Gastrointestinal parasite
diversity and relation to
climate in west Greenland
caribou
Chantal van Es
Student number: 3382222
Research Internship Master Companion Animal Medicine
Supervisors
Drs. Deborah van Doorn, Universiteit Utrecht
Dr. Karin Orsel, University of Calgary
Jillian Steele, BSc, University of Calgary
Calgary, June 8th 2012
Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Index
Summary…………………………………………………………………………………………………….3
Introduction………………………………………………………………………………………………. 4
Materials & Methods……………………………………………………………………………….. 6
Results……………………………………………………………………………………………… ………..8
Discussion………………………………………………………………………………………………….. 11
Acknowledgments…………………………………………………………………………………….. 13
References………………………………………………………………………………………………….14
Appendices………………………………………………………………………………………………. 17
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Summary
The aim of this study was to determine whether there is a relation between gastrointestinal parasite
diversity and climate in west Greenland. Two different caribou herds from two different areas, the
Kangerlussuaq-Sisimut (KS) area and the Akia-Maniitsoq (AM) area, were sampled and their
gastrointestinal nematodes were identified. Weather data on temperature, precipitation and wind
speed was obtained from the online database of the Danish Meteorological Institute and
the climate preferences of these nematodes, obtained from literature, were compared with the
climate data of the areas. These climate data show a continental climate in the KS area and a
maritime climate in the AM area. Our results show that parasites in western Greenland are
distributed following their preferred climatological factors. It, unfortunately, was not possible to say
that parasite distribution in this region is primarily influenced by climate, because there are many
other factors and weather conditions to take in consideration.
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Introduction
In Greenland, people partly depend on caribou for food and other economic aspects (Albon, 2002).
Caribou health and survival are affected by multiple factors, which include the presence of parasites
(Cuyler, 1999). Gastrointestinal parasites are negatively related to body condition, reproduction and
survival (Albon, 2002). Habitat and environment influence parasite distribution in many ways.
Human-mediated effects, such as environmental disruption, can lead hosts to contact new parasites
or change migratory patterns, which used to prevent, or decrease, infection with seasonal parasites
(Stromberg, 1997). One of the most important factors for development, survival and distribution of
the free-living stages of gastrointestinal nematodes are the weather conditions (Stromberg, 1997).
Parasites with free-living stages in the environment are likely to be affected by even the smallest
differences in temperature and precipitation (Kutz, 2012).
The two largest caribou herds in Greenland are believed to originate from the same ancestors
(Flagstad, 2003), but previous genetic research shows minimal gene transfer between these herds
(Jepsen, 2002). The herds are separated from each other by the Greenland ice cap, and have dealt
with different circumstances.
Besides contact with different animals, the herds also have different climates. The two areas are
separated from each other by the Greenland ice cap, causing a difference in air pressure (Cuyler
2010, Tamstorf 2005). The KS area has a continental climate caused by a high pressure system and it
is characterized by more extreme temperatures, less precipitation, and less wind than the AM area
(Tamstorf 2005, Cuyler 2005b, Tamstorf 2004). The AM area, which lies in a low pressure system, is
characterized by a more maritime climate with temperate temperatures, higher levels of
precipitation, and more wind (Tamstorf 2004, Cuyler 2005b, Tamstorf 2005 ).
The most common gastrointestinal nematodes of caribou are Ostertagia spp., Teladorsagia spp. and
Marshallagia spp. (Kutz, 2012), although caribou may also host parasites of muskoxen, sheep, cattle
and horses which are brought into contact with - (Norlund, 1936). These common parasites are all
members of the family Trichostrongylidae. They have the same basic life cycle, but show variability in
their sensitivity to environmental factors(Stien, 2002). Eggs are passed in the feces and, time
dependent on environmental factors, such as temperature and moisture conditions, develop to the
infective third stage larvae (L3) during summer. With the right moisture levels, the L5 will migrate
from the feces on to the herbage and be ingested during normal grazing behavior. After ingestion by
a compatible ungulate host species, the L5 becomes sexually mature in the abomasum (Taylor,
2007).The free living stages on the pasture are especially dependent on environmental conditions.
Developmental steps, such as hatching, larval development, and migration appear to depend on
weather conditions, such as temperature, moisture, and winds, although many others may play roles
(Stromberg, 1997).
