Bulk tank milk surveillance as a measure to detect Coxiella burnetii shedding dairy goat herds in the Netherlands between 2009 and 2014.

In the period from 2005 to 2009, Coxiella burnetii was a cause of abortion waves at 28 dairy goat farms and 2 dairy sheep farms in the Netherlands. Two years after the first abortion waves, a large human Q fever outbreak started mainly in the same region, and aborting small ruminants were regarded as most probable source. To distinguish between infected and noninfected herds, a surveillance program started in October 2009, based on PCR testing of bulk tank milk (BTM) samples, which had never been described before. The aim of this study was to analyze the effectiveness of this surveillance program and to evaluate both the effect of culling of pregnant dairy goats on positive farms and of vaccination on BTM results. Bulk tank milk samples were tested for C. burnetii DNA using a real-time PCR, and results were analyzed in relation to vaccination, culling, and notifiable (officially reported to government) C. burnetii abortion records. In spring and autumn, BTM samples were also tested for antibodies using an ELISA, and results were evaluated in relation to the compulsory vaccination campaign. Between October 2009 and April 2014, 1,660 (5.6%) out of 29,875 BTM samples from 401 dairy goat farms tested positive for C. burnetii DNA. The percentage of positive samples dropped from 20.5% in 2009 to 0.3% in 2014. In a multivariable model, significantly higher odds of being PCR positive in the BTM surveillance program were found in farms of which all pregnant dairy goats were culled. Additionally, the risk for C. burnetii BTM PCR positivity significantly decreased after multiple vaccinations. Bulk tank milk ELISA results were significantly higher after vaccination than before. The ELISA results were higher after multiple vaccinations compared with a single vaccination, and ELISA results on officially declared infected farms were significantly higher compared with noninfected farms. In conclusion, BTM surveillance is an effective and useful tool to detect C. burnetii shedding dairy goat herds and to monitor a Q fever outbreak, and thus the effect of implemented measures.

Dynamics of Salmonella inoculated during rearing of black soldier fly larvae (Hermetia illucens).

The black soldier fly is currently the most produced edible insect on industrial scale, with its larval stage being processed into animal feed as the main application. As this insect species enters the feed and food chain, good hygiene and monitoring practices are needed to avoid the entrance of foodborne pathogens via the larvae. However, insufficient data on the risk of such introductions via industrial larvae production are available. To address this gap, a range of rearing trials were conducted in which the substrate, chicken feed, was inoculated with different levels of Salmonella and in which total viable counts and Salmonella counts were determined during the following days. The outgrowth of Salmonella was slower in those experiments with a lower initial contamination level than in experiments with a higher level. No significant reducing effect originating from the larvae on the substrate Salmonella counts was observed, in contrast to previous studies using other substrates. Our study also revealed that airborne transmission of Salmonella is possible under rearing conditions corresponding to those applied at industrial production sites. Based on our results, we recommend insect producers to use substrate ingredients free of Salmonella, and not to count on the antimicrobial activities that BSFL may exert in some situations towards food pathogens. More inoculation studies using other Salmonella serotypes, other zoonotic bacteria, other substrates, larvae of other ages and including variations on rearing protocols are needed in order to obtain a general view on the dynamics of food pathogens in this insect species and to support comprehensive risk assessments.

Staphylococcus aureus in Substrates for Black Soldier Fly Larvae (Hermetia illucens) and Its Dynamics during Rearing.

