Chemoautotrophy, symbiosis and sedimented diatoms support high biomass of benthic molluscs in the Namibian shelf.

The molluscs Lucinoma capensis, Lembulus bicuspidatus and Nassarius vinctus are highly abundant in Namibian oxygen minimum zone sediments. To understand which nutritional strategies allow them to reach such impressive abundances in this extreme habitat we investigated their trophic diversity, including a chemosymbiosis in L. capensis, focussing on nitrogen biochemical pathways of the symbionts. We combined results of bulk nitrogen and carbon (δ13C and δ15N) and of compound-specific isotope analyses of amino acid nitrogen (AAs-δ15NPhe and δ15NGlu), with 16S rRNA gene sequencing of L. capensis tissues and also with exploratory results of ammonium, nitrate and nitrite turnover. The trophic position (TP) of the bivalve L. capensis is placed between autotrophy and mixotrophy, consistent with its proposed symbiosis with sulfur-oxidizing Candidatus Thiodiazotropha sp. symbionts. The symbionts are here revealed to perform nitrate reduction and ammonium uptake, with clear indications of ammonium host-symbionts recycling, but surprisingly unable to fix nitrogen. The TP of the bivalve L. bicuspidatus is placed in between mixotrophy and herbivory. The TP of the gastropod N. vinctus reflected omnivory. Multiple lines of evidences in combination with current ecosystem knowledge point to sedimented diatoms as important components of L. bicuspidatus and N. vinctus' diet, likely supplemented at times with chemoautotrophic bacteria. This study highlights the importance of benthic-pelagic coupling that fosters the dietary base for macrozoobenthos in the OMZ. It further unveils that, in contrast to all shallow water lucinid symbionts, deeper water lucinid symbionts rely on ammonium assimilation rather than dinitrogen fixation to obtain nitrogen for growth.

Microcapillary sampling of Baltic Sea copepod gut microbiomes indicates high variability among individuals and the potential for methane production.

The paradox of methane oversaturation in oxygenated surface water has been described in many pelagic systems and still raises the question of the source. Temora sp. and Acartia sp. commonly dominate the surface and subsurface waters of the central Baltic Sea. It is hypothesised that their gut microbiome at least partly contributes to the methane anomaly in this ecosystem. However, the potential pathway for this methane production remains unclear. Using a microcapillary technique, we successfully overcame the challenge of sampling the gut microbiome of copepods <1 mm. 16S rRNA gene amplicon sequencing revealed differences among the dominant bacterial communities associated with Temora sp. (Actinobacteria, Betaproteobacteria and Flavobacteriia) and Acartia sp. (Actinobacteria, Alphaproteobacteria and Betaproteobacteria) and the surrounding water (Proteobacteria, Cyanobacteria and Verrucomicrobia), but also intraspecific variability. In both copepods, gut-specific prokaryotic taxa and indicative species for methane production pathways (methanogenesis, dimethylsulfoniopropionate or methylphosphonate) were present. The relative abundance of archaea and methanogens was investigated using droplet digital polymerase chain reaction and showed a high variability among copepod individuals, underlining intra- and interspecific differences in copepod-associated prokaryotic communities. Overall, this work highlights that the guts of Temora sp. and Acartia sp. have the potential for methane production but are probably no hotspot.

The occurrence and genetic characterization of Cryptosporidium and Giardia species in foals in Belgium, The Netherlands, Germany and Greece.

