6 research outputs found
Co-existence and niche separation of two subspecies of Photorhabdus temperata associated with Heterorhabditis downesi in a dune grassland
Entomopathogenic nematodes belonging to the families Heterorhabditidae and
Steinernematidae form symbiotic relationships with entomopathogenic bacteria from
the genera Photorhabdus and Xenorhabdus, respectively. Previous studies showed
that Heterorhabditis downesi was associated with two different colour variants (pur
and yel) of its Photorhabdus bacterial symbiont in sand dunes on North Bull Island
in Dublin Bay, with evidence of the pur variant being more common to the front of
the dunes. The main objectives of this study were to confirm the distribution of both
colour variants of the nematode-bacterium complex on North Bull Island, identify
them, investigate the specificity of the nematode-bacterium relationship and
ascertain characteristics of the bacterial variants that might explain their distribution.
A section of the dune system on North Bull Island was surveyed in 2008 and resurveyed
in 2012 (Chapter 2). In 2008 the colour variants were spatially segregated,
with nematodes associated with the pur variant recovered from the front section of
the dunes only and nematodes associated with the yel variant recovered from the rear
section only. In 2012 nematodes associated with each colour variant were recovered
from the front section of the dunes and only nematodes associated with the yel
variant were recovered from the rear section of the dunes. The nematode-pur
complex was more abundant than the nematode-yel complex in sections where both
combinations co-occurred. Three laboratory lines of each nematode and its
associated bacterial colour variant were established for further study. A phylogenetic
analysis of the nematode ITS region showed that all the nematode isolates were
monophyletic, with no difference between the isolates carrying the pur or yel
bacterial symbionts. On the other hand, a phylogenetic analysis of a portion of the
gyrB gene of the bacterial isolates showed that the two colour variants were distinct
subspecies, Photorhabdus temperata subsp. temperata (yel) and Photorhabdus
temperata subsp. cinerea (pur). This confirms Photorhabdus temperata subsp.
temperata as the bacterial partner in the YEL H. downesi – Photorhabdus complex.
In addition, this is the first report of Photorhabdus temperata subsp. cinerea in
Ireland and also one of the few reports of the co-occurrence of two different
symbiotic bacteria subspecies with a single nematode species (Chapter 2). In
antibiosis tests against a range of organisms the P. temperata subsp. cinerea isolates
had a greater inhibitory effect than the P. temperata subsp. temperata isolates. Only
one pair of isolates, yel1 and pur2, showed any antibiotic activity against the other
subspecies and this pair were mutually inhibitory (Chapter 2). Phenotypic tests
indicated that the subspecies had different growth requirements as the pur isolates
grew better on MacConkey agar than on nutrient agar while there was no difference
between the two media for the yel isolates. Biochemical characterisation was carried
out using API 20E test strips. There was variability both within and between the
colour variants, however a notable difference was that only the yel isolates were able
to utilise trehalose, a major component of insect hemolymph (Chapter 2).
The cross-compatibility of the two different bacterial subspecies with the
heterogeneous nematode isolates was tested by injecting Galleria mellonella larvae
with 10,000 cells of either the pur1 or yel3 isolate or both and then infecting them
with H. downesi nematodes carrying either pur1 or yel3 bacteria. The emerging
nematodes carried the bacterial subspecies that had been injected into the host,
irrespective of which bacterial subspecies the infecting nematode had carried and
hybrid lines of nematode-bacterium complex were successfully maintained for the
duration of this study (Chapter 3). In the case of cadavers infected with both
bacterial types all the emerging nematodes tended to carry a single subspecies.
However, between 20 % and 30 % of the cadavers produced some nematodes
carrying the pur1 and some carrying the yel3 isolate, (indicating that both bacterial
subspecies were colonising the insect host and subsequently colonising the
developing nematodes prior to emergence) and where this occurred a higher
proportion of the nematodes carried the yel3 isolate. Furthermore, there was an
apparent shift in the colonisation of the nematodes within the host cadaver the
proportion of nematodes carrying the yel3 isolate was higher in later emerging
nematodes (Chapter 3).
