Comparison of swabbing and destructive methods for microbiological pig carcass sampling

Aims: To compare the Belgian swabbing sampling method for pig carcasses with the reference destructive method with regard to Escherichia coli and aerobic plate counts, Salmonella and Campylobacter prevalence and their relationship. Methods and Results: Recovery was significantly lower for the swabbing method and corresponded to a recovery of 36% for E. coli counts and 81% for aerobic plate counts in comparison with the destructive method. There was no significant difference between the swabbing and destructive sampling methods for the prevalence of Salmonella or Campylobacter. A higher median for E. coli counts was detected for samples where Salmonella or Campylobacter were detected. The same association was also observed between the median for aerobic plate counts and the presence of Campylobacter. Conclusions: The method of swabbing used, covering 600 cm² on each half‐pig carcass, is efficient for the sampling of pig carcasses in comparison with the reference destructive method. Significance and Impact of the Study: This study describes an efficient method for microbiological pig carcass sampling. The Belgian swabbing method should continue to be used to allow the follow up of bacterial contamination in the Belgian meat production chain.     

Denture contamination by yeasts in the elderly

Objectives: The aim of this study was to investigate yeast carriage in healthy denture wearers by swabbing and to evaluate the effect of denture hygiene habits. Materials and methods: Denture wearers (n = 87) without evidence of denture stomatitis or any other oral disease were investigated by separately swabbing the fitting surface of the upper denture and the corresponding palatal mucosa in contact with the appliance. In a group of volunteers, a gel without any active compound was spread on the palatal side of the denture once in every morning for 2 weeks. Results: Screening showed Candida colonisation of upper prosthesis in 75.9% of individuals. The most frequent species isolated were Candida albicans (77.9% of the positive cultures), Candida glabrata (44.1%) and Candida tropicalis (19.1%). Carriage of more than one yeast species was found in 48.5% of the contaminated dentures. There was a statistically significant association between denture contamination and palatal mucosa colonisation (chi‐squared test: p < 0.0001). Repeated swabbings after 1 week as well as during a weekly follow‐up for 1 month confirmed the denture contamination and its degree of severity. A daily gel application produced a yeast‐count decrease to 10% of the initial value after 2 weeks (chi‐squared test: p = 0.0134 and p = 0.2841 for prosthesis and palatal mucosa, respectively). Conclusion: This study documented the reliability of oral swabbing when investigating yeast carriage in healthy denture wearers. Moreover, just a diagnostic tool, sampling upper dentures for Candida could be the opportunity to verify the patient’s compliance to hygiene advice.     

Variation in carbon and nitrogen isotopic ratios of fin and muscle tissues of Longnose Gar (Lepisosteus osseus) and Smallmouth Buffalo (Ictiobus bubalus)

Fin clips have been proposed as a non-lethal and non-invasive alternative to dorsal muscle samples in stable isotope analysis. However, potential differences in elemental composition and turnover rates can bias inferences when different tissues are combined. Here, we tested the average difference and correlation of the isotopic signature of δ¹³C and δ¹⁵N between muscle and fin samples in two arge-bodied fishes: Longnose Gar (Lepisosteus osseus) and Smallmouth Buffalo (Ictiobus bubalus). We found that δ¹⁵N signatures of muscle and fin tissues were strongly and positively correlated for both species, although the muscle tissue for Smallmouth Buffalo was slightly more enriched in δ15N. δ¹³C signatures of both tissues were significatively different and not strongly correlated for Longnose Gar, but similar and strongly correlated for Smallmouth Buffalo. Our results suggest that fin and muscle tissue can be combined for analyses of δ¹⁵N, but correction for higher enrichment of muscle tissue may be necessary for Smallmouth Buffalo. Conversely, combining fin and muscle tissue for analysis of δ¹³C requires more caution due to their weaker correlation and dependence of species identity.     

