The  results  in  this  work  evaluated  the sensitivity and specificity of the olfactory capacity of the intervening dog in the  fungus  biodetection.  On  100  attempts,  a  sensitivity  and  a  specificity of  100%  (Table  1)  were  obtained  for the biodetection of the fungus Sclerotium rolfsii when the 4-year-old English Springer Spaniel female dog was faced to a straight line of 5 equally distanced Falcons.
The  post-inoculation  time  of  the samples,  used  in  the  sessions,  varied between  24  and  72H,  however  there was no difference in the detection response by the female dog Julieta. This is  also  a  noteworthy  additional  fact, given  the  potential  for  prevention  that can represent the detection of a fungus with such a short post-inoculation time. Further work is being established to assess the olfactory ability to detect the presence  of  the  fungus  in  inoculated turfgrass samples and finally to detect it in the field.info:eu-repo/semantics/publishedVersio

Coelho, Luísa

Dionísio, Lídia

Duarte, Amilcar

Guerrero, Carlos

Serrão, M.

English

Sapientia

7th ETS CONFERENCE 2020TURF SOLUTIONS for the  FUTURESelected papers (Part 2/3) for the 7th ETS Conference 2020, cancelled due to Covid-19Ausgewählte Fachbeiträge für die  aufgrund der Corona-Pandemie abgesagten  7. ETS-Konferenz in Amsterdam.In Abstimmung mit dem ETS-Board und unter fachlicher Leitung der Deutschen Rasengesellschaft e.V. veröffentlicht die  Köllen Druck + Verlag GmbH ausgewählte und „peer-reviewed 2-page-paper“ der ETS-Tagung.In drei Ausgaben der Zeitschrift „RASEN – European Journal of Turfgrass Science“ erscheinen Fachbeiträge  zu folgenden Schwerpunkt-Themen:)) Ausgabe 02/20: „Drought, Irrigation and Water consumption“)) Ausgabe 03/20: „Disease and Pest Management + Biostimulants“)) Ausgabe 04/20: „Maintenance and Nutrition + Impact for the Environment“Inhalte Ausgabe 03/20AutorenThema: „Disease and Pest Management + Biostimulants“Paper Nr. / SeiteStephens, C.M., J.P. Kerns and T.W. GannonTurfgrass management practices influence fungicide fate on golf course  putting greens  1 / 68Miller, G.L. and M.A. BrothertonCreeping bentgrass summer decline as influenced by  climatic conditions and cultural practices  2 / 70Coelho, L., L. Dionísio, M. Reis and  C. Guerrero Use of cork residues to control turfgrass diseases   3 / 72Espevig, T., K. Normann, M. Usoltseva,  K. Entwistle, J.A. Crouch and T.S. AamlidIn vitro screening of turfgrass species and cultivars  for resistance to Dollar spot isolates of different origin   4 / 74Rosenbusch, J., A. Floss and W. Praemassing Antagonist and soil additive to control Microdochium nivale disease   5 / 76Koch, P., D. Smith, C. Mattox, B. McDonald,  E. Braithwaite, A. Kowalewski, M. Sheridan  and E. NangleDevelopment of a logistic regression model for the prediction of  Microdochium patch  6 / 78De Luca, V. and D. Gómez de Barreda Effect of a Biostimulant on late season Bermudagrass  implantation   7 / 80Owen, A.G., T.I. Williams, and D. HiltzSeaweed (Ascophyllum nodosum) extraction method produces chemically  different formulations with contrasting effects on turfgrass rooting  8 / 82Williams, T.I., A.C. Gange and A.G. OwenAscophyllum nodosum extract use on plant parasitic nematode abundance and diversity on a golf green  9 / 84Serrão, M., L. Coelho, L. Dionísio, C. Guerrero and A. DuarteBiodetection of turfgrass fungal diseases using sniffer dogs 10 / 8686 RASEN · TURF · GAZON  3/2020Biodetection of turfgrass fungal diseases using sniffer dogs Serrão, M., L. Coelho, L. Dionísio, C. Guerrero and A. DuarteIntroductionThe Directive 2009/128/EC1 of the Eu-ropean Parliament and of the Council, of 21 October, establishes a framework, for Community action, to achieve the sustainable use of pesticides by redu-cing the risks and impacts of its use on human health and in the environment. This Directive also promotes the use of integrated pest management, as well approaches or techniques such as non-chemical alternatives to pesti-cides that enhanced the need to find new techniques for detection, identifi-cation and quantification of pathogens in plants or in soil in fields used for agri-culture.Fungi turfgrass pathogens cause eco-nomically important