Plumage colour variations in the Agapornis genus: A review

Henriëtte van der Zwan*1, Carina Visser2 & Rencia van der Sluis1
1
Centre of Human Metabolomics, North-West University, Potchefstroom, South Africa
2
Department of Animal and Wildlife Sciences, University of Pretoria, Pretoria, South Africa
*Corresponding author Email: agapornis.genome.study@gmail.com
ORCID ID: 0000-0001-5288-7609

ABSTRACT

The genus Agapornis consists of nine small African parrot species that are globally well known as pets, but are

also found in their native habitat. Illegal trappings, poaching and habitat destruction are the main threats these

birds face in the wild. In aviculture, Agapornis breeding is highly popular all across the globe. Birds are mainly

selected based on their plumage colour variations but very little molecular research has been conducted on this

topic. There are 30 known colour variations amongst the nine species and most of these are inherited as

Mendelian traits. However, to date none of the genes or polymorphisms linked to these variations have been

identified or verified. Due to unethical breeding practices the need for the development of molecular tests such

as identification verification tests or species identity tests is growing. Future research is paramount to ensure the

conservation of wild populations as well as aiding breeders in improving breeding strategies.

Keywords: Psittacidae, lovebirds, aviculture, molecular breeding tools

INTRODUCTION

Globally, there are 356 extant psittaciform, or parrot, species (Forshaw, 1989) of which twenty are native to the

African continent and Madagascar (Perrin, 2009). Amongst this order, the genus Agapornis (Selby, 1836) is

found and consists of nine species, namely A. roseicollis, A. taranta, A. canus, A. personatus, A. lilianae, A.

swindernianus, A. fischeri, A. pullarius and A. nigrigenis (Moreau, 1948; Dilger, 1960; Forshaw, 1989). These

species are more commonly known as lovebirds and are distributed across Madagascar (A. canus) and Africa

(the remaining eight species) (Dilger, 1960; Forshaw, 1989; Perrin, 2012). Table 1 lists the distribution of each

species, size of the birds, known sub-species, common name and popularity of each species in aviculture. In

addition to existing in their natural habitat, lovebirds are popular pets and eight of the nine species, A.

swindernianus being the exception, are kept and bred as domestic birds (Hayward, 1979; Silva & Kotlar, 1989;

Forshaw, 1989; Van den Abeele, 2016).

1
 Lovebirds have been kept as pets since the eighteenth century with reports of A. pullarius being bred in

Britain in the 1880’s (Parkes, 1982). Hayward (1979) states that lovebirds are popular in aviculture due to the

fact that they breed relatively easy in captivity, are hardy, active and have a range of plumage colours. They can

also be bred in a small aviary making them ideal birds for today’s modern lifestyle. As a consequence of their

popularity in aviculture, birds in their natural habitat are being threatened by poaching and trapping for export to

pet markets (Warburton & Perrin, 2005 & 2006; Mzumara et al., 2016 a & b). Plumage colour variations are the

main economic factor breeders select for and there are at least 30 colour variations amongst domestic lovebirds

that are accepted by shows and auctions (Hayward, 1978, Van den Abeele, 2016). Rare colour variants are sold

for up to 700 times as much as a wild type coloration bird of the same species in Europe (Personal

communication: Mr. Dirk Van den Abeele) and there are reports of mark-ups of up to 1300 times in Asia

(Diega, 2017). All of these colour variants are genetically inherited, but to date the only reports of the mode of

inheritance of these traits are breeders’ records. Due to many of these colour variants being recessively

inherited, it may lead to inbreeding of related birds in order to create a recessive plumage genotype. Since some

lovebird species can inter-mate, hybridization of different species is also of great concern in aviculture.

Despite their popularity, very little molecular genetic research has been conducted on these species, in

particular for plumage coloration inheritance. Currently the genes and polymorphisms linked to lovebird

coloration are not yet known and therefore breeders cannot screen young birds to confirm their colour genotype.

There is also no screening test to identify species nor a species-specific parentage panel or identity confirmation

test available. These tests could be useful not only in aviculture but also in conservation to monitor natural

populations.

This paper reviews the importance of the conservation of the lovebird species in their natural habitat as

well as the role these birds play in aviculture. The mechanisms of parrot plumage colour expression will be

discussed as well as the different colour variations that are found in this genus and the importance thereof as an

economic driving factor for these species. The lack of genetic screening tests makes genetic progress in

aviculture difficult and this paper briefly investigates the different diagnostic tests that could improve lovebird

breeding.

GENUS AGAPORNIS

Lovebirds have been domesticated since the 1880’s but surprisingly little molecular genetic research has been

conducted on these species. The first taxonomic classification of the species was done by Moreau (1948). The

2
Figure 1. The classification of the nine Agapornis species as made by Manegold &
Podsiadlowski (2014) (Adapted from Manegold & Podsiadlowski, 2014).

3
nine extant species were classified into two groups (Group A and B) with two sister taxon groups consisting of

A. roseicollis and A. swindernianus respectively, as can be seen in Figure 1. This classification was revised by

Dilger (1960) who classified them only as three different groups based on the presence or absence of the white

eye ring, whether they are monomorphic or not and an intermediate group.

Molecular genetic advances gave researchers the opportunity to study these birds on a more detailed

genetic level in the late 1990’s. Eberhard (1998) sequenced a portion of the cytochrome-b (cytb) mitochondrial

gene of eight Agapornis species (with the exception of A. swindernianus). The molecular evidence from this

study confirmed the classification by Dilger (1960) and categorized the species as the eye-ring group, sexually

dimorph group and transitional group. In a later study by Manegold & Podsiadlowski (2014) the cytb

mitochondrial gene of a museum sample from A. swindernianus was additionally analysed and A. swindernianus

was found to be a sister clade to all other Agapornis species (Figure 1). A. canus, A. pullarius, A. swindernianus

and A. roseicollis all have known subspecies (Table 1). Some of the sub-species are poorly differentiated and

researched (Forshaw, 1989; Van den Abeele 2016).