Blood and cloacal swab sampling for avian influenza monitoring has no effect on survival rates of free‐ranging ducks

Concerns about the spread of avian influenza viruses (AIVs) have led to cloacal swab sampling of hundreds of thousands of birds worldwide as part of AIV surveillance schemes, but the effects of cloacal swabbing have not been adequately evaluated. We tested for differences between swabbed, swabbed and bled, and non‐sampled wild ducks in terms of live re‐encounter and dead recoveries for Common Pochard Aythya ferina and Tufted Duck Aythya fuligula, and also determined re‐encounter and recovery rates for Mallard Anas platyrhynchos and Common Teal Anas crecca. No effects of sampling methods were detected, except in Teal. Re‐encounter rates were lower in sampled Teal than in controls, with annual re‐encounter probabilities being 25% and 35% lower in males and females, respectively. Teal possibly left or avoided sampling sites, or sought sites where they were less detectable after sampling. In general, no deleterious effects were found, suggesting that cloacal swabbing and blood sampling are suitable methods for conducting AIV surveillance in ducks.     

Validation of swabs as a non-destructive and relatively non-invasive DNA sampling method in fish

Non-destructive methods of collecting DNA from small fish species can be problematic, as fin clips can potentially affect behaviour or survivorship in the wild. Swabbing body mucus may provide a less invasive method of DNA collection. However, risk of contamination from other individuals in high density groups could give erroneous genotyping results. We compared multilocus microsatellite genotypes from the same individuals when collected at low and high density and compared this with fin clips. We found no differences between these categories, with a genotyping error rate of 0.42%, validating the use of body mucus swabbing for DNA collection in fish.     

Dream Clairvoyance Study II Using Dynamic Video-Clips: Investigation of Consensus Voting Judging Procedures and Target Emotionality

This partial replication study investigated whether individual versus small group consensus target judging procedures, and/or the emotionality of dynamic target video clips, would affect the frequency of correct identification of the target in a free-response dream ESP study. Two people located in Edinburgh (Scotland) and a third person located in Derby (England) acted both as experimenters and as participants and slept at their respective homes. On each of the 28 trial nights, a randomly-selected video clip was shown repeatedly between 3.00–4.30 am. The following morning the participants viewed four video clips (i.e., 3 decoys plus the target) and then judged the correspondences between the clips and records of their dream mentation. The Edinburgh participants obtained a greater number of direct hits using consensus as opposed to individual judgements. A discussion consensus procedure was marginally more successful than a more objective consensus procedure (12 hits, p = .0294, ES(h) = 0.38 vs. 11 hits, p = .0679, ES(h) = 0.30). Participants, both as a group and as individuals, obtained a greater proportion of direct hits when the target was emotionally negative than when it was either positive or neutral.     

基于红外相机技术的白冠长尾雉集群行为研究

白冠长尾雉(Syrmaticus reevesii)生性机警,行为隐蔽,在野外研究其集群行为较为困难。红外相机技术为解决这一问题提供了有益的工具。为研究白冠长尾雉集群行为及其地理变异,采用红外相机技术在湖北广水平靖关村和河南董寨自然保护区两个地点对其集群行为进行了调查。依据前期无线电遥测的结果,在两个研究地点分别设置了1 km×1 km的样地,在样地中分别放置了25台红外相机进行监测,野外累计放置了12412个相机日,获得视频24374段,其中有白冠长尾雉的视频有1361段。以有集群行为的独立视频数量(m)与独立视频数量(M)的比表示集群率,以拍摄到的白冠长尾雉个体数量(n)与有集群行为的独立视频数量(m)的比代表集群强度。结果表明,平靖关和董寨两地均是数量为2只的群体占比最大,在平靖关和董寨分别占68.23%、72.79%,Pearson卡方检验的结果表明群体大小在两地之间无显著差异(Pearson χ~2=6.522,df=5,P=0.259),而集群方式在两地之间则差异显著(Pearson χ~2=46.415,df=4,P0.001),两地均以单性集群为主要集群方式。单因素方差分析结果表明,集群率在季节间差异不明显。Neu方法分析发现,在平靖关,白冠长尾雉对灌丛和竹林显著偏好,对针阔混交林、杉木林和乔灌丛显著回避;而在董寨,其对针阔混交林和针叶林明显偏好,对杉木林和阔叶林明显回避。     

