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14. ESICM LIVES 2016: part three: Milan, Italy. 1–5 October 2016

Author

Velasquez, T., Mackey, G., Lusk, J., Kyle, U. G., Fontenot, T., Marshall, P., Shekerdemian, L. S., Coss-Bu, J. A., Nishigaki, A., Yatabe, T., Tamura, T., Yamashita, K., Yokoyama, M., Ruiz-Rodriguez, J. C., Encina, B., Belmonte, R., Troncoso, I., Tormos, P., Riveiro, M., Baena, J., Sanchez, A., Bañeras, J., Cordón, J., Duran, N., Ruiz, A., Caballero, J., Nuvials, X., Riera, J., Serra, J., Rutten, A. M. F., van Ieperen, S. N. M., Der Kinderen, E. P. H. M., Van Logten, T., Kovacikova, L., Skrak, P., Zahorec, M., Kyle, U. G., Akcan-Arikan, A., Silva, J. C., Mackey, G., Lusk, J., Goldsworthy, M., Shekerdemian, L. S., Coss-Bu, J. A., Wood, D., Harrison, D., Parslow, R., Davis, P., Pappachan, J., Goodwin, S., Ramnarayan, P., Chernyshuk, S., Yemets, H., Zhovnir, V., Pulitano’, S. M., De Rosa, S., Mancino, A., Villa, G., Tosi, F., Franchi, P., Conti, G., Patel, B., Khine, H., Shah, A., Sung, D., Singer, L., Haghbin, S., Inaloo, S., Serati, Z., Idei, M., Nomura, T., Yamamoto, N., Sakai, Y., Yoshida, T., Matsuda, Y., Yamaguchi, Y., Takaki, S., Yamaguchi, O., Goto, T., Longani, N., Medar, S., Abdel-Aal, I. R., El Adawy, A. S., Mohammed, H. M. E. H., Mohamed, A. N., Parry, S. M., Knight, L. D., Denehy, L., De Morton, N., Baldwin, C. E., Sani, D., Kayambu, G., da Silva, V. Z. M., Phongpagdi, P., Puthucheary, Z. A., Granger, C. L., Rydingsward, J. E., Horkan, C. M., Christopher, K. B., McWilliams, D., Jones, C., Reeves, E., Atkins, G., Snelson, C., Aitken, L. M., Rattray, J., Kenardy, J., Hull, A. M., Ullman, A., Le Brocque, R., Mitchell, M., Davis, C., Macfarlane, B., Azevedo, J. C., Rocha, L. L., De Freitas, F. F. M., Cavalheiro, A. M., Lucinio, N. M., Lobato, M. S., Ebeling, G., Kraegpoeth, A., Laerkner, E., De Brito-Ashurst, I., White, C., Gregory, S., Forni, L. G., Flowers, E., Curtis, A., Wood, C. A., Siu, K., Venkatesan, K., Muhammad, J. B. H., Ng, L., Seet, E., Baptista, N., Escoval, A., Tomas, E., Agrawal, R., Mathew, R., Varma, A., Dima, E., Charitidou, E., Perivolioti, E., Pratikaki, M., Vrettou, C., Giannopoulos, A., Zakynthinos, S., Routsi, C., Atchade, E., Houzé, S., Jean-Baptiste, S., Thabut, G., Genève, C., Tanaka, S., Lortat-Jacob, B., Augustin, P., Desmard, M., Montravers, P., de Molina, F. J. González, Barbadillo, S., Alejandro, R., Álvarez-Lerma, F., Vallés, J., Catalán, R. M., Palencia, E., Jareño, A., Granada, R. M., Ignacio, M. L., Cui, N., Liu, D., Wang, H., Su, L., Qiu, H., Li, R., Jaffal, K., Rouzé, A., Poissy, J., Sendid, B., Nseir, S., Paramythiotou, E., Rizos, M., Frantzeskaki, F., Antoniadou, A., Vourli, S., Zerva, L., Armaganidis, A., Riera, J., Gottlieb, J., Greer, M., Wiesner, O., Martínez, M., Acuña, M., Rello, J., Welte, T., Atchade, E., Mignot, T., Houzé, S., Jean-Baptiste, S., Thabut, G., Lortat-Jacob, B., Tanaka, S., Augustin, P., Desmard, M., Montravers, P., Soussi, S., Dudoignon, E., Ferry, A., Chaussard, M., Benyamina, M., Alanio, A., Touratier, S., Chaouat, M., Lafaurie, M., Mimoun, M., Mebazaa, A., Legrand, M., Sheils, M. A., Patel, C., Mohankumar, L., Akhtar, N., Noriega, S. K. Pacheco, Aldana, N. Navarrete, León, J. L. Ávila, Baquero, J. Durand, Bernal, F. Fernández, Ahmadnia, E., Hadley, J. S., Millar, M., Hall, D., Hewitt, H., Yasuda, H., Sanui, M., Komuro, T., Kawano, S., Andoh, K., Yamamoto, H., Noda, E., Hatakeyama, J., Saitou, N., Okamoto, H., Kobayashi, A., Takei, T., Matsukubo, S., Rotzel, H. B., Lázaro, A. Serrano, Prada, D. Aguillón, Gimillo, M. Rodriguez, Barinas, O. Diaz, Cortes, M. L. Blasco, Franco, J. Ferreres, Roca, J. M. Segura, Carratalá, A., Gonçalves, B., Turon, R., Mendes, A., Miranda, F., Mata, P. J., Cavalcanti, D., Melo, N., Lacerda, P., Kurtz, P., Righy, C., Rosario, L. E. de la Cruz, Lesmes, S. P. Gómez, Romero, J. C. García, Herrera, A. N. García, Pertuz, E. D. Díaz, Sánchez, M. J. Gómez, Sanz, E. Regidor, Hualde, J. Barado, Hernández, A. Ansotegui, Irazabal, J. M. Guergué, Spatenkova, V., Bradac, O., Suchomel, P., Urli, T., Lazzeri, E. Heusch, Aspide, R., Zanello, M., Perez-Borrero, L., Garcia-Alvarez, J. M., Arias-Verdu, M. D., Aguilar-Alonso, E., Rivera-Fernandez, R., Mora-Ordoñez, J., De La Fuente-Martos, C., Castillo-Lorente, E., Guerrero-Lopez, F., Lesmes, S. P. Gómez, Rosario, L. E. De la Cruz, Pertuz, E. D. Díaz, Hernández, A. Ansotegui, Romero, J. C. García, Sánchez, M. J. Gómez, Herrera, A. N. García, Ramírez, J. Roldán, Sanz, E. Regidor, Hualde, J. Barado, León, J. P. Tirapu, Navarro-Guillamón, L., Cordovilla-Guardia, S., Iglesias-Santiago, A., Guerrero-López, F., Fernández-Mondéjar, E., Vidal, A., Perez, M., Juez, A., Arias, N., Colino, L., Perez, J. L., Pérez, H., Calpe, P., Alcala, M. A., Robaglia, D., Perez, C., Lan, S. K., Cunha, M. M., Moreira, T., Santos, F., Lafuente, E., Fernandes, M. J., Silva, J. G., Rosario, L. E. de la Cruz, Lesmes, S. P. Gómez, Herrera, A. N. García, Romero, J. C. García, Pertuz, E. D. Díaz, Sánchez, M. J. Gómez, Sanz, E. Regidor, Echeverría, J. G. Armando, Hernández, A. Ansotegui, Hualde, J. Barado, Podlepich, V., Sokolova, E., Alexandrova, E., Lapteva, K., Kurtz, P., Shuinotsuka, C., Rabello, L., Vianna, G., Reis, A., Cairus, C., Salluh, J., Bozza, F., Torres, J. C. Barrios, Araujo, N. J. Fernández, García-Olivares, P., Keough, E., Dalorzo, M., Tang, L. K., De Sousa, I., Díaz, M., Marcos-Zambrano, L. J., Guerrero, J. E., Gomez, S. E. Zamora, Lopez, G. D. Hernandez, Cuellar, A. I. Vazquez, Nieto, O. R. Perez, Gonzalez, J. A. Castanon, Bhasin, D., Rai, S., Singh, H., Gupta, O., Bhattal, M. K., Sampley, S., Sekhri, K., Nandha, R., Aliaga, F. A., Olivares, F., Appiani, F., Farias, P., Alberto, F., Hernández, A., Pons, S., Sonneville, R., Bouadma, L., Neuville, M., Mariotte, E., Radjou, A., Lebut, J., Chemam, S., Voiriot, G., Dilly, M. P., Mourvillier, B., Dorent, R., Nataf, P., Wolff, M., Timsit, J. F., Ediboglu, O., Ataman, S., Ozkarakas, H., Kirakli, C., Vakalos, A., Avramidis, V., Obukhova, O., Kurmukov, I. A., Kashiya, S., Golovnya, E., Baikova, V. N., Ageeva, T., Haritydi, T., Kulaga, E. V., Rios-Toro, J. J., Perez-Borrero, L., Aguilar-Alonso, E., Arias-Verdu, M. D., Garcia-Alvarez, J. M., Lopez-Caler, C., De