Karyotyping of the A. roseicollis genome by Nanda et al. (2007) revealed that this species has a diploid

number of 48 macro-chromosomes. In the same study the budgerigar (Melopsittacus undulates) was found to

have 2n = 62 macro-chromosomes. The lower number of Agapornis chromosomes might indicate a fusion of

ancestral micro-chromosomes (Nanda et al., 2007). The budgerigar had a marginally larger genome size of 1.2

Gb (Ganapathy et al., 2013) compared to the A. roseicollis genome of 1.1 Gb (van der Zwan et al., 2018).

Crossbreeding between some of the species is possible and can produce viable hybrids (McCarthy, 2006; Van

den Abeele, 2016). The species’ habitat ranges do not overlap and therefore interbreeding is rarely found in the

wild. The only habitat overlap is between A. personatus and A. swindernianus (Moreau, 1948; Dilger, 1960;

Forshaw, 1989) but these species’ behaviour is so different that, even though not well researched, it is doubted

that interbreeding occurs. In aviculture, unfortunately, it is not uncommon to see hybridization of species.

Hybrids are not generally accepted at auctions and shows (Van den Abeele, 2016) and crosses between species

should not be encouraged in aviculture. McCarthy (2006) list the possible observed crosses between Agapornis

species that could produce viable young and a summary is given in Table 2 (viable young born depicted with an

“x”).

Viable offspring hatching from a mating between an A. canus male and Melopsittacus undulatus (Budgerigar)

female, A. nigrigenis male and M. undulates female, A. personatus male and M. undulates female and A.

roseicollis male and M. undulates female (McCarthy, 2006), have also been reported. It is well known that some

4
Table 1. A summary of Agapornis species

Species Distribution Size Sub-species Common name Aviculture References

A. canus Madagascar 14cm A. c. canus and Lavender head, grey Less common Forshaw, 1989; Haywards,

A. c. ablectaneus head, Madagascar 1979; Van den Abeele,

lovebird 2016

A. taranta Ethiopia, Eritrea 17cm None Black-winged or Less common Forshaw, 1989; Marez,

and Djibouti Abyssinian lovebird 2003; Tekalign & Bekele

2011, Van den Abeele,

2016

A. pullarius Across the African 15cm A. p. pullarius and Red-headed lovebird Uncommon Silvia & Kotlar, 1988;

equator from A. p. ugandae Forshaw, 1989; Van den

Kenya to Guinea Abeele, 2016; Egwumah &

over 26 countries Iboyi, 2017

A. roseicollis Angola, Namibia, 16cm A. r. roseicollis and Peach-faced or rosy- Very common Forshaw, 1989; Ndithia et

Botswana and A. r catumbella faced lovebird al., 2007; Van den Abeele,

South Africa 2016

A. Cameroon, Gabon, 13cm A. s swinderninus Black-collared lovebird Not found Forshaw, 1989; Manegold

swindernianus Congo, CAR, DRC A. s zenkeri & Podsiadlowski, 2013

and Uganda A. s emini

5
Species Distribution Size Sub-species Common name Aviculture References

A. personatus Tanzania 15cm None Masked or black- Very common Forshaw, 1989; Perrin

masked lovebird 2009, Van den Abeele,

2016

A. lilianae Zimbabwe, Zambia 14.5cm None Lilian’s lovebird Very common Forshaw, 1989; Mzumara

and Malawi et al., 2016 a & b; Van den

Abeele, 2016

A. fischeri Tanzania 15cm None Fischer’s lovebird Very common Forshaw, 1989;

Mwangomo et al. 2007;

Van den Abeele, 2016

A. nigrigenis Zambia 14cm None Black cheeked lovebird Very common Forshaw, 1989; Warburton

& Perrin, 2005 & 2006;

Van den Abeele, 2016

6
Table 2. Possible viable matings amongst domestic Agapornis species.

A. A. A. A. A. A. A. A.

canus fischeri nigrigenis lilianae personatus roseicollis pullarius taranta

A. canus x x x x x x x

A. fischeri x x x x x x

A. nigrigenis x x x x x

A. lilianae x x x x x

A. personatus x x x x x x x

A. roseicollis x x x x x x

A. pullarius x x x x

A. taranta x x x

7
plumage colour variations were introduced to other Agapornis species by intentional crossing of the species that

can intermate (van den Abeele, 2016).

Ecology and threats to natural populations

The behaviour and habitat of the nine species varies greatly - A. swindernianus are exclusive forest canopy

dwellers, A. lilianae prefer river beds and Mopane woodlands, A. taranta lives at high altitudes of 1600 to 3800

meters, A. pullarius are lowland tropical birds, A. roseicollis is found in semi-arid environments, A. fischeri and

A. personatus are lowland savanna and wood-land dwellers, A. canus prefers lightly wooded habitats and A.

nigrigenis can be found in mopane (Colophospermum mopane) woodlands (Foshaw, 1989; Perrin, 2009; Van

den Abeele, 2016). The main diet of most of the Agapornis species consist of grass seeds, however A.

swindernianus feed on ripened figs (Forshaw, 1989; Perrin, 2009). Moreau (1948) notes that A. swindernianus

cannot survive in captivity if not fed figs, which could explain why these birds are not kept as pets.

