Palavras-chave
biodiversity
leafy greens
food contamination
foodborne diseases
food safety
Como Citar
Resumo
Ciliated protozoa are an ubiquitous group of eukaryotes that have been poorly reported on leafy greens. The present study aimed to verify the occurrence and diversity of ciliated protozoa in the leaves and roots of three of the most commonly consumed leafy greens in Brazil – lettuce, rocket and coriander. The vegetable samples were washed by manual agitation (3 minutes) in two different media (mineral water and Page´s Amoeba Saline solution). After washing, the contents were incubated in Petri dishes and aliquots were removed for microscopic identification and in vivo observation. A total of 21 ciliated protozoa species were found, most of which were bacterivorous. Leafy greens have commonly been associated with foodborne outbreaks and ciliated protozoa, which although they are not a Public Health concern, can act as “Trojan Horses” harboring bacteria, viruses and other protozoa cysts and oocysts and can suggest a new route towards microbiological quality related to the food chain. This is the first report of ciliated protozoa on leafy greens consumed in Brazil.
Referências
1. Idahosa OT. Parasitic contamination of fresh vegetables sold in Jos markets. Global J Med Res. 2011;11(1):20-5.
2. US Department of Health and Human Services and US Department of Agriculture. 2015-2020 Dietary Guidelines for Americans. 8.ed. Washington (DC): USDA; 2015. [access 2019 dez 10] Avaliable in: https://health.gov/sites/default/files/2019-09/2015-2020_Dietary_Guidelines.pdf
3. Wu QJ, Wu L, Zheng LQ, Xu X, Ji C, Gong TT. Consumption of fruit and vegetables reduces risk of pancreatic cancer: evidence from epidemiological studies. Eur J Cancer Prev. 2016;25(3):196-205. https://dx.doi.org/10.1097/CEJ.0000000000000171
4. Vaerewijck MJM, Sabbe K, Baré J, Houf K. Occurrence and diversity of free-living protozoa on butterhead lettuce. Int J Food Microbiol. 2011;147(2):105-11. https://dx.doi.org/10.1016/j.ijfoodmicro.2011.03.015
5. Bennett SD, Sodha SV, Ayers TL, Lynch MF, Gould LH, Tauxe RV. Produce-associated foodborne disease outbreaks, USA, 1998–2013. Epidemiol Infect. 2018;146(11):1397-1406. https://dx.doi.org/10.1017/S0950268818001620
6. World Health Organization (WHO). WHO estimates of the global burden of foodborne diseases: foodborne disease burden epidemiology reference group 2007- 2015. WHO; 2015 [access 2017 abr 15]. Avaliable in: https://apps.who.int/iris/handle/10665/199350
7. Ministério da Saúde (BR). Secretaria de Vigilância em Saúde. Surtos de Doenças Transmitidas por Alimentos no Brasil; 2016. [access 2017 may 01]. Avaliable in: http://portalarquivos.saude.gov.br/images/pdf/2016/junho/08/Apresenta----o-Surtos-DTA-2016.pdf
8. Vaerewijck MJM, Baré J, Lambrecht E, Sabbe K, Houf K. Interactions of foodborne pathogens with free-living protozoa: potential consequences for food safety. Compr Rev Food Sci Food Saf. 2014;13(5):924-44. https://dx.doi.org/10.1111/1541-4337.12100
9. Siqueira-Castro ICV, Greinert-Goulart JA, Bonatti TR, Yamashiro S, Franco RMB. First report of predation of Giardia sp. cysts by ciliated protozoa and confirmation of predation of Cryptosporidium spp. oocysts by ciliate species. Environ Sci Pollut Res. 2016;23:11357-62. https://dx.doi.org/10.1007/s11356-016-6689-y
10. Chavatte N, Lambrecht E, Van Damme I, Sabbe K, Houf K. Abundance, diversity and community composition of free-living protozoa on vegetable sprouts. Food Microbiol. 2016;55:55-63. https://dx.doi.org/10.1016/j.fm.2015.11.013
11. Gourabathini P, Brandi MT, Redding KS, Gunderson JH, Berk SG. Interactions between food-borne pathogens and protozoa isolated from lettuce and spinach. Appl Environ Microbiol. 2008;74(8):2518-25. https://dx.doi.org//10.1128/AEM.02709-07
12. Denoncourt AM, Paquet VE, Charette SJ. Potential role of bacteria packaging by protozoa in the persistence and transmission of pathogenic bacteria. Front Microbiol. 2014;21(5):1-11. https://dx.doi.org//10.3389/fmicb.2014.00240
13. King BJ, Monis PT, Keegan AR, Harvey K, Saint C. Investigation of the survival of Cryptosporidium in environmental waters. Cooperative Research Centre for Water Quality and Treatment, Salisbury. Research Report 47; 2007. [access 2019 dez 06]. Avaliable in: https://www.waterra.com.au/publications/documentsearch/?download=81
14. King CH, Shotts Jr EB, Wooley RE, Porter KG. Survival of coliforms and bacterial pathogens within protozoa during chlorination. Appl Environ Microbiol. 1988; 54(12): 3023-33. https://dx.doi.org/10.1128/AEM.54.12.3023-3033.1988
15. Alhabbal AT. The prevalence of parasitic contamination on common sold vegetables in Alqalamoun region. Int J Pharm Sci Rev Res. 2015;30(1):94-7.
