Hansen, M. S.

Holm, E.

Jensen, T. K.

Larsen, G.

Wolf-Jäckel, Godelind

Infections are the most common cause of bovine abortion. Here we report recent diagnostic findings in bovine abortion material from Denmark, a country with a large dairy sector and high animal health standards. This study was conducted in order to gain in-depth knowledge on infectious causes of abortions i.e. to identify and localize infectious agents in placental and foetal tissues. The cultivation-independent methods Fluorescence in situ hybridization (FISH) and second generation sequencing were applied additionally to routine histopathology and bacterial cultivatio

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Downloaded from orbit.dtu.dk on: Mar 30, 2019Bovine Abortions and Stillbirths in Denmark 2015 to 2017Wolf-Jäckel, Godelind; Hansen, Mette Sif; Larsen, Gitte; Holm, Elisabeth; Jensen, Tim KårePublication date:2018Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Wolf-Jäckel, G., Hansen, M. S., Larsen, G., Holm, E., & Jensen, T. K. (2018). Bovine Abortions and Stillbirths inDenmark 2015 to 2017. Poster session presented at Annual Meeting of the European Society of VeterinaryPathology and the European College of Veterinary Pathology, Cluj-Napoca, Romania.Bovine Abortions and Stillbirths in Denmark 2015 to 2017Godelind A. Wolf-Jäckel, DVM, PhD-student; M.S. Hansen, G. Larsen, E. Holm, T.K. JensenBuilding 202, KemitorvetDK-2800 Kgs. Lyngby gowol@vet.dtu.dkFigure 4. Placenta. Multiple cotelydonaryvilli invaded by fungal septate and branching hyphae. Fungal structures have been detected within placental lesions in 3% of the investigated placentas usingFISH.Screening for bacteria with general probeFigure 3. Placenta. Example of a cotelydonary villus with extendend and necrotic trophoblasts containing large numbers of intracytoplasmatic bacteria(green). Bacterial localisation and morphology will be used to chooserelevant species-specific probes for furtheridentification of intralesional bacteria.Screening for fungi with general probeFISHBRUCELLA ABORTUS CULTIVATIONAll foetal organ pools were negative for Brucella abortus.HISTOPATHOLOGICAL SCREENING FOR NEOSPORA CANINUMNeosporosis was diagnosed in 30 out of 162 abortions (19%) based on findings in HE stained tissue sections of brain, heart, and liver.ELISAIn 90% of the cases, a blood sample of the dam was submitted. All samples were negative for maternal BVDV antibodies.Infections are the most common cause of bovine abortion. Here we report recent diagnostic findings in bovine abortion material from Denmark, a country witha large dairy sector and high animal health standards. This study was conducted in order to gain in-depth knowledge on infectious causes of abortions i.e. toidentify and localize infectious agents in placental and foetal tissues. The cultivation-independent methods Fluorescence in situ hybridization (FISH) andsecond generation sequencing were applied additionally to routine histopathology and bacterial cultivation.0.8% of reported abortions during study periodDanish Holstein 62%Danish Jersey 13% Crossbreed 9% Danish Red 7% Figure 1. The study population consisted predominantly of dairy cows (5% beef) from mainly conventionalfarms (9% organic) and originated from across the country matching the geographical distribution of dairyfarms in Denmark (map displaying dairy farms as grey dots and abortion submitting farms as red dots). Themajority of the abortions took place during mid to late gestation.STUDY POPULATION SAMPLE MATERIALLungLiverKidneyBrain HeartPlacentaplacental interfaceoral routesystemic routeRESULTSFigure 2. Organs collected at necropsy for histological and molecular analyses. Organs were chosen based onknown predeliction sites for abortogenic infections (e.g. neosporosis in brain, liver, heart; Leptospira interrogansin kidneys) and related to potential ports of entry into the foetus. Abortion material was submitted to DTU Vet forroutine abortion diagnostics from January 2015 until June 2017.Neosporosis was the most frequently diagnosed infection. No epizootic abortifacients were found on study population level, however, due to very few abortions submitted per herd, no conclusions can be drawn on herd level.Fungi seem to play a minor role as arbortogenic agent in Denmark.CONCLUSIONSSECOND GENERATION SEQUENCINGAmpliconhttps://i.stack.imgur.com/J80iz.jpgFigure 5. DNA was extracted from placenta, a liver and lung pool as well as kidney. Part of the total bacterial DNA per sample was amplified using universal bacterial primers binding adjacent to the variable regions V1 and V2 of the 16S rRNA gene. The resulting amplicon was sequenced on the llumina MiSeq platform. Species n Genus/Species n Genus/Species n Genus/Species nEscherichia coli 37 Aeromonas spp. 7 Listeria monocytogenes 3 Moellerella wisconsensis 1Acinetobacter spp. 15 Staphylococcus spp. 6 Vibrio spp. 3 Morganella morganii 1Aerococcus spp. 13 Trueperella pyogenes 5 Vagococcus spp. 3 Pantoea agglomerans 1Proteus 13 Streptococcus spp. 5 Lactobacillus spp. 2 Pasteurella spp. 1Lactococcus spp. 9 Bacillus licheniformis 5 Klebsiella spp. 2 Serratia fonticola 1Enterococcus spp. 8 Hafnia alvei 5 Citrobacter spp. 1 no bacteria isolated 30Table 1. Bacterial genuses/species isolated using routine aerobe cultivation.n = number of abortions from which genus/species was isolated7747 40 37 3653113 119 122 1250%10%20%30%40%50%60%70%80%90%100%placentitis pneumonia myocarditis encephalitis hepatitisnegativepositiveSECOND GENERATION SEQUENCINGAll abortion samples were negative for DNA from the following known abortifacients:Chlamydia/Parachlamydia spp.Brucella abortusCampylobacter fetusPajaroellobacter abortibovis (epizootic bovine abortion)Listeria ivanoviiLeptospira interrogans DNA was detected in one liver sample.Coxiella burnetii DNA was detected in samples from four abortions.Figure 6. Distribution of organ lesions diagnosed in HE tissue sections shown as number and percentage of abortions. 

Bovine Abortions and Stillbirths in Denmark 2015 to 2017

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Bovine Abortions and Stillbirths in Denmark 2015 to 2017

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