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S1 - P - 20 OVINE ABORTION CAUSED BY YERSINIA PSEUDOTUBERCULOSIS – A CASE REPORT Varpu Hirvelä-Koski, Minna Nylund Finnish Food Safety Authority, Research Department, Oulu, Finland Ovine abortion, pseudotuberculosis Introduction Yersinia pseudotuberculosis is a gram-negative organism that causes various syndromes including mesenteric lymphadenitis and septicaemia in wild and domestic animals as well as in humans. Wild birds and rodents are considered to be reservoirs of the bacterium. Sporadic abortions due to placentitis have been reported in sheep, goats and cows (1,2). We describe here a case of ovine abortion caused by Yersinia pseudotuberculosis. Material & methods The samples were obtained from an organic farm keeping about 300 ewes in central Finland. The ewes were traditional Finnish homebred sheep. Two of the ewes aborted before the lambing season with an interval of two weeks, approximately one month before the estimated lambing. Samples from the second abortion were sent to the laboratory, unfortunately frozen. Two fetuses and the afterbirth were obtained to the laboratory. The samples were examined using gross pathology and histopathology (HE stain). Lung, liver and contents of abomasum as well as the afterbirth were cultivated on the following media: trypticase soy agar incorporated with 5 % bovine blood (aerobic incubation), fastidious anaerobe agar (FAA) (microaerophilic), FAA (anaerobic), selective FAA for the detection of Campylobacter fetus (microaerophilic), Sabouraud agar (aerobic), salmonella enrichment media (pre-enrichment broth, modified semi-solid Rappaport Vassiliadis medium). STAMP stain was used for the detection of brucellosis using material from lung, liver and afterbirth. All plates were incubated at 37⁰+ 1⁰C. The identification of Yersinia pseudotuberculosis was based on colony morphology, gramstain, oxidase reaction, production of acid from glucose, typical growth on MacConkey agar and API 20 E profile. Wild rodents are considered to be one reservoir of Yersinia pseudotuberculosis. The population densities of wild voles has been high in Finland in recent years. It is possible that this phenomenon has increased the infection pressure caused by Yersinia pseudotuberculosis. References 1. Karbe, E, Erickson, ED (1984). Ovine abortion and stillbirth due to purulent placentitis caused by Yersinia pseudotuberculosis. Vet. Pathol. 21, 601-606 2. Juste, RA, Minguijón, E, Arranz, J, Fuertes, M, Beltrán de Heredia, I (2009). Lamb mortality in an outbreak of Yersinia pseudotuberculosis mastitis, as a collateral effect of colostrum feeding for Lentivirus-control. Small Ruminant Res. 86, 46-53. 3. Hallanvuo, S (2009). Foodborne Yersinia. Identification and molecular epidemiology of isolates from human infections. Acad. Diss., University of Helsinki, Finland. National Institute for Health and Welfare, Research series no 11, Helsinki University Print. Results The fetuses, a male and a female weighed 347,5 and 600,8 g, the fetal length being 21 and 26 cm, respectively. In gross pathology no specific lesions could be found. Both fetuses and the afterbirth had quite intense autolytic changes. Yersinia pseudotuberculosis was isolated as a pure growth from the afterbirth. Growth was observed on all non-selective media at all atmospheres. The bacterium was detected also from the inner organs of both fetuses. The growth was most abundant in cultivations from abomasum. Other pathogenic organisms were not detected. The bacterium was a gram-negative small rod, oxidase negative, acid but not gas was produced from glucose. There were small brown colonies on MacConkey. API 20 E profile suggested identification to Yersinia pseudotuberculosis. Histopathological examination revealed suppurative placentitis and non-suppurative conjunctivitis. In the villi of the cotyledons there were multiple necrotic foci with neutrophilic and mononuclear infiltrates. There were masses of coccobacilli and infiltrations of mononuclear and plasma cells with a few granulocytes in the lamina propria of the conjunctiva of both fetuses. In other internal organs the autolytic changes were quite intense and any specific lesions could not be found. Discussion & conclusions Yersinia pseudotuberculosis is often observed as the cause of septicemia in wild animals in Finland, e,g, hare. The bacterium has also caused several large outbreaks of food-borne illnesses in humans in Finland in the last decade. Yersinia pseudotuberculosis has been reported as a rare causative agent of sporadic abortion of sheep and goat. However, as far as we know, this is the first time that it was isolated from ovine abortion samples in Finland.