Black soldier fly larvae (BSFL; Hermetia illucens) are promising insects for the conversion of organic waste streams into valuable biomolecules. Such waste streams can contain foodborne pathogens. To assess this risk factor, this study evaluated the presence of Staphylococcus aureus in waste streams as a substrate ingredient for BSFL production as well as in the rearing process. First, the general microbiological quality and the occurrence of S. aureus were investigated for different waste streams. Staphylococcus aureus was abundantly present. Control of pH and water activity should avoid pathogens, which cannot grow in single-substrate ingredients, redeveloping when mixing streams for optimal substrate conditions for BSFL production. Next, it was investigated whether S. aureus present in the substrate was ingested and/or eradicated by BSFL. In inoculation trials, with S. aureus added to chicken feed as the substrate at 3 or 7 log CFU/g, the larvae showed a reducing effect on S. aureus. After 6 days, S. aureus counts were below the detection limit (2.0 log CFU/g) in all larvae samples and decreased in the substrate to <2.0 and <3.1 log CFU/g for inoculation levels of 3 and 7 log CFU/g, respectively. While this is promising, it is still recommended to monitor and control this pathogen in BSFL rearing. Intriguingly, screening of antimicrobial activity of dominant microorganisms associated with BSFL showed a clear activity of Trichosporon isolates against S. aureus. Future research should explore whether Trichosporon, which is frequently observed in BSFL, plays a role in controlling specific microorganisms, such as S. aureus. IMPORTANCE Given the increasing need for (more sustainable) methods to upcycle organic waste streams, the interest to rear insects, like black soldier fly larvae (BSFL), on such streams is increasing. This study reveals that S. aureus is abundantly present in such waste streams, which might be a point of attention for insect producers. At the same time, it reveals that when S. aureus was inoculated in chicken feed as the substrate, it was not detected in the larvae and was reduced in the substrate after 6 days. Future inoculation trials should investigate whether this reduction is substrate dependent or not. Toward the future, the role of the BSFL microbiota in controlling intestinal bacterial community homeostasis should be explored, because one of the dominant microorganisms associated with BSFL, Trichosporon spp., showed clear activity against S. aureus.

Aspects of the epidemiology, research, and control of lentiviral infections of small ruminants and their relevance to Dutch sheep and goat farming.

In 1862, the veterinarian Loman reported the first sheep in The Netherlands with symptoms associated with lentiviral infection, although at the time the symptoms were ascribed to ovine progressive pneumonia. In the following century, similar cases were reported by South African, French, American, and Icelandic researchers. Extensive research into the pathology, aetiology, and epidemiology of this slowly progressive and ultimately fatal disease was initiated in several countries, including the Netherlands. Studies of the causative agents--maedi visna virus (MVV) in sheep and caprine arthritis encephalitis virus (CAEV) in goats, comprising the heterogeneous group of the small ruminant lentiviruses (SRLV)--prompted the development of diagnostic methods and the initiation of disease control programmes in many European countries including the Netherlands, as a pioneer in 1982, and in the U.S.A. and Canada.

Premotor cortical ablations in monkeys: contralateral changes in visually guided reaching behavior.

In rhesus monkeys (Macaca mulatta), ablation of the premotor and supplementary motor areas and the adjoining rostral half of the precentral gyrus impairs the capacity of the contralateral arm to reach around a transparent obstacle to a visible food reward, and results in a tendency of this arm to reach around a transparent obstacle to a visible food reward, and results in a tendency of this arm to reach straight to where the food is visible. This may reflect a disinhibition of brainstem pathways which steer the arm and hand straight ot a visual target.

Presence and species identity of rumen flukes in cattle and sheep in the Netherlands.