Underestimate of PS2 efficiency in the field due to high leaf temperature resulting from leaf clipping and its amendment

Chlorophyll fluorescence parameter F_v/F_m, an indicator of the maximum efficiency of PS2, is routinely measured in the field with plant leaves darkened by leaf clips. I found that on a sunny day of subtropical summer, the F_v/F_m ratio was often underestimated because of a large F₀ value resulted from a high leaf temperature caused by clipping the leaf under high irradiance, especially for long (e.g. 20 min) duration. This phenomenon may overestimate the down-regulation of PS2 efficiency under high irradiance. When leaf temperature was lower than 40 °C, the F₀ level of rice leaves under clipping remained practically unchanged. However, F₀ increased drastically with leaf temperature rising over 40 °C. In most measurements, no significant difference in F_m was found between rice leaves dark-adapted by leaf clips for 10 min and for 20 min. Therefore, shading leaf clips to prevent a drastic increase of leaf temperature, using F₀ measured immediately after the leaf being darkened to calculate F_v/F_m, as well as shortening the duration of leaf clipping are useful means to avoid an underestimate of F_v/F_m.     

Buccal swab: A tissue sampling method for refinement of experimental procedures involving rainbow trout

Buccal swabbing is a minimally invasive method to obtain DNA and biological material from humans and animals, including fish. Reports on buccal swabbing in fish are few and only for a limited number of species. Rainbow trout (Oncorhynchus mykiss) is an important animal model and because the yield of DNA may vary among and within different species in individuals of different sizes, it was selected as useful to optimize the buccal DNA collection in this species. Different storage methods were evaluated, aimed at DNA preservation by limiting DNA degradation and bacterial growth, using commonly available and inexpensive reagents. DNA quality was also tested by amplification of a single‐copy nuclear gene and a mitochondrial gene. The results suggest that ethanol is the best storage choice for buccal swab sampling in fish genetic studies, as well as suitable for small‐bodied rainbow trout.     

Similarities and differences in 13C and 15N stable isotope ratios in two non‐lethal tissue types from shovelnose sturgeon Scaphirhynchus platorynchus (Rafinesque, 1820)

We tested the hypothesis that δ¹³C and δ¹⁵N signatures of pectoral spines would provide measures of δ¹³C and δ¹⁵N similar to those obtained from fin clips for adult shovelnose sturgeon Scaphirhynchus platorynchus. Thirty‐two shovelnose sturgeon (fork length [FL] = 500–724 mm) were sampled from the lower Mississippi River, USA on 23 February 2013. Isotopic relationships between the two tissue types were analyzed using mixed model analysis of covariance. Tissue types differed significantly for both δ¹³C (P < 0.01; spine: mean = ?23.83, SD = 0.62; fin clip: mean = ?25.74, SD = 0.97) and δ¹⁵N (P = 0.01; spine: mean = 17.01, SD = 0.51; fin clip: mean = 17.19, SD = 0.62). Neither FL nor the FL × tissue type interaction had significant (P > 0.05) effects on δ¹³C. Fin clip δ¹³C values were highly variable and weakly correlated (r = 0.16, P = 0.40) with those from pectoral spines. We found a significant FL‐tissue type interaction for δ¹⁵N, reflecting increasing δ¹⁵N with FL for spines and decreasing δ¹⁵N with FL for fin clips. These results indicate that spines are not a substitute for fin clip tissue for measuring δ¹³C and δ¹⁵N for shovelnose sturgeon in the lower Mississippi River, but the two tissues have different turnover rates they may provide complementary information for assessing trophic position at different time scales.     

