °C. Steps 2–4 were repeated for a total of 8 cycles completed. Again, the second PCR product was purified using AMPure XP using a similar procedure to that described above, with 40uL of AMPure XP and 37uL 10 mM Tris pH 8.5 Buffer. An evaluation of final products was completed using electrophoresis in 2% agarose gel, again observing for bands of the appropriate length.
When bands were not visible or of an inappropriate length, spectrophotometry was performed to identify nucleic acid quantity; adjustments to the volumes of KAPA Ready mix, DNA product, and sterile water were made based on the quantity of nucleic acids identified as needed to produce bands of the appropriate size.
Purified samples were normalized to a final concentration of 12.5 nM and submitted for sequencing to the University of Guelph's Advanced Analysis Centre. An Illumina MiSeq (San Diego, California, USA) and 2 × 250 chemistry were utilized for sequencing of the library pool.
Microbiota assessment and analyses
Analysis of the sequencing results was conducted using Mothur software (v1.35) [52]. Paired-end reads were aligned and sequences greater than 244 bp or less than 237 bp in length were removed from the data set. Any sequences containing long runs (> 8 bp) of holopolymers, as well as ambiguous base calls, sequences that did not align with the 16S rRNA gene V4 region, or that were identified as chimeras were also removed. Sequences from mitochondria, chloroplasts, Archaea, and eukaryotes were also removed. Sequences were binned into operational taxonomic units (OTUs) at the 3% dissimilarity level using nearest neighbour clustering using open OTU picking. Comparison of relative abundances of different taxa was conducted using one-way ANOVAs within the JMP 12 Response Screening Platform (SAS Institute Inc., Cary, NC, USA). Adjustment of p values to account for false discovery rate (FDR) was performed using the Benjamini-Hochberg technique. Analysis was performed both with facility (i.e., where the animal came from) named as a random effect and without. Follow-up post-hoc testing of taxa identified as significantly different between groups was conducted using the Steel-Dwass nonparametric test. Results were considered significant when adjusted P values were ≤ 0.05. Differences in microbiota composition based on age (fryers versus does) and season (summer versus winter), were calculated exclusively for commercial meat rabbits.
For subsequent analysis, subsampling was performed to normalize sequence numbers. Good's Coverage was used to assess coverage to ensure that the microbial population was adequately represented within the samples. Population richness was assessed using Chao1 to identify the number of different organisms present within each sample and the inverse Simpson's index was used to describe population diversity to determine the abundance of the different organisms present within each sample. The Jaccard index (which measures community membership by comparing the number of shared OTUs, but not their abundance) and Yue and Clayton measure of dissimilarity (which measures community structure by examining shared OTUs and their relative abundances) were used to assess population dissimilarity and to create dendrograms to compare how closely related the microbial communities of each sample were to one another. These were compared between groups for source, age and season using unweighted UniFrac (which measures the phylogenetic distance between communities based on membership), as well as parsimony tests (which examines community membership similarily). Figures were created using FigTree (v1.4.2) (http://tree.bio.ed.ac.uk) and JMP 12. Principal coordinate analysis (PCoA) was also performed to visualize data similarities and dissimilarities between microbial communities and figures were created in JMP 12. LEfse was also performed to identify OTUs that were differentially abundant between groups.
ANOVA:
FDR:
GI:
LDA:
Linear discriminant analysis
LEfSe:
Linear discriminant analysis effect size
OTU:
Operational taxonomic units
PCoA:
Principal coordinate analysis
REC:
Rabbit enteritis complex
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We thank Joyce Rousseau, Mackenzie Slifierz, Mohammed Jalai, and Jutta Hammermeuller for technical assistance, and Patrick Boerlin and Scott McEwen for editorial assistance.
This project was funded by a grant from the Ontario Ministry of Agriculture, Food, and Rural Affairs (200494), Farm and Food Care Ontario, Floradale Feed Mill, and BW Feeds. The funding sources had no influence on the study design, outcomes or interpretation of results.
Availability data and materials
The dataset used and analysed during the current study is available from the following site: Kylie, Jennifer; Turner, Patricia V.; Weese, Scott, 2017, "Prevalence of enteric disease agents in Ontario commercial rabbits: Zoonotic potential and impact on animal health", https://doi.org/10.5683/SP/BT7HN2, Scholars Portal Dataverse, V1.
Department of Pathobiology, University of Guelph, Guelph, ON, N1G 2W1, Canada
Jennifer Kylie
, J. Scott Weese
& Patricia V. Turner
Search for Jennifer Kylie in:
Search for J. Scott Weese in:
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PVT and JSW conceived of the work and prepared the grant; PVT, JSW, and JK recruited samples for the study, reviewed and analyzed the results, and prepared and reviewed the manuscript; and JK conducted the work. All authors read and approved the final manuscript.
Correspondence to Patricia V. Turner.
The study was approved by the University of Guelph Animal Care Committee (e1279). The facility is in compliance with the Animals for Research Act of Ontario and holds a Good Animal Practice certificate conferred by the Canadian Council for Animal Care.
All authors consent to publication of this original article.
Table S1. Relative abundance of predominant (≥ 1%) taxonomic classifications of bacteria isolated from the feces of domestic rabbits (n = 168). (DOCX 14 kb)
Table S2. Relative abundance and FDR p-values for significantly different bacterial phyla, classes, orders, families, and genera in the rabbit fecal microbiota when compared between commercial (n = 86), companion (n = 54), laboratory (n = 14), and shelter (n = 14) rabbit samples. (DOCX 24 kb)
Kylie, J., Weese, J.S. & Turner, P.V. Comparison of the fecal microbiota of domestic commercial meat, laboratory, companion, and shelter rabbits (Oryctolagus cuniculi). BMC Vet Res 14, 143 (2018). https://doi.org/10.1186/s12917-018-1464-6
Accepted: 18 April 2018
Gastrointestinal microbiota