La Fuente-Martos, C., Rodriguez-Fernandez, S., Sanchez-Orézzoli, M. Gomez, Martin-Gallardo, F., Nikhilesh, J., Joshi, V., Villarreal, E., Ruiz, J., Gordon, M., Quinza, A., Gimenez, J., Piñol, M., Castellanos, A., Ramirez, P., Jeon, Y. D., Jeong, W. Y., Kim, M. H., Jeong, I. Y., Ahn, M. Y., Ahn, J. Y., Han, S. H., Choi, J. Y., Song, Y. G., Kim, J. M., Ku, N. S., Shah, H., Kellner, F., Rezai, F., Mistry, N., Yodice, P., Ovnanian, V., Fless, K., Handler, E., Alejos, R. Martínez, Romeu, J. D. Martí, Antón, D. González, Quinart, A., Martí, A. Torres, Llaurado-Serra, M., Lobo-Civico, A., Ventura-Rosado, A., Piñol-Tena, A., Pi-Guerrero, M., Paños-Espinosa, C., Peralvo-Bernat, M., Marine-Vidal, J., Gonzalez-Engroba, R., Montesinos-Cerro, N., Treso-Geira, M., Valeiras-Valero, A., Martinez-Reyes, L., Sandiumenge, A., Jimenez-Herrera, M. F., Helyar, S., Riozzi, P., Noon, A., Hallows, G., Cotton, H., Keep, J., Hopkins, P. A., Taggu, A., Renuka, S., Sampath, S., Rood, P. J. T., Frenzel, T., Verhage, R., Bonn, M., Pickkers, P., van der Hoeven, J. G., van den Boogaard, M., Corradi, F., Melnyk, L., Moggia, F., Pienovi, R., Adriano, G., Brusasco, C., Mariotti, L., Lattuada, M., Bloomer, M. J., Coombs, M., Ranse, K., Endacott, R., Maertens, B., Blot, K., Blot, S., Amerongen, M. P. van Nieuw, van der Heiden, E. S., Twisk, J. W. R., Girbes, A. R. J., Spijkstra, J. J., Riozzi, P., Helyar, S., Cotton, H., Hallows, G., Noon, A., Bell, C., Peters, K., Feehan, A., Keep, J., Hopkins, P. A., Churchill, K., Hawkins, K., Brook, R., Paver, N., Endacott, R., Maistry, N., van Wijk, A., Rouw, N., van Galen, T., Evelein-Brugman, S., Taggu, A., Krishna, B., Sampath, S., Putzu, A., Fang, M., Berto, M. Boscolo, Belletti, A., Cassina, T., Cabrini, L., Mistry, M., Alhamdi, Y., Welters, I., Abrams, S. T., Toh, C. H., Han, H. S., Gil, E. M., Lee, D. S., Park, C. M., Winder-Rhodes, S., Lotay, R., Doyle, J., Ke, M. W., Huang, W. C., Chiang, C. H., Hung, W. T., Cheng, C. C., Lin, K. C., Lin, S. C., Chiou, K. R., Wann, S. R., Shu, C. W., Kang, P. L., Mar, G. Y., Liu, C. P., Dubó, S., Aquevedo, A., Jibaja, M., Berrutti, D., Labra, C., Lagos, R., García, M. F., Ramirez, V., Tobar, M., Picoita, F., Peláez, C., Carpio, D., Alegría, L., Hidalgo, C., Godoy, K., Bakker, J., Hernández, G., Sadamoto, Y., Katabami, K., Wada, T., Ono, Y., Maekawa, K., Hayakawa, M., Sawamura, A., Gando, S., Marin-Mateos, H., Perez-Vela, J. L., Garcia-Gigorro, R., Peiretti, M. A. Corres, Lopez-Gude, M. J., Chacon-Alves, S., Renes-Carreño, E., Montejo-González, J. C., Parlevliet, K. L., Touw, H. R. W., Beerepoot, M., Boer, C., Elbers, P. W. G., Tuinman, P. R., Abdelmonem, S. A., Helmy, T. A., El Sayed, I., Ghazal, S., Akhlagh, S. H., Masjedi, M., Hozhabri, K., Kamali, E., Zýková, I., Paldusová, B., Sedlák, P., Morman, D., Youn, A. M., Ohta, Y., Sakuma, M., Bates, D., Morimoto, T., Su, P. L., Chang, W. Y., Lin, W. C., Chen, C. W., Facchin, F., Zarantonello, F., Panciera, G., De Cassai, A., Venrdramin, A., Ballin, A., Tonetti, T., Persona, P., Ori, C., Del Sorbo, L., Rossi, S., Vergani, G., Cressoni, M., Chiumello, D., Chiurazzi, C., Brioni, M., Algieri, I., Tonetti, T., Guanziroli, M., Colombo, A., Tomic, I., Colombo, A., Crimella, F., Carlesso, E., Gasparovic, V., Gattinoni, L., Neto, A. Serpa, Schmidt, M., Pham, T., Combes, A., de Abreu, M. Gama, Pelosi, P., Schultz, M. J., Katira, B. H., Engelberts, D., Giesinger, R. E., Ackerley, C., Yoshida, T., Zabini, D., Otulakowski, G., Post, M., Kuebler, W. M., McNamara, P. J., Kavanagh, B. P., Pirracchio, R., Rigon, M. Resche, Carone, M., Chevret, S., Annane, D., Eladawy, S., El-Hamamsy, M., Bazan, N., Elgendy, M., De Pascale, G., Vallecoccia, M. S., Cutuli, S. L., Di Gravio, V., Pennisi, M. A., Conti, G., Antonelli, M., Andreis, D. T., Khaliq, W., Singer, M., Hartmann, J., Harm, S., Carmona, S. Alcantara, Almudevar, P. Matia, Abellán, A. Naharro, Ramos, J. Veganzones, Pérez, L. Pérez, Valbuena, B. Lobo, Sanz, N. Martínez, Simón, I. Fernández, Arrigo, M., Feliot, E., Deye, N., Cariou, A., Guidet, B., Jaber, S., Leone, M., Resche-Rigon, M., Baron, A. Vieillard, Legrand, M., Gayat, E., Mebazaa, A., Balik, M., Kolnikova, I., Maly, M., Waldauf, P., Tavazzi, G., Kristof, J., Herpain, A., Su, F., Post, E., Taccone, F., Vincent, J. L., Creteur, J., Lee, C., Hatib, F., Jian, Z., Buddi, S., Cannesson, M., Fileković, S., Turel, M., Knafelj, R., Gorjup, V., Stanić, R., Gradišek, P., Cerović, O., Mirković, T., Noč, M., Tirkkonen, J., Hellevuo, H., Olkkola, K. T., Hoppu, S., Lin, K. C., Hung, W. T., Chiang, C. C., Huang, W. C., Juan, W. C., Lin, S. C., Cheng, C. C., Lin, P. H., Fong, K. Y., Hou, D. S., Kang, P. L., Wann, S. R., Chen, Y. S., Mar, G. Y., Liu, C. P., Paul, M., Bougouin, W., Geri, G., Dumas, F., Champigneulle, B., Legriel, S., Charpentier, J., Mira, J. P., Sandroni, C., Cariou, A., Zimmerman, J., Sullivan, E., Noursadeghi, M., Fox, B., Sampson, D., McHugh, L., Yager, T., Cermelli, S., Seldon, T., Bhide, S., Brandon, R. A., Brandon, R. B., Zwaag, J., Beunders, R., Pickkers, P., Kox, M., Gul, F., Arslantas, M. K., Genc, D., Zibandah, N., Topcu, L., Akkoc, T., Cinel, I., Greco, E., Lauretta, M. P., Andreis, D. T., Singer, M., Garcia, I. Palacios, Cordero, M., Martin, A. Diaz, Pallás, T. Aldabó, Montero, J. Garnacho, Rey, J. Revuelto, Malo, L. Roman, Montoya, A. A. Tanaka, Martinez, A. D. C. Amador, Ayala, L. Y. Delgado, Zepeda, E. Monares, Granillo, J. Franco, Sanchez, J. Aguirre, Alejo, G. Camarena, Cabrera, A. Rugerio, Montenegro, A. Pedraza, Pham, T., Beduneau, G., Schortgen, F., Piquilloud, L., Zogheib, E., Jonas, M., Grelon, F., Runge, I., Terzi, N., Grangé, S., Barberet, G., Guitard, P. G., Frat, J. P., Constan, A., Chrétien, J. M., Mancebo, J., Mercat, A., Richard, J. C. M., Brochard, L., Soilemezi, E., Koco, E., Savvidou, S., Nouris, C., Matamis, D., Di