To test the hypothesis that the distribution of the colour variants was due to host
specialisation (H. downesi complex differs from front to back of the dune system)
the pathogenicity of the homogenous and heterogenous combinations against
Hylobius abietis larvae was investigated. The LD50 was significantly higher for the
homogenous YEL/yel combination compared to the heterogeneous YEL/pur
combination. There was no difference in reproduction rate of the nematodes carrying
either subspecies in either H. abietis or G. mellonella and this was backed up by tests
of all three isolates of each colour type. All six isolates from North Bull Island were
tested against insects from three orders (Lepidoptera, Coleoptera (2) and Diptera),
with no differences in mortality or reproduction rate. This is the first report of EPN
being able to use the novel host kelp fly (Coelopa spp.). While there was no
difference in mortality rates between the PUR and YEL strains in this host there did
appear to be a difference in the rate of progression of the infection with nematodes
carrying the yel bacteria emerging earlier than those carrying the pur bacteria
(Chapter 4).
To test the hypothesis that the bacteria influenced nematode survival in desiccated
hosts G. mellonella larvae were infected with the homogenous and heterogeneous
combinations and stored in dry sand or at 0% relative humidity. There was no
difference due to bacteria type in the number of cadavers with emergence from those
stored in dry sand for 28 days. There was a highly significant difference due to
bacteria type in the number of cadavers with emergence for cadavers stored at 0 %
relative humidity for 28 days, with emergence from more cadaver infected with the
pur1 isolate. Overall, more nematodes emerged from cadavers infected with the pur1
isolate following 28 days storage in dry sand or at 0 % relative humidity (Chapter 5).
Heterorhabditis downesi is associated with two different subspecies of
Photorhabdus temperata on North Bull Island: Photorhabdus temperata subsp.
temperata and Photorhabdus temperata subsp. cinerea and the association of the
nematode with a particular subspecies appears to vary with the soil conditions
prevalent in the front and the rear sections of the dunes: Under normal conditions P.t.
temperata appears to outcompete P.t. cinerea in forming a symbiotic association
with the nematode when both subspecies are present in the same insect host,
resulting in the H. downesi-P.t. temperata association predominating in the rear
section of the dunes, where the increased organic content of the soil favours moisture
retention. On the other hand the drier conditions that predominate in the sandier soil
towards the front of the dune system favour H. downesi-P.t. cinerea association
Co-existence and niche separation of two subspecies of Photorhabdus temperata associated with Heterorhabditis downesi in a dune grassland
Entomopathogenic nematodes belonging to the families Heterorhabditidae and
Steinernematidae form symbiotic relationships with entomopathogenic bacteria from
the genera Photorhabdus and Xenorhabdus, respectively. Previous studies showed
that Heterorhabditis downesi was associated with two different colour variants (pur
and yel) of its Photorhabdus bacterial symbiont in sand dunes on North Bull Island
in Dublin Bay, with evidence of the pur variant being more common to the front of
the dunes. The main objectives of this study were to confirm the distribution of both
colour variants of the nematode-bacterium complex on North Bull Island, identify
them, investigate the specificity of the nematode-bacterium relationship and
ascertain characteristics of the bacterial variants that might explain their distribution.
A section of the dune system on North Bull Island was surveyed in 2008 and resurveyed
in 2012 (Chapter 2). In 2008 the colour variants were spatially segregated,
with nematodes associated with the pur variant recovered from the front section of
the dunes only and nematodes associated with the yel variant recovered from the rear
section only. In 2012 nematodes associated with each colour variant were recovered
from the front section of the dunes and only nematodes associated with the yel
variant were recovered from the rear section of the dunes. The nematode-pur
complex was more abundant than the nematode-yel complex in sections where both
combinations co-occurred. Three laboratory lines of each nematode and its
associated bacterial colour variant were established for further study. A phylogenetic
analysis of the nematode ITS region showed that all the nematode isolates were
monophyletic, with no difference between the isolates carrying the pur or yel
bacterial symbionts. On the other hand, a phylogenetic analysis of a portion of the
gyrB gene of the bacterial isolates showed that the two colour variants were distinct
subspecies, Photorhabdus temperata subsp. temperata (yel) and Photorhabdus
temperata subsp. cinerea (pur). This confirms Photorhabdus temperata subsp.
temperata as the bacterial partner in the YEL H. downesi – Photorhabdus complex.