Specially designed and built in 2010, with attention to every detail this magnificent yacht has one master, one VIP and two guest cabins each with air conditioning and en suit. She welcomes up to 8 passengers on board. Complete with an exceptional team of 4 who will be at your service through out your holiday.


The Monaco Yacht Show 2012 saw 33,000 luxury yachting companies, yacht owners, wealthy customers and representatives of future purchasers networking at the yacht show's 500 stands and aboard the 100+ superyachts.
Yachting Pages delivering box set copies of its superyacht directories.
Monaco Yacht Show 2012 was a record year for attendance.
With yacht prices having fallen significantly in the past four years and a wider choice available, luxury yachting is a buyer's market, benefiting affluent customers provided they decide to buy quickly, and the amount of potential buyers at the show reinforced this.
In this competitive situation brokers and shipyards brought out their best selection of superyachts for the four days of the Monaco Show; 103 superyachts of an average 46.70m length, including six mega-yachts of more than 70m and 38 new 2012 models.
Again this year the showcase spread along the Port Hercule quayside. Offshore 120 superyachts were at anchor, directly linked by tenders to the exhibitors on the docks. The constant to and fro of tenders indicated that meetings with potential buyers were aplenty.
Bertrand Vogèle, CEO of agency Yachting Partners International commented, "The most important moment of the year; the Monaco Yacht Show brings together all the output of boat builders wanting to show their latest models, with important boats for sale. It's the superyachting Mecca".
Jonathan Beckett, CEO of brokerage agency Burgess shares his view, "We had eight yachts on show, between 47m and 77m, and two catalogue items presented by the their builders as new 2012 models; we also had 25 other yachts at anchor offshore."
The boat builders had an equally successful show according to Henk De Vries, CEO of Feadship who was interviewed in the MYS studio, "The 2012 Monaco show was a success for us; much better than last year. But I imagine you've been hearing the same thing all day. The 2011 show was itself the best there had been since the start of the crisis (…); the crisis is still with us, but serious people are keeping busy. We have to be fit and ready for the fierce competition but that's OK. If you sit complacently on your laurels, you're out of the race."
Theo Hooning, secretary general of SYBAss (Superyacht Builders Association) stressed the need to be at the Monaco Yacht Show to promote luxury yachting to a new clientele of non-initiates, "Once again this year the Monaco Yacht Show has proven itself the most important place to be if you want to share in the experience of superyachting. With such a variety of superyachts on show and events like our Monaco Sunset party, visitors had a host of opportunities to discover and understand the nature of these exceptional boats."
There were cocktail parties at the stands, press talks and onboard parties and gala dinners in Monte Carlo's grand hotels, totalling approximately 100 events this year; a record.
Patrick Hoffman, CEO of Ulysse Nardin, an official 2012 MYS sponsor commented, "Ulysse Nardin has a long history in watchmaking but also in yachting and the sea; Monaco is a platform for meeting people in a convivial setting to share our passion for yachts and timepieces. It really is the ideal place for meeting the ideal people in an ideal atmosphere".
German auto group BMW, sponsor and official courtesy car provider for the 2012 MYS, considered the show "the ideal stage for presenting the new BMW 7 and BMW 6 luxury models to a top-rank global audience".
The official visit of HSH Prince Albert II of Monaco to the Monaco Yacht Show was the occasion this year for three superyachts to be awarded trophies: The 2012 MYS design award went to sailing yacht Vertigo (67.2m, Alloy Yachts) for the outstanding work by French naval architects Philippe Briand and Christian Liagre. The trophy, a crystal glass sailing yacht, was made by MYS partner Baccarat. The Green Star Class 2012 certificate for boats built in compliance with rigorous environmental standards was awarded to Better Place (50.5m, Wally) and Aslec 4 (45m, Rossinavi) by Italian ship classification company RINA.
Although MYS can always be sure of coverage in the yachting press, the organisers' extended efforts to promote the show to the business and general media – and so raise the profile of the luxury yachting market among the general public. As well as fifty interviews with leading figures in yachting recorded in the MYS Breaking News Centre (TV studio) and fed daily to the international lifestyle media, more than 800 journalists and media professionals were accredited.
The portrayal of superyachting is no longer restricted to the specialist media; the sector is naturally of interest to the business media since it is a significant driver of the global economy.