The purpose of the study was to gain knowledge about the prevalence and identity of rumen flukes (RF) in cattle and sheep in the Netherlands. Routine faecal examinations of diagnostic submissions between May 2009 and September 2014 showed a mean annual herd or flock RF prevalence of 15.8% for cattle and 8.0% for sheep. Prevalence in cattle was higher after 2012 than before, which may reflect a change in detection method as well as an increase in true prevalence. During November and December 2014, an abattoir survey was conducted to allow for scoring of rumen fluke burden and to obtain specimens for molecular species characterization. Over 8 visits to 5 abattoirs in areas deemed to pose a high risk for trematode infection, 116 cows and 41 sheep from 27 herds and 10 flocks were examined. Prevalence of RF was higher in beef cattle than in dairy cattle and higher in cattle than in sheep. Median fluke burden was >100 specimens per animal for most positive animals. Using a semi-quantitative RF density score as a gold standard, sensitivity and specificity of a modified quantitative Dorsman egg counting method were estimated at 82.6% and 83.3%, respectively. Of 14 collected adult rumen flukes, twelve (8 bovine and 4 ovine specimens) were identified as Calicophoron daubneyi. The other two, of bovine origin, were identified as Paramphistomum leydeni, which was unexpected as in other European countries all recently collected rumen flukes in both cattle and sheep were identified as C. daubneyi. The findings implicate that multiple rumen fluke species, intermediate host species and transmission cycles may play a role in rumen fluke infections in the Netherlands.

Haemonchus contortus resistance to monepantel in sheep.

In a sheep farm in the Netherlands with a suspected Haemonchus contortus resistance to monepantel (Zolvix®, Novartis Animal Health), a fecal egg count reduction test was carried out in two groups of lambs, according to the method of the World Association for the Advancement of Veterinary Parasitology. Group 1 was the untreated control group, and group 2 was treated with monepantel at the manufacturer's recommended dose rate. Efficacy of treatment with monepantel was 0%. Larval identification of pre- and post-treatment coprocultures revealed 100% H. contortus larvae. On this farm, after a perceived reduction in efficacy of ivermectin and doramectin, the sheep farmer started using monepantel in July 2012, and since then, monepantel was used as the sole anthelmintic. Breeding sheep were treated twice each year in 2013 and 2014, and lambs two times in 2012, four times in 2013, and three times in 2014, before monepantel resistance was suspected, and confirmed three weeks later. Although the frequency of monepantel treatments on this farm was relatively high with treatments on thirteen separate occasions in two years time, possibly establishing favorable conditions for a competitive advantage for resistant H. contortus, it is remarkable that resistance to monepantel was established in such a very short period. This study confirms, to the best of our knowledge, the first case of H. contortus resistance to monepantel occurring in the field.

Scintigraphic demonstration of accessory hepatic duct leak following liver transplantation.

Few noninvasive methods are available to diagnose complications following liver transplantation. Hepatobiliary scintigraphy can differentiate rejection from primary biliary complications such as obstruction or extravasation in patients with nonspecific clinical findings such as fever and rising liver function studies. In the following case report, an unexpected biliary leak from a recipient accessory hepatic duct was demonstrated by [99mTc] DISIDA scintigraphy following liver transplantation.

Iodine-131 MIBG scintigraphy of neuroendocrine tumors other than pheochromocytoma and neuroblastoma.

Metaiodobenzylguanidine (MIBG) locates most pheochromocytomas and neuroblastomas. The tracer is concentrated in intracellular storage vesicles by an active process. Many other neuroendocrine tumors of the amine precursor uptake and decarboxylation (APUD) series have hormonal storage vesicles and, thus, the potential to take up [131I]MIBG. A variety of neuroendocrine tumors in 57 patients were studied 1, 2, and 3 days after 0.5 mCi [131I]MIBG. Views from skull to pelvis were obtained. Results of MIBG scans were compared with all available imaging modalities (including plain radiography, liver scan, ultrasound, computed tomography, and angiography) and surgical exploration. The neuroendocrine nature of the tumor was determined by histology, immunohistochemistry, electron microscopy, and the assay of appropriate biogenic amines and peptide hormones. Results were (positive/total cases): carcinoids (four of ten), nonsecreting paragangliomas (three of three), sporadic medullary carcinomas of the thyroid (MCT) (one of five), familial MCT (one of 26), chemodectomas (two of five), oat cell carcinomas (zero of four), choriocarcinoma (one of one), atypical schwannoma (with storage granules) (one of one), Merkel cell skin cancer (one of one), islet cell carcinoma (zero of one). We conclude that a wide range of neuroendocrine tumors show [131I]MIBG uptake; tumors other than pheochromocytomas and neuroblastomas are less often seen scintigraphically, but in certain cases (e.g., carcinoid and nonsecreting paragangliomas) scintigraphy may be useful in depicting the extent and location of disease and may indicate therapeutic potential. Iodine-131 MIBG shows promise in the diagnosis and staging of tumors of varied types.