In addition, this is the first report of Photorhabdus temperata subsp. cinerea in
Ireland and also one of the few reports of the co-occurrence of two different
symbiotic bacteria subspecies with a single nematode species (Chapter 2). In
antibiosis tests against a range of organisms the P. temperata subsp. cinerea isolates
had a greater inhibitory effect than the P. temperata subsp. temperata isolates. Only
one pair of isolates, yel1 and pur2, showed any antibiotic activity against the other
subspecies and this pair were mutually inhibitory (Chapter 2). Phenotypic tests
indicated that the subspecies had different growth requirements as the pur isolates
grew better on MacConkey agar than on nutrient agar while there was no difference
between the two media for the yel isolates. Biochemical characterisation was carried
out using API 20E test strips. There was variability both within and between the
colour variants, however a notable difference was that only the yel isolates were able
to utilise trehalose, a major component of insect hemolymph (Chapter 2).
The cross-compatibility of the two different bacterial subspecies with the
heterogeneous nematode isolates was tested by injecting Galleria mellonella larvae
with 10,000 cells of either the pur1 or yel3 isolate or both and then infecting them
with H. downesi nematodes carrying either pur1 or yel3 bacteria. The emerging
nematodes carried the bacterial subspecies that had been injected into the host,
irrespective of which bacterial subspecies the infecting nematode had carried and
hybrid lines of nematode-bacterium complex were successfully maintained for the
duration of this study (Chapter 3). In the case of cadavers infected with both
bacterial types all the emerging nematodes tended to carry a single subspecies.
However, between 20 % and 30 % of the cadavers produced some nematodes
carrying the pur1 and some carrying the yel3 isolate, (indicating that both bacterial
subspecies were colonising the insect host and subsequently colonising the
developing nematodes prior to emergence) and where this occurred a higher
proportion of the nematodes carried the yel3 isolate. Furthermore, there was an
apparent shift in the colonisation of the nematodes within the host cadaver the
proportion of nematodes carrying the yel3 isolate was higher in later emerging
nematodes (Chapter 3).
To test the hypothesis that the distribution of the colour variants was due to host
specialisation (H. downesi complex differs from front to back of the dune system)
the pathogenicity of the homogenous and heterogenous combinations against
Hylobius abietis larvae was investigated. The LD50 was significantly higher for the
homogenous YEL/yel combination compared to the heterogeneous YEL/pur
combination. There was no difference in reproduction rate of the nematodes carrying
either subspecies in either H. abietis or G. mellonella and this was backed up by tests
of all three isolates of each colour type. All six isolates from North Bull Island were
tested against insects from three orders (Lepidoptera, Coleoptera (2) and Diptera),
with no differences in mortality or reproduction rate. This is the first report of EPN
being able to use the novel host kelp fly (Coelopa spp.). While there was no
difference in mortality rates between the PUR and YEL strains in this host there did
appear to be a difference in the rate of progression of the infection with nematodes
carrying the yel bacteria emerging earlier than those carrying the pur bacteria
(Chapter 4).
To test the hypothesis that the bacteria influenced nematode survival in desiccated
hosts G. mellonella larvae were infected with the homogenous and heterogeneous
combinations and stored in dry sand or at 0% relative humidity. There was no
difference due to bacteria type in the number of cadavers with emergence from those
stored in dry sand for 28 days. There was a highly significant difference due to
bacteria type in the number of cadavers with emergence for cadavers stored at 0 %
relative humidity for 28 days, with emergence from more cadaver infected with the
pur1 isolate. Overall, more nematodes emerged from cadavers infected with the pur1
isolate following 28 days storage in dry sand or at 0 % relative humidity (Chapter 5).