Will Princess Charlotte Elizabeth Diana be a billion pound baby?
Princess Charlotte Elizabeth Diana named after Prince Charles-her grandfather, the Queen-her great grandmother and Diana-her grandmother is likely to become as much a fashion icon as her mother Kate.
It has been predicted by a retail expert that the Princess of Cambridge, who made her arrival on Saturday, 2nd May, 2015, will be worth around £150 million to the British economy, meaning that she could be worth around £1billion by the time she reaches her 7th birthday.
Already a range of royal baby memorabilia has been released, products available include a coffee mug (£19.95), a white velvet cradle Christmas decoration (£14.95) embroidered with pearls and a £95 limited edition teddy bear on a velvet cushion.
Have you already bought a royal baby souvenir? Get in touch and tell us what it was.
COMPETITION: Win My 1st Years Bathrobe – As Seen On Prince George!
Filed Under: Blog, Coffee With Mrs Claus, Gifts For Kids At Christmas - Gift Category, Gifts For Men At Christmas - Gift Category, Gifts For Women At - Gift Christmas, Uncategorized Tagged With: cambridge, charlotte elizabeth diana, date of birth, hrh, kate and will, kate middleton, princess, royals, Will Princess Charlotte Elizabeth Diana be a billion pound baby?


text:
7-year-old Kerala students made to stand in scorching heat for 3 hours for failing to pay school fees
The incident occurred in the Settlement School in Aluva, Ernakulam on Wednesday. The students did not pay Rs 990 for the month of March.
By Nandita Natrajkumar
[Representational image]Creative Commons.
Two 7-year-old students in a school in Ernakulam, Kerala were made to stand in the scorching heat for three hours as a punishment for not paying their monthly fees. In addition to this, they were not allowed to write their examinations.
The incident occurred in the Settlement School in Aluva, Ernakulam on Wednesday, March 27, reports The News Minute. The students did not pay Rs 990 for the month of March.
Unfortunately, the school did not take the soaring temperatures into account, nor did they pay any heed to the medical condition of the one student who suffered from a nervous condition and had problems in his eye as well. Their punishment ended at 12 noon.
Sariga, the mother of the boys who was punished, said that the school did warn them of dire consequences if the fees weren't paid. She went on to tell TNM that she did not expect the authorities to take such an action against 7-year-old children.
The distraught mother said that she initially thought the only punishment meted out to her son was not being allowed to write the exam. She said that only later on, her son went on to reveal what exactly happened. She contacted the school teacher and was told that the punishment was ordered by the principal of the institution.
"We could not meet the principal on Wednesday. By the time we came with the panchayat representatives to the school, she had left. We then talked to management authorities, but they talked very rudely to us," Sariga was quoted as saying by TNM.
The Kerala State Human Resources Commission issued a suo motu case against the school on Thursday. In addition to this, the Commission's Chairman, Justice Antony Dominic, asked the District Collector and Deputy Director of Education for a full report on the issue within three weeks.
A case has been filed by parents at the Aluva West police station against the school principal under Section 75 (punishment for cruelty to child) of Juvenile Justice Act.
Dozens Of Indian Schoolgirls Beaten After Fighting Off Sexual Harassers

