Maleo Research Articles

Spatial Study of Maleo Bird Habitat Area Change in Sulawesi

Maleo (Macrocephalon maleo), a unique Sulawesi endemic bird, is threatened by habitat loss. This study aims to analyse changes in maleo habitat areas in Sulawesi using spatial data and propose effective conservation measures. Spatial data in the form of maleo home range maps (IUCN) were used for overlay analysis. The results of the overlay analysis were compared with land cover data to identify changes in maleo habitat area. Of the total area of approximately 884,040 hectares, 14.13% or 124,887.63 hectares have been converted to agricultural land. The construction of new settlements has affected 0.46% or 4,087.5 hectares of the habitat. In addition, plantations have occupied 2.82% or 24,965.92 hectares of the maleo habitat area. Changes to maleo habitat areas in Gorontalo are significant, with agricultural land conversion, settlement development, and plantations being the main threats. Effective conservation efforts should focus on habitat restoration, law enforcement, community education, and the development of environmentally friendly economic alternatives.

A proxy for brain-to-endocranial cavity index in non-neornithean dinosaurs and other extinct archosaurs.

Although the brain fills nearly the entire cranial cavity in birds, it can occupy a small portion of it in crocodilians. The lack of data regarding the volumetric correspondence between the brain and the cranial cavity hampers thorough assessments of the degree of encephalization in non-neornithean dinosaurs and other extinct archosaurs and, consequently, informed inferences regarding their cognitive capacities. Existing data suggest that, across extant archosaurs, the degree of endocranial doming and the volume of intracranial nonneural components are inversely related. We build upon this information to develop an equation relating these two anatomical features in non-neornithean dinosaurs and other extinct archosaurs. We rely on measurements of the endocast doming and brain-to-endocranial cavity (BEC) index in extant relatives of non-neornithean dinosaurs, namely, the crurotarsans Caiman crocodilus, Crocodylus niloticus, and Crocodylus porosus; the paleognaths Struthio camelus and Apteryx mantelli; and the fowl Macrocephalon maleo, Gallus gallus, Meleagris gallopavo, Phasianus colchicus, and Anas platyrhynchos. Applying the equation to representative endocasts from major clades of dinosaurs, we found that BEC varies from about 0.6 in ceratopsians and thyreophorans to around 0.7 in ornithopods, pachycephalosaurians, sauropods, and theropods. We, therefore, warn against the use of a catch-all value, like 0.5, and instead encourage refinement in the adoption of BEC across archosaurs.

Modeling suitable habitats of maleo (Macrocephalon maleo sal. müller 1846) in Gorontalo

Maleo (Macrocephalon maleo) is one of the endangered bird species in Indonesia. This avifauna species is an endemic bird to Sulawesi Island. It is distributed from the south to the north of Sulawesi, including Gorontalo. Currently, information on suitable habitat models for M. maleo is very limited, while this information is required to support the conservation of M. maleo. This study aimed to model the potential habitat for M. maleo using species distribution modeling (SDM) with vegetation cover variables as predictors. The model was built based on the M. maleo occurrence points. The suitable habitat was then evaluated using area under the curve analysis and the receiver operating characteristic curve (AUCROC). Based on the model, the AUC is valued at 0.729, which is considered reasonable and indicates that the model can be used to depict the potential habitats for the species. In this study, most of the west and east parts of Gorontalo were considered not suitable for Maleo. While the coastal areas of Gorontalo were considered very suitable. This was confirmed for both the north and south coastal areas of Gorontalo. Then it is strongly recommended to conserve and protect most of those coasts to ensure the Maleo conservation.

Health status assessment of Maleo Bird (Macrocephalon maleo) nesting ground at Kadidia Village, Lore Lindu National Park

IUCN categorizes Maleo Bird (Macrocephalon maleo) as an endemic and endangered species meaning that the species is facing a high risk of extinction soon. One of the critical aspects determining the species^ existence is its habitat. The endangered species habitat not only plays a vital role in the species^ survival but also in its to reproduce. The research method used was the modified Environmental Health Index Determination developed. The research was at Kadidia Village, Lore Lindu National Park. The research aims to determine the habitat’s health status by identifying its Health Index. The research method used is a site survey by applying direct observation to habitat components, including conditions, species, and related socio-economic aspects. Variables measured are excellent, good, moderate and poor. The results showed that the health index of the habitat (nesting ground) is moderate/fair, with a Health Index of 0.55. Therefore, regular monitoring and conservation action should ensure that the habitat is adequately managed and conserved.