Faecal samples were collected from foals between the age of 1 week and 6 months in Belgium, The Netherlands, Germany and Greece. A quantitative direct immunofluorescence assay based on the commercial MERIFLUOR Cryptosporidium/Giardia kit was performed to evaluate the presence of (oo) cysts. Parasite positive samples were genotyped, based on the 18S ribosomal DNA gene and the heat shock protein (HSP70) gene for Cryptosporidium and on the ß-giardin gene and the triose phosphate isomerase (TPI) gene for Giardia. In total, 134 foals from Belgium, 44 foals from The Netherlands, 30 foals from Germany and 190 foals from Greece were examined. No Cryptosporidium oocysts were identified in faecal samples from foals in Germany and The Netherlands. In Belgium and Greece, 4.5% and 1.1% of the foals examined were Cryptosporidium positive, respectively, all with a low oocyst excretion ranging from 100 to 2450 oocysts per gram of faeces. For Giardia, 14.2%, 11.4%, 10.0% and 11.6% of the foals in Belgium, The Netherlands, Germany and Greece, respectively, were found to excrete cysts, with a range of 50 up to 4,000,000 cysts per gram of faeces. Younger animals secreted significantly more Giardia cysts than older horses (p<0.05), but no significant correlation between Giardia infection and diarrhoea was observed. Most Giardia positive samples belonged to assemblage AI and/or BIV, but also assemblage E was detected in two samples. Together with the identification of Cryptosporidium horse genotype, this suggests only a low risk for zoonotic transmission.

An integrated fish-plankton aquaculture system in brackish water.

Integrated Multi-Trophic Aquaculture takes advantage of the mutualism between some detritivorous fish and phytoplankton. The fish recycle nutrients by consuming live (and dead) algae and provide the inorganic carbon to fuel the growth of live algae. In the meanwhile, algae purify the water and generate the oxygen required by fishes. Such mechanism stabilizes the functioning of an artificially recycling ecosystem, as exemplified by combining the euryhaline tilapia Sarotherodon melanotheron heudelotii and the unicellular alga Chlorella sp. Feed addition in this ecosystem results in faster fish growth but also in an increase in phytoplankton biomass, which must be limited. In the prototype described here, the algal population control is exerted by herbivorous zooplankton growing in a separate pond connected in parallel to the fish-algae ecosystem. The zooplankton production is then consumed by tilapia, particularly by the fry and juveniles, when water is returned to the main circuit. Chlorella sp. and Brachionus plicatilis are two planktonic species that have spontaneously colonized the brackish water of the prototype, which was set-up in Senegal along the Atlantic Ocean shoreline. In our system, water was entirely recycled and only evaporation was compensated (1.5% volume/day). Sediment, which accumulated in the zooplankton pond, was the only trophic cul-de-sac. The system was temporarily destabilized following an accidental rotifer invasion in the main circuit. This caused Chlorella disappearance and replacement by opportunist algae, not consumed by Brachionus. Following the entire consumption of the Brachionus population by tilapias, Chlorella predominated again. Our artificial ecosystem combining S. m. heudelotii, Chlorella and B. plicatilis thus appeared to be resilient. This farming system was operated over one year with a fish productivity of 1.85 kg/m2 per year during the cold season (January to April).

Assessing resistance against macrocyclic lactones in gastro-intestinal nematodes in cattle using the faecal egg count reduction test and the controlled efficacy test.

The main objective of the present study was to evaluate the accuracy of the faecal egg count reduction test (FECRT) to assess the resistance status of ivermectin (IVM)-resistant isolates of the cattle nematodes Ostertagia ostertagi and Cooperia oncophora, using the controlled efficacy test (worm counts) as a reference. The second objective was to investigate whether both IVM-resistant isolates showed side-resistance against moxidectin (MOX) under controlled conditions. Thirty male Holstein calves were experimentally infected with 25,000 L3 of an IVM-resistant O. ostertagi isolate and 25,000 L3 of an IVM-resistant C. oncophora isolate. Twenty-eight days later the calves were randomly divided into 2 treatment groups and 1 untreated control group. Animals in groups 1 and 2 received MOX (Cydectin(®) 1%, Pfizer) and IVM (Ivomec(®) 1%, Merial) respectively, by subcutaneous injection at a dose rate of 0.2mg/kg bodyweight. Faecal samples were collected 7 and 14 days after treatment and animals were necropsied 14/15 days post-treatment. Both the FECRT and the controlled efficacy test demonstrated that the O. ostertagi and C. oncophora isolates were resistant against IVM, with efficacies below 90%. The IVM-resistant O. ostertagia isolate was still susceptible to MOX treatment, as shown by over 99% reduction in egg counts and worm burden. The FECRT suggested borderline resistance against MOX in the IVM-resistant C. oncophora isolate, with egg count reductions between 97% (95% CI: 76; 100) at day 7 and 86% (95% CI: 49; 96) at day 14. However, the controlled efficacy test clearly showed MOX-resistance, with a decrease of only 31% (95% CI: -12; 57) in C. oncophora worm numbers. After MOX treatment, a significantly lower number of eggs per female C. oncophora worms was counted compared to the control group (43% reduction). Due to this reduced fecundity, the FECRT may fail to detect MOX-resistance.