The current global situation shows a shift in climates around the world (Lafferty, 2009),which is
expected to be a major factor in changing endemic parasite faunas (Poulin, 2006). In what direction
the changes will go, is only guesswork at this moment, as it is possible that the geographical range of
some generalist parasites will expand, that previously unimportant pathogens will emerge as
destructive, or that some parasites will extinct by direct effects of higher temperatures on the
developmental rates and transmission, or by effects of climate on their hosts (Poulin, 2006). Before
we can begin to understand the directions of these changes, we must first accumulate knowledge of
how current climate systems effect parasite distribution, so we may begin to understand how this
may change in the future.
The research I undertook will begin to fill this knowledge gap, as investigated the relationship
between gastrointestinal parasite diversity and climate in western Greenland. For this I examined
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
what our current status of knowledge is regarding caribou gastrointestinal parasites and their
climatic preferences, and then examined the situation in west Greenland to see if the parasites in
those regions could be found were we would expect them
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Materials & Methods
This research is a part of a larger project on parasites of caribou in West-Greenland (Cuyler 2012;
Steele, 2011).
Study Area
In the springs of 2008 and 2009 caribou were sampled from two areas in West Greenland. The
caribou collected in March-April 2008 were from the southern herd of AM and in March 2009 from
the northern herd of KS.
Fig 1. Locations of the two largest West Greenland caribou populations, Kangerlussuaq-Sisimiut and
Akia-Maniitsoq (Cuyler, 2010).
Weather Conditions
There are many weather conditions which should be taken in consideration in regards to parasite
transmission and survival (Pandey, 1972). This paper reports monthly minimum and maximum
temperature, mean monthly temperature, mean monthly precipitation and mean monthly wind
speed. Temperature and precipitation are known for their important influence on parasite
development and survival (Taylor, 2007). The free-living stages are especially dependent on
temperature for their development and survival on the pasture (Stromberg, 1997). Moisture is also
important for development and survival, but plays a crucial role in allowing their movement from
fecal material to the herbage (Stromberg, 1997). Wind speed has influences on temperature and
precipitation and is therefore an important factor influencing parasite transmission and survival, and
in our understanding of these other features of climate (Ollerenshaw, 1969).The temperature is
influenced by wind chill; wind can increase or decrease the temperature. Also, wind influences the
amount of precipitation; wind brings precipitation or keeps it away (Ollerenshaw, 1969).
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
All data was obtained from the online database of the Danish Meteorological Institute (DMI 2012).
The weather stations were located at Nuuk in the AM area and at Kangerlussuaq in the KS area.
These stations were selected for this work because they were the closest to the locations of
sampling.
Because the found nematodes are all of the Trichostrongylidae family, the free-living stages are on
the pasture around the same period. The eggs are deposited on the pasture in the beginning of
summer, develop during the rest of summer and the most of L3 is ingested at the end of summer
(Taylor, 2007). However, the presence of parasites depends on weather conditions during its whole
life cycle, so in this report we selected a time-frame from the previous summer till the sample date of
the herds for the climate analysis. This means for the AM herd a time-frame from June 2007 up to
the end of April 2008 and for the KS herd we examined a time-frame from June 2008 up to the end of
March 2009.
Study Animals
Adult caribou were selected through convenience sampling, shot and examined as per CARMA
protocols (Gunn & Nixon, 2008). For our analysis we used the data collected from the 76 adult
animals; 41 animals from AM and 35 animals from KS.
For these animals, the gastrointestinal tracts were removed at death, frozen and then processed
according to protocol following thawing (Appendix 1). The abomasal washes were preserved in 70%
ethanol or 10% formalin and transported to the University of Calgary. They were analyzed following
an in lab protocol (Appendix 2), and the nematodes found were placed in cryovials and stored at -80
°C. These steps were already done prior to my involvement with the project.
Parasites
Identification of adult male nematodes was done morphologically following an in-lab key, focusing
on; body length, bursa pattern, spicules length and character, esophageal valve length and width and
synlophe pattern (Appendix 3). The key was developed for arctic nematodes species and those
species which could have been present due to the introduction of domestic species; nevertheless,
other species were not excluded from our search and were included in the analysis if identified.
Nematodes were cleared and mounted from compound microscopy using lactophenol. For the AM
samples reserved in 70% ethanol with more than 50 male nematodes, only 50 randomly selected
nematodes were mounted. The rest of the nematodes were reserved for molecular identification
later in the project. Of the samples kept in 10% formalin, all the male nematodes were identified
using microscopy as these samples aren’t ideal for DNA extraction techniques.