Mussi, R., Spadaro, S., Volta, C. A., Mariani, M., Colaprico, A., Antonio, C., Bruno, F., Grasso, S., Rodriguez, A., Martín-Loeches, I., Díaz, E., Masclans, J. R., Gordo, F., Solé-Violán, J., Bodí, M., Avilés-Jurado, F. X., Trefler, S., Magret, M., Reyes, L. F., Marín-Corral, J., Yebenes, J. C., Esteban, A., Anzueto, A., Aliberti, S., Restrepo, M. I., Larsson, J. Skytte, Redfors, B., Ricksten, S. E., Haines, R., Powell-Tuck, J., Leonard, H., Ostermann, M., Berthelsen, R. E., Itenov, T. S., Perner, A., Jensen, J. U., Ibsen, M., Jensen, A. E. K., Bestle, M. H., Bucknall, T., Dixon, J., Boa, F., MacPhee, I., Philips, B. J., Doyle, J., Saadat, F., Samuels, T., Huddart, S., McCormick, B., DeBrunnar, R., Preece, J., Swart, M., Peden, C., Richardson, S., Forni, L., Kalfon, P., Baumstarck, K., Estagnasie, P., Geantot, M. A., Berric, A., Simon, G., Floccard, B., Signouret, T., Boucekine, M., Fromentin, M., Nyunga, M., Sossou, A., Venot, M., Robert, R., Follin, A., Renault, A., Garrouste, M., Collange, O., Levrat, Q., Villard, I., Thévenin, D., Pottecher, J., Patrigeon, R. G., Revel, N., Vigne, C., Mimoz, O., Auquier, P., Pawar, S., Jacques, T., Deshpande, K., Pusapati, R., Wood, B., Pulham, R. A., Wray, J., Brown, K., Pierce, C., Nadel, S., Ramnarayan, P., Azevedo, J. R., Montenegro, W. S., Rodrigues, D. P., Sousa, S. C., Araujo, V. F., Leitao, A. L., Prazeres, P. H., Mendonca, A. V., Paula, M. P., Das Neves, A., Loudet, C. I., Busico, M., Vazquez, D., Villalba, D., Lischinsky, A., Veronesi, M., Emmerich, M., Descotte, E., Juliarena, A., Bisso, M. Carboni, Grando, M., Tapia, A., Camargo, M., Ulla, D. Villani, Corzo, L., dos Santos, H. Placido, Ramos, A., Doglia, J. A., Estenssoro, E., Carbonara, M., Magnoni, S., Donald, C. L. Mac, Shimony, J. S., Conte, V., Triulzi, F., Stretti, F., Macrì, M., Snyder, A. Z., Stocchetti, N., Brody, D. L., Podlepich, V., Shimanskiy, V., Savin, I., Lapteva, K., Chumaev, A., Tjepkema-Cloostermans, M. C., Hofmeijer, J., Beishuizen, A., Hom, H., Blans, M. J., van Putten, M. J. A. M., Longhi, L., Frigeni, B., Curinga, M., Mingone, D., Beretta, S., Patruno, A., Gandini, L., Vargiolu, A., Ferri, F., Ceriani, R., Rottoli, M. R., Lorini, L., Citerio, G., Pifferi, S., Battistini, M., Cordolcini, V., Agarossi, A., Di Rosso, R., Ortolano, F., Stocchetti, N., Lourido, C. Mora, Cabrera, J. L. Santana, Santana, J. D. Martín, Alzola, L. Melián, del Rosario, C. García, Pérez, H. Rodríguez, Torrent, R. Lorenzo, Eslami, S., Dalhuisen, A., Fiks, T., Schultz, M. J., Hanna, A. Abu, Spronk, P. E., Wood, M., Maslove, D., Muscedere, J., Scott, S. H., Saha, T., Hamilton, A., Petsikas, D., Payne, D., Boyd, J. G., Puthucheary, Z. A., McNelly, A. S., Rawal, J., Connolly, B., McPhail, M. J., Sidhu, P., Rowlerson, A., Moxham, J., Harridge, S. D., Hart, N., Montgomery, H. E., Jovaisa, T., Thomas, B., Gupta, D., Wijayatilake, D. S., Shum, H. P., King, H. S., Chan, K. C., Tang, K. B., Yan, W. W., Arias, C. Castro, Latorre, J., De La Rica, A. Suárez, Garrido, E. Maseda, Feijoo, A. Montero, Gancedo, C. Hernández, Tofiño, A. López, Rodríguez, F. Gilsanz, Gemmell, L. K., Campbell, R., Doherty, P., MacKay, A., Singh, N., Vitaller, S., Nagib, H., Prieto, J., Del Arco, A., Zayas, B., Gomez, C., Tirumala, S., Pasha, S. A., Kumari, B. K., Martinez-Lopez, P., Puerto-Morlán, A., Nuevo-Ortega, P., Pujol, L. Martinez, Dolset, R. Algarte, González, B. Sánchez, Riera, S. Quintana, Álvarez, J. Trenado, Quintana, S., Martínez, L., Algarte, R., Sánchez, B., Trenado, J., Tomas, E., Brock, N., Viegas, E., Filipe, E., Cottle, D., Traynor, T., Martínez, M. V. Trasmonte, Márquez, M. Pérez, Gómez, L. Colino, Martínez, N. Arias, Muñoz, J. M. Milicua, Bellver, B. Quesada, Varea, M. Muñoz, Llorente, M. Á. Alcalá, Calvo, C. Pérez, Hillier, S. D., Faulds, M. C., Hendra, H., Lawrence, N., Maekawa, K., Hayakawa, M., Ono, Y., Kodate, A., Sadamoto, Y., Tominaga, N., Mizugaki, A., Murakami, H., Yoshida, T., Katabami, K., Wada, T., Sawamura, A., Gando, S., Silva, S., Kerhuel, L., Malagurski, B., Citerio, G., Chabanne, R., Laureys, S., Puybasset, L., Nobile, L., Pognuz, E. R., Rossetti, A. O., Verginella, F., Gaspard, N., Creteur, J., Ben-Hamouda, N., Oddo, M., Taccone, F. S., Ono, Y., Hayakawa, M., Iijima, H., Maekawa, K., Kodate, A., Sadamoto, Y., Mizugaki, A., Murakami, H., Katabami, K., Wada, T., Sawamura, A., Gando, S., Kodate, A., Katabami, K., Wada, T., Ono, Y., Maekawa, K., Hayakawa, M., Sawamura, A., Gando, S., Andersen, L. W., Raymond, T., Berg, R., Nadkarni, V., Grossestreuer, A., Kurth, T., Donnino, M., Krüger, A., Ostadal, P., Janotka, M., Vondrakova, D., Kongpolprom, N., Cholkraisuwat, J., Pekkarinen, P. T., Ristagno, G., Masson, S., Latini, R., Bendel, S., Ala-Kokko, T., Varpula, T., Vaahersalo, J., Hoppu, S., Tiainen, M., Mion, M. M., Plebani, M., Pettilä, V., Skrifvars, M.B., Son, Y., Kim, K. S., Suh, G. J., Kwon, W. Y., Ko, J. I., Park, M. J., Cavicchi, F. Zama, Iesu, E., Nobile, L., Vincent, J. L., Creteur, J., Taccone, F. S., Tanaka, H., Otani, N., Ode, S., Ishimatsu, S., Martínez, L., Algarte, R., Sánchez, B., Romero, I., Martínez, F., Quintana, S., Trenado, J., Vondrakova, D., Ostadal, P., Kruger, A., Janotka, M., Malek, F., Neuzil, P., Yeh, Y. C., Chen, Y. S., Wang, C. H., Huang, C. H., Chao, A., Lee, C. T., Lai, C. H., Chan, W. S., Cheng, Y. J., Sun, W. Z., Kaese, S., Horstmann, C., Lebiedz, P., Mourad, M., Gaudard, P., Eliet, J., Zeroual, N., Colson, P., Ostadal, P., Mlcek, M., Hrachovina, M., Kruger, A., Vondrakova, D., Janotka, M., Mates, M., Hala, P., Kittnar, O., Neuzil, P., Jacky, A., Rudiger, A., Spahn, D. R., Bettex, D. A., Kara, A., Akin, S., Dos reis Miranda, D., Struijs, A., Caliskan, K., van Thiel, R. J., Dubois, E. A., de Wilde, W., Zijlstra, F., Gommers, D., Ince, C., Marca, L., Xini, A., Mongkolpun, W., Cordeiro, C. P. R., Leite, R. T., Lheureux, O., Bader, A., Rincon, L., Santacruz, C., Preiser, J. C., Chao, A., Chao, A. S., Chen, Y. S., Kim, W., Ahn, C., Cho, Y., Lim, T. 