COSEWIC Assessment and Status Report on the Nine-spotted Lady Beetle Coccinella novemnotata Canada - 2016

• Table of contents
• Assessment summary
• Executive summary
• Technical summary
• Wildlife species description and significance
  • Name and classification
  • Morphological description
  • Population spatial structure and variability
  • Designatable units
  • Special significance
• Distribution
  • Global range
  • Canadian range
  • Extent of occurrence and area of occupancy
  • Search effort
• Habitat
  • Habitat requirements
  • Habitat trends
• Biology
  • Life cycle and reproduction
  • Physiology and adaptability
  • Dispersal and migration
  • Interspecific interactions
• Population sizes and trends
  • Sampling effort and methods
  • Abundance
  • Fluctuations and trends
  • Rescue effect
• Threats and limiting factors
  • Invasive and other problematic species and genes (Threat 8.)
    • Invasive non-native/alien species (8.1)
      • Competition and intraguild predation:
      • Parasites, parasitoids, pathogens and fungi:
  • Pollution (Threat 9.)
    • Agricultural and forestry effluents (9.3)
  • Natural system modifications (Threat 7.)
    • Other ecosystem modifications (7.3)
  • Agriculture and aquaculture (Threat 2.)
  • Residential and commercial development (Threat 1.)
  • Number of locations
• Protection, status and ranks
  • Legal protection and status
  • Non-legal status and ranks
  • Habitat protection and ownership
• Acknowledgements and authorities contacted
• Information sources
• Biographical summary of report writer(s)
• Collections examined

• Figure 1. Nine-spotted Lady Beetle (Coccinella novemnotata).
• Figure 2. The geographic range of the Nine-spotted lady Beetle (Coccinella novemnotata).
• Figure 3. Extent of occurrence and index of area of occupancy for the Nine-spotted Lady Beetle based on museum collections and recent surveys (1897 - 2014).
• Figure 4. Extent of occurrence and index of area of occupancy for the Nine-spotted Lady Beetle based on museum collections (1995 - 2004).
• Figure 5. Recent extent of occurrence and index of area of occupancy for the Nine-spotted Lady Beetle based on museum collections and recent surveys (2005 - 2014).
• Figure 6. Search effort for the Nine-spotted Lady Beetle (Coccinella novemnotata).
• Figure 7. Search effort sites (orange dots) with an 18 km radius (blue dots), overlap with 287 known sites for the Nine-spotted Lady Beetle (Coccinella novemnotata) (black dots).
• Figure 8. Search effort sites (orange dots) with a 120 km radius (blue circles) overlap with 729 known sites for the Nine-spotted Lady Beetle (Coccinella novemnotata) (black dots).
• Figure 9. Changes in relative abundance of the native Nine-spotted Lady Beetle (Coccinella novemnotata) (white fill), and the non-native Seven-spotted Lady Beetle (Coccinella septempunctata) (grey fill) and Multi-coloured Asian Lady Beetle (Harmonia axyridis) (black fill) compared to all databased Coccinellidae in BC, AB, SK, MN, ON and QC.

• Table 1. There are ~1,061 Nine-spotted Lady Beetle specimens databased from 1897 - 2014 in Canada (see Collections Examined).
• Table 2. Targeted search effort 2013 - 2014. Total search effort of 262.4 hours across 230 sites detected four Nine-spotted Lady Beetles: one from Osoyoos (BC) and three from Medicine Hat (AB).
  • Table 2a. Province of British Columbia
  • Table 2b. Province of Alberta
  • Table 2c. Province of Ontario
  • Table 2d. Province of Quebec
  • Table 2e. Province of Northwest Territories
  • Table 2f. Province of Yukon
• Table 3a. Changes in relative abundance (RA) of the Nine-spotted Lady Beetle (NSLB) to native and non-native lady beetles (Coccinellidae) collected in British Columbia, Alberta, Saskatchewan, Manitoba, Ontario and Quebec.
• Table 3b. Changes in relative abundance (RA) of the Nine-spotted Lady Beetle (NSLB) compared to native lady beetles (Coccinellidae) collected in British Columbia, Alberta, Saskatchewan, Manitoba, Ontario and Quebec.
• Table 4a. Percent change in relative abundance over two decades of the Nine-spotted Lady Beetle (NSLB) to all native and non-native lady beetles (Coccinellidae) collected in British Columbia, Alberta, Saskatchewan, Manitoba, Ontario and Quebec.
• Table 4b. Percent change in relative abundance over two decades of the Nine-spotted Lady Beetle (NSLB) to all native lady beetles (Coccinellidae) collected in British Columbia, Alberta, Saskatchewan, Manitoba, Ontario and Quebec.

• Appendix 1. IUCN Threats calculation on the Nine-spotted Lady Beetle.

COSEWIC status reports are working documents used in assigning the status of wildlife species suspected of being at risk. This report may be cited as follows:

COSEWIC. 2016. COSEWIC assessment and status report on the Nine-spotted Lady Beetle Coccinella novemnotata in Canada. Committee on the Status of Endangered Wildlife in Canada. Ottawa. x + 57 pp.

(Species at Risk Public Registry website).

COSEWIC would like to acknowledge Paul Grant for writing the status report on the Nine-spotted Lady Beetle (Coccinella novemnotata) in Canada, prepared under contract with Environment Canada. This status report and was overseen and edited by Jennifer Heron, Co-chair of the COSEWIC Arthropods Specialist Subcommittee.

COSEWIC Secretariat
c/o Canadian Wildlife Service
Environment and Climate Change Canada
Ottawa, ON
K1A 0H3

Tel.: 819-938-4125
Fax: 819-938-3984
E-mail: COSEWIC E-mail
Website: COSEWIC

Également disponible en français sous le titre Ếvaluation et Rapport de situation du COSEPAC sur la Coccinelle à neuf points (Coccinella novemnotata) au Canada

Nine-spotted Lady Beetle - Photo by John Acorn.

The Nine-spotted Lady Beetle (Coccinella novemnotata Herbst) is a small beetle (4.7 – 7.0 mm) that is native to North America. Adults are readily identifiable by external morphological features: their wing covers are pale orange to red, with a dark line where the two wing covers meet. They generally have nine black spots on their wing covers, but the size and number of these spots can vary. Furthermore, the head and pronotum are black with white markings. This charismatic species was once one of the more common and widespread lady beetles in North America, playing an important role as a biological control agent of aphids and other insect pests.

The Nine-spotted Lady Beetle is a wide-ranging species occurring throughout most of southern Canada with a range that extends along the international border from Vancouver Island to southern Quebec; with northern range limits near: Quesnel, British Columbia; Edmonton, Alberta; Lake Athabasca, Saskatchewan and Roberval, Quebec. The Nine-spotted Lady Beetle also ranges across the continental United States southwards almost to the Mexican border.

Nine-spotted Lady Beetles are habitat generalists, known to consume a wide variety of prey across a wide range of habitats. They occur within agricultural areas, suburban gardens, parks, coniferous forests, deciduous forests, prairie grasslands, meadows, riparian areas and isolated natural areas. This broad habitat range reflects their ability to exploit seasonal changes in prey availability across different vegetation types.

Nine-spotted Lady Beetles have four life stages: egg, larva, pupa and adult, and can have two generations per year. Adults of the spring generation can undergo aestivation to avoid high summer temperatures and lay eggs in early autumn. Adults of the autumn generation congregate over winter and undergo diapause; becoming active and reproducing when temperatures warm in the early spring. This species occupies a wide ecological niche across a wide variety of habitats and temperature regimes in Canada. Little is known on the natural dispersal rates for the Nine-spotted Lady Beetle. In general, lady beetles are very mobile, display low site fidelity, and readily engage in short- and long-distance dispersal. Drivers of dispersal are a combination of prey density and environmental variables such as temperature, wind speed and rainfall. This species does not migrate. Both adult and larval stages are predatory and prey primarily on aphids. In turn, this species is also subject to predation by introduced lady beetles, other invertebrates and vertebrates, and susceptible to parasitoids and pathogens.