Immunolymphoscintigraphy in breast cancer: evaluation using 131I-labeled monoclonal antibody B72.3.

Noninvasive axillary lymph node staging was investigated using [131I]murine monoclonal antibody B72.3 in 16 patients with breast cancer scheduled for axillary dissection. [131I]B72.3 was injected into ipsilateral finger webs or around the breast biopsy. Scintigraphy to 72 h and gamma-counting/immunohistochemistry of nodes were performed. Specific antibody uptake (%ID/g) and the ratio of specific:nonspecific antibody uptake were not significantly different in tumor-positive versus tumor-negative nodes, suggesting that [131I]B72.3 is unsuitable to discriminate axillary node tumor involvement.

The effect of cobalt supplementation on the immune response in vitamin B12 deficient Texel lambs.

The effect of cobalt supplementation on the immune reactivity in vitamin B12 deficient lambs was measured by comparing the humoral and cell-mediated immune responses against bovine herpes virus type 1 and Mycobacterium paratuberculosis. In addition, faecal egg counts were performed after natural infection with gastrointestinal nematodes. The experiments were performed with registered Texel twin lambs of the same sex. One lamb of each twin received three cobalt pellets divided over the grazing period. The non-supplemented lambs had lower serum vitamin B12 levels than their supplemented brother or sister. Our results demonstrate significantly lower lymphoblastic responses against Mycobacterium paratuberculosis in non-supplemented lambs compared with supplemented lambs 4 weeks after paratuberculosis vaccination. Vitamin B12 deficient lambs in this study had significant higher faecal egg counts than their supplemented brother or sister after natural infection with gastrointestinal nematodes. No differences were found in total and differential white blood cell counts, in total protein, albumin, alpha-, beta- and gamma-globulin and in antibody production against bovine herpes virus type 1 and Mycobacterium paratuberculosis.

Fatal cerebral embolism in a young patient with an occult left atrial myxoma.

We report a young patient with a fatal cerebral embolism from an occult atrial myxoma. The patient died before echocardiography was performed and at autopsy the definite diagnosis was made. Our patient suffered from migraine of increasing frequency. The physical exercise of sexual intercourse was the precipitating factor of this fatal embolism. The importance of early echocardiography is stressed, especially in view of the recent tendency of early and aggressive stroke treatment.

Resistance of Fasciola hepatica against triclabendazole in cattle and sheep in The netherlands.

In the winter of 1998/1999, sheep on a farm in the province of North Holland, The Netherlands, died from subacute and chronic liver fluke disease despite four previous treatments with triclabendazole (TCBZ). Faecal examinations of sheep and cattle on the farm showed high number of liver fluke eggs. In a randomised clinical trial, the fluke egg output was monitored weekly for 3 weeks in sheep which were treated with TCBZ or with closantel; in dairy cows treated with TCBZ or with clorsulon; and in heifers treated with TCBZ or clorsulon. The results showed a significant reduction of 99.7, 98.1 and 99.2%, respectively, in fluke egg output at 21 days in all non-TCBZ treated animals. TCBZ treatment produced percentage decreases of 15.3, 4.3 and 36.6%, respectively. These results are highly indicative of the presence of TCBZ-resistant Fasciola hepatica in sheep and cattle on this farm.

An experimental study on triclabendazole resistance of Fasciola hepatica in sheep.