Heterorhabditis downesi is associated with two different subspecies of
Photorhabdus temperata on North Bull Island: Photorhabdus temperata subsp.
temperata and Photorhabdus temperata subsp. cinerea and the association of the
nematode with a particular subspecies appears to vary with the soil conditions
prevalent in the front and the rear sections of the dunes: Under normal conditions P.t.
temperata appears to outcompete P.t. cinerea in forming a symbiotic association
with the nematode when both subspecies are present in the same insect host,
resulting in the H. downesi-P.t. temperata association predominating in the rear
section of the dunes, where the increased organic content of the soil favours moisture
retention. On the other hand the drier conditions that predominate in the sandier soil
towards the front of the dune system favour H. downesi-P.t. cinerea association
An Entomopathogenic Nematode Extends Its Niche by Associating with Different Symbionts
Bacterial symbionts are increasingly recognised as mediators of ecologically important traits of their animal hosts, with acquisition of new traits possible by uptake of novel symbionts. The entomopathogenic nematode Heterorhabditis downesi associates with two bacterial symbionts, Photorhabdus temperata subsp. temperata and P. temperata subsp. cinerea. At one intensively studied coastal dune site, P. temperata subsp. cinerea is consistently more frequently isolated than P. temperata subsp. temperata in H. downesi recovered from under the bare sand/Ammophila arrenaria of the front dunes (where harsh conditions, including drought, prevail). This is not the case in the more permissive closed dune grassland further from the sea. No differences were detected in ITS1 (internal transcribed spacer) sequence between nematode lines carrying either of the two symbiont subspecies, nor did they differ in their ability to utilise insects from three orders. The two symbionts could be readily swapped between lines, and both were carried in equal numbers within infective juveniles. In laboratory experiments, we tested whether the symbionts differentially affected nematode survival in insect cadavers that were allowed to dry. We assessed numbers of nematode infective juveniles emerging from insects that had been infected with H. downesi carrying either symbiont subspecies and then allowed to desiccate for up to 62 days. In moist conditions, cadavers produced similar numbers of nematodes, irrespective of the symbiont subspecies present, while under desiccating conditions, P. temperata subsp. cinerea cadavers yielded more nematode progeny than P. temperata subsp. temperata cadavers. Desiccating cadavers with the same nematode isolates, carrying either one or the other symbiont subspecies, confirmed that the symbiont was responsible for differences in nematode survival. Moreover, cadavers harbouring P. temperata subsp. cinerea had a reduced rate of drying relative to cadavers harbouring P. temperata subsp. temperata. Our experiments support the hypothesis that H. downesi can extend its niche into harsher conditions by associating with P. temperata subsp. cinerea
Co-existence and niche separation of two subspecies of Photorhabdus temperata associated with Heterorhabditis downesi in a dune grassland
Entomopathogenic nematodes belonging to the families Heterorhabditidae and
Steinernematidae form symbiotic relationships with entomopathogenic bacteria from
the genera Photorhabdus and Xenorhabdus, respectively. Previous studies showed
that Heterorhabditis downesi was associated with two different colour variants (pur
and yel) of its Photorhabdus bacterial symbiont in sand dunes on North Bull Island
in Dublin Bay, with evidence of the pur variant being more common to the front of
the dunes. The main objectives of this study were to confirm the distribution of both
colour variants of the nematode-bacterium complex on North Bull Island, identify
them, investigate the specificity of the nematode-bacterium relationship and
ascertain characteristics of the bacterial variants that might explain their distribution.
A section of the dune system on North Bull Island was surveyed in 2008 and resurveyed
in 2012 (Chapter 2). In 2008 the colour variants were spatially segregated,
with nematodes associated with the pur variant recovered from the front section of
the dunes only and nematodes associated with the yel variant recovered from the rear
section only. In 2012 nematodes associated with each colour variant were recovered
from the front section of the dunes and only nematodes associated with the yel
variant were recovered from the rear section of the dunes. The nematode-pur
complex was more abundant than the nematode-yel complex in sections where both
combinations co-occurred. Three laboratory lines of each nematode and its
associated bacterial colour variant were established for further study. A phylogenetic
analysis of the nematode ITS region showed that all the nematode isolates were
monophyletic, with no difference between the isolates carrying the pur or yel
bacterial symbionts. On the other hand, a phylogenetic analysis of a portion of the
gyrB gene of the bacterial isolates showed that the two colour variants were distinct
subspecies, Photorhabdus temperata subsp. temperata (yel) and Photorhabdus
temperata subsp. cinerea (pur). This confirms Photorhabdus temperata subsp.
temperata as the bacterial partner in the YEL H. downesi – Photorhabdus complex.