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Results
Climate Data
Nematodes depend on climate during the whole life cycle, so I looked at the weather data of the year
before the sample date. If the weather was atypical for the known climates of these areas, I would
also look at other years, but, fortunately, the weather was typical for these areas.
Temperature
The KS area (Table 1) had extremer temperatures in comparison with the AM area (Table 2). The KS
area had a relatively cold winter and a warm summer, while the AM area had a relatively mild winter
and a mild summer.
Mean temperature
Minimum
Maximum
(°C)
temperature (°C)
temperature (°C)
June 2008
11.0
4.1
16.6
July
11.8
4.8
17.7
August
8.0
2.9
13.0
September
3.1
-0.8
6.7
October
-4.7
-9.0
-0.6
November
-9.6
-13.6
-5.0
December
-14.9
-19.5
-9.8
January 2009
-15.7
-20.6
-10.6
February
-14.3
-19.6
-9.3
March
-20.2
-26.4
-13.7
Table 1. Monthly maximum and minimum temperature and monthly mean temperature (°C) in
Kangerlussuaq-Sisimut
Mean temperature
Minimum
Maximum
(°C)
temperature (°C)
temperature (°C)
June 2007
5.7
2.9
9.4
July
8.5
5.1
12.5
August
9.2
6.5
13.0
September
3.5
1.3
6.3
October
-0.6
-3.2
2.3
November
-3.2
-6.0
-0.4
December
-8.3
-10.9
-6.0
January 2008
-11.1
-13.5
-8.6
February
-12.5
-15.0
-9.4
March
-8.2
-11.2
-4.8
April
-1.3
-4.2
2.2
Table 2. Monthly maximum and minimum temperature and monthly mean temperature (°C) in AkiaManiitsoq
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Precipitation
In both areas fell more precipitation in summer than in winter (Table 3). The KS area is more dry than
the AM area, which had relatively a large amount of precipitation.
Mean precipitation (mm)
KS
2008-2009
AM
2007-2008
June
July
August
September
6
10
55
42
53
132
105
116
October
November
17
18
27
111
December
8
60
January
February
6
2
47
56
March
2
76
April
N/A
14
Table 3. Monthly mean precipitation (mm) in Kangerlussuaq-Sisimut and in Akia-Maniitsoq
Wind speed
The KS area has relatively low wind speeds in comparison to the AM area. In both areas there is no
clear difference between the seasons.
Mean wind speed (m/s)
KS
2008-2009
AM
2007-2008
June
6
6
July
5
5
August
3
6
September
3
8
October
4
7
November
4
11
December
5
6
January
4
5
February
3
8
March
3
7
April
N/A
5
Table 4. Monthly mean wind speed (m/s) in Kangerlussuaq-Sisimut and in Akia-Maniitsoq
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Climate Summary
The weather conditions seen in Nuuk in 2007 and 2008 and in Kangerlussuaq in 2008 and 2009,
follow the expected climate patterns for those regions. The AM area has a maritime climate, which is
temperate, and therefore on average more moderate and with higher levels of precipitation and
winds than KS. The KS area has a continental climate, which has more extreme temperatures, hotter
summers and colder winters, along with less precipitation and wind. (Tamstorf 2004, Cuyler 2005b,
Tamstorf 2005)
Parasites
Samples from the AM herd were positive solely for Ostertagia gruehneri. KS samples, in contrast,
were positive for Teladorsagia spp. and Marshallagia marshalli. It was not possible to make a
difference between the Teladorsagia species under the microscope, so we consider that both T.
circumcincta and T. boreoarcticus are possible candidate species.
The preferential climate conditions from literature of these nematodes are cool, moist weather for
Ostertagia gruehneri and dry weather for Teladorsagia spp. and Marshallagia marshalli (Hoar 2012,
Nabavi 20110). According to the literature Teladorsagia spp. and Marshallagia marshalli prefer
higher temperatures, but they are also found in other Arctic regions, so they can survive in west
Greenland (Popiolek 2007).
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Discussion
In western Greenland parasites appear distributed following their preferred climatological factors.