The historically broad geographic range and prominence of the Nine-spotted Lady Beetle stands in stark contrast to its current distribution. Prior to 1975, this species was widely distributed across North America and was one of the more common lady beetles collected. This species has since declined and is rarely collected despite targeted searches. Over the last decade the Nine-spotted Lady Beetle has continued to decrease in relative abundance when compared to other lady beetles.

The specific causes of decline in the Nine-spotted Lady Beetle are unknown. Possible threats to this species include negative interactions with recently arrived non-native species, such as the Seven-spotted Lady Beetle and the Multi-coloured Asian Lady Beetle, through competition, intraguild predation or indirect effects through the introduction of pathogens. Other possible threats include direct and indirect effects of pesticide/chemical use associated with agriculture to control their main prey species aphids, and habitat loss through urban expansion, abandonment of farmland, and other human disturbances.

There are no laws in Canada that protect the Nine-spotted Lady Beetle, its residence or habitat. The NatureServe global conservation status rank is G2 (imperilled). The species has not been assigned a conservation status rank in Canadian provinces or territories. However, while this species is not currently listed in Québec, it is likely to be designated Threatened or Vulnerable in that province.

The Committee on the Status of Endangered Wildlife in Canada (COSEWIC) was created in 1977 as a result of a recommendation at the Federal-Provincial Wildlife Conference held in 1976. It arose from the need for a single, official, scientifically sound, national listing of wildlife species at risk. In 1978, COSEWIC designated its first species and produced its first list of Canadian species at risk. Species designated at meetings of the full committee are added to the list. On June 5, 2003, the Species at Risk Act (SARA) was proclaimed. SARA establishes COSEWIC as an advisory body ensuring that species will continue to be assessed under a rigorous and independent scientific process.

The Committee on the Status of Endangered Wildlife in Canada (COSEWIC) assesses the national status of wild species, subspecies, varieties, or other designatable units that are considered to be at risk in Canada. Designations are made on native species for the following taxonomic groups: mammals, birds, reptiles, amphibians, fishes, arthropods, molluscs, vascular plants, mosses, and lichens.

COSEWIC comprises members from each provincial and territorial government wildlife agency, four federal entities (Canadian Wildlife Service, Parks Canada Agency, Department of Fisheries and Oceans, and the Federal Biodiversity Information Partnership, chaired by the Canadian Museum of Nature), three non-government science members and the co-chairs of the species specialist subcommittees and the Aboriginal Traditional Knowledge subcommittee. The Committee meets to consider status reports on candidate species.

Class Insecta - insects

Subclass Pterygota - winged insects

Order Coleoptera - beetles

Suborder Polyphaga - lady beetles, longhorn beetles, weevils, click beetles, fireflies, scarab beetles, rove beetles

Superfamily Cucujoidea - lady beetles, bark beetles, fungus beetles, sap beetles

Family Coccinellidae - lady beetles

Subfamily Coccinellinae

Tribe Coccinellini

Genus Coccinella

Species Coccinella novemnotata Herbst, 1793 - Nine-spotted Lady Beetle

Scientific name: Coccinella novemnotata

English Common Names: Nine-spot ladybug, nine-spotted ladybug, nine-spotted lady bird beetle, Nine-spotted Lady Beetle

French Common Name: Coccinelle à neuf points

The family Coccinellidae contains about 6,000 species worldwide in about 360 genera (Vandenberg 2002; Giorgi and Vandenberg 2009). In Canada there are 60 genera containing 161 species, of which 9 are adventive and are now well established (Hodek et al. 2012; Bousquet et al. 2013). The taxonomy, identification and geographic distribution of these species in Canada are well known (Dobzhansky 1935; Watson 1956; Brown 1962; Brown and de Ruette 1962; Belicek 1976; Watson 1976; Larochelle 1979; Gordon 1985; Vandenberg 2002; Majka and McCorquodale 2006; Acorn 2007; Marriott et al. 2009; Majka and McCorquodale 2010; Hodek et al. 2012; Bousquet et al. 2013).

The genus Coccinella contains 15 species, found primarily in North America. Within Canada, 11 species are native, including Coccinella novemnotata, and 2 species have been introduced (ITIS 2015).

The Nine-spotted Lady Beetle (Coccinella novemnotata) (Figure 1) was first described as a distinct species by Herbst (1793). There has been no further taxonomic work on the species and this description is still considered valid. No subspecies are recognized.

Lady beetles are holometabolous insects. They have four developmental life stages (egg, larva, pupae and adult). Each stage is morphologically different from the next.

Adult Nine-spotted Lady Beetles (4.7 - 7.0 mm) have elytra (wing covers) that are pale orange to red, most commonly with nine variably sized black spots, four on each elytra, with one central spot. However, the number and size of spots can vary across individuals to the point where some lack spots entirely. In the Nine-spotted Lady Beetle, the suture (where wing covers meet) has a dark narrow line. The head is broad and black with a pale band between the eyes. The anterior margin of the pronotum is entirely pale and black posteriorly (Gordon 1985; Acorn 2007) (Figure 1). Adults do not show sexual dimorphism (Stellwag and Losey 2014). The pale anterior pronotal margin and blackish sutural margin of the elytra readily distinguish the Nine-spotted Lady Beetlefrom other lady beetles.

The Nine-spotted Lady Beetle has yellow- to orange-coloured elongate eggs, approximately 1 mm in length that are laid upright in tightly packed clusters of approximately eighteen (Hodek et al. 2012). The larvae are black with periodic orange/red markings at the sides, and are elongated diamond-shaped with stubby, sometimes prickly looking legs. Larvae terga, or dorsal segments, have mound-like projections bearing seta, or hair-like structures (Rees et al. 1994). For detailed descriptions and keys to larvae stages see Rees et al. (1994). The pupae are usually yellow to orange with black markings (Hodek et al. 2012).

In Canada, the spatial structure and variability of Nine-spotted Lady Beetle subpopulations have not been studied. Similarly, limited genetic studies have occurred on this species and there is currently no evidence of subspecies genetic structure.

Allozyme variation was investigated in non-native (n = 8) and native (n = 6) lady beetles in North America, including the Nine-spotted Lady Beetle (Krafsur et al. 2005). For this study 38 specimens of Nine-spotted Lady Beetlewere collected from three areas in North America (Iowa, New York, and Arkansas). This study determined allele diversities and heterozygosities were similar in non-native and native lady beetles and therefore no obvious relationship existed between successful colonization of new habitats and genetic diversity (Krafsur et al. 2005). This study also determined that there were high rates of gene flow within in all lady beetle subpopulations (Krafsur et al. 2005). In addition, all lady beetles showed a remarkable degree of dispersion with little detectable subpopulation subdivision (Krafsur et al. 2005).

The Nine-spotted Lady Beetle has one designatable unit within Canada. No subspecies are recognized. Although the species occurs across the multiple ecological areas, there is little detectable subpopulation subdivision (Krafsur et al. 2005).

Lady beetles are iconic species to the general public. Prior to significant declines, the Nine-spotted Lady Beetle was one of the more common lady beetle species in Canada. As a predator of a large variety of aphid species in addition to other pest herbivores, it had an important economic role as a biological control agent in gardens and agricultural crops (Wheeler and Hoebeke 1995; Hesler et al. 2012). The observed decline of this charismatic species has led to public interest in their conservation and in this species' role in ecosystem function (Evans 2004; Harmon et al. 2007; Losey et al. 2007; Gardiner et al. 2011; Gardiner et al. 2012; Losey et al. 2012; Bahlai et al. 2013; Turnipseed et al. 2014; Ugine and Losey 2014).

Initiatives such as the Lost Lady Bug Project, which enable citizen scientists to help find and document Nine-spotted Lady Beetles across North America, demonstrate significant public interest in this species and shifting trends in lady beetle composition across landscapes.