S1- P - 21 ANALYSIS OF THE EFFICIENCY OF THE MCMASTER METHOD IN RAYNAUD’S MODIFICATION IN DETECTION OF TOXOCARA SP. AND TRICHURIS SP. EGGS IN CARNIVORES FAECES Maciej Kochanowski, Joanna Dąbrowska, Jacek Karamon, Tomasz Cencek National Veterinary Research Institute, Departament of Parasitology and Invasive Disease, Pulawy, Poland McMaster method, efficiency, carnivores, Toxocara, Trichuris Introduction Carnivores animals are the hosts of the important zoonotic intestinal parasites. Faeces of these animals, containing eggs of dangerous parasites are a potential source of infection for humans. The particular importance in the spread of dangerous invasive parasitic forms may have a common use of this faeces as natural organic fertilizers. The aim of study was to determine the efficiency of McMaster method in Raynaud’s modification (Raynaund, 1970) in the detection of parasite eggs of the genera Toxocara and Trichuris present in the faeces of carnivores, using faeces samples enriched with known numbers of eggs. Materials & methods Four variants of this McMaster method (counting in one grid, two grids, the whole McMaster chamber and flotation slide) were used to examine dog faeces samples enriched with Toxocara sp., Trichuris sp. and Ascaris suum eggs. A. suum eggs were the control of influence of the parasite eggs properties on the results. One hundred and sixty enriched samples were prepared from dog faeces (20 repetitions for each eggs level) containing 15, 25, 50, 100, 150, 200, 250 and 300 eggs of the three types of eggs in 1 g of faeces. Moreover, to compare the influence of kind of faeces on the efficiency of the method 160 pig faecal samples were prepared and enriched with A. suum eggs.in the same way Results The highest (worst) limit of detection (Table 1) in all McMaster variants were obtained with eggs of Toxocara sp. (from 25 to 250 eggs / g faeces, depending on the variant) - about twice higher than for Trichuris sp. and A. suum eggs (from 15 to 100 eggs / g feces). The highest (worst) limit of detection for flotation in the tube were obtained for Trichuris sp. eggs (100 eggs / g) - it was 4-times higher than the results for other types of eggs (25 eggs / g). The best results of the limit of detection, sensitivity, the lowest coefficients of variation (providing about repeatability) were obtained with the use of whole McMaster chamber variant. There was no significant impact of faeces properties (dog and pig faeces) on the obtained results. Multiplication factors (to estimate the real number of eggs in 1 g of faeces) were calculated on the basis of the transformed equation of the trend line illustrating the relationship between number of eggs found by the examination and the number of eggs added to the sample. Multiplication factors for eggs of Toxocara sp. and Trichuris sp. were higher than planned by Raynaud, and for A. suum were comparable with them. The multiplication factor estimated for flotation in the tube (flotation slide variant) for all types of examined parasite eggs were significantly higher than that given by Raynaud (Table 2). Table 1: Limit of detection for all variants of the McMaster method in Raynaund’s modification Variants Dog faeces Pig faeces of the method Toxocara Trichuris Ascaris Ascaris one grid 250 100 100 50 two grids 100 50 25 25 whole chamber 25 15 15 15 flotation slide 25 100 25 25 Discussion & conclusions Most of the publications about detection of parasite eggs are based on material from natural or experimental infection. For that reason the real number of eggs in faeces can not be properly assessed (Vadlejch et al, 2011; Mes et al, 2007). Therefore, in order to obtain the most reliable results in our experiment the material enriched with a known number of parasite eggs was used. The results of experiments indicate that detection of parasite eggs of carnivores by McMaster method in Raynaud’s modification differs from the basic parameters of this modification. There were significant differences in the efficiency of the method according to the type of parasite eggs. In the case of Toxocara sp. eggs examined in the McMaster chamber there were observed significantly lower numbers of detected eggs in 1 g of faeces than obtained for Trichuris sp. and A. suum eggs. Most likely reason of the lowest detection Toxocara sp. eggs in the McMaster chamber is the specific shell structure of these eggs – namely, they have strong adhesive properties (Overgaauw, Knapen, 2008). It can result in higher losses of parasite eggs during performing of the method. The variant in the tube (flotaion slide) was characterized by the lowest number of detected eggs in the case of Trichuris sp. It is probably related to the high specific gravity of the Trichuris sp. eggs affects to less effective flotation process in tube. According to our study the detection of Toxocara sp. eggs in McMaster chamber is twice lower than planned by Raynaud. In the case of Trichuris sp. eggs it is slightly lower. However, A. suum eggs in pig and dog feces, were detected in number close to the assumptions of McMaster method in Raynaud’s modification. Detection of all three types parasite eggs as a result of the tube flotation (flotation slide variant) is several times lower than the assumptions of Raynaud's modification (which is at the level of detection obtained in the variant of two grids of McMaster chamber). Our results allowed us to assess the real efficiency of the McMaster method in Raynaud’s modification in carnivore feces samples enriched with a known number of parasite eggs. The investigation carried out with selected (important to the diagnostic and the zoonotic point of view) two types of nematodes (Toxocara sp. and Trichuris sp.) showed a significant effect of parasite genus on the detection efficiency. However, it is desirable to extend calibration of the method for other kinds of parasites whose eggs differ significantly in the morphology - in particular with regard to the eggs of tapeworms. References 1. Mes T., Eysker M., Ploeger H., 2007. A simple, robust and semiautomated parasite egg isolation protocol. Nature Protocols. 2. Overgaauw P., Knapen F., 2008. Toxocarosis, an important zoonosis. European Journal of Companion Animal Practice Vol. 18 Issue 3 (2008). 3. Raynaud J., 1970. Etude de l’efficacite d’une technique de coproscopie quantitative pour le diagnostic de routine et le controle des infestations parasitaires des bovins, ovins, equines et porcins. Annales de Parasitologie (Paris). 45 pp. 321-342 4. Vadlejch J., Petrtyl M., Zaichenko I., Cadkova Z., Jankovska I., Langrova I., Moravec M., 2011. Which McMaster egg counting technique is the most reliable? Parasitology Research 109(5): 1387-1394 (2011) Table 2: Multiplication factors for estimation of the number of eggs in 1g of faeces, for all variants of the McMaster method in Raynaund’s modification Variants Dog faeces Pig faeces of the method Toxocara Trichuris Ascaris Ascaris one grid 244 145 108 111 two grids 120 78 54 63 whole chamber 40 26 22 22 flotation slide 49 62 35 43
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2 nd CONGRESSOFTHEEUROPEAN ASSOCIAT
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Welcome Dear colleagues, Welcome to
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Day4 Wednesday,4July2012 4 th sessi
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antigens or attenuated vaccines can
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conditions, e.g. Staphylococcus aur
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S1 - O - 1 ORAL FLUIDS - SAMPLE MAT
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List of Abstracts (in order of numb
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S1-P-03 BALLAGI ANDREA S1-P-04 BENI
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S3-P-06 KELLER SELINA S3-P-07 KÖNI
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