Foci report on indigenous Dermacentor reticulatus populations in Belgium and a preliminary study of associated babesiosis pathogens.

The occurrence of autochthonous clinical cases of canine and equine babesiosis in Belgium during the last two decades suggests that the vector of the pathogens responsible for these diseases, Dermacentor reticulatus (Ixodida: Ixodidae), may be present in this country. Consequently, evidence for the presence of this tick species in different locations within Belgium was investigated. Four different locations were monitored by flagging in 2010; these included the locations at which D. reticulatus was previously found on a dog in 2009 and on two red deer in 2007. Two different species of tick were identified, Ixodes ricinus (Ixodida: Ixodidae) and D. reticulatus. A total of 282 D. reticulatus adult ticks (98 males, 184 females) were collected from the four sites. Ticks were found mainly from early March until the end of May and a peak in activity was apparent in March. A Babesia spp. (Piroplasmida: Babesiidae) genus-specific polymerase chain reaction test based on the amplification of a fragment of the 18S rRNA gene was used to investigate the potential presence of Babesia spp. All DNA extracts isolated from the total tick samples yielded negative results. Additional studies to accurately determine the distribution and vectorial capacity of this important tick species in Belgium are warranted.

Giardia duodenalis cyst survival in cattle slurry.

Giardia duodenalis is a protozoan parasite known to infect animals and humans. Zoonotic transmission of G. duodenalis can occur by the consumption of drinking water produced from surface water that is contaminated by runoff from manure-laden fields or pastures. Although it was previously reported that storing solid cattle manure decreases G. duodenalis cyst viability, no data are available on cyst survival in slurry waste from cattle. In this study the number and the viability of G. duodenalis cysts was determined in cattle slurry for up to 90 days. G. duodenalis cysts were counted in 30 slurry samples with a quantitative direct immunofluorescence assay. The geometric mean number of cysts was reduced by 77% after 90 days (P<0.0014), although there was substantial variability between samples. A fluorogenic dye staining using 4',6'-di-amino-2-phenylindole and propidium iodide showed a decreased viability from 45 days onwards, and after 90 days incubation, only 3% of the cysts were viable. Gerbils and lambs were artificially infected with 50 day-old and 90 day-old cysts and faecal excretion of G. duodenalis was monitored between 5 and 7 days after infection. Seven days after infection the gerbils were euthanized for Giardia trophozoite counts. Although one cyst was found in the faeces of one of the gerbils after infection with 50 day-old cysts, no trophozoites were recovered from the intestines of any gerbil (n=8). Experimental infection of lambs with 10(5)50 day-old and 90 day-old slurry cysts caused low cyst excretion in one out of two and one out of three lambs, respectively. Together, these data show that storage of cattle slurry for 90 days greatly reduces the number and viability of G. duodenalis cysts.

Work-related upper extremity disorders: one-year follow-up in an occupational diseases registry.