There is no available Aboriginal Traditional Knowledge specifically for the Nine-spotted Lady Beetle.

The Nine-spotted Lady Beetle is a wide-ranging species occurring through most of southern Canada and the continental United States to the Mexican border (Brown 1962; Gordon 1985) (Figure 2).

The Canadian range of the Nine-spotted Lady Beetle stretches from Vancouver Island, primarily through southern Canada and the prairies to southern Quebec (Brown 1962; Gordon 1985; Grant pers. data) (Figure 2). At the northernmost extent of its range the Nine-spotted Lady Beetle has been recorded near: Quesnel (BC); Edmonton (AB); Lake Athabasca (SK) and Roberval (QC). The range map for the Nine-spotted Lady Beetle in Gordon (1985) contains one record from Great Slave Lake in the Northwest Territories. This record could not be verified and is considered outside its known geographic range. It is possible Nine-spotted Lady Beetle could range within southern portions of NT and YT; however, there are no verified records as of the preparation of this report (2016). The Canadian range for this species is based on historical and current collection records, although there are gaps in survey coverage and some records are quite old (> 50 years).

Within the last ten years there have been thirteen records of Nine-spotted Lady Beetles in Canada from: two sites in Cranbrook (BC); one site in Kamloops (BC); one site in Osoyoos (BC); two sites in Williams Lake (BC); one site in Calgary (AB); one site in Cardston (AB); three sites in Medicine Hat (AB); one site in Steveville (AB); and one site in Mont St-Hilaire (QC).

Extent of occurrence (EOO) for the Nine-spotted Lady Beetle is based on databased museum collections and surveys. Based on a minimum convex polygon within the extent of Canada's jurisdiction, the EOO from 1897 - 2014 (all databased records) is 3,253,910 km² (Figure 3). The EOO calculated from 1995 - 2004 records is 559,510 km² (Figure 4). The EOO calculated from 2005 - 2014 records is 716,847 km² (Figure 5).

An index of area of occupancy (IAO) based on the databased museum collections and surveys from 1897 - 2014 (all databased records) is 1,308 km² (Figure 3). The IAO calculated from 1995 - 2004 records is 64 km² (Figure 4). The IAO calculated from 2005 - 2014 records is 40 km² (Figure 5).

Museum and collection records for the Nine-spotted Lady Beetle date from 1897 - 2014. A database of almost 23,000 lady beetle records (Coccinellidae), including 1,061 records for the Nine-spotted Lady Beetle have been compiled from 26 collections across Canada (see Collections Examined). While this database contains records of lady beetles across all Canadian provinces and territories, Nine-spotted Lady Beetles are only recorded from British Columbia, Alberta, Saskatchewan, Manitoba, Ontario and Quebec (Table 1).

Surveys have not been systematic or comprehensive over time and across the range of the Nine-spotted Lady Beetle. There are large areas and time periods with little data. In Canada most search effort has also been focused within agricultural systems or near urban centres, rather than in less disturbed and natural habitats (Acorn 2007; McCorquodale et al. 2011). While some collections across Canada currently do not have information databased, within numerous other insect collections, specimens have been reliably identified to assess the historical status of lady beetles in Canada (McCorquodale et al. 2011; Grant pers. data).

In preparation for this status report, sites that had recent historic records of the Nine-spotted Lady Beetle were revisited and targeted surveys were conducted within possible geographic survey gaps, including remote natural areas in northern British Columbia, Alberta, Yukon and Northwest Territories (Figure 6). There were 230 sites searched in 2013 and 2014 for a total search effort of 262.4 hours (Table 2). For an obvious, easily collected beetle, this represents a relatively large search effort per site. However, only four specimens were found in previously known sites for Nine-spotted Lady Beetles; three in Medicine Hat (AB) and one in Osoyoos (BC).

The dispersal ability of Nine-spotted Lady Beetle is unknown. However, based on potential dispersal ability (under ideal conditions) of other lady beetle species (see Dispersal and Migration) the species could potentially fly from 18 km to up to 120 km in a single flight (Jeffries et al. 2013). These potential dispersal distances were used to estimate overlap between search effort and databased sites of Nine-spotted Lady Beetles. An 18 km radius around the 230 search effort sites in 2013 and 2014 overlapped with 287 databased sites and 729 sites with a 120 km radius (Figure 7 and Figure 8). As this species is broadly distributed and highly mobile, this search effort represents relatively decent coverage of known sites for Nine-spotted Lady Beetles.

The Nine-spotted Lady Beetle is a habitat generalist and known to occur where there are areas of shrubs or small trees interspersed with open grassy areas, but not continuous closed canopy forests. This species has been recorded within agricultural areas, suburban gardens, parks, coniferous forests, deciduous forests, prairie grasslands, meadows, riparian areas and other natural open areas. Within agricultural crops it was one of the more dominant lady beetles found on alfalfa, potatoes, corn, soybean, and cotton (Wheeler and Hoebeke 1995; Harmon et al. 2007; Losey et al. 2007; Gardiner et al. 2011). It was also readily found on a wide variety of other crops in gardens and on grass, clover and weeds (Wheeler and Hoebeke 1995; Harmon et al. 2007; Losey et al. 2007; Gardiner et al. 2011). The Nine-spotted Lady Beetle can also be found in a wide variety of non-agricultural vegetation including birch, pine, spruce, maple, mountain ash, poplar, willow, sage, cherry, alder, thistles, grasslands, and scruff pea plants along the edge of sand dunes (Wheeler and Hoebeke 1995; Acorn 2007; Harmon et al. 2007; Losey et al. 2007).

Nine-spotted Lady Beetles move across different habitats and vegetation to exploit seasonal changes in prey availability and their distribution is therefore driven to a large extent by prey availability rather than habitat type (Hagen 1962; Hodek and Honěk 1996; Sloggett and Majerus 2000; Hodek et al. 2012).

Overwintering adults tend to aggregate in well ventilated microhabitats such as under stones, rock crevices, in grass tussock, in leaf litter, or in tree bark (Hodek et al. 2012). Larvae are generally located in habitat with an abundance of prey, and pupate in the same habitat.

The Nine-spotted Lady Beetle has a large range in Canada spanning numerous ecozones and habitat types (Gordon 1985). This species also readily disperses short and long distances to exploit changes in prey availability over the season and across vegetation types. No studies have specifically related habitat trends to declines in Nine-spotted Lady Beetle subpopulations. It is unknown if specific habitat trends have caused this particular lady beetle, with its wide diet and habitat range, to decline over much of its known range across Canada.

However, widespread and cumulative habitat conversion could have potentially led to subpopulation declines in some parts of its range. Expansion of major urban centres, including areas of greater Vancouver, Victoria and Calgary, intensive use of agricultural landscapes, and other industrial practices lead to cumulative habitat quality decline and habitat loss (Federal, Provincial and Territorial Governments of Canada 2010; Javorek and Grant 2011).

In recent decades, the capacity of agricultural landscapes to provide habitat for wildlife has declined significantly across Canada's ecozones (Federal, Provincial and Territorial Governments of Canada 2010; Javorek and Grant 2011). One of the causes for this is the more intensive use of agricultural land. This includes heavier reliance on chemicals for pest control, which presumably could negatively affect Nine-spotted Lady Beetles directly, or indirectly by impacting their prey.

Abandonment of managed lands and farms resulting in regrowth of trees could also potentially result in less favourable foraging for the Nine-spotted Lady Beetle (Harmon et al. 2007; Bucknell and Pearson 2007). This slow natural succession has mainly occurred in eastern Canada.

While large-scale changes in habitat and prey availability suggest a possible explanation, there are no data to demonstrate causality between a changing landscape and lady beetle densities (Elliott and Kieckheffer 1990; Elliott et al. 1999; Harmon et al. 2007).