destructive di-seases and the symptoms of infection are mainly recognized after the pa-thogen has invaded plant tissues. Pre-cocious detection of infection, precise identification, differentiation, and quan-tification of the pathogens in plants and/or in soil are essential for the de-velopment of strategies to reduce the incidence and spread of the diseases. On golf courses, maintaining a healthy turfgrass and identifying in vivo the presence of turfgrass diseases has become increasingly important in tur-fgrass management. An on-sight iden-tification of the presence of the disease is normally done when symptoms are present. However, it is important to establish adequate management prac-tices, or techniques, that may gather information of turfgrass health before disease symptoms appear. For fungal diseases, symptoms appear only a few days or even weeks after the fungus in-fection. On the other hand, fungi emit volatile substances that, although not noticeable by human smell, can be de-tected by dogs.Since the use of hunting dogs, 12 000 years ago2, several different applica-tions for sniffer dogs have emerged, including the most classics and well-known such as explosives, narcotics, people searching and tracking, and the search for corpses2,3. However, the continued study of the dog’s olfactory ability has determined that its olfac-tory detection threshold for a given volatile organic compound can be as low as 40 parts per billion to 1.5 parts per trillion4. This evidence reinforced the introduction of olfactory detection (biodetection), in different areas of science.  In human health, biodetection can be used for early detection of va-rious types of cancer5,6 and the specific localization of bacteria responsible for hospital-acquired infections7. It can be used also in the area of plant protec-tion8, to which this work refers.A biodetection of fungal disease has been carried out at the University of Algarve where it has been developed a project for the biodetection of the phytopathogenic fungus Sclerotium rolfsii, using a 4-year-old English Sprin-ger Spaniel female dog “Julieta”. This work is divided into 3 different phases. The first phase, already completed and to which this article refers, had two main objectives: (i) determination of dog’s ol-factory ability to detect and recognize the fungus odor in inoculated samples; (ii) analyze the ability to differentiate it when compared to control samples. The next two phases will assess the olfactory ability to detect the presence of the fungus in inoculated turfgrass samples and, finally, to detect it in field conditions. Preliminary results validate the potential use of canine biodetection in the early identification of turfgrass pathogens, achieving 100% sensitivity and 100% specificity in the identifica-tion of the fungus  Sclerotium rolfsii.Materials and MethodsThe phytopathogenic fungus was inoculated into 50 mL Falcon tubes containing 25 mL of Potato dextrose agar (PDA). For this purpose, PDA was autoclaved and solidified in an inclined position. In each Falcon, a 6 mm PDA disc containing Sclerotium rolfsii myce-lium was inoculated.The whole dog training protocol, from basic obedience to biodetection, uses only clicker positive reinforcement techniques. The first training stage was the direct association of the fungus odor. This 3-week stage, with a total 1  DIRECTIVE 2009/128/EC: Official Journal of the European Union, L 309, pp. 71-84.2  FURTON, K.G. and J.M. LAWRENCE, 2001: The scientific foundation and efficacy of the use of canines as chemical detectors for explosives.  Talanta, 54(3), 487–500.3  LAZAROWSKI, L. and D.C. DORMAN, 2014: Explosives detection by military working dogs: Olfactory generalization from components to mixtures.  