redpajama_set_name: RedPajamaCommonCrawl


text:
Young Visionaries Youth Leadership Academy Expands Programs
Terrance Stone started Young Visionaries Youth Leadership Academy in San Bernardino, Calif., 20 years ago as a way to give back to the community.
Today, he's the CEO and founder of the nonprofit, which has received more than $1 million in grants over the last few years to provide mentoring, job training, mental health services, and more to at-risk youth, the San Bernardino Sun reports.
"We're sending e-blasts and social media posts to let people know we're hiring and beefing up our administration," Stone tells the Sun.
"Given that amount of money, you have to have administrative staff to make sure all the paperwork and accounting systems are working properly."
Among Stone's recent grants: $400,000 to Loma Linda University for a violence prevention program, $1.2 million to the High Desert area for a mentoring program, and $135,000 to the IE Community Foundation to provide free movie nights in some of the area's "hard-to-reach community high-risk areas," according to a press release from Young Visionaries.
"We're going to start doing movies in some of these hard-to-reach community high-risk areas, and set up a giant 70-foot theater," Stone tells the Sun. Read the Entire Article
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Everything about the eruption of Mount Toba challenges the human mind. It is given an eight on the Volcanic Explosivity Index; that's the highest rating possible. For comparison, when Mount St. Helens blew its top in May 1980, it was given a rating of five.
However, the Index is logarithmic, meaning that Toba's eruption was several thousand times bigger than that of Mount St. Helens.
Volcanologists describe the Mount Toba event as "mega-colossal," chucking out somewhere around 2,800 cubic kilometres of debris. When it settled back down it covered the ground in a layer of ash over South Asia, the Indian Ocean, and the Arabian and South China Seas. This layer can still be seen.
Another measure of the scale of this eruption is that Mount Toba is now Lake Toba, a body of water 100 km long, 30 km wide, and more than 500 metres at its deepest.
India was in the direct line of fire and may have suffered a mass extinction of human and other life. Oppenheimer suggests nobody on the Indian subcontinent would have survived the cataclysm.
In a program aired on the Australian Broadcasting Corporation Science Show, Martin Williams, an emeritus professor at the University of Adelaide, spoke about the worldwide impact of the explosion.
It's been estimated that this plunge in temperatures killed off three quarters of the plant life in the Northern Hemisphere with equally catastrophic impacts in the southern half of the planet. It didn't help that the Earth was going through one of its periodic ice ages.
It was at least six years before the Sun's life-giving heat and light reached their normal intensity. This disrupted weather patterns such as seasonal rains causing plants bearing berries and nuts to die and game animal populations to plunge. Humans starved.
The loss of vegetation meant a shortage of food for all animals, including humans, and a resulting famine. This prompted anthropologist Stanley Ambrose of the University of Illinois to advance the notion that the human population came close to extinction in the aftermath of the Mount Toba eruption.
This is taken as tentative proof that at least some people in India survived the after effects of Mount Toba's bout of seismic indigestion, although Clarkson and Roberts are quick to point out more research is needed.
When Mount Tambora exploded in Indonesia in 1816 it caused a "Year Without a Summer" in the Northern Hemisphere. Mount Toba's eruption was 100 times greater than this.
Here's a cheery thought: Mount Toba may erupt again. Indonesia Geological Experts Association president Rovicky Dwi Putrohari says although the mountain disappeared 74,000 years its magma chamber remains intact. Researchers have located this large pool of pressurized liquid rock at between 20 and 100 kilometres below the surface.
According to The National Geographic, Mount Toba's big bang created enough lava to build two Mount Everests.
Some animals, dogs in particular, are thought to be able to detect infrasounds that give warning of an impending natural disaster such as a volcanic eruption, earthquake, or tsunami. These are noises that are inaudible to humans. Another theory is that some animals are sensitive to changes in electromagnetic fields that may presage a disaster.
About 80 percent of the world's land surface was created by volcanic action and today more than 500 volcanoes remain active.
"Ancient 'Volcanic Winter' Tied to Rapid Genetic Divergence in Humans." ScienceDaily, September 8, 1998.
"Humans Came 'Close to Extinction.' " Dr. David Whitehouse, BBC News, September 8, 1998.
"Volcano May not Have Blown it." Deborah Smith, Sydney Morning Herald, July 23, 2007.
"How Human Beings Almost Vanished From Earth In 70,000 B.C." Robert Krulwich, National Public Radio, October 22, 2012.
"North Sumatra's Mount Toba May Erupt Again, at any Time." Apriadi Gunawan, The Jakarta Post, November 4, 2013.
Well, nothing we can do about it so if you are a worshiper, get right with your Higher Power. If not, some kind of life should go on. Maybe you'll hit the lottery!
I'll take the former course. Not a gambler.


When this sword is out....sh** just got real.
What is the source of your avatar?
Hey what's going on, i have an aeon related question if you dont mind. Not sure if anyone knows of this tech trap or not: 4a-CE tech into fire left, right, and back leaves them stuned for 66b-BE k, tech left also leaves them in range for 3b into CE.... Alot of damage, close to 200. I tested this online and in training mode so it should work.
Playing offline all day everyday!
What? Was it something I did?
You just learn this? I got mad Yoshi troll technology.
Lets play tomorrow. Chaos K on here wants the match recorded...he asked me...I dunno how u feel about that, if u are ok i have to find somebody then cuz i can't record.
Sincerely Happy New Year to all of you Scrubs, Top Players, Friends and Trolls and I wish you the best for 2013.
Well I'm still here for 2 weeks or so. Just tell me when you feel like playing.
Hi. I was wondering if you would be up to play some games on XBL today? If you do, give me a timeline so I'll be there and we don't have to wait for each other.
Hum yeah. It's now or never so might as well! Give me your ID.
Ok I will send invite.
Sure is cold back home.
Just wanted to say an absolute well done at NEC!