The International Union of Conservation of Nature (IUCN) lists Agapornis nigrigenis as vulnerable, A.

fischeri and A. lilianae as near threatened and the other species as least concern. All species except for A.

roseicollis are classified by The Convention on International Trade in Endangered Species of Wild Fauna and

Flora (CITES) as CITES Appendix II species, indicating that they are not threatened with extinction, but trade

must be controlled to prevent their extinction. Several studies have been conducted on the natural lovebird

populations and the threats these birds are faced with. In conclusion Tekalign & Bekele (2011) (on A. taranta),

Mzumara et al. (2016 a & b) (on A. lilianae), Egwumah & Iboyi, 2017 (on A. pullarius) and Perrin (2012) (all

species) found that reduction of their natural habitat due to agricultural activity and trapping or poaching is the

main threats to these species. In addition, Mzumara et al., (2016a) also found that the poisoning of birds in

Malawi is a common practice as these birds destroy crops.

Worldwide, there is a keen interest in lovebird breeding. Membership of the world’s largest breeding

association - The Belgium Agapornis Breeders Association (BVA-International) - is approximately 5000

members (Personal communication, Mr Dirk Van den Abeele) and other societies across Europe, Africa,

Australia, the United States of America, South America and Asia (Van den Abeele, 2016) exists with various

member numbers. In a recent poll conducted by Ornitho Genetics VZW (2017), the most popular lovebird

species bred in captivity was A. fischeri, followed by A. personatus and A. roseicollis. The popularity of a

species in aviculture could drastically influence their numbers in the natural habitat as traders will trap these

birds in order to sell them. Perrin (2012) reports that A. roseicollis, personatus and fischeri are all heavily traded

8
whereas A. lilianae are traded moderately and A. nigrigenis to a lesser extent. Trade and trappings are banned in

several African countries which should have an effect on the natural populations’ growth.

Aviculturists have the responsibility to breed with purebred, healthy birds to ensure that if the wild

populations go extinct, these species will still survive in aviculture. With the use of molecular diagnostic tests,

breeders will be able to select birds with recessive colour variations without compromising the biodiversity of

the species. Unrelated birds that are heterozygous for a recessive colour trait, could be identified and mated,

rather than mating close relatives that are suspected heterozygotes. Breeders should be made aware of the

importance of conserving wild populations as well as ethical breeding practices. This could be attained by

promoting the importance of not selecting breeding pairs based on one trait (colour) only, but to keep

biodiversity and general genetic fitness in mind.

AGAPORNIS PLUMAGE COLORATION

As mentioned earlier, the main economic factor breeders select for is plumage coloration and many of these are

inherited as autosomal recessive traits (Table 3). This implies that an offspring with wildtype coloration has a

certain chance (depending on the parents’ genotypes) of being a heterozygote (thus a carrier or split) of this trait.

Breeders sell possible heterozygotes at a premium despite the fact that the buyer has neither proof that the bird is

an offspring of the colour variant nor that it is indeed a heterozygote for the colour variation. This could lead to

fraudulent transactions and emphasises the need for more research in the field of plumage colour genetics and

parentage verification testing for lovebirds.

Agapornis plumage colour and biological mechanisms controlling parrot coloration

Most of the Agapornis species has at least one colour variation that is found infrequently in wild populations

and, due to selective breeding, frequently in domestic populations. The main colour of all wild type Agapornis

birds is green with red, yellow, pink, black or grey on their heads, chest and neck and some blue in their tail

feathers, (Hayward, 1978, Silva & Kotlar, 1988, Forshaw, 1989; Van den Abeele, 2016), as seen in the photos

of the wildtype coloration of the eight domestic Agapornis species in Figure 2. In Figures 3 a- h some of the

colour variants of the different species are shown with the wildtype variant of that species. Reports on wild

lovebirds with colours other than the wildtype coloration is scarce but Warburton & Perrin (2005) observed one

yellow and three light-green A. nigrigenis birds in the mid Machile River region between May 1999 and May

2000. Van Den Abeele (2016) identified a wild-caught double factor green A. roseicollis in Namibia and in the

9
Table 3. Plumage colour variations and their inheritance patterns found amongst Agapornis species

A. roseicollis A. canus A. taranta A. pullarius A. personatus A. fischeri A. lilianae A. nigrigenis Inheritance pattern

Dark factor X X X X X AID

Blue X X X X AR

Aqua X X AR

Turquoise X X AR

Orange face X AR

Pale headed X AID

Opaline X X SLR

Ino (NSL#) X X X X AR

Ino (SL†) X SLR

Pastel X AR

Dark eyed clear X AR

Pallid X SLR

Cinnamon X SLR

Pale X X SLR

Marbled X AR

Dilute X X X X AR

Bronze fallow X X X AR

Pale fallow X X X AR

Dun fallow X AR

10
 Figure 2 a Figure 2 b Figure 2 c Figure 2 d

Figure 2 e Figure 2 f Figure 2 g Figure 2 h

Figure 2. Eight domestic Agapornis species with wildtype coloration.

11
Figure 3 a Figure 3 b

Figure 3 c Figure 3 d

Figure 3 e Figure 3 f

12
Figure 3 g Figure 3 h

Figure 3. Wildtype coloration and some colour variations of four Agapornis species
Figure 3a: A. fischeri: Wildtype
Figure 3b: A. fischeri: Turquoise
Figure 3c: A. personatus: Wildtype
Figure 3d: A. personatus: blue (blue left, DD blue middle – (homozygous dark factor), D blue
– (heterozygous dark factor - right),
Figure 3e: A. lilianae Wildtype
Figure 3f: A. lilianae Non-Sex linked ino green
Figure 3g: A. roseicollis: Wildtype
Figure 3h: A. roseicollis: Opaline d blue
(All Photos: Mr. Dirk Van den Abeele).