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Agudo, León, J. P. Tirapu, Irazabal, J. M. Guergue, Pérez, A. González, Fernández, P. Alvarez, Amor, L. Lopéz, Albaiceta, G. Muñiz, Lesmes, S. P. Gómez, Rosario, L. E. De la Cruz, Hernández, A. Ansotegui, Sanz, E. Regidor, Sánchez, M. J. Gómez, Calvo, S. Aldunate, Herrera, A. N. García, Hualde, J. Barado, Pascual, O. Agudo, León, J. P. Tirapu, Corona, A., Ruffini, C., Spazzadeschi, A., Marrazzo, F., Gandola, A., Sciurti, R., Savi, C., Catena, E., Ke, M. W., Cheng, C. C., Huang, W. C., Chiang, C. H., Hung, W. T., Lin, K. C., Lin, S. C., Wann, S. R., Chiou, K. R., Tseng, C. J., Kang, P. L., Mar, G. Y., Liu, C. P., Bertini, P., De Sanctis, F., Guarracino, F., Bertini, P., Baldassarri, R., Guarracino, F., Buitinck, S. H., van der Voort, P. H. J., Oto, J., Nakataki, E., Tsunano, Y., Izawa, M., Tane, N., Onodera, M., Nishimura, M., Ghosh, S., Gupta, A., De Gasperi, A., Mazza, E., Limuti, R., Prosperi, M., Bissenova, N., Yergaliyeva, A., Talan, L., Yılmaz, G., Güven, G., Yoruk, F., Altıntas, N. D., Mukherjee, D. N., Agarwal, L. K., Mandal, K., Palomar, M., Balsera, B., Vallverdu, M., Martinez, M., Garcia, M., Castellana, D., Lopez, R., Barcenilla, F., Kaminsky, G. E., Carreño, R., Escribá, A., Fuentes, M., Gálvez, V., Del Olmo, R., Nieto, B., Vaquerizo, C., Alvarez, J., De la Torre, M. A., Torres, E., Bogossian, E., Nouer, S. Aranha, Salgado, D. Ribeiro, Brugger, S. Carvalho, Jiménez, G. Jiménez, Torner, M. Miralbés, Vidal, M. Vallverdú, Garrido, B. Balsera, Casals, X. Nuvials, Gaite, F. Barcenilla, Cabello, J. Trujillano, Martínez, M. Palomar, Doganci, M., Izdes, S., Besevli, S. Guzeldag, Alkan, A., Kayaaslan, B., Ramírez, C. Sánchez, Balcázar, L. Caipe, Santana, M. Cabrera, Viera, M. A. Hernández, Escalada, S. Hípola, Vázquez, C. F. Lübbe, Penichet, S. M. Marrero, Campelo, F. Artiles, López, M. A. De La Cal, Santana, P. Saavedra, Santana, S. Ruíz, Repessé, X., Artiguenave, M., Paktoris-Papine, S., Espinasse, F., Dinh, A., El Sayed, F., Charron, C., Géri, G., Vieillard-Baron, A., Marmanidou, K., Oikonomou, M., Nouris, C., Dimitroulakis, K., Soilemezi, E., Matamis, D., Ferré, A., Guillot, M., Teboul, J. L., Lichtenstein, D., Mézière, G., Richard, C., Monnet, X., Pham, T., Beduneau, G., Schortgen, F., Piquilloud, L., Zogheib, E., Jonas, M., Grelon, F., Runge, I., Terzi, N., Grangé, S., Barberet, G., Guitard, P. G., Frat, J. P., Constan, A., Chrétien, J. M., Mancebo, J., Mercat, A., Richard, J. C. M., Brochard, L., Prīdāne, S., Sabeļņikovs, O., Mojoli, F., Orlando, A., Bianchi, I., Torriglia, F., Bianzina, S., Pozzi, M., Iotti, G. A., Braschi, A., Beduneau, G., Pham, T., Schortgen, F., Piquilloud, L., Zogheib, E., Jonas, M., Grelon, F., Runge, I., Terzi, N., Grangé, S., Barberet, G., Guitard, P. G., Frat, J. P., Constan, A., Chrétien, J. M., Mancebo, J., Mercat, A., Richard, J. C. M., Brochard, L., Kondili, E., Psarologakis, C., Kokkini, S., Amargianitakis, V., Babalis, D., Chytas, A., Chouvarda, I., Vaporidi, K., Georgopoulos, D., Trapp, O., Kalenka, A., Mojoli, F., Orlando, A., Bianchi, I., Torriglia, F., Bianzina, S., Pozzi, M., Iotti, G. A., Braschi, A., Lozano, J. A. Benítez, Sánchez, P. Carmona, Francioni, J. E. Barrueco, Ferrón, F. Ruiz, Simón, J. M. Serrano, Spadaro, S., Karbing, D. S., Gioia, A., Moro, F., Corte, F. Dalla, Mauri, T., Volta, C. A., Rees, S. E., Petrova, M. V., Mohan, R., Butrov, A. V., Beeharry, S. D., Vatsik, M. V., Sakieva, F. I., Gobert, F., Yonis, H., Tapponnier, R., Fernandez, R., Labaune, M. A., Burle, J. F., Barbier, J., Vincent, B., Cleyet, M., Richard, J. C., Guérin, C., Shinotsuka, C. Righy, Creteur, J., Taccone, F. S., Törnblom, S., Nisula, S., Vaara, S., Poukkanen, M., Andersson, S., Pettilä, V., Pesonen, E., Xie, Z., Liao, X., Kang, Y., Zhang, J., Kubota, K., Egi, M., Mizobuchi, S., Hegazy, S., El-Keraie, A., El Sayed, E., El Hamid, M. Abd, Rodrigues, N. J., Pereira, M., Godinho, I., Gameiro, J., Neves, M., Gouveia, J., e Silva, Z. Costa, Lopes, J. A., Mckinlay, J., Kostalas, M., Kooner, G., Dudas, G., Horton, A., Kerr, C., Karanjia, N., Creagh-Brown, B., Forni, L., Yamazaki, A., Ganuza, M. Sanz, Molina, J. A. Martinez, Martinez, F. Hidalgo, Freile, M. T. Chiquito, Fernandez, N. Garcia, Travieso, P. Medrano, Bandert, A., Frithiof, R., Lipcsey, M., Smekal, D., Schlaepfer, P., Durovray, J. D., Plouhinec, V., Chiappa, C., Bellomo, R., Schneider, A. G., Mitchell, S., Durrant, J., Street, H., Dunthorne, E., Shears, J., Caballero, C. Hernandez, Hutchison, R., Schwarze, S., Ghabina, S., Thompson, E., Prowle, J. R., Kirwan, C. J., Gonzalez, C. A., Pinto, J. L., Orozco, V., Patiño, J. A., Garcia, P. K., Contreras, K. M., Rodriguez, P., Echeverri, J. E., GETGAG Working Group, JSEPTIC (Japanese Society of Education for Physicians and Trainees in Intensive Care) Clinical Trial Group, CAPCRI Study, for the ReVA Research Network and the PROVE Network Investigators, from the FROG ICU Investigators, The WIND study group, Plug Working Group, GETGAG/SEMICYUC, AKI Research Group, St George’s University of London, IPREA Study Group, FINNRESUSCI Study Group, PICS- HCPA: Programa Intrahospitalar de Combate à Sepse do Hospital de Clínicas de Porto Alegre, ENVIN-HELICS Study Group, ARIAM registry of adult cardiac surgery, The Rapid Diagnosis of Infections in the Critically Ill Team, Tokyo Womens Medical University, PLUG working group, PLUG Working Group, On behalf of Okayama Research Investigation Organizing Network (ORION)investigators, PS-ICU Group, Japan Septic Disseminated Intravascular Coagulation (JSEPTIC DIC) study group, Student Research Committee - Shiraz University of Medical Sciences, ARIAM-ANDALUCIA, The WIND study group, PLUG Working Group, The WIND study group, PLUG Working Group, and Plug working group