Black leaf-clips increased minimum fluorescence emission in clipped leaves exposed to high solar radiation during dark adaptation

Tomato and pepper leaves were clipped with black leaf clips for dark adaptation under solar radiation in the late spring or early summer 2010 in southern Italy. The leaves showed highly variable maximum PSII quantum yield (F_v/F_m = 0.026−0.802) using a continuous-excitation fluorometer Pocket PEA. These results were confirmed using the modulated fluorometer FMS1 on tomato leaves in mid summer, with F_v/F_m as low as 0.222 ± 0.277 due to nearly equal minimum (F_o) and maximum (F_m) fluorescence emission. A significant clip effect on F_v/F_m occurred after only 12 (tomato) or 25 (pepper) min. Increasing the leaf temperature from 25 to 50°C reportedly induced an F_o increase and Fm decrease so that F_v/F_m approached zero. The hypothesis that black leaf clips overheated under intense solar irradiance was verified by shrouding the clipped leaves with aluminum foil. In clipped leaves of pepper, F_v/F_m with the black clip/Pocket-PEA was 0.769 ± 0.025 (shrouded) and as low as 0.271 ± 0.163 (nonshrouded), the latter showing a double F_o and 32% lower F_m. An 8% clip effect on F_v/F_m was observed with the white clip/FMS1. To avoid the clip effect in high irradiance environments, F_v/F_m measurements with black clip/Pocket PEA system required leaf dark adaptation with radiation-reflecting shrouds. It would be useful if manufacturing companies could develop better radiation-reflecting leaf clips for the Pocket PEA fluorometer.     

Characterization of microsatellite DNA markers in a critically endangered species,Mekong giant catfish,Pangasianodon gigas

Microsatellite DNA markers for a critically endangered Mekong giant catfish (Pangasianodon gigas Roberts and Vidthayanon, 1991) were developed from fin clips collected from captive fish using (GT)₁₅ probe. The number of alleles per locus ranged from two to four. The expected heterozygosities ranged from 0.13 to 0.68. Also, these primers were successfully amplified in four closely related species, Pangasius bocourti, Pangasius conchophilus, Pangasius larnaudii and Pangasius sanitwongsei with the number of alleles per locus ranged from 1 to 13, 1 to 16, 1 to 12 and 1 to 4, respectively. These markers should prove to be very useful for the evaluation of genetic diversity for this species and other related Pangasius species.     

Application of non‐lethal stable isotope analysis to assess feeding patterns of juvenile pallid sturgeon Scaphirhynchus albus: a comparison of tissue types and sample preservation methods

Traditional techniques for stable isotope analysis (SIA) generally require sacrificing animals to collect tissue samples; this can be problematic when studying diets of endangered species such as the pallid sturgeon Scaphirhynchus albus. Our objectives were to (i) determine if pectoral fin tissue (non‐lethal) could be a substitute for muscle tissue (lethal) in SIA of juvenile pallid sturgeon, and (ii) evaluate the influence of preservation techniques on stable isotope values. In the laboratory, individual juvenile pallid sturgeon were held for up to 186 day and fed chironomids, fish, or a commercially available pellet diet. Significant, positive relationships (r² ≥ 0.8) were observed between fin and muscle tissues for both δ¹⁵N and δ¹³C; in all samples isotopes were enriched in fins compared to muscle tissue. Chironomid and fish based diets of juvenile pallid sturgeon were distinguishable for fast growing fish (0.3 mm day^?1) using stable δ¹⁵N and δ¹³C isotopes. Frozen and preserved fin tissue δ¹⁵N isotopes were strongly related (r² = 0.89) but δ¹³C isotopes were weakly related (r² = 0.16). Therefore, freezing is recommended for preservation of fin clips to avoid the confounding effect of enrichment by ethanol. This study demonstrates the utility of a non‐lethal technique to assess time integrated food habits of juvenile pallid sturgeon and should be applicable to other threatened or endangered species.     

