Changes in Hepatic Metabolism of Rattus norvegicus Infected to Angiostrongylus cantonensis (Nematoda) and Exposed to Glyphosate-Based Herbicide

Special Article - Herbicides

Ann Agric Crop Sci. 2021; 6(6): 1092.

Changes in Hepatic Metabolism of Rattus norvegicus Infected to Angiostrongylus cantonensis (Nematoda) and Exposed to Glyphosate-Based Herbicide

Braga BV1,2, Garcia JS2*, Simões RO3, Silva JSP4, Castro LS4, Silva CH5 and Maldonado Júnior A2

1Programa de Pós-Graduação em Biologia Parasitária, Instituto Oswaldo Cruz-FIOCRUZ, Rio de Janeiro, RJ, Brazil

2Laboratório de Biologia e Parasitologia de Mamíferos Silvestres Reservatórios, Instituto Oswaldo Cruz-FIOCRUZ, Rio de Janeiro, RJ, Brazil

3Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, RJ, Brazil

4Laboratório de Fisiologia das Relações Parasitárias, Departamento de Ciências Fisiológicas, Instituto de Ciências Biológicas e da Saúde da Universidade Federal Rural do Rio de Janeiro, RJ, Brazil

5Instituto de Ciência e Tecnologia em Biomodelos-FIOCRUZ, Rio de Janeiro, RJ, Brazil

*Corresponding author: Gracia JS, Laboratório de Biologia e Parasitologia de Mamíferos Silvestres Reservatório, Instituto Oswaldo Cruz-FIOCRUZ, Av. Brazil 4365, 21040-900, Rio de Janeiro, RJ, Brazil

Received: July 27, 2021; Accepted: August 26, 2021; Published: September 02, 2021

Abstract

Helminth infection associated with exposure to pesticides has received little attention regarding its effect on the human population and on farm and wild animals. The aim of this study was to evaluate the effects a glyphosate-based herbicide on the hepatic and glycemic metabolism of Rattus norvegicus (Wistar) infected by Angiostrongylus cantonensis. Experimental groups were orally infected with 50 L3 larvae of A. cantonensis and exposed to the herbicide after and before the infection. Biochemical serum analyses were carried out to determine the levels of Aspartate Aminotransferase (AST), Alanine Aminotransferase (ALT), Alkaline Phosphatase (ALP), Total Bilirubin (TB), total protein, albumin, urea, creatinine, uric acid, glucose and hepatic glycogen. All exposed groups showed an increase in the concentration of glycogen, AST, ALT and TB, the last ones suggesting liver tissue damage. Exposure to the herbicide caused hyperalbuminemia as an antioxidant response to the herbicide. These findings contribute to a better understanding of how glyphosate-based herbicides can change the hepatic metabolism the vertebrate and to influence the parasite-host relationship.

Keywords: Biochemical hepatic parameters; Experimental Infection; Helminth; Roundup®

Abbreviations

ALT: Alanine Aminotransferase; AST: Aspartate Aminotransferase; ALP: Alkaline Phosphatase; TB: Total Bilirubin; TP: Total Protein; L3: Third-Stage Larvae; LABPMR: Laboratory of Biology and Parasitology of Wild Mammal Reservoirs; IOCFIOCRUZ: Oswaldo Cruz Institute-FIOCRUZ; C: Control Group; AI: Acute Infection; CI: Chronic Infection; pi: Post-Infection; IHE: Immediate Herbicide Effect, DHE: Delayed Herbicide Effect; I + E; E+I; CEUA-IOC: Ethics Committee on the Use of Animals of Oswaldo Cruz Institute; ANOVA: Analysis Of Variance

Introduction

The active ingredient glyphosate (N-(phosphonomethyl) glycine) is one of the most used herbicides in the world, in several commercial formulations [1-4]. Glyphosate-based products have been tested in conventional experimental models, such as rats and mice, and the results have confirmed their toxic potential and ability to cause metabolic changes [5-9]. Several helminth species have been used as experimental models in herbicide tests to check their effect on parasite biology and influence on the parasite-host relationship [10-13].