Published
2016
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20. Staphylococcus aureus From Goats Are Genetically Heterogeneous and Distinct to Bovine Ones

Author

Romano, A., Gazzola, A., Bianchini, V., Cortimiglia, Claudia, Maisano, A. M., Cremonesi, P., Graber, H. U., Vezzoli, F., Luini, M., Cortimiglia C. (ORCID:0000-0001-6884-091X), Romano, A., Gazzola, A., Bianchini, V., Cortimiglia, Claudia, Maisano, A. M., Cremonesi, P., Graber, H. U., Vezzoli, F., Luini, M., and Cortimiglia C. (ORCID:0000-0001-6884-091X)

Abstract

Staphylococcus aureus is one of the major pathogens responsible for intramammary infections in small ruminants, causing severe economic losses in dairy farms. In addition, S. aureus can contaminate milk and dairy products and produce staphylococcal enterotoxins, being responsible for staphylococcal food poisoning. Currently, data on the population structure and the virulence gene patterns of S. aureus strains isolated from goat milk is limited. Therefore, this study aimed at defining Ribosomal Spacer PCR (RS-PCR) genotypes, clonal complexes (CC), spa types, and virulence gene profiles of S. aureus isolated from goat milk samples from Lombardy region of Italy. A total of 295 S. aureus isolates from 65 goat bulk tank milk samples were genotyped by RS-PCR. spa typing and virulence gene patterns of a subgroup of 88 isolates were determined, and MLST was performed on a further subgroup of 39 isolates, representing all the spa types identified during the analysis. This study revealed 7 major genotypic clusters (CLR, CLAA, CLZ, CLAW, CLBW, CLS, and CLI), of which S. aureus CLR (19.8%) was the most common. A total of 26 different spa types were detected, the most prevalent types were t1773 (24%), t5428 (22.7%), and t2678 (12.5%). Overall, 44.3% of all isolates harbored at least one enterotoxin gene. The most prevalent was the combination of sec-sel genes (35.2%). Based on their MLST, isolates were assigned to 14 different CC, with majority grouped as CC133 (24%), CC130 (19.6%), and CC522 (19.6%). The caprine S. aureus population was depicted with a minimum spanning tree and an evolutionary analysis based on spa typing and MLST, respectively. Then, the variability of such strains was compared to that of bovine strains isolated in the same space-time span. Our results confirmed that S. aureus isolates from goats have wide genetic variability and differ from the bovine strains, supporting the idea that S. aureus from small ruminants may constitute a distinct population.

Published
2020

23. GENOTIPIZZAZIONE DI S. AUREUS ISOLATI DA LATTE BOVINO DI AZIENDE LOMBARDE

Author

Gazzola A. [1], Romanò A. [1, Maisano A.M. [1], Spelta C. [1], Bertocchi L. [2], Ferrara G [2], Finazzi G. [3], Filipello V. [3], Decastelli L. [4], Bergagna S. [5], Cremonesi P. [6], Vezzoli F. [1], Graber HU. [7], and Luini M.*[1]

Subjects
Staphylococcus aureus, adlb gene, RS-PCR
Abstract

Staphylococcus aureus strains (n=1559) isolated from North Italian dairy herds were genotyped by RS-PCR. A selection of these strains (84 from bulk tank milk and 91 from individual cow milk) was also analyzed for the presence of adlb gene, which is considered a possible marker of contagiousness. The adlb gene amplification was included in a multiplex PCR also targeting nuc and mecA genes. RS-PCR identified 83 different genotypes combined into 48 genotypic clusters, among which genotype B with his variants (GTB) was the most prevalent (n=491). A clonal diffusion of a single genotype was shown in most farms (373/458), especially when GTB was involved. Most of Staph. aureus belonging to GTB carried the adlb gene (38/43), as well as very few strains belonging to a different genotype. When investigated, all of them were associated with high prevalence in the herds, confirming that RS-PCR and adlb PCR could be useful tools in predicting the clinical properties of isolates.