Information is compiled from general lady beetle references (Acorn 2007; Hodek et al. 2012) and where applicable references are provided specifically for Nine-spotted Lady Beetles.

Lady beetles are holometabolous, meaning they have a complete metamorphosis and pass through egg, larva, pupa and adult life stages. Nine-spotted Lady Beetles can have two generations per year (McMullen 1967) but the life history of lady beetles often depends on regional climatic conditions (Hodek et al. 2012). Adult Nine-spotted Lady Beetles have life spans which shorten with increasing temperature (Hodek et al. 2012). Within laboratory settings Nine-spotted Lady Beetle adults have been recorded to live for 62, 48 and 21 days at 21°C, 27°C and 32°C, respectively (McMullen 1967). This data suggests adults may live longer in cooler regions. Adults of the spring generation can undergo aestivation to avoid high summer temperatures, and lay eggs in early autumn (McMullen 1967; Hodek et al. 2012). Adults of the autumn generation congregated overwinter and undergo diapause, only becoming active and reproducing when temperatures rise in the early spring (McMullen 1967; Hodek et al. 2012; Losey et al. 2012).

At 25°C the pre-oviposition period (number of days between eclosion or emergence from pupae, and first egg laying) for the Nine-spotted Lady Beetle female is approximately 5 days, followed by an oviposition period of approximately 31 days (Ugine and Losey 2014). During this period Nine-spotted Lady Beetles can lay upwards of 690 eggs (Ugine and Losey 2014). The eggs of Nine-spotted Lady Beetles are laid upright, in tightly packed clusters of approximately 18 eggs, on a range of plants that are likely to support subpopulations of aphids (Acorn 2007; Hodek et al. 2012). Many females also lay unfertilized eggs, along with the fertile eggs, as another food source for young larvae (Acorn 2007).

Development from egg to adult takes approximately 20 days for the Nine-spotted Lady Beetle, depending on temperature (Ugine and Losey 2014). Larvae of the Nine-spotted Lady Beetle hatch from eggs after approximately 3 days (Ugine and Losey 2014). Nine-spotted Lady Beetles then undergo 4 instars before pupating, metamorphosing and reaching adulthood (Losey et al. 2012). This species reaches its third instar in approximately 4 to 5 days and takes an additional 7 days to reach its fourth instar and pupate. After approximately 5 days as a pupa, Nine-spotted Lady Beetles emerge as adults (Ugine and Losey 2014). One day after emerging, the elytra on adult Nine-spotted Lady Beetles harden (Losey et al. 2012). Male lady beetles locate females based on chemical and visual cues, and both sexes are polygynandrous mating with multiple partners (Omkar and Srivastava 2002; Srivastava and Omkar 2004; Acorn 2007).

In many lady beetles, the sex ratio is close to 1:1 and the activity of the follicular tissue in the testes starts in the pupa, so mating can begin shortly after emerging (Acorn 2007; Hodek et al. 2012). In the Nine-spotted Lady Beetle the sex ratio is 56:44, with slightly more females to males. Female Nine-spotted Lady Beetles on average also weigh more than males (30.3 mg vs. 25.6 mg), yet have a fairly equal body size (Smith 1966). Weight and size of adult lady beetles are also positively correlated with increased availability of food, which in turn is correlated with their ability to survive over winter (Smith 1966). When food is scarce lady beetles will have smaller body sizes and weights and decreased survivorship over winter (Smith 1966).

The Nine-spotted Lady Beetle displays aposematism, or bright warning colours to deter predators (Acorn 2007). Although undocumented, this species (like other lady beetles), likely is able to reflex bleed, releasing defensive alkaloids from tibio-femoral joints, when provoked (Hodek et al. 2012). There are about 50 different alkaloids that have been identified in lady beetles (Laurent et al. 2005). The various alkaloid compositions across species also vary in respect to their effects on predators (Marples et al. 1989; Laurent et al. 2005; Hodek et al. 2012).

Nine-spotted Lady Beetles also occupy a wide ecological niche across a variety of temperature regimes in Canada, are cold-tolerant, and as adults are able to overwinter. This plasticity also enables this species to exploit seasonal changes in prey availability across different habitats and vegetation (Hodek et al. 2012). Its ability to adapt, however, may be limited. Competition with other introduced species of lady beetles may be a factor in recent decreases in body size of Nine-spotted Lady Beetles (Losey et al. 2012) (see Interspecific Interactions).

Little is known on the natural dispersal rates specifically for the Nine-spotted Lady Beetle. In general lady beetles are very mobile, display low site fidelity, and readily engage in short- and long-distance dispersal (van der Werf 2000; Acorn 2007; Hodek et al. 2012). The ability to disperse relatively long distances has resulted in high rates of gene flow between subpopulations (Krafsur et al. 2005) and enables lady beetles to exploit changes in prey availability (Hodek et al. 2012).

Drivers of dispersal are a combination of prey density and environmental variables such as temperature, wind speed and rainfall (Ives et al. 1993; Hodek and Honěk 1996; van der Werf 2000; Cardinale et al. 2006; Krivan 2008; Jeffries et al. 2013). Previous work has also shown that lady beetle emigration decreases with increasing prey abundance (Ives 1981; Ives et al. 1993; Elliott 2000; van der Werf 2000; Cardinale et al. 2006; Jeffries et al. 2013) and the density of adult lady beetles is positively correlated with aphid density (Turchin and Kareiva 1989; Hodek and Honěk 1996; Osawa 2000; Evans and Toler 2007).

Calculating dispersal rates over longer distances has been hampered by the difficulty of tracking the insects in the field. One study using vertical-looking entomological radars determined that the majority of lady beetles fly at 150 - 479 metres above ground level (m AGL) perhaps due to decreasing air temperatures and increasing energetic requirements of reaching higher altitudes (Jeffries et al. 2013). Mean flight speed of lady beetles ranged from 31 km/h at 150 m AGL to 59 km/h at 1500 m AGL (Jeffries et al. 2013). Using tethered flight experiments, this study also estimated a mean flight time of 36.5 minutes, with a maximum of over 2 hours (Jeffries et al. 2013). Extrapolating from these results it was estimated that with ideal meteorological conditions, lady beetles could fly 18 km in a single flight (30 km/h for 36.5 minutes) and a few individuals flying at high altitudes and speeds (59 km/h for two hours) could potentially fly 120 km in a single flight (Jeffries et al. 2013).

Nine-spotted Lady Beetles are generalists in food and habitat use, often tracking changes in aphid abundance across many types of habitats (Hagen 1962; Hodek and Honěk 1996; Sloggett and Majerus 2000). Both adult and larval stages of the Nine-spotted Lady Beetle prey primarily on a wide variety of aphids (Acorn 2007; Hodek et al. 2012). They also prey on other small insects and eggs including spider mites, alfalfa weevils, leafhoppers, scale insects, psyllids, lepidopteran eggs, in addition to sap, nectar and pollen (Wheeler and Hoebeke 1995; Acorn 2007; Hesler et al. 2012; Losey et al. 2012). Lady beetles in general can be attracted to aphid densities of below 10 individuals per square metre, and even volatiles produced by herbivore-injured plants (Hodek et al. 2012).

The Nine-spotted Lady Beetle itself is also subject to intraguild predation by other introduced lady beetles (Turnipseed et al. 2014). There is a broad coincidence between subpopulation decline for the Nine-spotted Lady Beetle and the introduction and spread of the Seven-spotted Lady Beetle (Coccinella septempunctata) and the Multi-coloured Asian Lady Beetle (Harmonia axyridis). A direct causal link is not obvious, though potential mechanisms include direct competition for food, intraguild predation, and spread of new parasitoids or pathogens. Competition with introduced lady beetle species over aphid prey and other food is also suspected to have led to declines in the body size of Nine-spotted Lady Beetles (Losey et al. 2012) (see Threats). Declines in body size through competition may also reduce their ability to survive over winter (Smith 1966; Losey et al. 2012).