In addition, this is the first report of Photorhabdus temperata subsp. cinerea in
Ireland and also one of the few reports of the co-occurrence of two different
symbiotic bacteria subspecies with a single nematode species (Chapter 2). In
antibiosis tests against a range of organisms the P. temperata subsp. cinerea isolates
had a greater inhibitory effect than the P. temperata subsp. temperata isolates. Only
one pair of isolates, yel1 and pur2, showed any antibiotic activity against the other
subspecies and this pair were mutually inhibitory (Chapter 2). Phenotypic tests
indicated that the subspecies had different growth requirements as the pur isolates
grew better on MacConkey agar than on nutrient agar while there was no difference
between the two media for the yel isolates. Biochemical characterisation was carried
out using API 20E test strips. There was variability both within and between the
colour variants, however a notable difference was that only the yel isolates were able
to utilise trehalose, a major component of insect hemolymph (Chapter 2).
The cross-compatibility of the two different bacterial subspecies with the
heterogeneous nematode isolates was tested by injecting Galleria mellonella larvae
with 10,000 cells of either the pur1 or yel3 isolate or both and then infecting them
with H. downesi nematodes carrying either pur1 or yel3 bacteria. The emerging
nematodes carried the bacterial subspecies that had been injected into the host,
irrespective of which bacterial subspecies the infecting nematode had carried and
hybrid lines of nematode-bacterium complex were successfully maintained for the
duration of this study (Chapter 3). In the case of cadavers infected with both
bacterial types all the emerging nematodes tended to carry a single subspecies.
However, between 20 % and 30 % of the cadavers produced some nematodes
carrying the pur1 and some carrying the yel3 isolate, (indicating that both bacterial
subspecies were colonising the insect host and subsequently colonising the
developing nematodes prior to emergence) and where this occurred a higher
proportion of the nematodes carried the yel3 isolate. Furthermore, there was an
apparent shift in the colonisation of the nematodes within the host cadaver the
proportion of nematodes carrying the yel3 isolate was higher in later emerging
nematodes (Chapter 3).
To test the hypothesis that the distribution of the colour variants was due to host
specialisation (H. downesi complex differs from front to back of the dune system)
the pathogenicity of the homogenous and heterogenous combinations against
Hylobius abietis larvae was investigated. The LD50 was significantly higher for the
homogenous YEL/yel combination compared to the heterogeneous YEL/pur
combination. There was no difference in reproduction rate of the nematodes carrying
either subspecies in either H. abietis or G. mellonella and this was backed up by tests
of all three isolates of each colour type. All six isolates from North Bull Island were
tested against insects from three orders (Lepidoptera, Coleoptera (2) and Diptera),
with no differences in mortality or reproduction rate. This is the first report of EPN
being able to use the novel host kelp fly (Coelopa spp.). While there was no
difference in mortality rates between the PUR and YEL strains in this host there did
appear to be a difference in the rate of progression of the infection with nematodes
carrying the yel bacteria emerging earlier than those carrying the pur bacteria
(Chapter 4).
To test the hypothesis that the bacteria influenced nematode survival in desiccated
hosts G. mellonella larvae were infected with the homogenous and heterogeneous
combinations and stored in dry sand or at 0% relative humidity. There was no
difference due to bacteria type in the number of cadavers with emergence from those
stored in dry sand for 28 days. There was a highly significant difference due to
bacteria type in the number of cadavers with emergence for cadavers stored at 0 %
relative humidity for 28 days, with emergence from more cadaver infected with the
pur1 isolate. Overall, more nematodes emerged from cadavers infected with the pur1
isolate following 28 days storage in dry sand or at 0 % relative humidity (Chapter 5).