This was already suggested in previous researches (Halvorsen, 1999, Korsholm, 1991, Stromberg,
1997), but our results confirm this and that parasite distribution follows expected known
“preferences” in regards to temperature, precipitation and wind speed.
The preferential climate conditions from literature of these nematodes are cool, moist weather for
Ostertagia gruehneri and dry weather for Teladorsagia spp. and Marshallagia marshalli (Hoar 2012,
Nabavi 20110). According to the literature Teladorsagia spp. and Marshallagia marshalli prefer
higher temperatures, but they are also found in other Arctic regions, so they can survive in west
Greenland (Popiolek, 2007).
Challenges
Parasite distribution in this region may be influenced by climate, but we can’t say for sure because
there are many other factors to take in consideration. The nematodes identified in these herds are
generalist species and there is no reason based solely on climate that would stop them from being
able to survive, although perhaps not thrive (Levine, 1963, Taylor, 2007). This means there could be
other reasons for the current distribution of parasite species.
The Akia-Maniitsoq (AM) and the Kangerlussuaq-Sisimut (KS) herds have had different contact with
other species over the years. The AM area was colonized by vikings and used for agriculture, which,
most likely, brought the caribou into contact with domestic cattle, sheep, goats and horses (Norlund,
1936). More recently, there was also an introduction of semi-domesticated Norwegian reindeer to
supplement the numbers of animals in the caribou herd (Cuyler, 1999). The KS area has not been
used for agriculture, but the caribou have been in contact with other ungulate species too. To make
hunting more accessible muskoxen were introduced to the area in 1962 and have spread across the
majority of the caribou range (Clausen, 1993). Because the herds are isolated from each other,
parasite transfer between the herds is unlikely. So parasites which may have been introduced along
with domestic ungulates into one herd, may not be able to transfer to the other.
Of course, it is a possibility that a combination of climate, isolation and different contact with other
species lead to the current distribution of the nematodes.
Future Steps
For further research it is necessary to have more information about the development of the
parasites, such as growth rate information, the exact time required at each stage for develop and
how long the free-living stages need to stay on the pasture. Also it is useful to know exactly how the
nematodes depend on environmental conditions, so these can be taken into consideration.
Also, it would be useful to have more information about the climate of Greenland. Survival and
transmission of nematodes depend on a whole range of weather conditions (Pandey, 1972). If you
have more detailed information on the conditions used in this project, it would be possible to look at
day-by-day data instead of monthly means. This is useful, because even little differences in weather
between the days could make big differences for parasite transmission and survival.
It would also be better to have more information about other weather conditions, such as snowfall
and, because many weather conditions have an influence on parasite transmission and survival and
should be taken in analysis.
Also, it could be useful to be critical about the samples and techniques used in this project. It may be
useful to have more samples, so you have more evidence. But in this project, it does not have to be
necessary to have more samples, because I would not expect to find other nematodes in these herds.
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
The key used for identification could also be discussed. It may be possible to make a clearer
differentiation between the species.
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
Acknowledgements
I want to thank Deborah van Doorn for her assistance and for giving me the opportunity to fulfill my
research project on the subject of parasitology. I really got a nice opportunity to see how the work in
this field looks like.
I want to thank Dr. Karin Orsel and Jillian Steele from University of Calgary for the opportunity to do
my research project at their university. Their effort to make this project a success made it very
instructive to me. Their hospitality was great and therefore I had a very nice time working and living
in Calgary.
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Appendix 1
SOP for GI tract removal, wash, and fecal collection in field conditions
1. Procedure Title
Method for GI tract removal, wash, and fecal collection in field conditions
2. Introduction
This technique is used to perform a GI tract wash in field conditions and obtain
1) abomasal wash samples
2) small intestinal samples
3) fecal samples
3. Definition
N/A
4. Personnel
SOP issued under authority of:
Persons authorized to perform the SOP:
Dr. S. Kutz, Jian Wang
All lab staff
5. Safety
Animals, and their GI tracts, are of unknown pathogenicity (bacterial, fungal, viral and parasitic
pathogens are all possible). Proper biosafety procedures should be followed for this process.
These include:
- Always wearing PPE (latex or nitrile gloves) in exposure situations
o Eye goggles are recommended
- Wash hands and forearms after performing the diagnostic test
As this procedure also involves using cutting equipment (carving knifes/scalpels), care should also be
taken not to cut oneself. If a wound is caused with the equipment proper first aid and biosafety
procedures for such an incident should be followed.
