PMID- 23062689
OWN - NLM
STAT- MEDLINE
DCOM- 20130628
LR  - 20181202
IS  - 1873-2550 (Electronic)
IS  - 0304-4017 (Linking)
VI  - 191
IP  - 3-4
DP  - 2013 Jan 31
TI  - Fasciola hepatica: histological demonstration of apoptosis in the reproductive 
      organs of flukes of triclabendazole-sensitive and triclabendazole-resistant 
      isolates, and in field-derived flukes from triclabendazole-treated hosts, using 
      in situ hybridisation to visualise endonuclease-generated DNA strand breaks.
PG  - 240-51
LID - S0304-4017(12)00471-2 [pii]
LID - 10.1016/j.vetpar.2012.09.014 [doi]
AB  - Investigation of the triclabendazole (TCBZ) resistance status of populations of 
      Fasciola hepatica in field cases of fasciolosis, where treatment failure has been 
      reported, can be supported by histological examination of flukes collected from 
      recently treated hosts. In TCBZ-sensitive flukes (TCBZ-S) exposed to TCBZ 
      metabolites for 1-4days in vivo, but not in TCBZ-resistant flukes (TCBZ-R), 
      morphological changes suggestive of apoptosis occur in cells undergoing meiosis 
      or mitosis in the testis, ovary and vitelline follicles. In order to verify or 
      refute the contention that efficacy of TCBZ treatment is associated with 
      apoptosis in the reproductive organs of flukes, histological sections of TCBZ-S 
      (Cullompton isolate) flukes and TCBZ-R (Sligo isolate) flukes were subjected to 
      the TdT-mediated dUDP nick end labelling (TUNEL) in situ hybridisation method, a 
      commercially available test specifically designed to label endonuclease-induced 
      DNA strand breaks associated with apoptosis. Additionally, sections of in 
      vivo-treated and untreated flukes originating from field outbreaks of suspected 
      TCBZ-S and TCBZ-R fasciolosis were labelled by the TUNEL method. It was found 
      that in treated TCBZ-S flukes, strong positive labelling indicating apoptosis was 
      associated with morphologically abnormal cells undergoing mitosis or meiosis in 
      the testis, ovary and vitelline follicles. Background labelling in the positive 
      testis sections was attributed to heterophagy of cell debris by the sustentacular 
      tissue. The triggering of apoptosis was probably related to failure of spindle 
      formation at cell division, supporting the contention that TCBZ inhibits 
      microtubule formation. In treated TCBZ-R (Sligo Type 1) flukes, and in treated 
      flukes from field outbreaks of suspected TCBZ-R fasciolosis, no significant 
      labelling was observed, while sections of fluke derived from a field case of 
      fasciolosis where TCBZ resistance was not suspected were heavily labelled. Light 
      labelling was associated with the testis of untreated Cullompton (TCBZ-S) and 
      Sligo Type 2 (TCBZ-R) flukes, which exhibit abnormal spermatogenesis and 
      spermiogenesis, respectively. This was attributed to apoptosis and to heterophagy 
      of effete germ line cells by the sustentacular tissue. It is concluded that 
      demonstration of apoptosis by in situ hybridisation using the TUNEL method on 
      sections of 1-4days in vivo TCBZ-treated F. hepatica can contribute to the 
      diagnosis of TCBZ resistance in field outbreaks of fasciolosis.
CI  - Copyright (c) 2012 Elsevier B.V. All rights reserved.
FAU - Hanna, R E B
AU  - Hanna RE
AD  - Veterinary Sciences Division, Agri-Food and Biosciences Institute, Stormont, 
      Belfast BT4 3SD, United Kingdom. bob.hanna@afbini.gov.uk
FAU - Forster, F I
AU  - Forster FI
FAU - Brennan, G P
AU  - Brennan GP
FAU - Fairweather, I
AU  - Fairweather I
LA  - eng
PT  - Journal Article
DEP - 20120918
PL  - Netherlands
TA  - Vet Parasitol
JT  - Veterinary parasitology
JID - 7602745
RN  - 0 (Anthelmintics)
RN  - 0 (Benzimidazoles)
RN  - 4784C8E03O (Triclabendazole)
RN  - EC 3.1.- (Endonucleases)
SB  - IM
MH  - Animals
MH  - Anthelmintics/*pharmacology
MH  - Apoptosis
MH  - Benzimidazoles/*pharmacology
MH  - *DNA Breaks
MH  - Drug Resistance/*drug effects
MH  - Endonucleases/*metabolism
MH  - Fasciola hepatica/*cytology/*drug effects/genetics
MH  - Female
MH  - Genitalia/cytology/drug effects
MH  - In Situ Nick-End Labeling
MH  - Male
MH  - Sheep
MH  - Triclabendazole
EDAT- 2012/10/16 06:00
MHDA- 2013/07/03 06:00
CRDT- 2012/10/16 06:00
PHST- 2012/04/04 00:00 [received]
PHST- 2012/09/05 00:00 [revised]
PHST- 2012/09/06 00:00 [accepted]
PHST- 2012/10/16 06:00 [entrez]
PHST- 2012/10/16 06:00 [pubmed]
PHST- 2013/07/03 06:00 [medline]
AID - S0304-4017(12)00471-2 [pii]
AID - 10.1016/j.vetpar.2012.09.014 [doi]
PST - ppublish
SO  - Vet Parasitol. 2013 Jan 31;191(3-4):240-51. doi: 10.1016/j.vetpar.2012.09.014. 
      Epub 2012 Sep 18.