text:
Raspberry Pi review
Gareth Halfacree finds out whether the £29 credit-card sized board with a 700MHz CPU and 256MB RAM can be used as an everyday PC.
by: Gareth Halfacree
1. Raspberry Pi review
Having examined the potential business benefits of the Raspberry Pi, we see if the tiny little board is capable of being used as a general-purpose computing machine.
The Raspberry Pi Model B comes with a Broadcom BCM2835 system-on-chip processor running at 700MHz and a VideoCore IV GPU running at 250MHz. A single 256MB module of Hynix LPDDR memory running at 400MHz provides RAM for the CPU and GPU, with the typical split leaving around 186MB of memory available for the user.
To get a feel for how the Pi performs as an actual PC, rather than as a development board, we installed the Debian Linux distribution with the lightweight LXDE desktop environment. Debian boots into a text-based terminal in just 13 seconds from a cold start, using a Transcend Class 10 SD card.
From here, it takes approximately eight seconds to load the LXDE desktop - at which point the system is ready for use. Loading the Midori web browser from the desktop takes around six seconds, plus a further eight seconds to load the IT Pro homepage.
It is possible to browse the web with the Pi, but Adobe Flash is not supported at this time
If you're not impressed by a 12 second load time for a lightweight web browser, you should remember this is being performed by a device running on a 5V 1A power supply and taking up no more room than a short stack of business cards.
Once loaded, the lack of hardware accelerated video starts to bite. Scrolling on complex pages is painful, taking a good couple of seconds to get started and then skipping down the page in an extremely jerky fashion. Once hardware accelerated drivers are bundled with the distribution, we expect this to improve dramatically.
Loading resource hungry programs such as the The Gimp picture editor, really stretches the Pi's CPU. The first load took 87 seconds and subsequent loads, which skipped the 'querying plug-ins' step - took an average of 43 seconds. Not too shabby, but a far cry from the sub-ten seconds the same program takes to load on a desktop machine.
The Pi isn't ideal for running resource hungry programs, but it wasn't designed to do so
It's when loading The Gimp and other more complex programs, you realise why the various Linux distributions for the Pi have a CPU monitor somewhere on the taskbar. Without a graph showing the CPU maxing out, it's all too easy to wonder if the delay is because you didn't double-click fast enough or whether the system has hung altogether.
In short: we found the Pi it perfectly possible to use the Raspberry Pi as a day-to-day machine, so long as you're fairly patient.
Raspberry Pi review - currently reading


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Victoria Beckham: 'It's time to bring sexy back. I'm sick of comfort dressing'
The designer is embracing body-con for the first time in a decade
By Tamara Abraham, Acting Senior Fashion Editor 14 January 2022 • 10:09am
Victoria Beckham in dresses from her VB Body capsule collection Credit: Getty Images
If you didn't already know that Victoria Beckham had spent a lot of time in Florida over the past year or so, you only need to glance at her latest collection (and year-round tan) to work it out.
The punchy apple greens, tangerine and warm terracotta which run through the pre autumn/winter 2022 collection on the rails lining her west London office are inspired by her time in Miami and Palm Beach, and a favourite restaurant in particular: St Ambroeus. "I was really taken by the colour palette," she says. "The walls with this gorgeous orange colour and then the banquettes were this mossy green. And just the whole vibe of this restaurant just felt really fresh and a different take on colour. So that was the starting point, the interior of that restaurant. I was going, 'David, take a picture of me here so that I can send it off to the atelier.'"
Certainly, she knows how to mix colours in an interesting way. When we meet, she is wearing a purple blouse with a tie neck (it's a sample she's road testing; her feedback will be that the neckline is a little low) along with a pair of chartreuse cords, chunky black heeled boots and a grey checked blazer draped over her shoulders.
It helps that she has a fresh Miami tan - her most recent visit was immediately after Christmas, when the whole family flew out to spend time with her eldest son Brooklyn, his fiancee Nicola Peltz and his future in-laws. She's hoping that travel restrictions and Covid won't affect the couple's upcoming wedding in April.
The palette borrows from the decor of Victoria's favourite Palm Beach restaurant. Versatility is also key, with a reversible trench coat
Bronzed skin aside, the key to carrying off bold colours like this, she says, is to keep the pieces themselves classic with a great cut (making each item as flattering as possible is a priority) with the interest coming from the combination of hues rather than any trend-led fashion frippery.
Now, more than ever, she wants to give her customers clothes they can wear year after year. A reversible trench coat is one example of her efforts to achieve this; a roll-neck body is a layering piece that has the potential to make summer dresses year-round wardrobe workhorses.
Perhaps the biggest Miami influence on this collection though, and on Victoria's style in general, is the return of body-con - a look she hasn't embraced since her WAG days - and later the figure-hugging silhouettes with which she first launched her label over a decade ago.
"I used to call them 'sucky sucky' dresses," she says of those early designs. Now she feels they have a place in her wardrobe again. "If I were to go out in Miami in a midi-length skirt and a pussy bow blouse, everyone would look at me like I'm completely crazy. They're going out with not a lot of clothing on. That's why I came up with the idea. I actually want to wear tight dresses again, I haven't dressed like that for a long, long time. The more I got into fashion, the more clothes I was wearing, the more layers, the bigger the clothes, the lower the shoes… [Now] I'm ready to make a change. I want to celebrate being a woman and having a feminine body."
A VB monogram dress styled with one of Victoria's roll-neck bodies underneath
These new pieces are designed to be comfortable, but they have nothing to do with lounging around. "It really was about bringing sexy back," she says of the design process. "I was sick of comfort dressing. I want to celebrate being a woman, I don't want to hide under baggy clothes, I don't want to talk about being in lockdown and wanting to dress comfortably. I think it's time to start dressing properly and get to the office."
Although the VB Body capsule of densely knit dresses tops and leggings will live under the umbrella of the main collection (the Victoria Victoria Beckham line has been absorbed into it too), it will be relatively affordable, with a bandeau starting at £90. "It's very inclusive," she adds. "Whether you are a size six, or the complete other end of the scale, you're going to look amazing in these dresses. They make every woman look curvy and really, really feminine."
Demand already appears to be strong for VB Body. Victoria wore a red knit dress from the line in New York towards the end of last year, and since then, Selfridges has been inundated with enquiries about them. This will be good news for Victoria Beckham CEO Marie Leblanc de Reynies - the business was still operating at a loss in December, although those losses have halved since she took the reins.
"The business is in the best shape it's ever been," Victoria says. "Marie has been the most wonderful CEO, she really has got things shipshape. Business is still challenging for everyone in this climate because we dress women to go out, to go to work, to go to parties to travel, all of the above have been very difficult. That being said, I think that my partner has just been wonderful. It's what I needed to get the business back on track."
Victoria's shift towards body-conscious femininity has nothing to do with pleasing a man, though one person who will be delighted by it is Mr Beckham. "My husband said to me once, 'Why do you all wear so many clothes? Why is everything so big?' At the time, we wanted to dress like that," she says. "Now I am ready to not dress like that anymore. Just celebrate being a woman, right?"
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Armenia is the excellent opportunity to enjoy the very natural and amazing organic food and cuisine. Historically located in the cross line of the silk road Armenians always used spices and vegetables and meat combination in his cuisine. And the complete set of this is armenian unique wine and brandy. Enjoy!!!