13
1932 Proceedings of the Zoological Society of London a wild-caught A. personatus male was the first blue A.

personatus documented. Some of these variations have been known to breeders for almost a century. Toerien

(1950) reports a lutino A. roseicollis bred from a wild caught population from Namibia, whereas Delacour

(1942) reports the same variation in A. fischeri and Moreau (1948) in A. lilianae.

In most avian species, plumage coloration is affected by genetically controlled dietary intake of

carotenoids (Sage, 1962; Guay et al., 2012), but in the case of parrots, diet has little or no influence on coloration

as it is mainly due to genetic variation (Hill & McGraw, 2006). Black, brown and red-brown plumage, beak,

claw and eye colours in all bird species are caused by the pigment melanin, whereas red, yellow, pink and orange

colours found in parrot plumage are caused by psittacofulvin pigments (Mundy, 2006; Hill & McGraw, 2006).

Parrots are the only group of organisms known to synthesize psittacofulvin pigments in their plumage cells

(Stradi et al., 2001; Hudon & Brush, 1990; McGraw & Nogare, 2005, Hill & McGraw, 2006). Pigments

(melanin and psittacofulvins) and structural changes in the feather barbs are the two mechanisms that produce

the various parrot colours (Stoddard & Prum, 2011). Physical interactions of light waves with β-keratin rods in

the feather barbs produces purple, blue and green colours (Dyck, 1971 a & b; Prum et al., 1994; Prum, 2006).

The two mechanism work together as follows: pigments (melanins and psittacofulvins) absorb and reflect light

selectively (D’Alba et al., 2012) and due to the structural changes in the feather barbs the light is reflected

irregularly (Dyck, 1971 a & b; Hill & McGraw, 2006; Berg & Bennett, 2010). The barb of a green feather, for

example, contains a cortex filled with yellow psittacofulvin pigment. Light is partially absorbed by the

eumelanin pigment in the feather, and blue light is reflected back through the air vacuoles. It then passes through

the yellow psittacofulvin pigment and through coherent scattering of the quasi-ordered β-keratin rods, the light

(and ultimately the feather) appears green (Dyck 1971 a & b). Dark green feathers reflect about half the light

compared to lighter green feathers (Dyck, 1971 a & b). Mundy (2006) stresses the fact that these two

mechanisms (pigment and structural changes) are most probably under independent genetic control but that no

candidate genes have been associated with structural coloration control.

Agapornis plumage variations

Buckley (1987) defines a plumage colour polymorphism as the presence of a plumage colour aberration in an

interbreeding population of individuals of the same sex and age. There are at least 30 known naturally occurring

plumage colour variations amongst six of the domestic Agapornis species (Van den Abeele, 2016). The

inheritance pattern of these variations have mainly been determined by test matings performed by breeders, and

14
very little scientific literature is available on this subject. MUTAVI, a parrot breeding research and advice group

located in The Netherlands (www.MUTAVI.info) conducted research trials on the structures of Agapornis

feathers of various colorations. In Table 3, a summary of the different plumage colour variations per species

plus their inheritance patterns, is given. Given that none of the polymorphisms linked to these colour variations

have been identified, the colour patterns will be referred to as "plumage colour variations" and not “plumage

colour polymorphisms”.

There are four main categories of colour variations in Agapornis and a discussion with examples

thereof follows below. These include variations due to psittacofulvin reduction or modification, variations due

to a change in melanin, change in feather structure and changes in eumelanin. It is important to note that many

of these plumage colour variations can be combined that will result in a range of plumage colours (Hayward,

1979; Van den Abeele, 2016). Berg & Bennett (2010) as well as Mundy (2018) highlights the lack of research

on the genetic control of aberrant colours in caged parrots.

Aberrant plumage coloration due to psittacofulvin reduction or modification

Some of the most dramatic colour variations amongst the psittaciform order, is caused by the reduction or

modification of psittacofulvin within the feather cortex. In the 1932 Proceedings of the Zoological Society of

London a wild-caught A. personatus male was described as “the yellow coloring was absent, the areas of the

body normally yellow, being whitish and those normally green, blue.” Cooke et al. (2017) reported that since

the 19th century budgerigars bred in captivity has displayed a loss of psittacofulvin pigmentation causing birds to

have blue, instead of the wildtype green, plumage coloration. Yellow pigment in the cortex of the feather is

normally combined with blue light and results in visibly green feathers. Cooke et al. (2017) found that due to a

T>C substitution mutation in the MuPKS gene in budgerigars, no yellow psittacofulvin pigment is produced and

therefore blue light is reflected so that the feather appears blue. Other areas of the body that would normally be

red, yellow or orange due to psittacofulvins are white due to the absence of the psittacofulvin pigment (Prum,

2006; Van den Abeele, 2016). The same phenotype is observed amongst lovebirds and it is also inherited as an

autosomal recessive trait, as in budgerigars. Research done at MUTAVI (unpublished, personal communication

Mr Dirk Van den Abeele) has found that no yellow psittacofulvin pigment was present in blue lovebirds

compared to pigment being present in wildtype green birds. More research on the molecular basis of this gene in

lovebirds is needed before conclusions can be made for this genus.

15
 In contrast to the Blue phenotype where no psittacofulvin is produced, birds with Aqua and Turquoise

phenotypes produce a reduced amount of psittacofulvin. There are no reports of studies where the actual

pigment is measured, but visibly about 50% of the normal amount of psittacofulvin is produced and only about

40% of the normal levels of psittacofulvin is visible in birds with the Turquoise variation (Van den Abeele,

2016). It remains to be confirmed if a MuPKS polymorphism is causative to these phenotypes

Orange face and Pale headed are two popular phenotypes in lovebird aviculture and are only found

amongst A. roseicollis (Van den Abeele, 2016). The mechanism behind these phenotypes are poorly understood

(Personal communication, Mr Dirk Van den Abeele) but for Orange face individuals, red plumage is changed to

orange, whereas in pale headed individuals the psittacofulvin change from red to pink. Green plumage remains

unchanged for both phenotypes. Research by McGraw & Nogare (2005) indicated that the red, orange and pink

coloration are all due to psittacofulvin pigment in different concentrations and therefore these colour variations

could be caused by a change in psittacofulvin pigment concentration, but this remains to be confirmed.