Applied Animal Behaviour Science, 151, 84–93.4  CONCHA, A.R., C.M. GUEST, R. HARRIS, T.W. PIKE, A. FEUGIER, H. ZULCH and D.S. Mills, 2019: Canine Olfactory Thresholds to Amyl Acetate in a Biomedical Detection Scenario. Frontiers in Veterinary Science, 5.5  GUIRAO MONTES, Á., L. MOLINS, L. LÓPEZ-RODÓ, I. RAMÓN RODRÍGUEZ, G. SUNYER DEQUIGIOVANNI, N. VIÑOLAS SEGARRA, R.M. MARRADES  SICART, … and Á. AGUSTÍ GARCÍA-NAVARRO, 2017: Lung cancer diagnosis by trained dogs†. European Journal of Cardio-Thoracic Surgery, 52(6), 1206–1210.6  TAVERNA, G., L. TIDU, F. GRIZZI, V. TORRI, A. MANDRESSI, P. SARDELLA, … and P. GRAZIOTTI, 2015: Investigative Urology Olfactory System of Highly  Trained Dogs Detects Prostate Cancer in Urine Samples. Journal of Urology, 193, 1382–1387.7  CHARLES, M.K., Y. WANG, T. ZURBERG, J. KINNA and E. BRYCE, 2019: Detecting Clostridioides (Clostridium) difficile using canine teams:  What does the nose know? Infection Prevention in Practice, 1(1), 100005.8  ANGLADA, L.P., M.D. ÀNGELS and C. TORRAS, 2016: Detection of Verticillium dahliae in Olive Groves Using Canine Detection Units.  Agricultural Sciences, 7(7), 225–229.of 1000 repetitions, consisted of the establishment of a positive direct odor conditioning, since each repetition in which the dog “smelled” the inoculated sample, she was rewarded with its fa-vorite food. At the same time, the fo-cus position was also trained (the dog remains standing with its muzzle close to the inoculated sample). The second and third training stages consisted of introducing odor discrimination, where the dog gradually learned to discri-minate between inoculated samples and empty Falcons up to discrimina-tion between inoculated samples and control samples (1 inoculated sample and 4 control samples). At the end of the training stages, 10 sessions with 10 repetitions each were performed where the dog was faced to a straight line of 5 equally distanced Falcons (Figure 1), to ignore the control samples and fo-cus the inoculated one. In each ses-sion new inoculated sample and new control samples were used. All ses-sions were recorded.For this work it was registered the nu-mber of (1) True positives: the dog cor-rectly focuses the inoculated sample; (2) True negatives: the dog does not focus on the control samples; (3) False positives: the dog focuses on a control sample; (4) False negatives: the dog does not focus on the inoculated sample. Ten sessions of 10 repetitions each, were performed, to obtain the ex-pected results. The position of the ino-culated sample in each repetition was randomly determined with the roll of a die. During this random process, the dog and her guide waited in a different room, thus ensuring that the biodetec-tion performance wasn’t influenced by her guide.Results and DiscussionThe results in this work evaluated the sensitivity and specificity of the olfac-tory capacity of the intervening dog in the fungus biodetection. On 100 at-tempts, a sensitivity and a specificity of 100% (Table 1) were obtained for the biodetection of the fungus Sclero-tium rolfsii when the 4-year-old English Springer Spaniel female dog was faced to a straight line of 5 equally distanced Falcons (Table 1).The post-inoculation time of the samples, used in the sessions, varied between 24 and 72H, however there was no difference in the detection res-ponse by the female dog Julieta. This is also a noteworthy additional fact, given the potential for prevention that can represent the detection of a fungus with such a short post-inoculation time. Further work is being established to as-sess the olfactory ability to detect the presence of the fungus in inoculated turfgrass samples and finally to detect it in the field.Authors:M. Serrão,Associação Kokua –  Cães de Ajuda Social  (Social Aid Dogs),  Tavira, Portugal,L. Coelho, L. Dionísio,  C. Guerrero* and A. Duarte,Universidade do Algarve,  Faculdade de Ciências e Tecnologia,MedtiBio, Centre for Mediterranean Bioresources and Food,  Faro, Portugal.* E-Mail: cguerre@ualg.pt Selected papers for the 7th ETS Conference 2020, cancelled due to Covid-19 87Fig. 1: “Julieta” in one of the 100 attempts to face a straight line of 5 equally dis-tanced Falcons. The guide is signaling a positive response of biodetectionNumber of sessionsNumber of repeti-tionsTrue positives True negatives False positives False negatives10 10 100 400 0 0Tab. 1: Results of the olfactory capacity of the English Springer Spaniel female dog. Total attempts performed: 100.

Biodetection of turfgrass fungal diseases using sniffer dogs

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Biodetection of turfgrass fungal diseases using sniffer dogs

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