General predation on lady beetles by vertebrates such as birds is reduced by aposematic warning colours and distasteful defensive alkaloids excreted by reflex bleeding from the tibio-femoral joints (Laurent et al. 2005; Acorn 2007; Hodek et al. 2012). Despite these defences, lady beetles have been reported to be eaten by a wide range of vertebrate and invertebrate predators (Acorn 2007; Hodek et al. 2012). Web-building spiders are also frequently reported preying on lady beetles (Nentwig 1983; Richardson and Hanks 2009; Sloggett 2010).

Lady beetles, in general suffer from parasitism by various tachinid flies, phorid flies, chalcidoid wasps, parasitic mites, nematodes, sporazoans, fungi and bacteria (Wheeler and Hoebeke 1995; Acorn 2007; Bjornson 2008; Roy and Cottrell 2008; Hodek et al. 2012).

The braconid wasp (Dinocampus coccinellae) is the main parasitoid of numerous lady beetle species, including the Seven-spotted Lady Beetle and the Multi-coloured Asian Lady Beetle and can cause substantial reductions in subpopulations of the Nine-spotted Lady Beetle (Ceryngier and Hodek 1996; Abassi et al. 2001; Acorn 2007; Hodek et al. 2012). The braconid wasp currently has a cosmopolitan distribution covering all continents except Antarctica, and many islands (Hodek et al. 2012). The natural geographic range of the braconid wasp is difficult to reconstruct as it is believed this species arrived in some parts of its present distribution with lady beetles released for biological control purposes (Hodek et al. 2012).

Other interspecific interactions include parasitic mites (i.e., Coccipolipus hippodamiae), fungal pathogens (i.e., Beauveria bassiana), microsporidia (Nosematidae) and bacteria, which can all negatively impact lady beetle fitness and reduce survival over winter (Cali and Briggs 1967; Hurst et al. 1995; Barron and Wilson 1998; Webberley and Hurst 2002; Webberley et al. 2004).

Insect collections are important sources for information on geographic distribution of species (Wiggins et al. 1991). Specimens within Canadian collections have been collected by a mix of professional entomologists, students and keen amateurs during biodiversity inventories, general collections, taxon specific collections, ecological studies and applied studies on crops and forests. Data from collections have helped delineate geographic ranges of lady beetles and can be used to assess temporal changes in distribution and abundance if the strengths and weaknesses of collection data are understood and considered (McCorquodale et al. 2011).

Due to associated biases, accurately documenting changes in the geographic distribution of a species is a difficult task (Fortin et al. 2005; Elith et al. 2006; Koch and Strange 2009). Maps of geographic distribution may show a decrease in geographic range when in fact they reflect a decrease in subpopulation size, because with a reduced subpopulation there is a decrease in probability of collection (McCorquodale et al. 2011). In addition, collections can potentially be time series biased and may not reflect the true abundance of a species as experts may not continue to collect specimens of common lady beetles (McCorquodale et al. 2011). Conversely, newly introduced and invasive species might be collected out of proportion to the actual relative abundance of the species (McCorquodale et al. 2011).

Trends in absolute abundance are also biased by search effort. Therefore, relative abundance or the percent composition of a particular species relative to the total number of species is a common approach used to measure insect populations and reduce bias with search effort. For the Nine-spotted Lady Beetle, collection records are compared to all lady beetles (Coccinellidae) collected across similar time periods and geographic range as a proxy of abundance. In addition, collection records are also compared to only native lady beetles collected. As non-native species can potentially experience rapid subpopulation expansion and growth, inclusion of non-native species may produce artificially inflated declines. Conversely, as many species of native lady beetles are in decline across Canada, their use in measures of relative abundance may underestimate declines.

Multiple datasets from collections across Canada (see Collections Examined) were used to assess overall patterns of change in geographic distribution and relative abundance of the Nine-spotted Lady Beetle. The collated dataset contains almost 23,000 records of Coccinellidae from 1895 to 2014, including 1,061 Nine-spotted Lady Beetle specimens dated from 1897 - 2014. McCorquodale et al. (2011) visited numerous collections to identify and verify Coccinellidae specimens, before specimen label information was databased. Subsequently, additional museum and specimen data were compiled from surveys and collections during the preparation of this status report (Grant pers. data). Localities were georeferenced so that species could be mapped using geographic information system (GIS) software. Latitude and longitude were taken from labels when available, but for others, the latitude and longitude of the town centre on the label was used, unless a more specific locality could be determined. In 2013 and 2014 there were over 262.4 hours of field surveys conducted across 230 sites within this database (Table 2).

The following methods were used to characterize changes in the distribution of the Nine-spotted Lady Beetle over time and characterize search effort coverage:

1. Changes in the COSEWIC extent of occurrence (EOO) within the last ten years (2005 - 2014) compared to the previous decade (1995 - 2004) and all databased records (1897 - 2014) (Figures 3 - 5).
2. Changes in the COSEWIC index of area of occupancy (IAO) within the last ten years (2005 - 2014) compared to the previous decade (1995 - 2004) and all databased records (1897 - 2014) (Figures 3 - 5).
3. Search effort was combined with potential dispersal distances of 18 km and 120 km (Jeffries et al. 2013) to estimate overlap with databased Nine-spotted Lady Beetles sites (Figures 6 - 8).
4. The relative abundance of Nine-spotted Lady Beetles within museum collections, in ten-year increments across each jurisdiction where it is found. Relative abundance of the Nine-spotted Lady Beetle was calculated against all native and non-native lady beetles collected, in addition to all native lady beetles collected (Figure 9; Tables 3 and 4).

These data were supplemented by published research and expert opinion documenting subpopulation and range declines of the Nine-spotted Lady Beetle in North America.

Estimating abundance for wide-ranging insects such as the Nine-spotted Lady Beetle is not possible with current available data. As described above, changes in extent of occurrence (EOO), index of area of occupancy (IAO), and relative abundance will be used to measure conservation status.

Based on all databased records and surveys (1897 - 2014), the Nine-spotted Lady Beetle has an EOO of 3,253,910 km² and IAO of 1,308 km² (Figure 3). During 1995 - 2004 the EOO was calculated as 559,510 km² with an IAO of 64 km² (Figure 4). During the last decade (2005 - 2014) the EOO increased to 716,847 km², but IAO decreased to 40 km² (Figure 5). This is an estimated 28% increase in EOO and 37.5% reduction in IAO from the previous decade. The Nine-spotted Lady Beetle is a broadly distributed species across Canada and is highly mobile; surveys have not been complete over its entire range or time. Trends in this species' geographic distribution could reflect issues with survey coverage or detection rather than expansion or retraction of its range.

Historically the Nine-spotted Lady Beetle was widely distributed, occurring in southern British Columbia, Alberta, Saskatchewan, Manitoba, Ontario and Quebec. Within these areas, it was one of the more common lady beetles collected before 1975 (Brown 1940; Gordon 1985) but subsequently declined significantly (Acorn 2007; McCorquodale et al. 2011). During the previous decade (1995 - 2004) there were 65 Nine-spotted Lady Beetle records with a relative abundance of 0.027 compared to all native and non-native lady beetles (Coccinellidae) collected (Table 3a). During the last ten years (2005 - 2014), the number of records decreased to 13 from British Columbia, Alberta, and Quebec (Table 3a). During this decade it was not detected in Saskatchewan, and remained undetected in Manitoba, and Ontario, where it was found historically. The relative abundance of the Nine-spotted Lady Beetle therefore declined by -0.019 to only 0.008, which represents a national decline of 70.7% over the last ten years (Table 4a).