Published
2018

24. Indagini epidemiologiche, biomolecolari e di antibiotico-resistenza in aziende di bovine da latte con infezione da Streptococcus agalactiae

Author

Maisano AM, Romanò A, Maraschi F, Morandi S, Orlandelli C, Spelta C, Vezzoli F, Bertocchi L, and Luini M

Subjects
Streptococcus agalactie, MIC, Bovine da latte, Mastite
Abstract

Lo scopo di questo lavoro è stato quello di approfondire l'epidemiologia di Streptococcus agalactiae in aziende con recenti positività, in particolare a riguardo: delle condizioni d'allevamento, della prevalenza di bovine infette, della suscettibilità agli antibiotici e delle caratteristiche genotipiche dei ceppi. I risultati hanno dimostrato la presenza del patogeno in 13 allevamenti su 15 (1794 campioni di latte individuale), con prevalenza di bovine infette molto variabile (10,4-54,7%). Le condizioni d'allevamento (check-list) correlate con la prevalenza d'infezione sono risultate significative per: la preparazione igienica della mammella, l'esecuzione del post-dipping, la routine di mungitura, le procedure di lavaggio e disinfezione dell'impianto/tank, la somministrazione di latte infetto ai vitelli, l'igiene delle mammelle e la conta sia delle cellule somatiche che delle UFC/mL di S. agalactiae nel latte di massa. I risultati della minima concentrazione inibente su un ceppo per stalla (n. 13), hanno evidenziato resistenze alla Tetraciclina (n. 5), Eritromicina (n. 1) e Pirlimycina (n. 1); tutti i ceppi sono risultati sensibili ai ?-lattamici. La RAPD-PCR su almeno 4 ceppi per stalla, ha evidenziato una certa variabilità genetica (6 profili diversi su 55 ceppi), ed in 11 stalle la circolazione dei ceppi ha mostrato un profilo clonale, mentre in 2 stalle si sono evidenziati ceppi differenti. 18 ceppi rappresentativi di tutte le stalle sono risultati tutti positivi per il gene specie specifico e per i fattori di virulenza, neuC e cpsL (PCR-RT). I nostri risultati confermano il carattere contagioso di questa infezione e sottolineano la necessità di un controllo costante delle corrette procedure-igiene di mungitura, del management degli animali e del latte di massa.

Published
2018

25. INDAGINI AMBIENTALI IN ALLEVAMENTI BOVINI CON INFEZIONI INTRAMAMMARIE DA MRSA

Author

Romanò A. [1], Maisano A.M. [1], Spelta C. [1], Cremonesi P. [3], Garbarino C. [2], Arrigoni N. [2], Vezzoli F. [1], and Luini M.*[1]

Subjects
animal diseases, Environmental sampling, Dairy herds, MRSA
Abstract

The present study was conducted in dairy herds were MRSA presence was previously demonstrated with the aim to (i) evaluate the prevalence of MRSA intra-mammary infections (IMI) and (ii) identify the specific environmental contamination. MRSA IMI's were detected at low prevalence (less than 5%), except for one herd where it reached the 24,8%. The circulating MRSA genotypes were different for each herd and in two herds two different genotypes were found with one of them predominant. Four and 11 out of 18 cows, among those with MRSA IMI, were positive for nasal and udder skin swabs, respectively. The environmental analyses showed that milking parlor was systematically contaminated in different sites as well as the young calves mangers and buckets, whereas the heifers pens and mangers were not. Thus, in MRSA positive herds, especially milkers and personnel to young calves should be considered at risk of occupational infections.

Published
2016

27. Development of a droplet digital polymerase chain reaction for rapid and simultaneous identification of common foodborne pathogens in soft cheese

Author

Cremonesi, P., Cortimiglia, Claudia, Picozzi, C., Minozzi, G., Malvisi, M., Luini, M., Castiglioni, B., Cortimiglia C. (ORCID:0000-0001-6884-091X), Cremonesi, P., Cortimiglia, Claudia, Picozzi, C., Minozzi, G., Malvisi, M., Luini, M., Castiglioni, B., and Cortimiglia C. (ORCID:0000-0001-6884-091X)

Abstract

Dairy products can harbor various microorganisms (e.g., Campylobacter spp., Salmonella spp., Listeria monocytogenes, verocytotoxin-producing Escherichia coli) arising from animal reservoirs, and which can become important sources of foodborne illness. Therefore, early detection of food pathogens is crucial to prevent diseases. We wished to develop an accurate quantitative protocol based on a droplet digital polymerase chain reaction (ddPCR) involving eight individual TaqManTM reactions to detect simultaneously, without selective enrichment, Listeria spp., L. monocytogenes, Salmonella spp., verocytotoxin-producing E. coli and Campylobacter spp. in cheese. ddPCR (a "third-generation PCR") provides absolute quantification of target DNAs without requirement of a standard curve, which simplifies experimentation and data comparability. The accuracy, specificity and sensitivity of the developed ddPCR system were assessed using purified DNA from 50 reference pathogenic and non-pathogenic strains from international or Italian collections and analyzing soft cheese samples artificially contaminated with serial dilutions (from 4 × 106 to 4 × 101 CFU/g) of pure cultures from the American Type Culture Collection. Finally, the performance of our ddPCR system was compared by parallel testing with quantitative PCR: it gave higher sensitivity (102 CFU/g for the Listeria spp. assay) without the necessity of a standard curve. In conclusion, this is the first ddPCR system developed for simultaneous detection of common foodborne pathogens in cheese using a single set of amplification conditions. As such, it could become a useful strategy for high-throughput screening of microorganisms to evaluate the quality and safety of food products.

Published
2016

28. Molecular approaches for the identification of genetic subtypes of Staphylococcus aureus isolated from mammary infection

Author

Cremonesi P., Capra E., Pozzi F., Tilola M., Drigo I., Graber H.U., Luini M., Vezzoli F., Piccinini R., and Castiglioni B.

