The Influence of Pesticides on the Biology and Physiology of the Land
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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-4, Issue-5, Sep-Oct- 2019 https://dx.doi.org/10.22161/ijeab.45.23 ISSN: 2456-1878 The Influence of Pesticides on the Biology and Physiology of the Land Snail Bulimulus tenuissimus (Orbigny, 1935) Iza Patrício1, Florence Gonçalves Martins2, Tatiane Cristina dos Santos Bonfim2, Mauricio Carvalho de Vasconcellos1, Jairo Pinheiro3, Marta Julia Faro4, Clélia Christina Mello-Silva1 1Grupo de pesquisa em Saúde e Educação Ambiental Crítica com Ênfase nas Relações Parasitárias do Laboratório de Avaliação e Promoção da Saúde Ambiental, IOC, FIOCRUZ, Av. Brasil 4365, Manguinhos, 21040-900, RJ, Brazil Email: [email protected], [email protected], [email protected] 2Curso de Pós-Graduação em Ciências Veterinárias, Instituto de Veterinária, UFRRJ, BR 465, km 47, 23897-000, Seropédica, RJ, Brazil Email: [email protected], [email protected] 3Departamento de Ciências Fisiológicas, Instituto de Ciências Biológicas e da Saúde, UFRRJ, BR 465, km 47, 23897-000, Seropédica, RJ, Brazil Email: [email protected] 4Laboratório de Biologia e Parasitologia de Mamíferos Silvestres Reservatórios, IOC, FIOCRUZ, Av. Brazil 4365, Manguinhos, 21040- 900 RJ, Brazil Email: [email protected] Corresponding author: [email protected] Abstract— Some terrestrial gastropods are agricultural pests and are exposed to toxic products capable of causing physiological and biological alterations. The action of some herbicides and molluscicides has been studied on freshwater snails, but there are few studies of terrestrial species. This study analyzed the biological and physiological responses of Bulimulus tenuissimus exposed to the natural molluscicidal latex of Euphorbia milii var. hislopii and the synthetic herbicide Roundup® Original. A total of 210 snails were divided and exposed for 24 hours to the products (latex or herbicide in concentrations of 1,000 ppm and 100,000 ppm) and type 2 water. Of this total, 150 exposed animals were analyzed biochemically after 24 hours, and the others were observed during 30 days for analysis of the mortality rate. After 24 hours, some snails presented escape mechanisms such as epiphragm formation, cephalopedal mass retraction and burial. The latex did not cause metabolic alterations but the herbicide altered the carbohydrate metabolism of B. tenuissimus, reducing energy substrates in the sites analyzed (hemolymph, digestive gland and cephalopedal mass). Regarding the protein metabolism, there was no alteration after exposure to both analyzed substances. After 30 days of exposure, higher mortality was recorded for the groups exposed to the herbicide. These findings suggest that B. tenuissimus is resistant to the molluscicidal latex of Euphorbia milii var. hislopii more than the herbicide Roundup® Original. Keywords— Bulimulus tenuissimus, carbohydrate metabolism, Euphorbia milii var. hislopii, Roundup® Original. I. INTRODUCTION Institute of Geography and Statistics (IBGE) [3], with Agrotoxics are substances used to increase agricultural numerous active principles and commercial formulations, productivity through pest and disease control [1]. which can be classified according the action on the target However, their indiscriminate use can cause serious organism and the chemical group to which they belong [2]. damage to the environment and to human health, as well as Of the agrotoxics used in Brazil, 56% are herbicides [4]. putting selective pressure for development of resistance by Among the commonly used Roundup® stands out, which pests [2]. has as its active ingredient Glyphosate (N- Brazil has been the largest consumer of agrotoxics in the (phosphonomethyl) glycine) combined with a surfactant world since 2008 according to data from the Brazilian that facilitates transport by the plant. It is a non-selective www.ijeab.com Page | 1433 International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-4, Issue-5, Sep-Oct- 2019 https://dx.doi.org/10.22161/ijeab.45.23 ISSN: 2456-1878 herbicide, effective in the total destruction of weeds [5], of B. tenuissimus exposed to the natural molluscicide latex and in the control of plants in reforestation, gardens and of E. milii var. hislopii and the synthetic herbicide aquatic plants in fish farming tanks and in lakes [6]. From Roundup® Original. the Roundup® herbicide several formulations were commercialized by the Monsanto Company, one of them is II. MATERIAL AND METHODS Roundup® Original, which presents polyoxyethylene 2.1. Determining concentrations of E. milii var. hislopii amine surfactant (POEA), which helps the herbicide to latex and Roundup® Original penetrate the leaves of plants [7]. Some herbicides have The