As non-native species can potentially experience rapid subpopulation expansion and growth, inclusion of non-native species may produce artificially inflated declines. Therefore, relative abundance calculations for the Nine-spotted Lady Beetle were also compared to only native species (non-native records removed). However, as many species of native lady beetles are in decline across Canada, their use in measures of relative abundance may underestimate declines. Relative abundance of Nine-spotted Lady Beetles, compared to native lady beetles declined from 0.057 (1995 - 2004) to 0.022 (2005 - 2014) (Table 3b), which represents a national decline of 62% over the last ten years (Table 4b). Therefore, national trends of decline are likely greater than 62% and potentially as high as 70.7%. Over the last decade and across the vast majority of its range this species is now absent or continues to decline and likely only persists in extremely low numbers.

The decline in relative abundance of Nine-spotted Lady Beetles is concurrent with an increase in collection of non-native species such as the Seven-spotted Lady Beetle and the Multi-coloured Asian Lady Beetle (Figure 9).

McCorquodale et al. (2011) also reviewed evidence from literature and collection data from Quebec and Ontario to look at relative abundance and geographic ranges of a subset of 10 species of native and non-native lady beetles over time. This study focused on regions with high quality data, complete over the time period non-natives arrived in Canada. Collections used in this study were made by university students and broad surveys and are a reasonable reflection of local relative abundance. Within this study McCorquodale et al. (2011) inferred that Nine-spotted Lady Beetles declined in geographic range and relative abundance from about 18% prior to 1960 to <0.05% after 1980, concurrent with an increase in collection of non-native species. However, the relative abundance of other native species such as the Spotted Lady Beetle (Coleomegilla maculata lengi) also increased after the arrival of non-native species (McCorquodale et al. 2011). Importantly, this contrast in trends suggest declines of the Nine-spotted Lady Beetle are not explained by collecting bias (McCorquodale et al. 2011).

Combined, data on changes in relative abundance and maps of changes in geographic range in Canada suggest that collecting effort has been comprehensive enough to observe lady beetle patterns and these declines are real, not artifacts of inadequate sampling and that the Nine-spotted Lady Beetle, while managing to persist in very low numbers, has continued to decline over the last ten years across its range in Canada.

In the northeastern United States, the decline of the Nine-spotted Lady Beetle was documented by Wheeler and Hoebeke (1995). They highlighted studies that showed it was a common species in many areas of the northeast from the 1950s through 1970s, yet rarely encountered after 1985. Intensive surveys of lady beetles in Iowa show that Nine-spotted Lady Beetles were common and widespread prior to 1980, but are now very rare or extirpated (Hesler 2009). In Minnesota Nine-spotted Lady Beetles were abundant prior to 1980 but recent search effort suggests this species is absent from or below the detection threshold across the majority of the state (Koch 2011). Harmon et al. (2007) reviewed published literature as well as United States Department of Agriculture (USDA) records and concluded the relative abundance of Nine-spotted Lady Beetle subpopulations have declined significantly in the United States and Canada since the 1970s.

In general, trends in the relative abundance of native to non-native lady beetle assemblages of Canada and the United States declined by 68% after 1986 (Harmon et al. 2007). A similar study conducted in Michigan over 24 years from 1989 to 2012 found lady beetle assemblages became increasingly non-native dominated with 71% of lady beetles collected being non-native (Bahlai et al. 2013). Gardiner et al. (2009) found that non-native Seven-spotted Lady Beetles and Multi-coloured Asian Lady Beetles accounted for up to 90% of lady beetle communities in soybean fields in Michigan, Wisconsin and Iowa. Tumminello et al. (2015) suggest declines of the Nine-spotted Lady Beetle can be attributed to the establishment, spread and subpopulation increase of the Seven-spotted Lady Beetle. While reasons for the decline in native lady beetles remain unclear, there is a very clear and real trend in declines of native lady beetles across their range, including the Nine-spotted Lady Beetle.

In summary, this once-common lady beetle now appears to be very rare or below detection thresholds in many parts of its range. Continued declines in relative abundance and geographic range have been documented in numerous studies, throughout the Nine-spotted Lady Beetle's range across Canada and the United States (Staines et al. 1990; Wheeler and Hoebeke 1995; Marshall 1999; Stephans 2002; Acorn 2007; Harmon et al. 2007; Hesler and Kieckhefer 2008; Fothergill and Tindall 2010; Skinner and Domaine 2010; Evans et al. 2011; Koch 2011; McCorquodale et al. 2011).

Natural population fluctuations in lady beetle subpopulations are related to dispersal, prey availability, climatic conditions and overwinter survivorship. However, lady beetles, including the Nine-spotted Lady Beetle, do not experience extreme fluctuations (Acorn 2007; Harmon et al. 2007; McCorquodale et al. 2011). The recent and steady decline of this species across its entire global range, in many disparate areas, suggests that this trend is not likely a natural fluctuation.

The Nine-spotted Lady Beetle is broadly distributed and its range extends into the United States from coast to coast. As this species is highly mobile and readily disperses, subpopulations could potentially disperse and recolonize areas where the Nine-spotted Lady Beetle has declined, provided suitable habitat was available. However, as this species has also declined in the United States, and the reasons for the decline remain unknown, it is unlikely that rescue effect is possible.

The International Union for the Conservation of Nature - Conservation Measures Partnership (IUCN-CMP) threats calculator (Salafsky et al. 2008; Master et al. 2009) was used to classify and list threats to the Nine-spotted Lady Beetle. The results of the threats calculator show an overall threat impact of very high to high (Appendix 1). Threats below are listed in order of highest to lowest threat.

It has been widely reported that the accidental and intentional introduction of non-native species can negatively impact flora and fauna (New 1995; Cottrell and Shapiro-Ilan 2003; Evans 2004; Snyder and Evans 2006; Finlayson et al. 2008; Kenis et al. 2008; Kajita and Evans 2009; Crowder and Snyder 2010; Smith and Gardiner 2013; Ugine and Losey 2014; Tumminello et al. 2015). Insect generalist predators have been introduced outside their native range inadvertently or intentionally as biocontrol agents during the last century (Obrycki and Kring 1998; Evans et al. 2011). In North America alone, at least 179 non-native lady beetle species have been introduced, leading to nine non-native species becoming well established in Canada, including the Seven-spotted Lady Beetle and the Multi-coloured Asian Lady Beetle (Gordon 1985; Gordon and Vandenberg 1991; Harmon et al. 2007; Evans et al. 2011; McCorquodale et al. 2011; Bousquet et al. 2013). These non-native species continue to be widely available and released for biocontrol.

Significant declines in geographic range and abundance of native lady beetles are frequently due to changes in habitat or interactions with non-native species (New 1995; Cottrell and Shapiro-Ilan 2003; Evans 2004; Snyder and Evans 2006; Finlayson et al. 2008; Kenis et al. 2008; Kajita and Evans 2009; Crowder and Snyder 2010; Smith and Gardiner 2013; Ugine and Losey 2014; Tumminello et al. 2015).

The invasion of the Seven-spotted Lady Beetle and Multi-coloured Asian Lady Beetle into North America has been implicated in an overall reduction in Nine-spotted Lady Beetle and other native lady beetle subpopulations (Wheeler and Hoebeke 1995; Elliott et al. 1996; Marshall 1999; Ellis et al. 1999; Brown 2003; Cottrell and Shapiro-Ilan 2003; Turnock et al. 2003; Hesler et al. 2004; Acorn 2007; Harmon et al. 2007; Hesler and Kieckhefer 2008; Fothergill and Tindall 2010; Skinner and Domaine 2010; Evans et al. 2011; Losey et al. 2012; Comont et al. 2013; Turnipseed et al. 2014; Ugine and Losey 2014; Tumminello et al. 2015). Most explanations for this reduction in native subpopulations focus on negative interactions through competition, intraguild predation or indirect effects such as the introduction of pathogens (Schaefer et al. 1987; Ehler 1990; Cottrell and Shapiro-Ilan 2003; Louda et al. 2003; Evans 2004; Lucas 2005; Snyder and Evans 2006; Lucas et al. 2007; Kenis et al. 2008; Riddick et al. 2009; Evans et al. 2011; Turnipseed et al. 2014; Ugine and Losey 2014; Tumminello et al. 2015).