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Case Study – Euro Disney, the First 100 Days Essay
1342 Words Apr 17th, 2012 6 Pages
Case Study #1 – Euro Disney, The First 100 Days
1.0	Introduction
Ever since the first Disneyland was founded in 1955 in Anaheim, California, the Walt Disney Company had experienced nothing but success in the theme park business until its second oversea Disneyland – Euro Disney was opened in France in 1992.
Following the success of the company's first oversea Disneyland in Tokyo, Japan, on April 12, 1992, within its $4.4 billion budget, Euro Disney was opened in Marne-la-Vallee, France on a site that is one-fifth the size of Paris and just 20 miles to its west. Euro Disney was much like previous Disney theme parks with various of rides, attractions, hotels, restaurants, entertainment facilities, a campground, and even a championship …show more content…
"Another cornerstone of the Disney theme park franchise was the rich heritage of the Disney cartoon characters" (Loveman et al., 1992). The Disney cartoon characters had been a part of the childhood memories for many Americans. Each character is unique for the Americans. In the theme parks, those cartoon characters were brought to live. Visitors could always seek chances to fulfill their childhood dreams by taking photos with their favorite costumed characters in the theme parks.
"A third element of Disney's success was the unique role that visitors played in the theme" (Loveman et al., 1992). Visitors were more than just spectators but were considered important participants in the theme parks. They were always encouraged to join plays and to have frequent interactions with staff members.
3.0	Tokyo Disneyland
Disney's native success made the company started to expand overseas with the opening of Tokyo Disneyland on April 15, 1983. Tokyo Disneyland was owned and operated by the Oriental Land Company. Disney company designed the park and licensed the use of its characters in return for 10% of admissions revenues and 5% of food and souvenir revenues (Loveman et al., 1992). The tremendous success of Tokyo Disneyland started from its begining. Each year of operation, there were more than 10 million visitors coming to Tokyo Disneyland.
So why could Disney succeed in its expansion to Japan, which is a typical Eastern country that has many cultural differences
The Not-so-Wonderful World of Eurodisney-Things Are Better Now at Paris Disneyland
France was chosen to be the new location for which would be the biggest and most lavishing theme park that Disney had built around the world. Disney Management thought that because they had successfully opened Disneyland-Tokyo and surpassed their expectations, the European market was expected to succeed with the same business model and projected to generate over $100 million during the first year of operation which became a loss of $900 million by the summer of 1994. EuroDisney failed to attract…
Disney Case Essay
Ulset Title: International Modes of Entry Subtitle: The Case of Disney By, Carlos Gonzalez Hernandez This thesis was written as a part of the master program at NHH. Neither the institution, the supervisor, nor the censors are -through the approval of this thesis- responsible for neither the theories and methods used, nor results and conclusions drawn in this work. International Modes of Entry: The Case of Disney 1 Abstract The case of Disney's theme parks represents an opportunity to test…
Words: 19111 - Pages: 77
A Rose by Any Other Name Essay
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REAL MADRID CASE STUDY Summary Real Madrid is one of the most storied sports franchises in the world. It is one of the few sports teams with international appeal to a very broad fan base. This case delves into how Real Madrid transitioned from a good soccer team with a winning history into the global sports juggernaut we know today. We first examined Real Madrid's revenue streams and found that from their inception through the 1970s their business model was based almost entirely on their…
Essay Disney & Channel Business Proposal
@1.0 Executive Summary Disney characters including the Princess series, Pooh, Mickey Mouse and Minnie Mouse characters are designed collaborated with Chanel handbags. These collections are valuable luxury items and will launch in Japan with only 200 limited editions designed by Disney designers and produced by Chanel handmade systems. These items will be sold only at the Chanel Tower which is the largest Chanel boutique in the world located on Ginza Street in Tokyo. Japan has been a huge market…
Ford Motor Company Essay