crude latex extract was collected in the Ilha do deleterious effects on limnic mollusks, interfering at the Governador district (22°48´09´´S/ 43°12´35´´W) of the cellular and physiological, molecular level to reduce the city of Rio de Janeiro, Brazil. Determination of the survival rate and reproductive activity [8, 9, 10]. concentrations of the two substances was based on the Molluscicide is the classification of agrotoxic for control procedures described by Vasconcellos and Amorim [21], of mollusks present in gardens, fields and greenhouses, and based on pre-tests, concentrations of 1,000 ppm (parts including those that are intermediate hosts of parasites in per million) (0.1%) and 100,000 ppm (10%) for both aquatic environments, such as snails that transmit substances were chosen (mg/L for latex and ml/L for schistosomiasis. Poisoning by such substances can cause herbicide). osmotic imbalance, and as a consequence two mechanisms that cause mortality may occur: retraction of the 2.2. Formation of groups cephalopedal mass with the release of hemolymph and/or The parents of the snails used were obtained in a garden abnormal projection out of the shell [11], called “distress located in the municipality of Seropédica, Rio de Janeiro, syndrome”. Brazil (about: 22º 44' 38" S; 43º 42' 27" W). A total of 210 Molluscicides can be synthetic or natural. Chemical hatched mollusks were kept in the Laboratory for control of terrestrial species occurs mainly through Evaluation and Promotion of Environmental Health of synthetic molluscicides. Although effective, these Fiocruz in Rio de Janeiro, where they were monitored until compounds also act on non-target organisms, causing reaching sexual maturity (110 days). significant changes in ecosystems [12]. As a consequence, For the 24-hour and 30-day follow-up, the snails were there is increasing demand for readily biodegradable divided into five groups, with 30 and 12 specimens each, substances as an alternative to synthetic molluscicides respectively: two groups exposed to latex (L) at [13]. concentrations of 1,000 ppm and 100,000 ppm (L1,000 The latex of Euphorbia milii var. hislopii (N. E. B.) and L100,000, respectively); two groups exposed to (Euphorbiaceae), popularly known as crown of thorns, Roundup® Original (R) (R1,000 and R100,000, Christ plant, or Christ thorn, is considered one of the most respectively); and one control group sprayed with type 2 promising Brazilian molluscicides. In laboratory and field water. conditions it complies with the recommendations of the 2.3. Exposure World Health Organization (WHO) for use as a natural For the 24-h follow-up, 150 mollusks were exposed molluscicide, in addition to being biodegradable and less individually in disposable flasks with a capacity of 150 ml hazardous to non-target organisms [14], with effective containing 50 g of sterile soil moistened with water. results on limnic mollusks and amphibians [15]. Addition of the compounds was done by spraying the The land snail Bulimulus tenuissimus (Orbigny, 1935) is concentrations prepared for each product using 5 ml glass present in many Brazilian regions [16], where it is hand sprayers. Each mollusk received 2 ml of the product considered an agricultural pest. It is also the only species corresponding to its group, under a fume hood. Twenty- in the family Bulimulidae cited as an intermediate host of four hours after exposure, the specimens were observed some parasites of poultry [17]. Recently, studies showed and dissected for the collection of biological samples. that this mollusk is host of the nematode Angiostrongylus For the 30-day follow-up, the exposure occurred plastic cantonensis (Chen, 1935) [18,19], a rodent lung parasite terrariums with 12 cm in diameter and 8 cm in depth, capable of infecting humans and causing meningitis, containing 3 cm of moistened sterilized soil as substrate. characterized by eosinophilic inflammation [20]. Each terrarium received 24 ml of the substance Studies of synthetic or natural substances for the control of corresponding to its group through glass sprayers: 2 ml of terrestrial species are scarce, as well as the physiological the corresponding substance was sprayed onto each changes induced by these substances. For this reason, this mollusk, under a fume hood. They were observed every study analyzed the biological and physiological responses www.ijeab.com Page | 1434 International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-4, Issue-5, Sep-Oct- 2019 https://dx.doi.org/10.22161/ijeab.45.23 ISSN: 2456-1878 other day for 30 days (four weeks) after exposure to III. RESULTS determine the mortality rate. After 24 hours of exposure, exhaust mechanisms, In both cases, the control groups were sprayed with 2 ml of characterized by increased mucus