Competition and intraguild predation:

The geographic distribution of Nine-spotted Lady Beetles has declined rapidly across North America closely following the establishment, spread and subpopulation growth of non-native Seven-spotted Lady Beetles and Multi-coloured Asian Lady Beetles (Turnipseed et al. 2014; Ugine and Losey 2014; Tumminello et al. 2015). Before the 1980s Nine-spotted Lady Beetles were a fairly common lady beetle in North America (Gordon 1985; Tumminello et al. 2015). However, declines of Nine-spotted Lady Beetles were not widely recognized until the mid-1990s, almost 20 years after the arrival and subsequent spread and establishment of non-native lady beetles (Wheeler and Hoebeke 1995; McCorquodale et al. 2011).

Hodek and Michaud (2008) argued that the Seven-spotted Lady Beetle is a good competitor under a wide variety of conditions. Its ability to compete for food, mate, and lay eggs under a variety of conditions result in its doing well overall, rather than its being the best under a particular set of conditions (Hodek and Michaud 2008; McCorquodale et al. 2011). Losey et al. (2012) determined that scramble competition with Seven-spotted Lady Beetles has resulted in limited prey availability and decreased body size of Nine-spotted Lady Beetles. In support of scramble competition, where a finite resource is accessible to all competitors, Hoki et al. (2014) showed that Seven-spotted Lady Beetles were more voracious, had a higher aphid attack rate and lower aphid handling time compared with Nine-spotted Lady Beetles. Tumminello et al. (2015) also investigated scramble competition and intraguild predation, concluding that the displacement of the Nine-spotted Lady Beetle from its native range was likely driven by the Seven-spotted Lady Beetle, based on its faster development times, higher attack rate, larger body size and high rate of intraguild predation of Nine-spotted Lady Beetle.

Other studies have also shown that Nine-spotted Lady Beetle larvae are more likely to survive to become adults when reared with native larvae than with Seven-spotted Lady Beetle larvae, due to low predation rates on their eggs and larvae by native species (Turnipseed et al. 2014). Similar results were found for other native and introduced lady beetle species (Obrycki et al. 1998; Michaud 2002; Sato et al. 2004; Snyder et al. 2004; Lucas et al. 2007; Pell et al. 2008; Gardiner et al. 2011; Hodek et al. 2012; Smith and Gardiner 2013). Intraguild predation also plays a major role in preventing recolonization by native lady beetles, and females also avoid oviposition sites where intraguild predators are present (Ruzicka 1997; Hodek et al. 2012). Establishment of Seven-spotted Lady Beetles in agricultural landscapes has also resulted in documented declines of native lady beetles and aphid density (Alyokhin and Sewell 2004; Evans 2004).

Despite documented declines in subpopulations of native species of lady beetles in Canada (e.g., Turnock et al. 2003) and the arrival and range expansion of non-native lady beetles in North America (e.g., Wheeler and Stoops 1996; Lucas et al. 2007), the links between the non-native species and causes of the declines are not clear. For example, Acorn (2007) and Harmon et al. (2007) argued that there is little direct evidence that competition or other interactions with recently arrived non-native species have caused the declines in native species. While trends are consistent with expectations if Seven-spotted Lady Beetles and Multi-coloured Asian Lady Beetles negatively impact Nine-spotted Lady Beetles through scramble competition and intraguild predation, other potential mechanisms include introduction of parasitoids or pathogens (Losey et al. 2012).

Parasites, parasitoids, pathogens and fungi:

Non-native species may also affect native lady beetles indirectly through the introduction and transmission of new natural enemies such as parasites and pathogens (Bjornson 2008). Lady beetles are hosts to a variety of parasitoids (i.e., braconid wasp), parasitic mites (i.e., Coccipolipus hippodamiae), nematodes, protozoans, fungal pathogens (i.e., Beauveria bassiana), microsporidia (Nosematidae), and bacteria can all negatively impact lady beetle fitness and reduce survivorship overwinter (Cali and Briggs 1967; Hurst et al. 1995; Ceryngier and Hodek 1996; Barron and Wilson 1998; Webberley and Hurst 2002; Cottrell and Shapiro-Ilan 2003; Webberley et al. 2004; Bjornson 2008; Roy and Cottrell 2008; Riddick et al. 2009; Bjornson et al. 2011). Although the effect of these natural enemies on the Nine-spotted Lady Beetle is uncertain, native species often have a greater susceptibility to exotic pathogens (Cottrell and Shapiro-Ilan 2003). Obrycki (1989) reported on greater susceptibility of native lady beetles to the braconid wasp parasitoid compared to non-native species, such as the Multi-coloured Asian Lady Beetle. Cottrell and Shapiro-Ilan (2003) also reported on greater susceptibility of native lady beetles to a fungal pathogen (Beauveria bassiana) compared to the Multi-coloured Asian Lady Beetle. Greater susceptibility to exotic pathogens may therefore provide an intraguild advantage to non-native lady beetles and could have been a contributing factor in declines of Nine-spotted Lady Beetles.

While lady beetles can be more tolerant of pesticides than their prey (Gesraha 2007), pollution via agrochemicals to reduce insect pests can impact non-target lady beetles directly through topical contact; residual contact; inhalation of volatiles; and ingestion of insecticide-contaminated prey, nectar or pollen (Smith and Krischik 1999; Youn et al. 2003; Singh et al. 2004; Moser et al. 2008; Moser and Obrycki 2009; Eisenback et al. 2010) and indirectly through eliminating their food supply (Hodek et al. 2012; Bahlai et al. 2015). Zoophytophagy, omnivorous feeding behaviour that occurs when plant material (pollen, nectar, leaf tissue) is consumed by primarily predaceous species, increases fecundity and reduces development time (Coll 1998; Patt et al., 2003; Moser and Obrycki 2009). However, zoophytophagy can also be harmful if the plant material is chemically protected by insecticides (Moser and Obrycki 2009). Lady beetle susceptibility to insecticides varies with the species and the type of pesticide and can range from acute lethal effects to reduction in fecundity, behaviourally or reproductively by non-lethal concentrations of insecticides (Theiling and Croft 1988). Many insect predators exposed to more than one compound suffer synergistic detrimental effects, even for compounds that were equitably harmless when tested separately (Petersen 1993).

In urban and agricultural landscapes, lady beetle subpopulations may be threatened by a variety of pesticides including neonicotinoids, insect growth regulators and broad-spectrum pyrethroids, which tend to be more destructive to lady beetles than organophosphates (Kumar and Bhatt 2002; Moser and Obrycki 2009). Insect growth regulators such as buprofezen and pyriproxyfen generally lack acute toxicity to lady beetles, but may impair development and fecundity (Olszak et al. 1994). Neonicotinoids are a class of systemic pesticides that travel and accumulate throughout the plant, including in pollen and nectar. While very effective against plant pests, especially aphids, these pesticides have proven to be detrimental to insects at concentrations in the parts per billion (ppb) (Smith and Krischik 1999; Marletto et al. 2003). Neonicotinoids can also be applied to seeds prior to planting to protect seedlings from early-season root and leaf-feeding. In one study 72% of Multi-coloured Asian Lady Beetle larvae (Harmonia axyridis) exposed to seedlings treated with neonicotinoids developed neurotoxic symptoms (trembling, paralysis, and loss of coordination) from which only 7% recovered (Moser and Obrycki 2009). Therefore, the use of neonicotinoids may have negative effects on non-target species especially if zoophytophagy occurs.

Abandonment of managed lands and farms, specifically in eastern Canada, could potentially be a factor in the decline of the Nine-spotted Lady Beetle (Bucknell and Pearson 2007; Harmon et al. 2007). Urban expansion and abandonment of farmland may mean less favourable foraging for the Nine-spotted Lady Beetle (Harmon et al. 2007). While these large-scale changes in habitat and prey availability suggest a possible explanation, there are no data to demonstrate causality between a changing landscape and lady beetle densities (Elliott and Kieckheffer 1990; Elliott et al. 1999; Harmon et al. 2007).