PURPOSE: To study the course and consequences of work-related upper extremity disorders in the registry of the Netherlands Centre for Occupational Diseases (NCvB). METHODS: A follow-up study was performed in a sample of consecutive cases of work-related upper extremity disorders notified to the NCvB. Perceived severity was measured with VAS (0-100), quality of life with VAS (0-100) and SF-36, functional impairment with DASH and sickness absence with a questionnaire. Measurements took place directly after notification (T0) and after 3, 6 and 12 months (T1-T3). A linear mixed model was used to compare scores over time. RESULTS: Average age of the 48 consecutive patients (89% response) was 42 years; 48% were men. Perceived severity, functional impairment and sickness absence decreased statistically significant during the follow-up period, and quality of life scores improved. Patients older than 45 years scored worse on perceived severity of the disease, functional impairment and quality of life than did younger patients. CONCLUSIONS: The role of registries of occupational diseases for preventive policy can be extended by creating longitudinal data in sample projects. In the sample from our registry, work-related upper extremity disorders had a favourable course.

Giardia and other intestinal parasites in different dog populations in Northern Belgium.

The objectives of this study were to obtain data on the prevalence of intestinal parasites in different dog populations in northern Belgium, to estimate the zoonotic risk associated with these infections and to identify potential risk factors. Between 2004 and 2007 a total of 1159 faecal samples were collected from 451 household dogs, 357 dogs from breeding kennels and 351 dogs with gastrointestinal disorders. The samples from dogs with gastrointestinal disorders were sent to the diagnostic Laboratory for Parasitology at Ghent University by veterinary practitioners. In household dogs the prevalence of intestinal parasites was relatively low. Giardia was the most commonly found parasite (9.3%, CI 5.5-13.1), followed by Toxocara canis (4.4%, CI 2.7-6.8). Much higher infection rates were observed in kennel dogs, especially for Giardia spp. (43.9%, CI 37.8-50.0); T. canis (26.3%, CI 21.8-31.2) and Cystoisospora spp. (26.3%, CI 21.8-31.2). Also in dogs with gastrointestinal problems, Giardia spp. (18.1%, CI 13.1-23.1), Cystoisospora spp. (8.8%, CI 6.1-12.3) and T. canis (7.4%, CI 4.9-10.7) were the most frequently detected parasites. In all dog populations pups were more frequently infected with Cystoisospora (P<0.0001 to P<0.05), Giardia (P<0.001 to P<0.05), and T. canis (P<0.0001 to P<0.001) than adult dogs, except for T. canis in household dogs, where this correlation was not significant. A significant association of anthelmintic treatment with T. canis infections was only observed within the household population. Household dogs with a higher number of treatments per year were more frequently infected with T. canis (P<0.05). There was a significant difference between the different breeding kennels for the occurrence of Cystoisospora, Giardia and T. canis (P<0.0001) and large kennels harboured relatively more infected animals than smaller breeding facilities (P<0.05). However, this was not significant for Giardia spp. Breed and gender did not affect the risk of an infection in any of the study populations. Toxocara and Giardia present a zoonotic risk, especially in household dogs, where the majority of Giardia positive samples (80%) belonged to the zoonotic assemblage A. In kennel dogs and clinically affected dogs the host-specific Giardia assemblages C and D were most prevalent (94% and 80%, respectively).

Notification of occupational skin diseases by dermatologists in The Netherlands.

BACKGROUND: A voluntary surveillance scheme of occupational skin diseases (OSDs) in The Netherlands starting in 2001 aimed to improve insight in the incidence of OSD especially occupational contact dermatitis (OCD), risk professions and causal agents. This paper presents the results of this scheme during 2001-05. METHODS: Reports of new cases of OSD received from the participating dermatologists on a monthly basis were analysed. Data evaluated included information on diagnosis, sex, age, sickness, absenteeism, profession and causal agents. Relative differences in incidence rates between industries or branches were estimated by calculating incidence rate ratios. RESULTS: About 80% of the notifications concerned OCD. The highest number of notifications was recorded in the first year of the scheme. This was probably due to reporting of a mixture of incident and prevalent cases. During the following 5 years, the number of yearly notifications of OSD declined. Hairdressers, nurses, metalworkers, mechanics and cleaners were the most commonly affected professions. Wet work and irritating substances were the most frequently reported causal agents. Most patients with OCD were not absent from work. CONCLUSIONS: A voluntary surveillance scheme with dermatologists provides valuable data about the distribution of OCD in risk professions and the causal agents. However, it has certain limitations in assessing trends in incidence. Active medical surveillance in populations at risk should be encouraged not only to improve secondary prevention but also to obtain more reliable information about the incidence of OCD.