Subjects
molecular characterization, Staphylococcus aureus, mastitis
Abstract

AIM: S. aureus is one of the most important causative agents of bovine mastitis and the clinical outcome can vary in relation to the strains involved in the mammary infection. The aim of this study, funded by Lombardy Region, was to develop a diagnostic protocol for the identification of genetic subtypes of S. aureus responsible for bovine mastitis, with particular virulence and diffusiveness. MATERIAL AND METHODS: S. aureus strains associated with intra-mammary infections were isolated from 8 "control" (< 4% of infected cows) and 8 "case" (> 28% of infected cows) herds for a total of 2015 cows included in the study. Milk samples were cultured with standard methods and the DNA was extracted from S. aureus strains using a protocol described in literature (Cremonesi et al., 2006). Twenty-four and 626 strains isolated from single quarters of "control" and "case" herds respectively, were analysed by both RS-PCR (Fournier et al., 2008) and multiplex PCR. The first method is based on the amplification of the 16S-23S rRNA intergenic spacer region, the multiplex PCR targets the genes encoding for enterotoxins and other virulence genes such as leukocidins and leukotoxins. A subset of 55 strains representative of "control" and "case" isolates was also analysed by ribotyping (automatic Riboprinter), by Identibac array tube system and by protein profiling using MALDI-TOF MS. RESULTS: The RS-PCR analysis of the 650 strains revealed 10 different profiles. In 5 out of 8 "case" herds this method identified the genotype GTB and GTB-variant indicated in literature (Graber et al., 2009) as characteristic of highly diffusive and pathogenic strains, while GTK, GTR_VI and a new genotype named GTbM circulated in the remaining 3 herds. Seven out of the 8 "control" herds, revealed only one S. aureus profile with a predominance of the GTS genotype. Except for 4 out of the 55 representative strains analysed, ribotyping gave a response similar to RS-PCR. Moreover, MALDI-TOF and Identibac analyses proved to be able to integrate the molecular profiling of S. aureus isolated in field. CONCLUSION: Our results confirmed that RS-PCR could be used as a rapid test for molecular typing of S. aureus strains isolated from bovine mastitis and could identify genetic subtypes with particular virulence. Ribotyping, MALDI-TOF and Identibac analyses showed their capability for molecular strains characterization, integrating RS-PCR data. Multivariate analysis by combining data from different techniques could be used to discover a simple panel of markers associated to S. aureus virulence and diffusiveness. BIBLIOGRAFY Cremonesi P., et al. (2006). J. Dairy Sci., 89:163-169; Fournier C., et al. (2008). Res. Vet. Sci., 85: 439-448; Graber HU., et al. (2009). J. Dairy Sci., 92(4) 1442-1451

Published
2013

29. Impact of genetic variation and geographic distribution of porcine reproductive and respiratory syndrome virus on infectivity and pig growth

Author

Badaoui B., Grande R., Calza S., Cecere M., Luini M., Stella A., and Botti

Subjects
Male, Glycosylation, 040301 veterinary sciences, Swine, viruses, animal diseases, Porcine Reproductive and Respiratory Syndrome, Virulence, Biology, Polymerase Chain Reaction, Virus, law.invention, 0403 veterinary science, 03 medical and health sciences, Open Reading Frames, law, Semen, Genetic variation, Animals, Porcine respiratory and reproductive syndrome virus, Genetic variability, Polymerase chain reaction, Phylogeny, 030304 developmental biology, 2. Zero hunger, Genetics, Infectivity, 0303 health sciences, General Veterinary, Genetic Variation, virus diseases, 04 agricultural and veterinary sciences, General Medicine, Sequence Analysis, DNA, respiratory system, Porcine reproductive and respiratory syndrome virus, biology.organism_classification, ORF5, Virology, veterinary(all), Breed, 3. Good health, ORF7 sequences, Animals, Newborn, Italy, PRRSV, Female, Sequence Alignment, Research Article
Abstract

Background The porcine reproductive and respiratory syndrome (PRRS) is a devastating disease for the pig industry. In this study, we analysed the genetic variability of PRRS virus (PRRSV) as well as the relationship between the genetic variability, the geographical and temporal distribution of the PRRSV strains. Moreover, we investigated the association between the glycosylation patterns in PRRSV sequences and pigs growth. Results The data highlight that PRRSV strains evolve rapidly on individual farms, and temporal evolution of PRRSV is an important factor of genetic variability. Analysis of glycosylation sites in the glycoprotein 5 (GP5) ectodomain revealed that PRRSV isolates had seven combinations of putative N-linked glycosylation sites of which the N37/46/53 sites was found in 79% of the sequences. No significant relationship was found between the genetic variation of the PRRSV strains and the geographic distance. A significant relationship was found between the genetic variation and time of sampling when farm was considered as a factor in the analysis. Furthermore, the commercial semen from artificial insemination centres was not a source of PRRS transmission. The PRRSV having the glycosylation site at position N46 (N46+) were observed to have higher burden on pigs and accordingly the corresponding infected pigs had lower average daily gain (ADG) compared with those infected with PRRSV lacking the glycosylation at N46 (N46-) position site. This study showed that the number of piglets by litter infected by PRRSV was lower for the Landrace breed than for the other studied breeds (Large White, Duroc and Pietrain). Conclusions The PRRSV genetic variability which is determined by a local and temporal evolution at the farm level could be considered in a perspective of prevention. Moreover, the association between the PRRSV glycosylation patterns and its virulence could be of interest for vaccine development. The differences of resistance to PRRSV infections among pig breeds might open new horizons for the genetic selection of robustness against PRRSV infection.

Published
2013
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30. Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Italian dairy herds

Author

Piccinini R., Malvisi M., Cremonesi P., Capra E., Bignoli G., Castiglioni B., Valentino L., Pozzi F., Vezzoli F., and Luini M.

Subjects
Staphylococcus aureus, diffusiveness, MRSA, biochemical phenomena, metabolism, and nutrition, bacterial infections and mycoses
Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) has been reported in human medicine as a cause of nosocomial and community-associated infections. In veterinary medicine, the microorganism has been identified in a wide range of animals and diseases, including dairy cows mastitis. Therefore, MRSA is considered an emerging threat with a high zoonotic potential. AIM - In the present study, we investigated the diffusion of MRSA in Italian dairy herds and the prevalence of infected animals at herd level. MATERIALS AND METHODS Quarter milk samples were aseptically taken from all lactating cows of 83 S. aureus infected herds and bacteriological analysis was performed by culture on blood-agar plates. The hemolytic, Gram-positiv and coagulase-positive colonies of growth were identified as S. aureus. At least four isolates morphologically representative for each herd were tested by oxacillin disk-diffusion method to detect methicillin-resistance. DNA of oxacillin resistant isolates was extracted and the presence of mecA gene was investigated by PCR. RESULTS - All oxacillin-resistant isolates were confirmed as MRSA by mecA positivity. Prevalence of S. aureus infected cows and frequency of MRSA isolation were registered at herd level. Overall prevalence of mammary infections by S. aureus ranged 0,7 - 62%: in 28 herds (33.7%) the prevalence was higher than 25%, while in 27 herds (32.5%) it was lower than 4.7%. Methicillin-resistant strains were detected in 11 farms (13,25%), all belonging to the low-prevalence group. In 5 out of such 11 herds, all S. aureus isolates were MRSA, while in the remaining 6 herds both MRSA and methicillin-susceptible S. aureus (MSSA) were present. CONCLUSIONS - Our data confirmed the potential risk of subclinical mastitis caused by MRSA and suggest a low diffusiveness of these strains in infected herds. This characteristic could reduce the zoonotic potential of bovine MRSA, but the real prevalence in dairy farms could be underestimated, since bacteriological analysis of milk is usually performed only when a clear problem of mammary infections affects the herd.

Published
2013

31. Prevalence of Staphylococcus aureus and MRSA in bulk tank milk of lombardy dairy herds

Author

Cortimiglia C., Marzagalli L., Vezzoli F., Avisani D., Cremonesi P., Franco A., Battisti A., and Luini M.

