MicroRNAs as serum biomarker for Senecio brasiliensis poisoning in cattle

https://doi.org/10.1016/j.etap.2022.103906Get rights and content

Highlights

  • MiR-30a, −378, −21, −885, and −122 were more expressed in intoxicated animals.

  • MiR-122, −885 and −21 signatures demonstrated high sensitivity and specificity.

  • MiRNAs can be used as biomarkers for minimally invasive seneciosis diagnostic.

Abstract

Senecio spp. is one of the most frequent plant-related poisonings in cattle. Its ingestion generates the disease seneciosis, characterized by hepatic damages. Liver biopsies and serum markers dosage are tools used in diagnosis; however, many breeding cattle are undiagnosed. MicroRNAs are non-coding RNA, stable in biological fluids. Their difference in expression levels may indicate the presence of the poisoning. We analyzed the miRNA profiling to identify potential diagnostic biomarkers for Senecio brasiliensis poisoning. The expression of miR-21, miR-885, miR-122, miR-181b, miR-30a, miR-378, and let-7 f were evaluated in the serum of exposed cattle. At least one histological change was found in liver and lower quantity of albumin and high AST and ALP were also detected. MiRNAs miR-30a, miR-378, miR-21, miR-885, and miR-122 presented significantly higher expression in intoxicated animals than in healthy animals. Furthermore, miR-122, miR-885, and, especially, miR-21 signatures demonstrated high sensitivity and specificity, with potential application for detecting poisoning.

Introduction

Poisonous plants and their secondary metabolites are responsible for many cases of poisoning in cattle worldwide. It is established that Senecio spp. is one of the most frequent plant-related poisonings in animal production. It is an important problem in countries such as Brazil (Molossi et al., 2021) and Uruguay (García et al., 2020), as well as in Europe (Cortinovis and Caloni, 2013, Cortinovis and Caloni, 2015a), Australia, and North America (Gardner et al., 2006). The Senecio genus (Compositae family) is composed of more than 1200 species distributed worldwide, 25 of which have been confirmed to be poisonous. These plants contain a series of pyrrolizidine alkaloids (PAs) which become cumulative hepatotoxins after they are bioactivated to pyrrole metabolites in the liver (Cortinovis and Caloni, 2015a).

The clinical, anatomopathological, and histopathological profiles of Senecio spp. are quite characteristic in acute and subacute intoxications. Liver function tests and histological studies of liver biopsies are used to establish a prognosis in animals when there is mistrust of seneciosis (Barros et al., 2007, Giaretta et al., 2014b). Although hepatic biopsy is considered the optimal test for diagnosing seneciosis, it is an invasive method that must be performed by an experienced professional, and tissue processing is time-consuming, thus delaying clinical decision (Barros et al., 2007, Emanuelli et al., 2016).

An important aspect of public health is the transfer of toxins to humans through the ingestion of meat, milk, and other derivatives. The PAs can be eliminated in milk (Hoogenboom et al., 2011). A study showed that goats' milk poisoned with Crotalaria spectabilis, which contain PAs, induced toxicity in rats (MEDEIROS et al., 1999). The mutagenic and teratogenic effects of Senecio spp. alkaloids in milk should be considered, although there are no reports of diseases caused in humans due to their consumption (Cheeke, 1988). This problem becomes more specific when poisoned cattle do not present clinical signs.

In this context, microRNAs (miRNA) detection has aroused interest by playing an important role in veterinary medicine (Chodkowska et al., 2017, Do et al., 2021, Dong et al., 2017, Farrell et al., 2015, Mukiibi et al., 2020, Shaughnessy et al., 2020). The miRNAs are small 19–23 nucleotide non-coding RNA molecules that modulate approximately 30% of mRNA, with fundamental roles in proliferation, differentiation, and apoptosis mechanisms (Ambros, 2004, Chi et al., 2009). According to information from the largest miRNA database (miRBase), up to now, 1025 miRNA have been described in cattle (Griffiths-Jones, 2006). Currently, miRNAs have already been studied as biomarkers for the rapid diagnosis of mastitis in dairy cows (Li et al., 2015b) and as markers for hepatic function in anabolic use (Becker et al., 2011b), early gestation (Ioannidis and Donadeu, 2017), milk quality (Wang et al., 2016a), early detection of Mycobacterium avium (Britton et al., 2016), among other applications. A feature of great interest in using miRNA as diagnostic biomarkers is their presence in biological fluids such as serum, urine, milk, and saliva in a stable manner, allowing a less invasive detection (Sempere, 2012, Wang et al., 2016b).