Sentinel surveillance of occupational diseases: a quality improvement project.

BACKGROUND: Occupational diseases are generally underreported. The aim of this study was to evaluate whether a sentinel surveillance project comprising motivated and guided occupational physicians would provide higher quality information than a national registry for a policy to prevent occupational diseases. METHODS: A group of 45 occupational physicians participated in a sentinel surveillance project for two years. All other occupational physicians (N = 1,729) in the national registry were the reference group. We compared the number of notifications per occupational physician, the proportion of incorrect notifications, and the overall reported incidence of occupational diseases. RESULTS: The median number of notifications per occupational physician during the project was 13.0 (IQR, 4.5-31.5) in the sentinel group versus 1.0 (IQR, 0.0-5.0) in the reference group (P < 0.001). The proportion of incorrect notifications was 3.3% in the sentinel group and 8.9% in the reference group (P < 0.001). The overall reported occupational disease incidence was 7 times higher (RR = 6.9, 95% CI: 6.5-7.4) in the sentinel group (466 notifications per 100,000 employee years) than in the reference group (67 notifications per 100,000 employee years). CONCLUSIONS: A sentinel surveillance group comprising motivated and guided occupational physicians reported a substantially higher occupational disease incidence and a lower proportion of incorrect notifications than a national registry.

Outpatient angioplasty and stenting facilitated by percutaneous arterial suture closure devices.

AIM: To review our practice of outpatient percutaneous vascular interventions facilitated by an arterial suture device. MATERIALS AND METHODS: A retrospective review of all patients attending this tertiary centre for iliac or femoral intervention was undertaken between February 2001 and December 2004. All patients who underwent angioplasty or stenting had their puncture sites closed using a Perclose suture. Patients were kept flat for 15min and allowed to fully mobilize at 60min. Puncture sites were scored for visible bruising, haematoma and pain at discharge and on outpatient follow-up. Patient preference for future outpatient treatment was assessed. RESULTS: Fifty-seven outpatients underwent 81 punctures. Forty-eight (84%) patients underwent iliac angioplasty; of those 42% underwent stent placement. Six patients (10%) required inpatient admission, five secondary to failed suture deployment. One patient had a non-closer-related puncture site intimal flap occlusion successfully repaired at surgery. Fifty-one (90%) patients discharged with a mean time of 157min (60-280min). Forty-six (92%) patients had no visible bruising or palpable haematoma on discharge. No patient had a haematoma greater than 2.5cm. No discharged patient required readmission. Thirty percent reported a moderate to severe groin pain score (2-5/5) at discharge, increasing to 40% at follow-up. Forty-seven (98%) of the 48 patients, who expressed a preference, would be happy to undergo outpatient treatment again. CONCLUSION: Outpatient treatment is feasible, well tolerated and preferable to patients, but 10% will require inpatient admission. A planned post-procedure analgesia regimen or advice should be considered.

Conservative treatment for repetitive strain injury.