Habitat loss and declines in habitat quality are ongoing throughout the species' range (Federal, Provincial and Territorial Governments of Canada 2010; Javorek and Grant 2011). Homogenization of agricultural landscapes and changing agricultural practices such as intensive reliance on fertilizers and pesticides may also be contributing to local declines in native species (Wheeler and Hoebeke 1995; Bianchi et al. 2007; Evans et al. 2011). This is discussed in the pollution section (Threat 9).

Planting of genetically modified (GM) insect-resistant crops, e.g., GM maize engineered to express Bacillus thuringiensis (Bt) toxins was considered a potential risk to lady beetles because the toxin was present in pollen (Harwood et al. 2007), but not present in aphids (Hodek et al. 2012). While most studies have found no effect of Bt corn pollen consumption on fitness parameters of lady beetles (Duan et al. 2002; Lundgren and Wiedenmann 2002; Porcar et al. 2010), others have detected reduced fecundity and developmental delays (Moser et al. 2008).

Habitat loss and declines in habitat quality from expansion of residential and commercial developments may be contributing to local declines of this species. Green areas and local gardens within smaller urbanized area, however, may also still provide habitat for the Nine-spotted Lady Beetle.

It is not possible to calculate the number of locations for this species. The term 'location' defines a geographically or ecologically distinct area in which a single threatening event can rapidly affect all individuals of the taxon present. This species has a very broad geographic range, it is highly mobile and threats to it remain unclear. In absence of clearly defined threats over its range, the term 'location' cannot be used and the subcriteria that refer to the number of locations will not be met.

In regard to the number of sites, within the last ten years there have been thirteen records of the Nine-spotted Lady Beetle in Canada from nine sites: two sites in Cranbrook (BC); one site in Kamloops (BC); one site in Osoyoos (BC); two sites in Williams Lake (BC); one site in Calgary (AB); one site in Cardston (AB); three sites in Medicine Hat (AB); one site in Steveville (AB); and one site in Mont St-Hilaire (QC). Given its broad geographic range and dispersal ability, it is likely this species occurs at additional sites throughout its range.

There are no federal or provincial laws that protect the Nine-spotted Lady Beetle, mitigate threats to this group of insects or protect the species' nest sites or habitat.

In Quebec, this species is not currently listed as Threatened or Vulnerable (LEMV 2015). However, it is integrated on the list of Threatened or Vulnerable species (LEMV 2015) and therefore considered an at-risk species and afforded protection under sections 22 and 31.1 of the "Loi sur la qualité de l'environnement" (RLRQ, c. Q-2) (Environment Quality Act) (CQLR, c. Q-2).

The global status rank is G2 (imperilled) and the national status rank is unranked in Canada and the United States (NatureServe 2014). It is also unranked in most states, provinces and territories, but considered possibly extirpated in Alberta (SNR), Ontario (SH), Connecticut (SH), and Florida (SH).

The Canada National Status Ranks (Canadian Endangered Species Conservation Council [CESCC 2010]): Sensitive in Canada and provincially in NT, BC, AB and SK; Maybe At Risk in MN; Extirpated in ON and QC.

The IUCN Red list (2013): None

The species has not been reviewed or listed under the USA - federal Endangered Species Act.

Given the expansive range and broad habitat niche of the Nine-spotted Lady Beetle across Canada, several suitable areas of habitat occur within privately owned, urban and agricultural land, public land and protected areas.

The Canadian range of Nine-spotted Lady Beetle spans numerous provincial and national parks and protected areas.

The writer wish to thanks Jennifer Heron for supervising this report, as well Angele Cyr (COSEWIC Secretariat), in addition to David McCorquodale (Cape Breton University); John Acorn (University of Alberta); Isabelle Gauthier (Ministère des Forêts, de la Faune et des Parcs); John Losey (Cornell University); Cory Sheffield (Royal Saskatchewan Museum); Suzanne Carriere, Danny Allaire, Nicholas Larter (Northwest Territories Government); Mary Sabine (New Brunswick Government); Barb Sharanowski (University of Manitoba); Gilles Boiteau (Agriculture Canada); Ken Millard, Lisa Ott (Galiano Conservancy Association); Claudia Copley (Royal British Columbia Museum), Darren Copley, Rob Cannings (Royal British Columbia Museum); Syd Cannings (Canadian Wildlife Service); Gary Anweiler, Heather Leibel, Mattias Buck, Heidi Gartner, Robb Bennett, Erica McClaren, Berry Wijdeven, Mark Weston, Lynn Westcott, Sandy Cessellie, Bill Ramey, Bev Ramey, Michael Dunn, Geoff Lynch, Nick Burdock, Jeevan Sandu, Sylvia Latay, Rick Howie, Lea Gelling, Kathy Coot, Tom Foot, Karen Durovich, Sara Kalnay-Watson, Al Harris, Rob Foster, Vincent Bereczki, Bruce Bennett, and the Lost Ladybug Project.

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Acorn, J. 2007. Ladybugs of Alberta: Finding the spots and connecting the dots. The University of Alberta Press, Edmonton, Alberta.

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Wheeler, A.G., and E.R. Hoebeke. 1995. Coccinella novemnotata in northeastern North America: historical occurrence and current status (Coleoptera: Coccinellidae). Proceedings of the Entomological Society of Washington 97:701-716.

Wheeler, A.G., and C.A. Stoops. 1996. Status and spread of the Palaearctic lady beetles Hippodamia variegata and Propylea quatuordecimpunctata (Coleoptera: Coccinellidae) in Pennsylvania, 1993-1995. Entomological News 107:291-298.

Wiggins, G.B., S.A. Marshall, and J.A. Downes. 1991. The importance of research collections of terrestrial arthropods. Brief for the Biological Survey of Canada (Terrestrial Arthropods)

Youn, Y.N., M.J. Seo, J.G. Shin, C. Jang, and Y.M. Yu. 2003. Toxicity of greenhouse pesticides to multicolored Asian lady beetles, Harmonia axyridis (Coleoptera: Coccinellidae) Biological Control 28:164-170.

Dr. Paul Grant is an avid entomologist who has worked with many insect groups including dragonflies, butterflies, katydids and beetles. Currently, his research focus involves insect bioacoustics, specifically, the limitations and solutions for effective communication, predator-prey relationships, and utilizing insect calls to monitor species and habitats. Paul is also a Grasshopper Specialist Group member (GSG) for the International Union for Conservation of Nature & Species Survival Commission (IUCN / SSC) and Co-Chair for the Arthropod Species Specialist Committee (SSC) for the Committee on the Status of Endangered Wildlife in Canada (COSEWIC).

• Acadia University ^Footnote1
• Atlantic Forestry Centre, Fredericton ^Footnote1
• Canadian Forestry Service, Newfoundland ^Footnote1
• Canadian National Collection, Ottawa
• Canadian Museum of Nature ^Footnote1
• Claude Chantal Collection, Quebec
• Collection d'insecte du Québec
• Collection Ouellet-Robert, Quebec
• Great Lakes Forestry Centre ^Footnote1
• Greg Pohl pers. comm. 2014
• Insectarium Montréal
• IRM Collection, Quebec
• John Acorn pers. comm. 2014
• Laurentian Forestry Centre ^Footnote1
• Lost Lady Bug Project
• Musée d'Entomologie Lyman
• Nova Scotia Agricultural College ^Footnote1
• Royal Alberta Museum
• Royal BC Museum
• Royal Ontario Museum ^Footnote1
• Université Laval
• University of Alberta
• University of British Columbia
• University of Guelph ^Footnote1
• University of Manitoba
• University of New Brunswick ^Footnote1