It was once said, "Those who do not study the past are deemed to repeat it." On the brink of the new 21st century it is important for us at the Ford Motor Company to take a look at our past to see what has worked and what has not in order to set the standards for the automotive industry. It is also imperative to take a close look at what our competitors have done because we can also learn from their mistakes as well as improve on some of their ideas that have worked for them. It is important to…
Case Study Essay
PART II INSTRUCTOR'S NOTES ON TEXT CASES CASE GUIDE CHAPTER CASE | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10	| 11	| 12	| 13 | 14 | 15 | 16	| 17	| 18	| 19	| 1–1 Starbucks – Going Global Fast | X	| X	| | X	| X	| | | | | | X | X | | | | | | | | 1–2 Nestlé – The Infant Formula Incident | | X	| X	| X	| X	| | | X	| | | X | X | | | | | | | | 1–3 Coke and Pepsi Learn to Compete in India | | | | X	| X	| X	| | |…
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Fundamentals of Corporate Finance Essay
Copeland/Weston/Shastri Financial Theory and Corporate Policy Cox/Rubinstein Options Markets Dietrich Financial Services and Financial Institutions: Value Creation in Theory and Practice Dorfman Introduction to Risk Management and Insurance Dufey/Giddy Cases in International Finance Eakins Finance in .learn Eiteman/Stonehill/Moffett Multinational Business Finance Emery/Finnerty/Stowe Corporate Financial Management Fabozzi Bond Markets: Analysis and Strategies Fabozzi/Modigliani Capital Markets: Institutions…
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Essay Four Seasons Goes to Paris
meant providing high-quality, truly personalized service to enable guests to maximize the value of their time, however the guest defined doing so. In 1999, Four Seasons opened the Four Seasons Hotel George V Paris (hereafter, "F. S. George V"), its first French property, by renovating and operating the Hotel George V, a historic Parisian landmark. Doing so was, according to John Young, executive vice president, human resources, "one of our great challenges and triumphs." Young mused on what Four Seasons…
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THIRTEENTH EDITION Strategic Management CONCEPTS AND CASES Fred R. David Francis Marion University Florence, South Carolina Prentice Hall Boston Columbus Indianapolis New York San Francisco Upper Saddle River Amsterdam Cape Town Dubai London Madrid Milan Munich Paris Montreal Toronto Delhi Mexico City Sao Paulo Sydney Hong Kong Seoul Singapore Taipei Tokyo Strategic Management CONCEPTS AND CASES Editorial Director: Sally Yagan Editor in Chief: Eric Svendsen Acquisitions Editor: Kim Norbuta…
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Future New Album "Save Me" Download & Free Stream, Review, Cover Art
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Nicki Minaj's Boyfriend Is a Convicted Sex Offender Rapist – The Details!
Nicki Minaj's Boyfriend – Kenneth Petty – Has Been a Convicted Sex Offender Since 1994
When He Copped a Plea Deal for Attempted Rape & Served 4 Years in Prison
A post shared by Barbie® (@nickiminaj) on Dec 10, 2018 at 12:43pm PST
Nicki Minaj has caught a lot of flack in the past two weeks after she revealed who her new boo is. Internet sleuths quickly revealed that Kenny Petty is a registered sex offender and was previously convicted for killing a man. While Nicki's brushed off the criticism, The Blast has obtained the victim's detailed account of the incident and it is truly horrific.
We Break Down the Details Below!
Court documents reveal the victim's account in Kenneth Petty's rape case from 1994. The 16-year-old victim explained to police that she was allegedly kidnapped before being raped. The victim told police that Petty approached her from behind in Queens, NY before he "placed a hard circular object" to her back and "made a clicking sound." The victim, convinced it was a handgun, and took his instructions when he told her to "keep walking."
She claimed that he took her to a place down the street and forced her into an upstairs bedroom with the gun still pointed to her back. She alleges that he "laid his weight on top … pinched her sides and squeezed her neck causing her to sustain bruises and pain" before he "forced his penis" into her vagina.
The victim claimed that she made an attempt to leave the scene, but Petty pulled out a knife, put it against her stomach and told her that she "could not leave." She said that she eventually escaped after striking Petty with a plastic bottle.
Police claimed that he questioned his arrest when they went to his home, asking "what am I being arrested for" and claiming that he didn't do anything." He later admitted that he slept with the girl before.
Petty was convicted of attempted rape in the first degree after copping a plea deal. He served four years behind bars and is currently registered as a sex offender.
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text:
class Configurable < ActiveRecord::Base
  serialize :value