16. Foissner W, Agatha S, Berger H. Soil ciliates (Protozoa, Ciliophora) from Namibia (Southwest Africa), with emphasis on two contrasting environments, the Etosha Region and the Namib Desert. Denisia. 2002;5:1-1459. [access 2019 nov 20]. Avaliable in: http://www.wfoissner.at/data_prot/Foissner_etal_2002_1-1063.pdf
17. Dieckmann J. An improved protargol impregnation for ciliates yielding reproducible results. Europ J Protistol. 1995;31(4):372–82. https://dx.doi.org/10.1016/S0932-4739(11)80449-9
18. Berger H. Monograph of the Oxytrichidae (Ciliophora, Hypotrichia). In: Dumont HI, Werger MJA (editors). Monographiae Biologicae, vol. 78. Springer Science; 1999.p.1-1080. https://doi.org/10.1007/978-94-011-4637-1
19. Berger H. Monograph of the Gonostomatidae and Kahliellidae (Ciliophora, Hypotricha). In: Monographiae Biologicae, vol. 90, Springer Science; 2011.p.1-741. https://10.1007/978-94-007-455-8
20. Foissner W, Berger H. A user-friendly guide to ciliates (Protozoa, Ciliophora) commonly used by hydrobiologists as bioindicators in rivers, lakes and waste water, with notes in their ecology. Freshw Biol. 1996;35(2):375-482. https://doi.org/10.1111/j.13652427.1996.tb01775.x
21. Foissner W, Blatterer H, Berger H, Kohmann F. Taxonomische und ökologische revision der ciliaten des Saprobiensystems. Band I: Cyrtophorida, Oligotrichida, Hypotrichia, Colpodea. Informationsberichte des Bayerischen Landesamtes für Wasserwirtschaft; 1991.p.1478 [access 2019 mai 09]. Avaliable in: https://www.protozoology.com/ciliateatlas-I/
22. Foissner W, Berger H, Kohmann F. Taxonomische und ökologische revision der ciliaten des Saprobiensystems. Band II: Peritrichia, Heterotrichida, Odontostomatida. Informationsberichtedes Bayer. Landesamtes für Wasserwirtschaft; 1992.p.502. [access 2019 maio 09]. Avaliable in: https://www.protozoology.com/ciliateatlas/
23. Foissner W, Berger H, Kohmann F. Taxonomische und ökologische revision der ciliaten des Saprobiensystems – Band III: Hymenostomata, Prostomatida, Nassulida. Informationsberichtedes Bayer. Landesamtes für Wasserwirtschaft; 1994.p.548.[access 2019 mai 09]. Avaliable in: https://www.protozoology.com/ciliateatlas-III/
24. Lynn DH, Small EB. Phylum Ciliophora. In: Lee JJ, Bradbury PC, Leedale GF (editors.). An illustrated guide to the protozoa. Lawrence (KS): Society of Protozoologists; 2002.p.371-656.