Thus, the study of new forms of diagnosis is necessary and of great interest to the area. Therefore, this work aimed to identify a miRNA profile in cattle serum intoxicated to Senecio brasiliensis. The study of miRNA in the serum of animals poisoned by Senecio spp. may improve the knowledge of the disease, organs involved, degree of poisoning, and prognosis, allowing a greater understanding of the clinical signs.

Section snippets

Animals

This study analyzed naturally exposed animals that were assisted by veterinary doctors of the university (Adriano T. Ramos and André L.F. Goetten). Animals were kept in native pasture, not receiving ration. No experimental protocol with animals were performed, they received medical attention and diagnosis was done afterwards.

Toxicosis event

The cattle consisted of Aberdeen Angus cows (36−48 months old), reared in Curitibanos, SC, Brazil. The extensive grazing of beef cattle in these areas, is the most used

Animals

Animals presenting at least one histopathological alteration were considered intoxicated (20 animals), and those with no hepatic damage were grouped as control (eight healthy animals).

Serum miRNA expression

Relative expression of miR-30a, mir-378, mir-21, mir-122, and miR-885 were higher in intoxicated animals than in healthy animals (Fig. 1 B-E, G). No changes were observed in let-7f and miR-181b levels (Fig. 1 A, F).

Diagnostic value of the miR-30a, miR-378, miR-21, miR-885 and miR-122 in serum

Receiver operating characteristic (ROC) analyses were performed to explore the diagnostic value of

Discussion

Seneciosis is one of the most frequent plant-related poisonings in animal production that result in hepatotoxicity (Cortinovis and Caloni, 2015b, Molossi et al., 2021). Poisonous plants like Senecio spp. can induce the death of animals. However, its effects often remain unmeasured and are not easily identified. James et al. (2019) reports that big economic losses are associated with undiagnosed intoxications resulting in low animal performance, immunodeficiency, reduction in reproductive

Conclusion

In summary, our study analyzed miRNAs expressed differently between intoxicated and healthy animals and detected five upregulated miRNAs in the serum animals, allowing significant correlations. Furthermore, the sensitivity and specificity of the miRNAs by ROC analysis indicated that the miR-122, miR-885, and, especially, miR-21 signatures have potential application for detecting cattle poisoning by Senecio spp. Although the present study has some limitations, including the small sample size, it

CRediT authorship contribution statement

Evelyn Winter: Conceptualization, Writing – original draft, Supervision, Project administration. Julia Cisilotto: Investigation. André L.F. Goetten: Investigation. Ângela Veiga: Investigation. Adriano T. Ramos: Investigation. Francielli C. Zimermann: Investigation. Carolina Reck: Investigation. Tânia B. Creczynski-Pasa: Resources, Writing – review & editing.

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

This study was supported by FAPESC (Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina) grants ID 2017TR1702, by CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico), Brazil, which provided grant to Tânia B. Creczynski-Pasa, as well as by CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), Brazil, which provided PNPD-fellowships for Júlia Cisilotto.

References (105)

  • Y.C. Lai et al.

    Bovine serum miR-21 expression affected by mastitis

    Res Vet. Sci.

    (2021)
  • D. Löffler et al.

    Interleukin-6 dependent survival of multiple myeloma cells involves the Stat3-mediated induction of microRNA-21 through a highly conserved enhancer

    Blood

    (2007)
  • L. Ma et al.

    miRNA-21-3p targeting of FGF2 suppresses autophagy of bovine ovarian granulosa cells through AKT/mTOR pathway

    Theriogenology

    (2020)
  • R.T. Marquez et al.

    Correlation between microRNA expression levels and clinical parameters associated with chronic hepatitis C viral infection in humans

    Lab. Investig.

    (2010)
  • T. Mohri et al.

    Human CYP2E1 is regulated by miR-378

    Biochem. Pharmacol.

    (2010)
  • M.Z. Nasser et al.

    Circulating microRNAs (miR-21, miR-223, miR-885-5p) along the clinical spectrum of HCV-related chronic liver disease in Egyptian patients

    Arab J. Gastroenterol.

    (2019)
  • H. Tong et al.

    bta-miR-378 promote the differentiation of bovine skeletal muscle-derived satellite cells

    Gene

    (2018)
  • P. Vegh et al.

    Profiling microRNA expression in bovine alveolar macrophages using RNA-seq

    Vet. Immunol. Immunopathol.

    (2013)
  • B. Wang et al.

    miR-181b promotes hepatic stellate cells proliferation by targeting p27 and is elevated in the serum of cirrhosis patients

    Biochem. Biophys. Res. Commun.

    (2012)
  • E.A. Afanasyeva et al.

    MicroRNA miR-885-5p targets CDK2 and MCM5, activates p53 and inhibits proliferation and survival

    Cell Death Differ.

    (2011)
  • W. Al-Husseini et al.