  after_save    :invalidate_cache, if: -> { ConfigurableEngine::Engine.config.use_cache }
  after_destroy :invalidate_cache, if: -> { ConfigurableEngine::Engine.config.use_cache }

  validates :name, presence: true, uniqueness: { case_sensitive: true }

  validate :type_of_value
  before_save :serialize_value

  def self.defaults
    @defaults ||= HashWithIndifferentAccess.new(
      YAML.load_file(
        Rails.root.join('config', 'configurable.yml')
      )
    )
  end

  def self.keys
    self.defaults.collect { |k,v| k.to_s }.sort
  end

  def self.[]=(key, value)
    exisiting = find_by_name(key)
    if exisiting
      exisiting.update_attribute(:value, value)
    else
      create {|c| c.name = key.to_s; c.value = value}
    end
  end

  def self.cache_key(key)
    "configurable_engine:#{key}"
  end

  def self.[](key)
    return parse_value key, defaults[key][:default] unless table_exists?

    value, found = [false, false]
    database_finder = -> do
      config = find_by_name(key)
      found = !!config
      value = config.try(:value)
    end

    if ConfigurableEngine::Engine.config.use_cache
      found = Rails.cache.exist?(cache_key(key))
      if found
        value = Rails.cache.fetch(cache_key(key))
      else
        database_finder.call
        if found
          Rails.cache.write cache_key(key), value
        end
      end
    else
      database_finder.call
    end

    if found
      value
    else
      parse_value(key, defaults[key][:default]).tap do |val|
        if ConfigurableEngine::Engine.config.use_cache
          Rails.cache.write cache_key(key), val
        end
      end
    end
  end

  def value
    self.class.parse_value name, super
  end

  def self.parse_value key, value
    case defaults[key][:type]
    when 'boolean'
      [true, 1, "1", "t", "true"].include?(value)
    when 'decimal'
      value ||= 0
      BigDecimal(value.to_s)
    when 'integer'
      value.to_i
    when 'list'
      value ||= []
      if value.is_a? Array
        value
      else
        value.split("\n").collect { |v| v =~ /,/ ? v.split(',') : v }
      end
    when 'date'
      if value.is_a? Date
        value
      else
       Date.parse(value) if value.present?
      end
    else
      value
    end
  end

  def self.serialized_value key
    value_for_serialization key, self[key]
  end

  def self.value_for_serialization(key, value)
    if defaults[key][:type] == 'list' && value.is_a?(Array)
      if value.all? {|entry| entry.is_a? Array}
        value = value.collect {|entry| entry.join ','}
      end
      value.join("\n")
    else
      value.to_s
    end.gsub("\r\n","\n")
  end

  def self.method_missing(name, *args)
    name_stripped = name.to_s.sub(/[\?=]$/, '')
    if self.keys.include?(name_stripped)
      if name.to_s.end_with?('=')
        self[name_stripped] = args.first
      else
        self[name_stripped]
      end
    else
      super
    end
  end

  private

  def type_of_value
    return unless name
    valid = case Configurable.defaults[name][:type]
    when 'boolean'
      [true, 1, "1", "true", false, 0, "0", "false"].include?(value)
    when 'decimal'
      BigDecimal(value) rescue false
    when 'integer'
      Integer(value) rescue false
    when 'list'
      value.is_a?(Array)
    when 'date'
      value.is_a?(Date)
    else
      true
    end
    errors.add(:value, I18n.t("activerecord.errors.messages.invalid")) unless valid
  end

  def serialize_value
    self.value = Configurable.value_for_serialization name, value
  end

  def invalidate_cache
    Rails.cache.delete(Configurable.cache_key self.name)
  end
end

redpajama_set_name: RedPajamaGithub


The poll President Donald Trump has publicly touted isn't bringing him much good news lately. The latest survey released by Rasmussen Reports Thursday found Trump's approval rating his nearing his all-time low.
Just 44 percent of respondents approved of Trump's job performance in the survey, while 56 percent disapproved. That's just one percentage point better than the president's lowest approval ever in the Rasmussen Reports tracking poll: He hit 43 percent approval on multiple occassions. But Trump had risen to 46 percent at the end of June, and he also tweeted out the results of the poll earlier in the month—when he rose to 50 percent approval.
"Great news!" he tweeted at the time. Now: Not so much. The Rasmussen Reports survey interviewed 1,500 likely voters and had a margin of error of plus or minus 2.5 percentage points.
To be sure, the 44 percent approval rating cited by Rasmussen—which has typically found better results for the president and is generally considered to be right-leaning—remains markedly better than the results of other polling firms. In the latest Gallup survey Thursday, for instance, Trump's approval rating was 37 percent, with 57 percent disapproving.
The weighted average from data-focused website FiveThirtyEight, meanwhile, pegged Trump's approval at 39 percent and his disapproval at 55 percent. The FiveThirtyEight average adjusts for a given poll's quality, recency, sample size and partisan lean.
Trump, who was visiting Poland Thursday, has seen his approval rating trend steadily downward since taking office in January, his popularity waning under the weight of the investigations into his campaign's possible ties with Russia and a widely unpopular Republican health care bill. For context's sake, at about this point in Barack Obama's first term, the former president had an approval rating of about 58 percent.
While the Russia investigations and the health care debate have been the most serious strains on the president's popularity, he recently did himself no favors by getting into an internet spat with two TV hosts. Referencing Morning Joe'sMike Brzezinksi and Joe Scarborough, Trump said in a series of tweets last week: "I heard poorly rated @Morning_Joe speaks badly of me (don't watch anymore). Then how come low I.Q. Crazy Mika, along with Psycho Joe, came to Mar-a-Lago 3 nights in a row around New Year's Eve, and insisted on joining me. She was bleeding badly from a face-lift. I said no!"
A poll this week found that 65 percent of voters felt such online behavior was unacceptable, and 55 percent said they would describe Trump as sexist.