25. Mohamed MA, Siddig EE, Elaagip AH, Edris AMM, Nasr AA. Parasitic contamination of fresh vegetables sold at central markets in Khartoum state, Sudan. Ann Clin Microbiol Antimicrob. 2016;15:17. https://doi.org/10.1186/s12941-016-0133-5
26. Bichai F, Dullemont Y, Hijnen W, Barbeau B. Predation and transport of persistent pathogens in GAC and slow sand filters: A threat to drinking water safety? Water Res. 2014;64:296-308. https://doi.org/10.1016/j.watres.2014.07.005
27. Chavatte N, Baré J, Lambrecht E, Van Damme I, Vaerewijck M, Sabbe K et al. Co-occurrence of free-living protozoa and foodborne pathogens on dishcloths: implications for food safety. Int J Food Microbiol. 2014;191:89-96. https://doi.org/10.1016/j.ijfoodmicro.2014.08.030
28. Nadhanan RR, Thomas CJ. Colpoda secrete viable Listeria monocytogenes within faecal pellets. Environ Microbiol. 2014;16(2),396-404. https://doi.org/10.1111/1462-2920.12230
29. Fayer R, Trout JM, Walsh E, Cole R. Rotifers ingest oocysts of Cryptosporidium parvum. J Eukaryot Microbiol. 2000;47(2):161-3. https://doi.org/10.1111/j.1550-7408.2000.tb00026.x
30. Stott R, May E, Matsushita E, Warren A. Protozoan predation as a mechanism for the removal of Cryptosporidium oocysts from wastewaters in constructed wetlands. Water Sci Technol. 2001;44(11-12):191-8. https://doi.org/10.2166/wst.2001.0828
31. Stott R, May E, Ramirez E, Warren A. Predation of Cryptosporidium oocysts by protozoa and rotifers: implications for water quality and public health. Water Sci Technol. 2003;47(3):77-83. https://doi.org/10.2166/wst.2003.0166
32. Chen L, Zhao X, Shao C, Miao M, Clamp JC. Morphology and phylogeny of two new ciliates, Sterkiella sinica sp. nov. and Rubrioxytricha tsinlingensis sp. nov. (Protozoa, Ciliophora, Hypotrichia) from north-west China. Syst Biodivers. 2017;15 (2):131-42. http://dx.doi.org/10.1080/14772000.2016.1219426
33. Pushkareva VI, Ermolaeva SA. Listeria monocytogenes virulence factor Listeriolysin O favors bacterial growth in co-culture with the ciliate Tetrahymena pyriformis, causes protozoan encystment and promotes bacterial survival inside cysts. BMC Microbiol. 2010;10:26. https://doi.org/10.1186/1471-2180-10-26
34. McNealy T, Newsome AL, Johnson RA, Berk SG. Impact of amoebae, bacteria, and Tetrahymena on Legionella pneumophila multiplication and distribution in an aquatic environment. In: Marre R, Abu Kwaik Y, Bartlett C, Cianciotto NP, Fields BS, Frosch M et al. (editors). Legionella. Washington (DC): ASM Press; 2001.p.170-5.[access 2019 out15]. https://doi.org/10.1128/9781555817985.ch30
35. Voß HJ. Morphology and morphogenesis of Parentocirrus hortualis nov. gen., nov. spec: a new genus within the redefined family Kahliellidae sensu Eigner 1995 (Ciliophora, Hypotrichida). Eur J Protistol. 1997;33:30-47. https://doi.org/10.1016/S0932-4739(97)80019-3
36. Blatterer H, Foissner W. Morphological and ontogenetic comparison of two populations of Parentocirrus hortualis Voss 1997 (Ciliophora, Hypotrichida). Linzer Biol Beitr. 2003;35:831-54. [acesso 2018 abr 20]. Avaliable in: https://www.zobodat.at/pdf/LBB_0035_2_0831-0854.pdf
37. Paiva TS, Silva-Neto, I. Description of Parentocirrus brasiliensis sp. n. (Ciliophora: Spirotrichea), a new ciliate protist present in activated sludge. Zootaxa. 2004;504(1):1-10. https://doi.org/10.11646/zootaxa.504.1.1
38. Paiva TS, Silva-Neto I. Morphology and divisional morphogenesis of Nudiamphisiella interrupta Foissner, Agatha & Berger, 2002 (Ciliophora: Stichotrichia) based on a Brazilian strain. Eur J Protistol. 2009;45(4):271-80. https://doi.org/10.1016/j.ejop.2009.04.001