Colour variations due to a change in melanin

Mundy (2006) and McGraw (2006) noted that most colour variations are caused due to changes in melanin

distribution. However, little to no molecular research has been conducted on parrot species regarding the genetic

basis of these variations. Most of the common types of bird plumage colour variations can be greatly attributed

to a lack of melanin in the feathers. Examples of plumage coloration variations due to a change in melanin in

Agapornis is briefly given in Table 4.

Colour variations due to a change in the feather structure

The “dark factor” genotype causes the spongy zone of the feather to decrease in size. Due to this reduction only

about half the light is reflected throughout the visible spectrum compared to light green feathers (Dyck, 1971 a

& b). The inheritance of this phenotype is incomplete dominance and three phenotypes are observed.

Heterozygote birds (also called “single dark factor”) are slightly darker (a khaki green) than the wildtype and

birds that are homozygous for the mutation (“double dark factor”) are found to be an olive green colour

(Hayward, 1979; Van den Abeele, 2016; Unpublished data, MUTAVI). This phenotype is observed in five of

the Agapornis species as seen in Table 3. It is also found amongst the budgerigar resulting in the same colours

(Harris, 1979).

16
 Research projects conducted at MUTAVI, discovered that lovebirds with the slaty phenotype had a

change in the keratin in the feathers from a horn colour to a glassy see-through colour (Personal communication:

Mr. Dirk Van den Abeele) and plumage colour change to a steel blue colour. A similar phenotype, called slate,

is commonly found amongst budgerigars (Elliott, 2013) and is caused by changes in the feather structure and

inherited as a sex-linked recessive trait. Through test matings it was determined that this phenotype is inherited

as an incomplete dominant trait. However, there is visually no difference between a heterozygote and a

homozygote for the mutation and therefore genotypes cannot be determined based on phenotype alone. (Van

den Abeele, 2016). Further research is necessary to identify the gene and polymorphism linked to these

phenotypes and confirmation whether the slaty and slate phenotypes are caused by the same polymorphism.

The violet phenotype develops due to a structural change in the spongy zone of the feather. It is

however only visible when it is combined with the blue phenotype. If psittacofulvin is absent in the cortex (thus

a blue bird), combined with the structural change in the spongy zone, the bird appears violet. The same

phenotype is found in budgerigars with the same underlying inheritance pattern (Harris, 1979; Van den Abeele,

2016). There is a difference in appearance between a heterozygote and homozygote for the mutation in both of

the species.

Coloration due to changes in eumelanin

In 2017 the “Jade” phenotype was officially accepted as a plumage colour aberration in A. roseicollis, and is

inherited as an autosomal recessive trait. Interestingly the breeders of these birds observed that females are

lighter than males. This phenotype has not been described in the literature and all observations are from personal

communication with Mr. Dirk Van den Abeele. Birds with this variation appears to have a visible reduction of

about half of the eumelanin in their wing feathers and more so over their body, resulting in a yellow colour with

a darker hue. There is a greater eumelanin reduction in the core of the wing feathers which results in a slight

edge on the feathers. The bird’s rump is much lighter and the legs are of normal coloration, but the nails are

darker. The mask of these birds are of normal coloration.

The Euwing pattern is a colour variation only found amongst A. fischeri causing darker wings and an

overall different body colour. The cause of the phenotype is still unclear but is inherited as a dominant trait with

incomplete penetrance (Van den Abeele, 2016). Veeckmans (2016) notes that the euwing mutation “dulls” the

black eumelanin pigment in the body feathers and on the mantle in heterozygote birds. In homozygous affected

birds, the effect is much more prominent with additional eumelanin found in the wing covert feathers, making

17
the bird appear darker. By combining a bird with both the euwing mutation as well as the opaline mutation,

breeders can achieve spectacular colorations.

PARENTAGE VERIFICATION

Applying molecular techniques to confirm parentage has been well documented for various domesticated animal

species including cattle (Matukumalli et al., 2009); goats (Visser et al., 2011) and dogs (DeNise et al., 2003).

Marker panels for parentage verification in birds are also common, however these panels are applied more in

conservation than breeding (Jan & Fumagalli, 2016; Coetzer et al., 2017). Most of these studies used

microsatellite markers but in recent years the focus has shifted towards the use of Single Nucleotide

Polymorphisms (SNPs) (see Weinman et al. (2015) for a comparison). Identifying incorrect pedigrees are of

importance in livestock populations as it adversely influences genetic gains (Israel & Weller, 2000). Although

Agapornis species are not under directional selection for increased genetic gain, incorrect pedigrees will still

negatively affect the populations’ general fitness. Despite breeders’ efforts to separate breeding pairs and

recording the offspring, misidentification of parents still occurs. Due to the fact that many of the colour

variations are inherited as autosomal recessive traits, breeders may deliberately sell a young as an offspring of a

bird with the colour variation knowing there is no way of confirming either parentage or the colour genotype.

This creates a serious loophole that could be exploited by unethical breeders. Currently there is no species or

genus specific parentage and individual identification panel available for lovebirds. The development of such a

panel could benefit breeders, buyers and conservationists in not only conserving these species but also ensuring

ethical breeding practices.