Subjects
Staphylococcus aureus, MRSA
Abstract

AIMS S. aureus is the most important causative agent of subclinical mastitis in cattle. In the affected farms the clinical outcome and the consequent economic losses can vary in relation to the strains involved in the mammary infection. Methicillin-resistant strains (MRSA) have zoonotic importance, especially for farm workers and were also isolated from bovine mastitis in Italy and other countries[1,2]. The aim of our study was to evaluate the prevalence of S. aureus and MRSA in dairy herds and to identify the main circulating genotypes. MATERIALS AND METHODS We examined 509 samples of bulk tank milk from eight provinces of Lombardy region. Samples were plated directly on Blood agar and the S. aureus count was determined on Baird Parker Agar + RPF. For each sample, at least 5 colonies referable to S. aureus were tested for susceptibility to oxacillin by disk diffusion test. The same milk samples were examined by subsequent enrichment in MH broth + 7.5% NaCl and TSB + 5mg/l of oxacillin and plating on Brilliance MRSA agar (Oxoid). The presence of the mecA gene was confirmed by PCR [3]. Genotyping was performed by RS-PCR (16S-23S intergenic spacer PCR) [4] and Multilocus Sequence Typing (MLST) [5]. RESULTS A total of 211 out of 509 samples were positive for S. aureus (41.4%), with a prevalence in the different provinces from 9.1% to 56.8%. The counts of S. aureus showed values ranging between 10 and > 30000 cfu/ml with a median value of 100 cfu/ml. MRSA were isolated from 26 samples (5.1%). Five of these were demonstrated by both direct plating and plating after enrichment, 10 and 11 by enrichment or direct plating only, respectively. The RS-PCR identified several different profiles, including the so-called genotype B (GTB) [4] that was detected in 45 samples (21.3%). The characterization by MLST of 18 of the 26 samples positive for MRSA identified ten ST398, four ST1, three ST97 and one ST5. DISCUSSION The high prevalence of positive bulk milk samples confirms the importance of the S. aureus infection in Lombardy dairy farms. The analysis of circulating genotypes showed considerable variability, but it should be noted that a fairly significant rate is represented by GTB genotype which is associated with high infectivity and pathogenicity. MRSA were demonstrated in more than 10% of S. aureus positive samples, confirming the data obtained previously in the province of Lodi (personal data). The genotyping of MRSA strains showed a prevalence of typical LA-MRSA (ST398 and ST97) and ST1, the latter associated with community-acquired human infection. In the case of MRSA the application of more stringent control plans to reduce the risk of transmission to humans is recommended. REFERENCES 1) Benedetti V et.al (2010) Large Animal Review, 16:67-70; 2) Holmes MA and Zadocks RN (2011) J. Mammary Gland Biol. Neoplasia 16, 373-382; 3) McClure JA., et al. (2006) J. Clin. Microbiol., 44:1141-44; 4) Graber HU et al. (2009). J. Dairy Sci., 92(4) 1442-1451; 5) Enright MC et al (2000) J. Clin. Microbiol., 38, 1008-1015

Published
2013

38. Short communication: Prevalence of Staphylococcus aureus and methicillin-resistant S. aureus in bulk tank milk from dairy goat farms in Northern Italy

Author

Cortimiglia, Claudia, Bianchini, V., Franco, A., Caprioli, A., Battisti, A., Colombo, L., Stradiotto, K., Vezzoli, F., Luini, M., Cortimiglia C. (ORCID:0000-0001-6884-091X), Cortimiglia, Claudia, Bianchini, V., Franco, A., Caprioli, A., Battisti, A., Colombo, L., Stradiotto, K., Vezzoli, F., Luini, M., and Cortimiglia C. (ORCID:0000-0001-6884-091X)

Abstract

Staphylococcus aureus is regarded as a leading cause of mastitis in goats. However, few data are available on the presence of methicillin-resistant S. aureus (MRSA) in this species. The aim of this study was to assess the prevalence of S. aureus and MRSA in bulk tank milk samples from dairy goat farms in Northern Italy. Eighty-five out of 197 samples (43.1%) tested positive for S. aureus with counts ranging from 10 to more than 1.5×104 cfu/mL. The MRSA was screened by both direct plating followed by a disk diffusion test to evaluate methicillin resistance and a selective enrichment method. Methicillin-resistance was confirmed by mecA-specific PCR. Methicillin-resistant S. aureus was identified in 4 samples (2.0%) and multilocus sequence typing (MLST) showed the presence of livestock-associated MRSA belonging to lineages ST398 (n=3) and ST1 (n=1). In one case we demonstrated that the same MRSA strain was able to persist over time on the farm, being isolated from both bulk tank milk and the udder of 3 goats 1 yr after the first isolation. The high prevalence of S. aureus-positive herds detected in this study and the presence of MRSA strains belonging to livestock-associated genotypes is of concern, and represents a novel finding in the Italian dairy goat production system. The application of stringent measures for the control of S. aureus mastitis at the farm level seems appropriate to reduce the economic losses, and to minimize the risk of foodborne illness and the transmission of MRSA to humans by occupational exposure.

Published
2015

40. Genetic Loci Involved in Antibody Response to Mycobacterium avium ssp. paratuberculosis in Cattle

Author

Minozzi G., Buggiotti L., Stella A., Strozzi F., Luini M., and Williams J. L.

Abstract

Mycobacterium avium subsp. paratuberculosis (MAP) causes chronic enteritis in a wide range of animal species. In cattle, MAP causes a chronic disease called Johne's disease, or paratuberculosis, that is not treatable and the efficacy of vaccine control is controversial. The clinical phase of the disease is characterised by diarrhoea, weight loss, drop in milk production and eventually death. Susceptibility to MAP infection is heritable with heritability estimates ranging from 0.06 to 0.10. There have been several studies over the last few years that have identified genetic loci putatively associated with MAP susceptibility, however, with the availability of genome-wide high density SNP maker panels it is now possible to carry out association studies that have higher precision. Methodology/Principal Findings The objective of the current study was to localize genes having an impact on Johne's disease susceptibility using the latest bovine genome information and a high density SNP panel (Illumina BovineSNP50 BeadChip) to perform a case/control, genome-wide association analysis. Samples from MAP case and negative controls were selected from field samples collected in 2007 and 2008 in the province of Lombardy, Italy. Cases were defined as animals serologically positive for MAP by ELISA. In total 966 samples were genotyped: 483 MAP ELISA positive and 483 ELISA negative. Samples were selected randomly among those collected from 119 farms which had at least one positive animal. Conclusion/Significance The analysis of the genotype data identified several chromosomal regions associated with disease status: a region on chromosome 12 with high significance (P

Published
2010

46. Expression of Class II Major Histocompatibility Complex Molecules in Chronic Pulmonary Mycoplasma bovis Infection in Cattle

Author

Radaelli, E, Luini, M, Domeneghini, C, Loria, G, Recordati, C, Radaelli, P, Scanziani, E, Loria, GR, Scanziani, E., RADAELLI, PAOLO, Radaelli, E, Luini, M, Domeneghini, C, Loria, G, Recordati, C, Radaelli, P, Scanziani, E, Loria, GR, Scanziani, E., and RADAELLI, PAOLO

Abstract

Pulmonary inflammation often results in expression of the class II major histocompatibility complex (MHCII) by both professional antigen-presenting cells (APCs; histiocytes and lymphocytes) and non-professional APCs (respiratory epithelium and endothelium). In this study lesions from 17 cases of bovine chronic pneumonia, associated with Mycoplasma bovis infection, were examined immunohistochemically for M. bovis antigen and MHCII expression. Ten cases of chronic necrosuppurative bronchopneumonia (NBP) were shown to be characterized by abundant perinecrotic M. bovis antigen associated with scant MHCII expression by degenerate leucocytes. Seven cases of chronic catarrhal bronchointerstitial pneumonia (CBP) showed prominent MHCII expression by both professional APCs and respiratory epithelium, in the absence of intralesional M. bovis immunolabelling. The results suggest that prominent MHCII expression by both professional and non-professional APCs plays a role in the pathogenesis of M. bovis-induced CBP. Conversely, the role of MHCII expression in necrosuppurative foci typical of M. bovis-associated NBP can be considered negligible.

Published
2009

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