Various conservative treatment options for repetitive strain injury are widely used, despite questionable evidence of their effectiveness. This systematic review evaluates the effectiveness of these treatment options for relieving symptoms of repetitive strain injury and improving activities of daily living. Searches in Medline and Embase, with additional reference checking resulted in 15 eligible trials for this review. Methodological quality was assessed, and data-extraction was performed. With the use of a "best-evidence synthesis", no strong evidence was found for the effectiveness of any of the treatment options. There is limited evidence that multidisciplinary rehabilitation, ergonomic intervention measures, exercises, and spinal manipulation combined with soft tissue therapy are effective in providing symptom relief or improving activities of daily living. There is conflicting evidence for the effectiveness of behavioral therapy. In conclusion, little is known about the effectiveness of conservative treatment options for repetitive strain injury. To establish strong evidence, more high-quality trials are needed.

Time reproduction, working memory, and behavioral inhibition in children with ADHD.

Barkley (1997a) proposed that the central deficit in Attention Deficit Hyperactivity Disorder (ADHD) is in behavioral inhibition, resulting in deficits both in working memory and sense of time. To test this theory, 21 children with ADHD and a matched sample (ages 6-13) completed a variety of tasks that measured behavioral inhibition, working memory, attention, and time reproduction. Children with ADHD performed significantly below controls on measures of inhibition, attention, and time reproduction. They did not differ significantly from controls, however, on tasks of working memory. In addition, performance on the working memory tasks was not correlated with the time reproduction task. We discuss the relevance of these results in the context of current theories of ADHD.

WMS-III performance in patients with temporal lobe epilepsy: group differences and individual classification.

The utility of the WMS-III in detecting lateralized impairment was examined in a large sample of patients with temporal lobe epilepsy. Methods of analysis included evaluation of group means on the various indexes and subtest scores, the use of ROC curves, and an examination of Auditory-Visual Index discrepancy scores. In addition, performance on immediate and delayed indexes in the auditory and the visual modality was compared within each group. Of the WMS-III scores, the Auditory-Visual Delayed Index difference score appeared most sensitive to side of temporal dysfunction, although patient classification rates were not within an acceptable range to have clinical utility. The ability to predict laterality based on statistically significant index score differences was particularly weak for those with left temporal dysfunction. The use of unusually large discrepancies led to improved prediction, however, the rarity of such scores in this population limits their usefulness. Although the utility of the WMS-III in detecting laterality may be limited in preoperative cases, the WMS-III may still hold considerable promise as a measure of memory in documenting baseline performance and in detecting those that may be at risk following surgery.

The effect of cardiopulmonary bypass on circulating megakaryocytes.

Megakaryocytes (Mks) are found in the lungs and the blood stream as well as in the bone marrow. We modified a whole blood filtration method for Mks by immunostaining for CD61 using biotin streptavidin, and used this technique to study Mks and their morphology in the central venous and arterial circulations before, during and after cardiopulmonary bypass (CPB) in haematologically normal patients undergoing routine cardiac surgery. Blood samples were taken immediately after the insertion of central venous (V) and arterial (A) catheters and after thoracotomy, immediately before bypass. Further samples were taken after 60-90 min on-CPB and 180-240 min post-bypass. In comparison with the steady state before bypass, circulating Mk levels in blood on bypass increased dramatically, from (V) 10.93 +/- 3.94/ml (mean +/- SD) to 36.48 +/- 11.52/ml and from (A) 8.37 +/- 4.39/ml to 38.65 +/- 20.68/ml. This effect was still present, to a lesser extent, 180-240 min post-bypass. Circulating levels of Mks were consistently lower in the arterial circulation than in the venous circulation off bypass, but levels in the two circulations were comparable during CPB, confirming previous suggestions that the lungs are net removers of Mks from the circulation. Type 4 Mks, the largest and most normal morphologically, were rarely seen in arterial blood, but increased significantly during CPB, indicating that the lungs selectively remove large Mks. The lungs appear to play an active role in the regulation of Mk levels. This is lost during CPB and despite the extracorporeal 40 microm arterial line filter, large Mks enter the systemic circulation. More effective extracorporeal filtration of large Mks might reduce the neurological impairment seen in some patients who have undergone CPB.