SPECIES IDENTIFICATION

Rheindt & Edwards (2011) reports that hybridization, as well as the resulting genetic introgression, is not

uncommon in birds. In aviculture the crossing of two species might result in the introduction of a novel colour

variation in a different species. Since birds from some of the different Agapornis species can inter-mate (see

Table 2), many of the variations were transferred through genetic introgression from one species to another. For

example, the “dark factor” phenotype was first observed in A. roseicollis in 1968 and in the 1980s the first dark

factor A. personatus was identified. From there the phenotype was transferred to A. fischeri, A. lilianae and A.

nigrigenis by interbreeding of the species (Van den Abeele, 2016). Responsible aviculturists should take care

not to interbreed species just to develop “new” colours in a species or to develop a “new species” of Agapornis.

18
Hybrids are not generally accepted by breeding societies, but unknowing buyers might purchase a hybrid and

breed further with this bird, leading to an even larger hybrid population. In addition, conservationists and

custom officials can benefit from a species identification test to confirm species from a tissue sample. Genetic

testing to infer if a species or if an individual is a hybrid between two species has been applied frequently in

aquaculture (Rasmussen & Morrissey, 2008; Hohenlohe et al., 2011). Von Holdt et al. (2011) reports the

application of SNPs to distinguish between dog and wolf hybrids and Koutsogiannouli et al. (2010) applied

Restriction Fragment Length Polymorphisms (RFLPs) to identify hybrids between wild and domestic pigs. The

development of a similar panel of SNPs could aid breeders, breeding societies, buyers, conservationists and

custom officials in identifying from which species an individual or a tissue sample originated and whether the

individual is a purebred or hybrid bird.

THE ROLE OF AVICULTURE IN CONSERVATION OF AGAPORNIS SPECIES

As with many other domestic parrot species, some of the wild Agapornis species are under pressure due to,

amongst others, illegal trade to the pet market. The development of the genetic screening tests such as the

identity verification and species identification tests could aid conservationists in preventing extinction of these

species. Fortunately, none of the nine species are extinct yet but should the wild populations become extinct,

captive breeding populations could be used in programs to reintroduce these species to the wild. This, however

might not be a simple task due to genetic and phenotypic changes that occur in captivity (Snyders et al., 1996).

Domestication and tameness are additional problems since behaviour changes set in fairly quickly when a

species is selected for tameness and these birds could struggle to adapt to a natural lifestyle, especially in a

habitat with predators (Snyders et al., 1996). Conservationists and breeders should therefore combine their

efforts to firstly ensure the survival of the wild populations and secondly breeding for show or pet market. This

can be achieved by making breeders aware of the threat wild populations face and the importance of not only

selecting for one trait (colour) but to keep genetic diversity, inbreeding level and species in mind when a mating

is planned. Conservationists should continue monitoring wild population growth and should the need for a

captive breeding program with the aim to release domesticated birds to the wild arise, utilise genetic screening

tests to plan these programs.

19
CONCLUSION

The popularity of lovebirds in aviculture coupled with their reduced habitat and poaching has placed the natural

populations of these birds under strain. It is the aviculturists’ duty to breed the birds responsibly to ensure the

survival of these species should the wild population become extinct. More research is needed in the field of avian

genetics to determine the causative genes and mutations linked to colour variations and to develop genetic

screening tests to determine the genotype, species and pedigree of the bird.

ACKNOWLEDGMENTS

We would like to thank Mr Dirk Van den Abeele for sharing his insight, years of knowledge and photographs

with us. The study was funded by The Technology and Human Resources for Industry Programme (THRIP)

which forms part of the Department of Trade and Industry of the South African government project ID number

TP13082831254), Lumegen Laboratories as well as The Technology Innovation Agency, SABDI grant (407/01

SABDI 16/1016).

REFERENCES

Berg, M.L. & Bennett, A.T.D. (2010) The evolution of plumage colouration in parrots: a review. Emu, 110, 10 –

20.

Buckley, P.A. (1987) Avian Genetics: A population and ecological approach. Academic press, Harcort Brace

Jovanovich, Publishers.

Coetzer WG, Downs CT, Perrin MR, Willows-Munro S. (2017) Testing of microsatellite multiplexes for

individual identification of Cape Parrots (Poicephalus robustus): paternity testing and monitoring trade

PeerJ 5.

Cooke, T.F., Fischer, C.R., Wu, P., Khosla, C., Chuong, C-M., Bustamante, C.D. (2017) Genetic mapping and

biochemical basis of yellow feather pigmentation in budgerigars. Cell. 171, 427 – 439.

D’Alba, L., Kieffer, L., Shawkey, M.D. (2012) Relative contributions of pigments and biophotonic

nanostructures to natural color production: a case study in budgerigar (Melopsittacus undulatus) feathers.

Journal of Experimental Biology. 215, 1272 – 1277.

Delacour, J. (1942) Bird breeding in France. Avic. Mag. 1.

20
DeNise, S., Johnston, E., Halverson, J., Marshall, K., Rosenfeld, D., McKenna, S., Sharp, T., Edwards, J. (2003)

Power of exclusion for parentage verification and probability of match for identity in American kennel

club breeds using 17 canine microsatellite markers. Animal Genetics, 35, 1, 14 – 17.

Diega, A. (2017) Bird flu to increase foreign demand for African lovebirds. Business Mirror.

https://businessmirror.com.ph/bird-flu-to-increase-foreign-demand-for-african-lovebirds/

Dilger, W. (1960) The comparative ethology of the African parrot genus Agapornis. International Journal of

Behavioral Biology Ethology. 17, 6, 649-685.

Dyck, J. (1971 a) Structure and colour-production of the blue barbs of Agapornis roseicollis and Cotinga

maynana. Z. Zellforsch. 115, 17 – 29.

Dyck, J. (1971 b) Structure and spectral reflectance of green and blue feathers of the rose-faced lovebird

(Agapornis roseicollis). Det. Kongelige Danske Videnskabernes Selskab, Bilogiske Skrifter 18, 2.