Heterorhabditis downesi is associated with two different subspecies of
Photorhabdus temperata on North Bull Island: Photorhabdus temperata subsp.
temperata and Photorhabdus temperata subsp. cinerea and the association of the
nematode with a particular subspecies appears to vary with the soil conditions
prevalent in the front and the rear sections of the dunes: Under normal conditions P.t.
temperata appears to outcompete P.t. cinerea in forming a symbiotic association
with the nematode when both subspecies are present in the same insect host,
resulting in the H. downesi-P.t. temperata association predominating in the rear
section of the dunes, where the increased organic content of the soil favours moisture
retention. On the other hand the drier conditions that predominate in the sandier soil
towards the front of the dune system favour H. downesi-P.t. cinerea association
Co-existence and niche separation of two subspecies of Photorhabdus temperata associated with Heterorhabditis downesi in a dune grassland
Entomopathogenic nematodes belonging to the families Heterorhabditidae and
Steinernematidae form symbiotic relationships with entomopathogenic bacteria from
the genera Photorhabdus and Xenorhabdus, respectively. Previous studies showed
that Heterorhabditis downesi was associated with two different colour variants (pur
and yel) of its Photorhabdus bacterial symbiont in sand dunes on North Bull Island
in Dublin Bay, with evidence of the pur variant being more common to the front of
the dunes. The main objectives of this study were to confirm the distribution of both
colour variants of the nematode-bacterium complex on North Bull Island, identify
them, investigate the specificity of the nematode-bacterium relationship and
ascertain characteristics of the bacterial variants that might explain their distribution.
A section of the dune system on North Bull Island was surveyed in 2008 and resurveyed
in 2012 (Chapter 2). In 2008 the colour variants were spatially segregated,
with nematodes associated with the pur variant recovered from the front section of
the dunes only and nematodes associated with the yel variant recovered from the rear
section only. In 2012 nematodes associated with each colour variant were recovered
from the front section of the dunes and only nematodes associated with the yel
variant were recovered from the rear section of the dunes. The nematode-pur
complex was more abundant than the nematode-yel complex in sections where both
combinations co-occurred. Three laboratory lines of each nematode and its
associated bacterial colour variant were established for further study. A phylogenetic
analysis of the nematode ITS region showed that all the nematode isolates were
monophyletic, with no difference between the isolates carrying the pur or yel
bacterial symbionts. On the other hand, a phylogenetic analysis of a portion of the
gyrB gene of the bacterial isolates showed that the two colour variants were distinct
subspecies, Photorhabdus temperata subsp. temperata (yel) and Photorhabdus
temperata subsp. cinerea (pur). This confirms Photorhabdus temperata subsp.
temperata as the bacterial partner in the YEL H. downesi – Photorhabdus complex.
In addition, this is the first report of Photorhabdus temperata subsp. cinerea in
Ireland and also one of the few reports of the co-occurrence of two different
symbiotic bacteria subspecies with a single nematode species (Chapter 2). In
antibiosis tests against a range of organisms the P. temperata subsp. cinerea isolates
had a greater inhibitory effect than the P. temperata subsp. temperata isolates. Only
one pair of isolates, yel1 and pur2, showed any antibiotic activity against the other
subspecies and this pair were mutually inhibitory (Chapter 2). Phenotypic tests
indicated that the subspecies had different growth requirements as the pur isolates
grew better on MacConkey agar than on nutrient agar while there was no difference
between the two media for the yel isolates. Biochemical characterisation was carried
out using API 20E test strips. There was variability both within and between the
colour variants, however a notable difference was that only the yel isolates were able
to utilise trehalose, a major component of insect hemolymph (Chapter 2).
The cross-compatibility of the two different bacterial subspecies with the
heterogeneous nematode isolates was tested by injecting Galleria mellonella larvae
with 10,000 cells of either the pur1 or yel3 isolate or both and then infecting them
with H. downesi nematodes carrying either pur1 or yel3 bacteria. The emerging
nematodes carried the bacterial subspecies that had been injected into the host,
irrespective of which bacterial subspecies the infecting nematode had carried and
hybrid lines of nematode-bacterium complex were successfully maintained for the
duration of this study (Chapter 3). In the case of cadavers infected with both
bacterial types all the emerging nematodes tended to carry a single subspecies.