6. Procedure
6.1
Cut open animal and tie off esophagus, entry to abomasa and start of small intestine to
prevent mixing of contents. Remove GI tract.
6.2.1 Extract abomasa and first 3 feet of small intestine from GI tract and place onto separate
dissecting trays.
6.2.2 At this point, open the colon of the animal and remove 5g of feces. Place into a labeled
sample bag.
6.3
Taking the abomasa, remove the excess fat from the exterior. This will make it easier to
manipulate.
6.4
Cut the abomasa open along the greater curvature, and dispose of the loose contents by
scrapping gently with fingers.
- As the animal is freshly killed the majority of the nematodes will be attached to the
mucosal wall.
6.5
Check mucosa for lesions and remove a histology sample (1”x1”) from both the cardia and
pyloric areas. Place the samples into sample cups, labeled externally and with an internal
waterproof label, filled with 70% ethanol.
6.6
Vigorously rinse the abomasum, at least three times, into a large bucket with water. Also
rinse the tray into the bucket.
6.7
Bring the volume of the bucket up to the nearest 0.5L and remove two 10% aliquots.
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Gastrointestinal parasite diversity and relation to climate in west Greenland caribou
6.8
6.9
6.10
6.11
Drain each aliquot, separately, through a #100 sieve and rinse sieve into a labeled sample
cup, external and internal waterproof label, with 70% ethanol. Bring the volume of the
sample cup up to 100ml with 70% ethanol.
- Wash the sieve between uses with liquid soap and high pressure water to remove fat
deposits.
Examine small samples of the remaining wash on a white dissecting tray and look for
nematodes which should be reddish-brown coloured and moving. Go through remainder of
the wash in this fashion and place nematodes into heated saline to kill them. Then remove
them into labeled cryovials filled with 70% ethanol.
Washed abomasum can be discarded.
Small intestine (first 3 feet) should be treated as abomasum.
7. Equipment or Materials Required
Materials
7.1
#100 Standard sieve (x1)
7.2
Rubber policeman (x1)
7.3
Large dissecting trays (x2)
7.4
Small dissecting tray (white) (x1)
7.5
Dissecting probe or needle nosed tweezers (x1)
7.6
Carving knife (x1)
7.7
1x whirl-pack specimen bag (small)
7.8
Sample containers (x2)
7.9
Cryotubes (x1)
7.10 Freezer proof cryotube labels
7.11 Pencil
7.12 Large plastic bucket, at least 2L
7.13 Water squirt bottle
7.14 Small Petri-dish
7.15 Camp stove
7.16 Saline solution
7.17 Liquid dish soap (preferably dawn)
7.18 Measuring tape
Reagents
7.19
70% Ethanol
8. Highlights / Critical Control Points
8.1
As nematodes cannot be dried out for identification, great care should be taken to ensure
that all nematodes kept hydrated for work
9. Reporting
To be recorded in a blue lab book
9.1
Sample notes (i.e. Animal information, location)
9.2
Concerns during washing
- If no concerns; “Sample washed as per SOP – No Concerns”
9.3
Amount reserved in each 10% aliquot.
9.4
Total # of nematodes removed from the remainder of wash
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Appendix 2
SOP for the analysis of abomasal washes in a quantitative manner
1. Procedure Title:
Method for the analysis of abomasal washes in a quantitative manner
2. Introduction
This technique is used to analyse abomasal washes to quantify and isolate both larval nematodes and
adult GI parasites.
3. Definition
N/A
4. Personnel
SOP issued under authority of:
Persons authorized to perform the SOP:
Dr. S. Kutz, Jian Wang
All lab staff
5. Safety
As all materials are preserved in 70% ethanol or formalin the pathogenicity of the sample is vastly
reduced, however standard laboratory safety procedures should still be followed.
- Standard PPE (Lab coat and gloves) should be worn at all times
- Wash hands and forearms after performing the analysis
Formalin is highly carcinogenic and great care should be taken not to inhale or have skin contact with
the preservative. If direct exposure occurs, laboratory procedures for chemical contact should be
followed.