Eberhard, J.R. (1998) Evolution of Nest-Building Behavior in Agapornis Parrots. The Auk, 115, 2 455-464.

Egwumah, F.A. & Iboyi, M.O. (2017) Paramount factors influencing the breeding performance of lovebird

Agapornis pullaria. IJLST, 10, 7, 69-74.

Elliot, F.S. (2013) Budgerigar matings and colour expectations. Read Books Ltd. United Kingdom.

Forshaw, J. (1989) Parrots of the World. Princeton University Press, United States of America.

Ganeshkumar, G. Howard, J.T., Ward, J.M., Li, J., Li, B., Li, Y., Xiong, Y., Zhang, Y., Zhou, S., Schwartz,

D.C., Schatz, M., Aboukhalil, R., Fedrigo, O., Bukovnik, L., Wang, T., Wray, G., Rasolonjatovo, I.,

Winer, R., Knight, J.R., Koren, S., Warren, W.C., Zhang, G., Phillippy, A.M., Jarvis, E.D. (2014) High-

coverage sequencing and annotated assemblies of the budgerigar genome. GegaScience, 3, 11.

Guay, P.-J., Potvin, D.A. & Robinson, R.W. (2012) Aberrations in plumage coloration in birds. Australian field

ornithology, 29, 23 – 30.

Gunnarsson, U., Hellström, A.R., Tixier-Boichard, M., Minvielle, F., Bed’hom, B., Ito, S., Jensen, P., Rattink,

A., Vereijken, A., Andersson, L. (2007) Mutations in SLC45A2 cause plumage color variation in chicken

and Japanese quail. Genetic 175, 857 – 577.

Harris, R.L. (1979) Genetics and the budgerigar. AFA Watchbird Journal of the American Federation of

Aviculture, 6, 2.

Hayward, J. (1979) Lovebirds and their Colour Mutations.Blandford Press Ltd, United Kingdom.

Hill, G.E. & McGraw, K.J. (2000) Bird Coloration Mechanisms and measurements, Volume I. Harvard

University Press, United States of America.

21
Hohenlohe, P.A., Amish, S.J. Catchen, J.M., Allendorf, F.W., Luikart, G. (2011) Next-generation RAD

sequencing identifies thousands of SNPs for assessing hybridization between rainbow and westslope

cutthroat trout. Molecular ecology resources, 11, (Suppl. 1), 117 – 122.

Hudon, J. & Brush, A.H. (1990) Identification of carotenoid pigments in birds. FASEB J, 4, 2969.

Hume, J.P., & van Grouw, H. (2014) Colour aberrations in some extinct and endangered birds. Bull. B.O. C.

134, 3, 168 – 193.

Israel, C. & Weller, J.I. (2000) Effect of misidentification on genetic gain and estimation of breeding value in

dairy cattle populations. Journal of Dairy Science, 83, 1, 181 – 187.

Jan C, Fumagalli L. (2016) Polymorphic DNA microsatellite markers for forensic individual identification and

parentage analyses of seven threatened species of parrots (family Psittacidae). PeerJ 4.

Koutsogiannouli, E.A., Moutou, K.A., Sarafidou, T., Stamatis, C., Mamuris, Z. (2010) Detection of hybrids

between wild boars (Sus scrofa scrofa) and domestic pigs (Sus scrofa f. domestica) in Greece, using the

PCR-RFLP method on melanocortin-1 receptor (MC1R) mutations. Mammalian Biology, 75, 1, 69 – 73.

Manegold, A. & Podsiadlowski, L. (2014) On the systematic position of the Black-collared Lovebird

Agapornis swindernianus (Agapornithinae, Psittaciformes). J Ornithol, 155, 581 – 589.

Marez, C. (2003) The Abyssinian lovebird Agapornis taranta. AFA Watchbird Journal of the American

Federation of Aviculture, 30, 3.

Matukumalli, L.K., Lawley, C.T., Schnabel, R.D., Taylor, J.F., Allan, M.F., Heaton, M.P., O’Connoll, J.,

Moore, S.S., Smith, T.P.L., Sonstegard, T.S., van Tassell, C.P. (2009) Development and characterization

of a high density SNP genotyping assay for cattle. PLOS One, 4, 4.

McCarthy, E. (2006) Handbook of Avian Hybrids of the World. Oxford University Press, United Kingdom.

McGraw, K.J. & Nogare, M.C. (2005) Distribution of unique red feather pigments in parrots. Biol Lett. 1, 38 –

43.

McGraw, K.J. (2006) Mechanics of uncommon colors: pterins, porphyrins, and psittacofulvins. In: Bird

Coloration Mechanisms and measurements, Volume I. Harvard University Press, United States of

America.

Moreau, R. (1948) Aspects of evolution in the parrot genus Agapornis. IBIS, 90, 2, 206-239.

Mundy, N.I. (2006) A window on the genetics of evolution: MC1R and plumage colouration in birds. Proc. R.

Soc. B. 272, 1633 – 1640.

Mundy, N.I. (2018) Colouration genetics: Pretty polymorphic parrots. Current biology, 26, 3, R113-R114.

22
Mwangomo, E.A., Hardesty, L.H., Sinclair, A.R.E., Mduma, S.A.R., Metzger, K.L. (2007) Habitat selection,

diet and interspecific associations of the rufous-tailed weaver and Fischer’s lovebird. Afr. J. Ecol, 46, 267

– 275.

Mzumara, T.I., Perrin, M.R. & Downs, C.T. (2016 a). The drinking habits of the Lilian's lovebird and incidents

of poisoning at waterholes. African Journal of Ecology. 54, 3, 289 – 298.