However, between 20 % and 30 % of the cadavers produced some nematodes
carrying the pur1 and some carrying the yel3 isolate, (indicating that both bacterial
subspecies were colonising the insect host and subsequently colonising the
developing nematodes prior to emergence) and where this occurred a higher
proportion of the nematodes carried the yel3 isolate. Furthermore, there was an
apparent shift in the colonisation of the nematodes within the host cadaver the
proportion of nematodes carrying the yel3 isolate was higher in later emerging
nematodes (Chapter 3).
To test the hypothesis that the distribution of the colour variants was due to host
specialisation (H. downesi complex differs from front to back of the dune system)
the pathogenicity of the homogenous and heterogenous combinations against
Hylobius abietis larvae was investigated. The LD50 was significantly higher for the
homogenous YEL/yel combination compared to the heterogeneous YEL/pur
combination. There was no difference in reproduction rate of the nematodes carrying
either subspecies in either H. abietis or G. mellonella and this was backed up by tests
of all three isolates of each colour type. All six isolates from North Bull Island were
tested against insects from three orders (Lepidoptera, Coleoptera (2) and Diptera),
with no differences in mortality or reproduction rate. This is the first report of EPN
being able to use the novel host kelp fly (Coelopa spp.). While there was no
difference in mortality rates between the PUR and YEL strains in this host there did
appear to be a difference in the rate of progression of the infection with nematodes
carrying the yel bacteria emerging earlier than those carrying the pur bacteria
(Chapter 4).
To test the hypothesis that the bacteria influenced nematode survival in desiccated
hosts G. mellonella larvae were infected with the homogenous and heterogeneous
combinations and stored in dry sand or at 0% relative humidity. There was no
difference due to bacteria type in the number of cadavers with emergence from those
stored in dry sand for 28 days. There was a highly significant difference due to
bacteria type in the number of cadavers with emergence for cadavers stored at 0 %
relative humidity for 28 days, with emergence from more cadaver infected with the
pur1 isolate. Overall, more nematodes emerged from cadavers infected with the pur1
isolate following 28 days storage in dry sand or at 0 % relative humidity (Chapter 5).
Heterorhabditis downesi is associated with two different subspecies of
Photorhabdus temperata on North Bull Island: Photorhabdus temperata subsp.
temperata and Photorhabdus temperata subsp. cinerea and the association of the
nematode with a particular subspecies appears to vary with the soil conditions
prevalent in the front and the rear sections of the dunes: Under normal conditions P.t.
temperata appears to outcompete P.t. cinerea in forming a symbiotic association
with the nematode when both subspecies are present in the same insect host,
resulting in the H. downesi-P.t. temperata association predominating in the rear
section of the dunes, where the increased organic content of the soil favours moisture
retention. On the other hand the drier conditions that predominate in the sandier soil
towards the front of the dune system favour H. downesi-P.t. cinerea association
Higher relatedness mitigates mortality in a nematode with lethal male fighting
According to kin selection theory, individuals show less aggression towards their relatives. Limited dispersal promotes interactions among relatives but also increases competition among them. The evolution of cooperation in viscous populations has been subject of mainly theoretical exploration. We investigated the influence of relatedness on aggression in males of entomopathogenic nematode Steinernema longicaudum that engage in lethal fighting. In a series of in vitro experiments, we found that both competitor male group size and relatedness influence male mortality rates. Higher relatedness led to progressively lower rates of male mortality. In experimentally infected insects, wherein large numbers of males and females interact, the proportion of dead and paralysed (= terminally injured) males was higher when infection was established by infective juveniles originating from a mixture of three lines than in those infected by a single line. The results collectively show that Steinernema longicaudum males recognize their kin and consequently male mortality rates are lower in groups consisting of more related males. Furthermore, this monotonic negative relationship between aggression and relatedness suggests that kin selection benefits are still substantial even under extreme competition. Our experiments also suggest that kin recognition in entomopathogenic nematodes has a genetic basis rather than being strictly based on environmental cues. We discuss our findings within the theoretical context of the evolution of altruistic/cooperative behaviour in structured populations