    Characterization and profiling of liver microRNAs by RNA-sequencing in cattle divergently selected for residual feed intake

    Asian-Australas. J. Anim. Sci.

    (2016)
  • V. Ambros

    The functions of animal microRNAs

    Nature

    (2004)
  • C.L. Andersen et al.

    Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets

    Cancer Res.

    (2004)
  • C.S.L. Barros et al.

    Biópsia hepática no diagnóstico da intoxicação por Senecio brasiliensis (Asteraceae) em bovinos

    Pesqui. Veter. Bras.

    (2007)
  • P.R Giaretta et al.

    Seneciosis in cattle associated with photosensitization

    Pesq. Vet. Bras.

    (2014)
  • P.R. Giaretta et al.

    Clinical and pathological aspects of chronic Senecio spp. poisoning in sheep. Pesq.

    Vet. Bras.

    (2014)
  • C. Becker et al.

    Changes in the miRNA profile under the influence of anabolic steroids in bovine liver

    Analyst

    (2011)
  • C. Becker et al.

    Changes in the miRNA profile under the influence of anabolic steroids in bovine liver

    Analyst

    (2011)
  • V. Bihrer et al.

    Serum miR-122 as a biomarker of necroinflammation in patients with chronic hepatitis C virus infection

    Am. J. Gastroenterol.

    (2011)
  • T. Buza et al.

    Computational prediction of disease microRNAs in domestic animals

    BMC Res. Notes

    (2014)
  • S. Cermelli et al.

    Circulating microRNAs in patients with chronic hepatitis C and non-alcoholic fatty liver disease

    PloS One

    (2011)
  • L. Chang et al.

    miRNet 2.0: network-based visual analytics for miRNA functional analysis and systems biology

    Nucleic Acids Res.

    (2020)
  • P.R. Cheeke

    Toxicity and metabolism of pyrrolizidine alkaloids

    J. Anim. Sci.

    (1988)
  • S.W. Chi et al.

    Argonaute HITS-CLIP decodes microRNA-mRNA interaction maps

    Nature

    (2009)
  • K.A. Chodkowska et al.

    MicroRNA function in domestic animal physiology and diseases: a promising diagnostic tool for veterinary use

    Med. Weter.

    (2017)
  • S.D. Clark et al.

    Abnormal expression of miR-21 in kidney tissue of dogs with X-Linked hereditary nephropathy: a canine model of chronic kidney disease

    Vet. Pathol.

    (2019)
  • C. Cortinovis et al.

    Alkaloid-containing plants poisonous to cattle and horses in Europe

    Toxins

    (2015)
  • C. Cortinovis et al.

    Alkaloid-containing plants poisonous to cattle and horses in Europe

    Toxins

    (2015)
  • C. Coulouarn et al.

    Loss of miR-122 expression in liver cancer correlates with suppression of the hepatic phenotype and gain of metastatic properties

    Oncogene

    (2009)
  • A.M. Craig et al.

    Serum liver enzyme and histopathologic changes in calves with chronic and chronic-delayed Senecio jacobaea toxicosis

    Am. J. Vet. Res.

    (1991)
  • K. Dirksen et al.

    Use of serum MicroRNAs as biomarker for hepatobiliary diseases in dogs

    J. Vet. Intern. Med.

    (2016)
  • Do, D.N., Dudemaine, P.L., Mathur, M., 2021. miRNA Regulatory Functions in Farm Animal Diseases, and Biomarker...
  • H. Dong et al.

    Circulating microRNAs as potential biomarkers for veterinary infectious diseases

    Front. Vet. Sci.

    (2017)
  • D. Driemeier et al.

    Intoxicação experimental por Senecio oxyphyllus (Compositae) em bovinos

    Pesq. Vet. Bras.

    (1992)
  • W. Panziera et al.

    Senecio brasiliensis poisoning in calves: pattern and evolution of hepatic lesions

    Pesq. Vet. Bras.

    (2017)
  • Elsaeid Elnour, I., Dong, D., Wang, X., Zhansaya, T., Khan, R., 2020. Bta-miR-885 promotes proliferation and inhibits...
  • S.R. Eman et al.

    Evaluation of hepatocyte-derived microRNA-122 for diagnosis of acute and chronic hepatitis of dogs

    Vet. World

    (2018)
  • M.P. Emanuelli et al.

    What is your diagnosis? Liver from a cow

    Vet. Clin. Pathol.

    (2016)
  • D. Farrell et al.

    The identification of circulating MiRNA in Bovine serum and their potential as novel biomarkers of early mycobacterium avium subsp paratuberculosis infection

    PloS One

    (2015)
  • A. Fatima et al.

    The miRNAome of the postpartum dairy cow liver in negative energy balance

    BMC Genom.

    (2014)
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