Development of a Double Glass Mounting Method Using Formaldehyde Alcohol Azocarmine Lactophenol (FAAL) and its Evaluation for Permanent Mounting of Small Nema¬todes

  • Farzaneh ZAHABIUN Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
  • Seyed Mahmoud SADJJADI Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran AND Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
  • Farideh ESFANDIARI Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Keywords: Nematodes, Double Glass Mount-ing, Permanent Mounting, Formaldehyde Alcohol Azocarmine Lactophe-nol

Abstract

Background: : Permanent slide preparation of nematodes especially small ones is time consuming, difficult and they become scarious margins. Regarding this prob­lem, a modified double glass mounting method was developed and com­pared with classic method.Methods: A total of 209 nematode samples from human and animal origin were fixed and stained with Formaldehyde Alcohol Azocarmine Lactophenol (FAAL) followed by double glass mounting and classic dehydration method us­ing Canada balsam as their mounting media. The slides were evaluated in differ­ent dates and times, more than four years. Different photos were made with differ­ent magnification during the evaluation time.Results: The double glass mounting method was stable during this time and compa­rable with classic method. There were no changes in morphologic struc­tures of nematodes using double glass mounting method with well-defined and clear differentiation between different organs of nematodes in this method.Conclusion: Using this method is cost effective and fast for mounting of small nematodes comparing to classic method. 

References

Ashford RW and Crewe W. The parasites of Homo sapiens – an annotated checklist of the protozoa, helminths and arthropods for which we are home. 2nd ed. London: Taylor and Francis; 2003.

Zibaei M, Sadjjadi SM, Jahad-Hosseini SH. Toxocara cati larvae in the eye of a child: a case report. Asian Pac J Trop Biomed. 2014; 4(1): 53-5.

Sadjjadi SM, Ardehali SM, Shojaei A. A case report of Linguatula serrata in human pharynx from Shiraz, Southern Iran. Med J Islam Re-pub Iran.1988; 12 (2):193-4.

Meamar AR, Kia EB, Zahabiun F, Jafari-Mehr A, Moghadam A, Sadjjadi SM. The occurrence of severe infections with Rhabditis axei in AIDS patients in Iran. J Helminthol. 2007; 81(4): 351-2.

Seinhorst JW. A rapid method for the transfer of nematodes from fixative to anhydrous glyc-

erin. Nematologica. 1959; 4(1): 67-9.

Baker AD. Rapid method for mounting nema-todes in glycerin. Can Ent.1953; 85(2): 77-8.

Hooper DJ. Handling, fixing, staining and mounting nematodes. In: Southey, JF. (Ed.) Laboratory methods for work with plant and soil nematodes. London: MAFF; 1986. P. 59-80.

Sadjjadi SM. A polyvinyl alcohol (PVA)-containing medium for permanent mounting of helminth eggs. Trans Roy Soc Trop Med Hyg. 2002; 96(1):104.

Sabu L, Devada K, Sreekrishnan R. Single step processing of nematodes in FAAL solution. Vet Parasitol. 2012; 26(1): 91 – 2.

Al-Amura MFA, Hassen ZA, AL-Mhanawi BH. Staining technique for helminth parasites by use red beet (beta vulgaris l.) Extract. Bas J Vet Res. 2012;11(1):283-92.

Khrustalev AV, Hoberg EP. Silver staining for elucidation of the synloph in trichostrongyle nematodes. J Parasitol.1995; 81(6): 1016-18.

Khrustalev AV, Hoberg EP. Carmine-propi-onic acid stain for elucidation of fine cellular structure in nematodes. J Parasitol. 1996; 82(1): 176--8.

Naem S, Pagan C, Nadler SA.Structural resto-ration of nematodes and acanthocephalans fixed in high percentage alcohol using DESS solution and rehydration. J Parasitol. 2010; 96(4): 809–11.

Sadjjadi SM, Massoud J. Helminth parasites of wild rodents in Khuzestan province, south west of Iran. J Vet Parasitol. 1999; 13:55-6.

Zibaei M, Sadjjadi SM, Sarkari B. Prevalence of Toxocara cati and other intestinal helminths in stray cats in Shiraz, Iran. Trop Biomed. 2007; 24(2):39-43.

Tanideh N , Sadjjadi SM , Mohammadzadeh T. Helminthic infections of laboratory animals in animal house of Shiraz University of Medical Sciences and the potential risks of zoonotic in-fections for researchers. Iran Red Crescent Med J. 2010; 12(2):151-7.

Yamaguti S. Systema Helminthum. New York: Interscience Publishers; 1958-1963. Vol.I-IV.

May HG. Killing, staining and mounting para-sitic nematodes. Trans Am Microsc Soc. 1922; 41(2): 103-5.

Grewal PS, Richardson PN, Wright DJ. Effects of killing, fixing and mounting methods on taxonomic characters of parthenogenetic adult female Caenorhabditis elegans (Nematoda: Rhab-ditidae). Revue Nématol. 1990; 13 (4): 437-44.

Grewal PS. The use of agar as a cover-glass support for mounting nematodes. Revue Né-matol. 1990; 13 (1): 121-2.

Sepulveda MS, Kinsella JM. Helminth collec-tion and identification from wildlife. J Vis Exp. 2013;(82):e51000.

Ryss AY. Express technique to prepare perma-nent collection slides of nematodes. Zoosyst Rossica. 2003; 11: 257-60.

Kumagai M, Inaba T, Makioka A, Ishiwata K,

Onishi K, Watanabe N. An improved glycerin- jelly mounting procedure for permanent prepa-rations of helminth eggs. J Parasitol. 2010 ; 96(2):440-1.

Bird AF. Changes associated with parasitism in nematodes. I. Morphology and physiology of preparasitic and parasitic larvae of Meloidogyne ja-vanica. J Parasitol. 1967; 53(4): 768-76.

Bird AF. Further observations on the structure of nematode cuticle. Parasitol. 1958; 48(1-2): 32-7.

Published
2015-12-14
How to Cite
1.
ZAHABIUN F, SADJJADI SM, ESFANDIARI F. Development of a Double Glass Mounting Method Using Formaldehyde Alcohol Azocarmine Lactophenol (FAAL) and its Evaluation for Permanent Mounting of Small Nema¬todes. IJPA. 10(4):617-24.
Section
Original Article(s)