The efficacy of triclabendazole in sheep experimentally infected with Fasciola hepatica was studied. Two groups of 12 lambs were infected with a susceptible (S) or a resistant (R) strain of F. hepatica. Eight weeks after infection, six lambs of each group (ST and RT) were treated with triclabendazole (10mg/kg). The other lambs were used as untreated controls (SC and RC). The parameters studied were: GLDH, gamma-GT, ELISA measuring antibodies against recombinant cathepsin-L(1) and eggs per gram faeces (epg). The lambs were slaughtered 16 weeks after infection and the number of flukes counted. The GLDH, gamma-GT levels and the OD value of the ELISA decreased as a result of the treatment in group ST. Patent infections were observed in all animals of groups SC, RT and RC. In group ST, occasionally a few eggs were found in five lambs. The percentage of flukes was 31.3 in SC and 37.6 in RC. In the treated groups ST and RT, the percentage of flukes was 0.06 and 33.6, respectively. These results corresponded to efficacies of 99.8% in the susceptible and 10.8% in the resistant strain. Since the resistant strain was isolated from a mixed cattle and sheep farm, it confirms the presence of triclabendazole resistance in the Netherlands.

Dual photon absorptiometry. The importance of clinical correlation.

Dual photon absorptiometry is a reliable method of assessing bone mineral density. However, distortions of bony architecture, which may not be evident from the low-resolution images provided, can complicate the evaluation and lead to misleading findings. A case is presented which describes the importance of clinical and radiographic correlation.

Effect of cobalt supplementation on serum vitamin B12 levels, weight gain and survival rate in lambs grazing cobalt-deficient pastures.

The effect of cobalt supplementation on serum vitamin B12, growth rate and survival rate was measured in controlled field experiments with Texel twin lambs of the same sex, grazing cobalt-deficient pastures. The non-supplemented lambs had lower serum vitamin B12 concentrations than their supplemented brothers or sisters. During the experiments more lambs died in the non-supplemented than in the supplemented group. At the end of the experiments supplemented lambs weighed (mean live weight) 7.2, 9.5, and 11.0 kg more than non-supplemented lambs in 1991, 1992, and 1993, respectively. Sex-related differences in weight gain and survival rate were observed.

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Demography of Q fever seroprevalence in sheep and goats in The Netherlands in 2008.

At the end of 2007, the first year of what later turned out to be one of the largest Q fever outbreaks in the world with ultimately almost 3500 human patients notified in three years time, dairy goats were suspected to be the possible cause. However, current information on the Q fever prevalence in small ruminants in The Netherlands was lacking. A serological survey, using an indirect ELISA, was carried out in 15,186 sheep and goats in The Netherlands in 2008. In total, 2.4% (95% CI: 2.2-2.7) of the sheep and 7.8% (95% CI: 6.9-8.8) of the goats was seropositive for antibodies against Coxiella burnetii. In 14.5% (95% CI: 12.5-16.5) of the sheep flocks and 17.9% (95% CI: 14.2-21.5) of the goat herds at least one seropositive animal was found. In sheep flocks with at least one seropositive sheep, the within herd seroprevalence was 14.8% (95% CI: 12.6-17.0). In goat herds with at least one seropositive goat, the within herd seroprevalence was 29.0% (95% CI: 24.6-33.3). The seropositive sheep were equally distributed across the country. The seroprevalence in goats in the south-eastern part of The Netherlands, the area where most of the human Q fever cases were notified, was significantly higher than the seroprevalence in goats in the rest of The Netherlands. Dairy sheep and dairy goats had a significantly higher chance of being seropositive than non-dairy sheep and goats. During pregnancy and in the periparturient period, small ruminants tested significantly more often seropositive than in the early- or non-pregnant period. The seroprevalence as well as the true prevalence among small ruminants in The Netherlands were lower than prevalences reported elsewhere. The seroprevalence among sheep was also lower than reported in an earlier Dutch study in 1987. The Q fever seroprevalence was highest in pregnant and periparturient dairy goats in the south-eastern part of The Netherlands, which coincides with the region with the highest human incidence of Q fever.