Characterization of protein profile and evaluation of immune-peptide of crude and Excretory/Secretory antigens of some haematophagus leeches of India

P  PATHAK; S  ISLAM; J  BAM; D J  KALITA; S  BORAH; G  DAS

doi:10.56093/ijans.v95i8.160151

Authors

P PATHAK Lakhimpur College of Veterinary Science, Assam Agricultural University, Joyhing, North Lakhimpur, Assam, India
S ISLAM College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, Assam
J BAM ICAR-Research Centre for North-Eastern Hill Region Arunachal Pradesh Centre, Basar 791 101, Arunachal Pradesh
D J KALITA College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, Assam
S BORAH Lakhimpur College of Veterinary Science, Assam Agricultural University, Joyhing, North Lakhimpur, Assam
G DAS Subject Matter Specialist, Krishi Vigyan Kendra, Bongaigaon, Assam Agricultural University, Assam

https://doi.org/10.56093/ijans.v95i8.160151

Keywords:

Antigen, Excretory/ Secretory, Haematophagous leech, Protein

Abstract

Leech infestation leads to significant blood loss compared to other parasites, causes irritation in humans and predisposes livestock to secondary fly attack and maggot infestation. During blood feeding, leeches inject saliva containing bioactive proteins at the bite site that facilitate continuous blood flow and provoke inflammatory responses. The present study was undertaken to characterize the crude and excretory/secretory (E/S) antigens of three common leech species of Northeast India- two aquatic (Hirudinaria manillensis and H. granulosa) and one terrestrial (Haemadipsa sylvestris). Protein quantification revealed higher concentrations of crude antigens (3.60–4.14 g/dL) compared to E/S antigens (3.39–3.44 g/dl). Electrophoretic analysis showed 9 peptide bands in crude antigens (12.5-96 kDa) and five bands in E/S antigens (14-96 kDa), with no major differences in overall banding patterns. Periodic acid–Schiff (PAS) staining revealed the presence of two glycoproteins in the crude extracts of H. manillensis and H. granulosa and three in H. sylvestris (12.5–26 kDa). Glycoproteins could not be detected in E/S products of any of the three species. Hyperimmune serum raised in rabbits against both crude and E/S antigen was used for Enzyme immunotransfer blot (EITB) analysis, which detected two immune-reactive peptides in the crude antigens of H. manillensis and H. granulosa, and one in the E/S product of H. sylvestris.

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References

Abdualkader AM, Ghawi AM, Alaama M, Awang M and Merzouk A. 2013. Characterization and optimization of lyophilization and storage conditions of leech saliva extract from the tropical leech Hirudinaria manillensis. Pakistan Journal of Pharmaceutical Sciences 26(3): 525–35.

Alaama M, Alnajjar M, Abdualkader AM, Mohammad A and Merzouk A. 2011. Isolation and analytical characterization of local Malaysian leech saliva extracts. IIUM Engineering Journal 12(4): 51–59. DOI: https://doi.org/10.31436/iiumej.v12i4.156

Baskova I P, Kostrjukova E S, Vlasova M A, Kharitonova O V, Levitskiy S A, Zavalova L L, Moshkovskii S A and Lazarev V N. 2008. Proteins and peptides of the salivary gland secretion of medicinal leeches Hirudo verbana, H. medicinalis and H. orientalis. Biochemistry (Moscow) 3: 315–20. DOI: https://doi.org/10.1134/S0006297908030127

Baskova I P, Zavalova L L, Basanova A V, Moshkovskii S A and Zgoda VG 2004. Protein profiling of the medicinal leech salivary gland secretion by proteomic analytical methods. Biochemistry (Moscow) 69(7): 770–75. DOI: https://doi.org/10.1023/B:BIRY.0000040202.21965.2a

Fredric R and Govedich WE (2004). Euhirudinea (leeches). In: Freshwater Fauna of Malaysia and Borneo (YH Sen ed.). Academy of Sciences Malaysia, Kuala Lumpur, pp 175–90.

Harlow E and Lane D (1988). Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, New York, p 726.

Huang J and Honda W (2006). CED: A conformational epitope database. BMC Immunology 7: 7. DOI: https://doi.org/10.1186/1471-2172-7-7

Kose A (2008). Leech bites: Massive bleeding, coagulation profile disorders, and severe anemia. The American Journal of Emergency Medicine 26(9): 1067.e3–e6. DOI: https://doi.org/10.1016/j.ajem.2008.03.022

Laemmli UK 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 222: 680–85. DOI: https://doi.org/10.1038/227680a0

Lemke S, Muller C, Lipke E, Uhl G and Hildebrandt JP 2013. May salivary gland secretory proteins from hematophagous leeches (Hirudoverbana) reach pharmacologically relevant concentrations in the vertebrate host? PLoS One 8(9): e73809. DOI: https://doi.org/10.1371/journal.pone.0073809

Lok U, Bozkurt S, Okur M, Gulacti U and Hatipoglu S (2013). A rare case of adverse effects caused by leech bite. American Journal of Case Reports 14: 191–93. DOI: https://doi.org/10.12659/AJCR.883936

Markwardt F 1957. Die isolation and chemical characterization of hirudin. Hoppe-Seyler’s Zeitschriftfür Physiologische Chemie 308(2-4): 147–56. DOI: https://doi.org/10.1515/bchm2.1957.308.1.147

Najjari M, Solgi R and Kareshk AT 2022. Cellulitis caused by Hirudoorientalis bites that lead to an allergic reaction. Case Reports in Infectious Diseases 5493057. DOI: https://doi.org/10.1155/2022/5493057

Pathak P, Islam S, Bam J, Bhattacharya D, Das M, Kalita D J, Saikia GK, Das G and Bordoloi G 2020. Distribution of aquatic and terrestrial leeches in Assam, India. Journal of Entomology and Zoology Studies 8(4): 605–09.

Sawyer RT 1986. Leech Biology and Behaviour: Volume II: Feeding Biology, Ecology, and Systematics. Oxford University Press, New York.

Singh AP 2010. Medicinal leech therapy (Hirudotherapy): A brief overview. Complementary Therapies in Clinical Practice 16(4): 213–15. DOI: https://doi.org/10.1016/j.ctcp.2009.11.005

Soulsby EJL 1982. Helminths, Arthropods and Protozoa of Domesticated Animals. 7th Edn, ELBS and BaillièreTindall, London.

Wallis RB 1996. Hirudins: From leeches to man. Seminars in Thrombosis and Hemostasis 22(2): 185–96. DOI: https://doi.org/10.1055/s-2007-999007