Short communicationA revised method of examining fish for infection with zoonotic nematode larvae

- Visual examination is not sufficient to estimate abundance of anisakids in a fish.
- Overnight incubation of fish organs combined with visual examination is recommended.
- Number of larvae found in the recommended method is significantly higher.
- The recommended method is simple, cost effective and fast.
- The recommended method is not efficient on frozen fish.

The infection of fish with zoonotic nematodes, particularly anisakid nematodes is of great interest to many researchers who study food safety, human or animal health or who use them as biological tags for stock assessment studies. Accurate examination of fish for infection with anisakid larvae is crucial in making accurate estimates of their occurrence, abundance and prevalence in their fish hosts. Here we describe a new method of examining fish for infection with these parasites. In 2015, a total of 261 fish were purchased from a fish market in New South Wales, Australia. All fish were first examined by routine visual examination for infection with zoonotic nematode larvae and all data were recorded. Subsequently all internal organs were placed in a container and filled with water and incubated in the room temperature overnight. The prevalence, mean intensity and mean abundance of anisakids were significantly higher (p < 0.05) when the revised method of examination, i.e., combining visual examination and overnight incubation in room temperature, was employed (63.98, 8.23 and 5.27, respectively) compared to routine visual examination with or without the aid of a microscope (8.81, 3.78 and 0.33, respectively). The proposed method is effective and has several advantages, such as: not using UV or HCl for fish examination, allowing the examination of a larger number of fish in shorter time; larval specimens collected being suitable for both morphological and DNA sequencing; and being simple and inexpensive. The disadvantages would be the odour of the specimens after overnight incubation as well as not being suitable for use with frozen fish. We suggest that results, conclusions or recommendations made in studies that claim no anisakid/ascaridoid larvae were found in a fish should be approached carefully if it is only based on visual examination of the fish.