New information on the evolution of mating behaviour in Sepsidae (Diptera) and the cost of male copulations in <Emphasis Type="Italic">Saltella sphondylii</Emphasis>

Here we describe the hitherto unknown details of the highly unusual mating behaviour of Saltella sphondylii—a widely cited model for male longevity costs caused by multiple copulations. When compared to the known mating behaviour of 28 sepsid species, we find five unique behavioural elements based on frame-by-frame analyses of video-recordings. These new behaviours are documented with video clips. We suggest that the male longevity costs could be due to copulation bouts that involve multiple insertions of a comparatively membranous phallus into the female. We compare the phallus of the Saltella sphondylii to those from three other species (Themira putris, Parapaleosepsis plebeia, Sepsis punctum).     

A comparative study of the biomass properties of Erianthus and sugarcane: lignocellulose structure,alkaline delignification rate,and enzymatic saccharification efficiency

A comprehensive understanding of the structure and properties of gramineous lignocelluloses is needed to facilitate their uses in biorefinery. In this study, lignocelluloses from fractionated internode tissues of two taxonomically close species, Erianthus arundinaceus and sugarcane (Saccharum spp.), were characterized. Our analyses determined that syringyl (S) lignins were predominant over guaiacyl (G) or p-hydroxyphenyl (H) lignins in sugarcane tissues; on the other hand, S lignin levels were similar to those of G lignin in Erianthus tissues. In addition, tricin units were detected in sugarcane tissues, but not in Erianthus tissues. Distributions of lignin inter-monomeric linkage types were also different in Erianthus and sugarcane tissues. Alkaline treatment removed lignins from sugarcane tissues more efficiently than Erianthus tissues, resulting in a higher enzymatic digestibility of sugarcane tissues compared with Erianthus tissues. Our data indicate that Erianthus biomass displayed resistance to alkaline delignification and enzymatic digestion.     

Heterogeneity of chromosomal breakage levels in epithelial tissue of ataxia-telangiectasia homozygotes and heterozygotes

Summary The objective of this study was to obtain an estimate of the frequency distribution of spontaneous chromosomal breakage occurring in vivo in oral epithelia of 20 ataxiatelangiectasia patients (A-T homozygotes) and 26 parents (A-T obligate heterozygotes). Samples of exfoliated cells were obtained from each individual by swabbing the oral cavity and preparing air-dried slides. The percentage of exfoliated cells with micronuclei (MEC frequency) was used as an in vivo indicator for the amount of chromosomal breakage occurring in the tissue. As a population group, MEC frequencies of the A-T patients differed significantly from controls (mean for A-T patients, 1.51; for controls, 0.29; P<0.01). However, the values observed in individual patients ranged from MEC frequencies 10- to 12-fold above control values, to frequencies overlapping the upper values observed in the controls. Similarily, MEC frequencies observed among the A-T heterozygotes differed significantly from controls (mean for A-T heterozygotes, 1.02, mean for controls, 0.29; P<0.01). However, only 16 of the 26 individuals sampled had MEC frequencies >0.5%, the 90th percentile for controls (compared with 16 of the 20 A-T patients examined). Of the A-T patients 11 had been previously assigned to complementation groups on the basis of sensitivity to x-irradiation. Seven of the patients belonged to group A and had MEC frequencies ranging from 0.3% to 1.9% with the remaining patients belonging to group C with MEC frequencies of 0.2% to 0.9%. The data presented in this paper suggest that although levels of spontaneous breakage in epithelial tissues of A-T patients and A-T obligate heterozygotes are often significantly elevated, this is not the case in all individuals.     