Mzumara, T.I., Perrin, M.R. & Downs, C.T. (2016 b) Comparative performance of four survey methods for

assessing Lilian’s Lovebird abundance in Liwonde National Park, Malawi. Ostrich. 87, 3, 247 – 253.

Nanda, I., Karl, E.K Griffin, D.K., Schartl, M., Schmid, M. (2007) Chromosome repatterning in three

representative parrots (Psittaciformes) inferred from comparative chromosome painting. Cytogenet

Genome Res 117, 43 – 53.

Ndithia, H., Perrin, M.R., Waltret, M. (2007) Breeding biology and nest site characteristics of the Rosy-faced

Lovebird Agapornis roseicollis in Namibia. Ostrich, 78, 1, 13 – 20.

Parkes, C. (1982) Avian Biology, Volume 6. Elsevier Inc.

Perrin, M.R. (2009) Niche separation in African parrots. pp. 29–37. In: Harebottle, D.M., Craig, A.J.F.K.,

Anderson, M.D., Rakotomanana, H. & Muchai, M. (eds). Proceedings of the 12th Pan-African

Ornithological Congress, 2008. Cape Town, Animal Demography Unit.

Perrin, M. (2012) Parrots of Africa, Madagascar and the Mascarene islands. Biology, ecology and conservation.

Wits University Press, South Africa.

Proceedings of the Zoological Society of London. 1932. Mutant of A. personata. 1418.

Prum, R.O., Morrison, R.L. & Ten Eyck, G. R. (1994) Structural color production by constructive reflection

from ordered collagen arrays in a bird (Philepitta castanea: Eurylaimidae). Journal of Morphology, 222,

61 – 72.

Prum, R.O. (2006) Anatomy, physics, and evolution of structural colours. In: Bird Coloration Mechanisms and

measurements, Volume I. Harvard University Press, United States of America.

Rasmussen, R.S. & Morrissey, M.T. (2008) DNA-based methods for the identification of commercial fish and

seafood species. Comprehensive reviews in food science and food safety, 7.

Rheindt, F.E. & Edwards, S.V. (2011) Genetic introgression: an integral but neglected component of speciation

in birds. The Auk, 128, 4, 620 – 632.

Sage, B.L. (1962) Albinism and melanism in birds. British Birds, 55, 6.

Silva T & Kotlar, B. (1988) Breeding Lovebirds. T.F.H. Publications, Inc., United States of America.

23
Snyders, N. F., Derrickson, S. R., Beissinger, S. R., Wiley, J. W., Smith, T. B., Toone, W. D., Miller, B. (1996)

Limitations of captive breeding in endangered species recovery. Conservation biology, 10, 2, 338-348.

Stoddard, M.C. & Prum, R.O. (2011) How colorful are birds? Evolution of the avian plumage color gamut.

Behavioral Ecology, 22, 5, 1042 – 1052.

Stradi, R., Pini, E., Celentano, G. (2001) The chemical structure of the pigments in Ara macao plumage.

Comparative Biochemistry and Physiology Part B, 130, 57-63.

Tekalign, W. & Bekele, A. (2011) Status of and threats to the Black-winged Lovebird (Agapornis taranta) in

Entoto Natural Park and Bole sub-city, Addis Ababa, Ethiopia. Chinese Birds, 2, 4, 174 – 182.

Toerien, M.J. (1950) On the change of colour in a rosy-faced lovebird. Ostrich, 21, 1, 19 – 21.

Tsudzuki, M. (2008) Mutations of Japanese Quail (Coturnix japonica) and recent advances of molecular

genetics for this species. The Journal of Poultry Science, 45, 159 – 179.

Van den Abeele, D. (2016) Lovebirds Compendium, Genus Agapornis. About Pets, The Netherlands.

Van der Zwan, H., van der Westhuizen, F., Visser, C & van der Sluis, R. (2018) Draft de novo genome sequence

of Agapornis roseicollis for application in avian breeding. Animal Biotechnology. In press.

Van Grouw, H. (2013) What colour is that bird? The causes and recognition of common colour aberrations in

birds. British Birds, 106, 17 – 29.

Van Holdt, B.M., Pollinger, J.P., Earl, D.A., Parker, H.G., Ostrander, E.A., Wayne., R.K. (2013) Identification

of recent hybridization between gray wolves and domesticated dogs by SNP genotyping. Mamm genome,

24, 80 – 88.

Visser, C., van Marle-Köster, E., & Friedrich, H. (2011) Parentage verification of South African Angora goats,

using microsatellite markers. South African Journal of Animal Science, 41(3), 250-255.

Veeckmans, D. (2016) New mutation of Fischer’s lovebird – Agapornis fischeri: Opaline Euwing. Parrots Daily

News. http://www.parrotsdailynews.com/new-mutation-of-fischers-lovebird-agapornis-fischeri-opaline-

euwing/

Warburton L.S. & Perrin, M.R. (2005) Nest-site characteristics and breeding biology of the Black-cheeked

Lovebird Agapornis nigrigenis in Zambia. Ostrich. 76, 3&4, 162 – 174.

Warburton L.S. & Perrin, M.R. (2006) The Black-cheeked Lovebird (Agapornis nigrigenis) as an agricultural

pest in Zambia. Emu 160, 4, 321 – 328.

24
Weinman, L.R., Solomon, J.W., Rubenstein, D.R. (2015) A comparison of single nucleotide polymorphism and

microsatellite markers for analysis of parentage and kinship in a cooperatively breeding bird. Molecular

Ecology Resources. 15, 502 – 511.

Yakovlev AA, Kudryavtsev IV and Pigareva MD. (1975) A mutant Japanese quail (Coturnix coturnix japonica)

with marble plumage. Soviet Genetics, 11: 1225 – 1232.

25