39. Benčaťová S, Tirjaková E, Vďačný P. Resting cysts of Parentocirrus hortualis Voß, 1997 (Ciliophora, Hypotrichia), with preliminary notes on encystation and various types of excystation. Eur J Protistology. 2016;53:45-60. https://doi.org/10.1016/j.ejop.2015.12.003
40. Zou K, Thébault E, Lacroix G, Barot S. Interactions between the green and brown food web determine ecosystem functioning. Funct Ecol. 2016;30(8):1454-65. https://doi.org/10.1111/1365-2435.12626
41. Sladecek V. System of water quality from the biological point of view. Adv Limnol. 1973;7:1-218. https://doi.org/10.1002/iroh.19740590412
42. Madoni P. Ciliated protozoan communities and saprobic evaluation of water quality in the hilly zone of some tributaries of the Po River (northern Italy). Hydrobiologia. 2005;541(1):55-69. https://doi.org/10.1007/s10750-004-4667-8
43. Bonatti TR, Siqueira Castro ICV, Franco RMB. Checklist of ciliated protozoa from surface water and sediment samples of Atibaia River, Campinas, São Paulo (Southeast Brazil). Rev Bras Zoocienc. 2016;17(1):63-76. Avaliable in: https://periodicos.ufjf.br/index.php/zoociencias/article/view/24631
44. Yamashiro S. Removal of Giardia and Cryptosporidium, occurrence of microsporidia and characterization of microfauna from a combined system for wastewater treatment. [tese de doutorado].Campinas (SP): Universidade Estadual de Campinas; 2016. Avaliable in: http://repositorio.unicamp.br/jspui/handle/REPOSIP/332149
45. Siqueira-Castro ICV, Greinert-Goulart JA, Rosseto R, Guimarães JR, Franco RMB. Ciliated protozoa community of a combined UASB activated sludge system in southeastern Brazil. Environ Sci Pollut Res. 2016;23:23804-14. https://doi.org/10.1007/s11356-016-7591-3
46. Macarisin D, Bauchan G, Fayer R. Spinacia oleracea L. leaf stomata harboring Cryptosporidium parvum oocysts: a potential threat to Food Safety. Appl Environ Microbiol. 2010;76(2):555-9. https://doi.org/10.1128/AEM.02118-09
47. Machado ER, Maldonade IR, Riquette RFR, Mendes VS, Gurgel-Gonçalves R, Ginani VC. Frequency of enteroparasites and bacteria in the leafy vegetables sold in brazilian public wholesale markets. J Food Prot. 2018;81(4):542-8. https://doi.org/10.4315/0362-028X.JFP-17-358
48. Ministério da Saúde (BR). Secretaria de Vigilância em Saúde. Vigilância Epidemiológica das Doenças Transmitidas por Alimentos. Alves, R. III Encontro Nacional de Vigilâncias Sanitárias; agosto de 2014; São Paulo, SP. [access 2017 may 01]. Avaliable in: http://www.anrbrasil.org.br/new/pdfs/2014/3_PAINEL_ApresentacaoRejaneAlvesVigilanciaEpidemiologica-VE-DTA_Agosto_2014_PDF.pdf
49. Decol LT, Casarin LS, Hessel CT, Batista ACF, Allende A, Tondo EC. Microbial quality of irrigation water used in leafy green production in Southern Brazil and its relationship with produce safety. Food Microbiol. 2017;65:101-13. https://doi.org/10.1016/j.fm.2017.02.003
50. Cotterill FPD, Al-Rasheid K, Foissner W. Conservation of protests: is it needed at all? Biodivers Coserv. 2008: 17:427-43. https://doi.org/10.1007/s10531-007-9261-8
51. Robertson LJ, Van der Giessen JWB, Batz MB, Kojima M, Cahill S. Have foodborne parasites finally become a global concern? Trends Parasitol. 2013;29 (3):101-3. https://doi.org/10.1016/j.pt.2012.12.004
Este trabalho está licenciado sob uma licença Creative Commons Attribution 4.0 International License.
Copyright (c) 2020 Taís Rondello Bonatti, Sandra Yamashiro, Pedro Ferreira de Castro Santana e Freitas, Isabel Cristina Vidal Siqueira-Castro, Regina Maura Bueno Franco