The effect of clipping frequency on the competitive interaction between two perennial grass species

Summary The effect of clipping frequency on competition between Lolium perenne and Agrostis tenuis was investigated. The yield of clippings of both species increased and then declined during the 12-week period of the experiment, but the clip yield of Lolium was always significantly greater than that of Agrostis. Lolium was clearly the better competitor in unclipped controls. The proportion of the biomass contributed to the mixture by Agrostis increased as the interval between clips decreased. Tiller production was unaffected by increased clipping frequency in Lolium but was increased in Agrostis. Total yield was much more drastically reduced by frequent clipping in Lolium than in Agrostis, where yield was practically unaffected by wide variations in clipping frequency.These results are in agreement with the field distributions of the two species. They also suggest that differences in height and response to clipping are likely to confound any attempt to monitor the progress of competition experiments by measuring the yield of clippings.     

Parity Influences the Demeanor of Sows in Group Housing

Across the globe, producers are moving from individual housing to group housing for sows during gestation. Producers typically group sows of a range of parities together, although the impacts are largely unknown. This study examined the behavioral expression at mixing for young, midparity, and older sows. Ten mixed-parity groups were filmed at mixing on a commercial piggery. One-minute clips were edited from continuous footage where focal sows of known parity could be identified, and scored for qualitative behavioral expression. Parity 2 and 6 sows were more calm/tired than Parity 4 sows, who were more active/energetic. Parity 2 sows were more curious/inquisitive than Parity 4 and 6 sows, who were more anxious/frustrated. Correlations between qualitative behavioral expression and activity indicated sows scored as more calm/tired spent a greater proportion of time standing, while sows scored as more active/energetic spent more time performing avoidance behavior. Different body language is likely to reflect physical or affective differences in how sows cope with mixing.     

Subclinical psoroptic otocariasis in Brazilian sheep with comments on a technique for mite collection

Subclinical psoroptic otocariasis associated withPsoroptes cuniculi (Delafond) was diagnosed in four out of ten herds of sheep. Transmission of mites between sheep and goats and vice versa was detected in herds kept on the same pastures for over 2 years. Flushing the ear canals of sheep and goats with approximately 50 ml of water appreared to be more efficient than swabbing or otoscopic examination for diagnosis and/or mite collection.     

Public Awareness of Melioidosis in Thailand and Potential Use of Video Clips as Educational Tools

Background

Melioidosis causes more than 1,000 deaths in Thailand each year. Infection occurs via inoculation, ingestion or inhalation of the causative organism (Burkholderia pseuodmallei) present in soil and water. Here, we evaluated public awareness of melioidosis using a combination of population-based questionnaire, a public engagement campaign to obtain video clips made by the public, and viewpoints on these video clips as potential educational tools about the disease and its prevention.

Methods

A questionnaire was developed to evaluate public awareness of melioidosis, and knowledge about its prevention. From 1 March to 31 April 2012, the questionnaire was delivered to five randomly selected adults in each of 928 districts in Thailand. A video clip contest entitled “Melioidosis, an infectious disease that Thais must know” was run between May and October 2012. The best 12 video clips judged by a contest committee were shown to 71 people at risk from melioidosis (diabetics). Focus group interviews were used to evaluate their perceptions of the video clips.

Results

Of 4,203 Thais who completed our study questionnaire, 74% had never heard of melioidosis, and 19% had heard of the disease but had no further knowledge. Most participants in all focus group sessions felt that video clips were beneficial and could positively influence them to increase adherence to recommended preventive behaviours, including drinking boiled water and wearing protective gear if in contact with soil or environmental water. Participants suggested that video clips should be presented in the local dialect with simple words rather than medical terms, in a serious manner, with a doctor as the one presenting the facts, and having detailed pictures of each recommended prevention method.

Conclusions

In summary, public awareness of melioidosis in Thailand is very low, and video clips could serve as a useful medium to educate people and promote disease prevention.

Presented in Part

World Melioidosis Congress 2013, Bangkok, Thailand, 18–20 September 2013 (abstract OS VII-04).