6. Procedure
6.1
Take the sample to be examined and drain it through a #400 standard sieve
- Ethanol can be drained into the sink directly; if the sample is preserved in formalin it
must be drained into a metal hazardous liquid container and disposed of in the
appropriate liquid waste container
6.2
With water, rinse the sample container and label into the sieve and then rinse the sample
repeatedly to remove any traces of ethanol/formalin
- This can be done directly into the sink
6.3
Scrape the contents using a rubber policeman and/or rinse back into the sample container
with water and fill to 100mL
6.4
Taking a 5mL syringe with the tip cut off, stir the sample and remove 10mL (10%) into a petridish. Rinse the syringe vigorously into the dish and check for adult worms which may be stuck
to the syringe walls. Close the sample container.
- If samples are very thick 5mL (5%) can be aliquoted instead
6.5
Using a 3mL bulb-pipette with the tip cut off, aliquot small samples from the petri-dish onto a
grid petri-dish and dilute with water.
- Be prepared to dilute a lot!
6.6
Examine the grid petri-dish on a dissecting microscope under 3.2 power. Look for and remove
adult GI parasites and larval nematodes.
6.6.1 Adult nematodes should be counted (using a multi-count desk counter) and placed into small
petri-dishes filled ~1/2 way with 70% ethanol. Separate counts for males, females and heads.
- Nematodes should always be scooped from the sample and never tweezered.
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6.6.2
6.7
6.8
6.9
6.10
6.11
6.12
6.13
Larva should be counted and removed into a small cryotube filled halfway with 70% ethanol
using a 10μL pipette
- Once having ejected a larva into the cryotube dip the pipette tip into water so as to
not agitate the sample by adding ETOH to the petri-dish
Once whole petri-dish has been examined, rinse dish into a large plastic beaker and return to
Repeat until first 10% sample is complete.
Remove another 10% sample aliquot as per 6.4, and examine as above.
Once two 10% aliquots have been examined, the rest of the sample can be observed
following the same pattern under 1.2 power and only adult nematodes should be
counted/reserved.
- If sample is particularly dense, it can be rinsed through a #100 standard sieve at this
point to remove small particulates
Once the full sample has been examined, check bulb-pipette to ensure no adults are trapped
on the walls. Cryotubes filled with 1.5mL 70% ethanol should be prepared for female, male
and nematode bits. The reserved nematodes should be recounted and then placed in these.
Each tube should be labeled (as below) and then placed into -80°C freezer
Wash waste can be discarded down sink, and all equipment should be washed as per lab
protocols.
7. Equipment or Materials Required
Materials
7.1
#400 and #100 Standard sieves
7.2
Rubber policeman
7.3
5mL syringe with tip cut off (should be melted to ensure no rough edges)
7.4
3mL bulb-pipette with tip cut off
7.5
Dissecting probe or needle nosed tweezers for removing adult nematodes
7.6
10μL pipette and tips
7.7
1x Grid petri-dish
7.8
1x Petri-dish large, 3x petri-dish small
7.9 5 cryotubes
7.10 Freezer proof labels
7.11 Pencil
7.12 Large plastic beaker
7.13 Water squirt bottle
Reagents
7.14 70% Ethanol
8. Highlights / Critical Control Points
8.1
As nematodes cannot be dried out for identification, great care should be taken to check the
syringe and bulb-pipette to ensure that no nematodes are trapped on walls
8.2
During the second stage of the analysis for adult nematodes;
- Males include fractured portions with a copulatory bursa
- Females include fractured portions with a vulva
- Bits include heads and middle portions
8.3
Care should be taken to not count immature adults in adult count
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8.4
Care should be taken not to press/tweeze adult nematodes as this can cause them to
explode/break. Always scoop out adults.
9. Reporting
To be recorded in a blue lab book
9.1
Sample notes (i.e. Date, UCID, AID, SAMPLE TYPE (i.e. Abowash), TYPE OF PRESSERVATIVE
and APPROX. VOLUME OF CONTENTS) should be recorded before start of procedure
9.2
Concerns during washing
- If no concerns; “Sample washed as per SOP – No Concerns”
9.3
Counts of Aliquot I (Larva, Males, Females, Bits), Aliquot II (Larva, Males, Females, Bits), 80%
Remainder (Males, Females, Bits).
9.4
Total adult counts (as adults from larval aliquots and 90% remainder can be stored together
9.5
Number of vials reserved
To be recorded on record sheet
9.6
Separate “Abomasal wash examination sheet” should be filled in as analysis occurs
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Appendix 3 Key to the males of common abomasal nematode species of Rangifer spp.
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