Angiostrongylus cantonensis (Chen, 1935) is an endemic nematode in South and Southeast Asia and the Pacific Islands [14-15], and it is now observed in North and South America, the Caribbean, Africa, and Australia [16]. This wide distribution is particularly due to the presence in these locations of several species of gastropod molluscs susceptible to infection and that act as intermediate hosts [17-19], along with the presence of synanthropic rodents (Rattus rattus Linnaeus, 1758, and Rattus norvegicus Berkenhout, 1769) as definitive host. Under experimental conditions, infection by A. cantonensis can compromise the health of animals, depending mainly on the parasite burden and the animals’ immunological condition [20-22].

Humans become infected through ingestion of raw or undercooked foods contaminated with third-stage larvae (L3) [16,23,24]. However, they are considered accidental hosts because helminths do not develop to the adult stage and are retained in the meninges. This parasite causes eosinophilic neuromeningitis in humans, the main clinical symptom of the infection, which can lead to death if not properly treated. It is thus considered a public health problem in endemic areas [16,25,26].

Based on data on the toxic effect of glyphosate on rodents and helminths, Kelly et al. [27] suggested that animals exposed to this herbicide can become more susceptible to infection due to their lower resistance. To test this hypothesis, we evaluated the effects of the commercial formulation Roundup® on biochemical parameters of Wistar rats (R. norvegicus) infected by A. cantonensis.

Materials and Methods

Parasites and experimental infection

The strain of A. cantonensis used in experimental infections was isolated from the specimens of the snail Achatina fulica collected in the municipality of São Gonçalo (state of Rio de Janeiro, Brazil) (22°49’37’’S, 43°03’14”W) in 2014. Since then, the biological cycle has been maintained under experimental conditions at the Laboratory of Biology and Parasitology of Wild Mammal Reservoirs (LABPMR) of Oswaldo Cruz Institute (IOC/FIOCRUZ), using specimens of R. norvegicus (Wistar) and Biomphalaria glabrata as definitive and intermediate hosts, respectively. For the experimental infection, individual 3-month-old female rodents weighing 260g were infected by orogastric gavage with 50 third-stage larvae (L3) of A. cantonensis, previously recovered from B. glabrata specimens, using the sedimentation technique of Baermann & Moraes [28].

Roundup® concentration and rodent exposure

The glyphosate-based herbicide used in the assay was the commercial formulation Roundup® (480g/L isopropylamine salt N-(phosphonomethyl) glycine; 360g/L equivalent acid N-(phosphonomethyl) glycine; 684g/L inert ingredients) produced by Monsanto do Brasil Ltda. The animals were exposed daily, by orogastric gavage, to 500mg/kg body weight, for 15 days [7]. This concentration is considered not to cause adverse effects [29].

Experimental groups

Seven groups of 10 R. norvegicus (Wistar) females were formed: 1-control (C): not infected and not exposed, treated with 500μl of dechlorinated water; 2 and 3- Acute Infection (AI) and Chronic Infection (CI) both groups orally infected with 50 L3 of A. cantonensis, in a volume of 500μl of dechlorinated water, and analyzed 15 and 50 days Post-Infection (pi), respectively; 4 and 5- groups exposed daily to the herbicide for 15 days and analyzed those same 15 days (Immediate Herbicide Effect-IHE) and 65 days later (Delayed Herbicide Effect-DHE); 6-infected with 50 L3 of A. cantonensis in 500uL and exposed daily to the herbicide for 15 days after infection (I+E); 7- exposed to the herbicide daily for 15 days and 24 hours after the last administration, infected with 50 L3 of A. cantonensis in 500 uL of dechlorinated water (E+I). These last two groups (E+I and I+E) were analyzed 50 pi (Figure 1).

Citation:Braga BV, Garcia JS, Simões RO, Silva JSP, Castro LS, Silva CH, et al. Changes in Hepatic Metabolism of Rattus norvegicus Infected to Angiostrongylus cantonensis (Nematoda) and Exposed to Glyphosate-Based Herbicide. Ann Agric Crop Sci. 2021; 6(6): 1092.