Osteological comparison of casque ontogeny in palaeognathous and neognathous birds: insights for selecting modern analogues in the study of cranial ornaments from extinct archosaurs

Abstract Birds, along with their dinosaurian precursors, possess a variety of bony cranial expansions. A deep understanding of the phenotypic complexity of these structures would be useful for addressing the development, evolution, and function of hard-tissue cranial ornamentation. Yet, the evolutionary significance and function of these structures have gone largely unaddressed because no unifying conceptual framework for interpreting bony cranial expansions currently exists. To provide such a framework, we examine osseous ornament variation in modern birds, using µ-CT imaging to examine the cranial casque components, structural composition, and developmental changes of two neognathous (Numida meleagris, Macrocephalon maleo) and one palaeognathous species (Casuarius casuarius) and survey the avian osteology literature of the 11 orders containing members with osseous cranial ornamentation. Our anatomical analyses suggest two broad configuration categories: (i) geminal, in which ornaments consist of paired elements only (i.e. within Neognathae) and (ii) disunited, in which ornaments consist of unpaired, midline elements along with paired bones (i.e. within Palaeognathae). Ornament bones contribute to casque elevation (proximal ornament support), elaboration (distal ornament shape), or both. Our results hold utility for unravelling the selection processes, particularly in difficult-to-decipher display roles, that shaped modern avian casques, as well as for the use of extant avians as comparative analogues of non-avian dinosaurs with ornamental head structures.

Characteristics of Maleo bird spawning nests (Macrocephalon maleo) in Lake Towuti, South Sulawesi, Indonesia

Abstract. Karim HA, Najib NN, Ayu SM, Fidel. 2023. Characteristics of Maleo bird spawning nests (Macrocephalon maleo) in Lake Towuti, South Sulawesi.Biodiversitas 24: 670-676. Maleo birds (Macrocephalon maleo) have a habitat in the forest near the beach or in the middle of the forest near the hot springs. However, in Pekaloa Village, East Luwu District, Maleo birds are found in Lake Towuti, South Sulawesi, Indonesia. This, of course, shows that the habitat of Maleo birds on the shore of the lake is new information, especially on the island of Sulawesi. This research was conducted in Pekaloa Village, Towuti Sub-district, East Luwu District, South Sulawesi. This study was conducted to collect data on the characteristics of Maleo Senkawor (Macrocephalon maleo) nests. Secondary data collection began from scientific journals and works, as well as discussions with the surrounding community and the TWA manager of Lake Towuti, while primary data was obtained from measurements and direct observation in the field, as well as laboratory analysis. In this study, five nest plots with 13 nest holes were used, and the vegetation analysis used example plots measuring 175 m2. The characteristics of the nest pit have an average depth of 0.6 m, a surface diameter of 0.41 m, a temperature of 33.6oC, and a pH of 6.2. Vegetation analysis found eight species around the Maleo Senkawor bird's nest. Vegetation analysis used the density formula: highest frequency density 19.5 and highest frequency 0.57, lowest density 0.02 and lowest frequency 0.14. This study forms the basis of new information in the conservation of Maleo birds with the characteristics of the habitat conditions for nesting Maleo birds, especially in the coastal area of the lake.

The Population Study of Maleo Bird (Macrocephalon maleo) in The Nesting Ground of Saluki Village, Lore Lindu National Park

Sulawesi Island is an island that has a diversity of flora and fauna, some of which are endemic animals. One of the protected endemic fauna species is the maleo bird. Maleo was designated as a protected animal because the number of populations in the wild was decreasing, and its conservation status was in the endangered category. Concrete steps that could be taken to increase the maleo population were through various activities, including conducting ongoing and periodic studies of the maleo population. In particular, this study aims to examine the current population of the maleo bird in the nesting ground of Saluki Village, Lore Lindu National Park. In this study, the estimation of maleo population density uses two approaches. The first approach was direct observation of the maleo population outside the nesting habitat using the transect method. The second approach was using the active nest count approach at nesting sites. Based on the results, it was found that in the egg-laying locations, there were 32 active nests and 31 inactive nests. Based on these results, the maleo population density in the egg-laying locations was 29.7 individuals/Ha. While outside the egg-laying habitat, there were eight maleo individuals consisting of four juvenile individuals and four young adults (sub-adults). These results showed that the population density outside the egg-laying habitat was 1.7 individuals/Ha. Four maleo predators were found at the study site, namely Varanus salvator, Spilornis rufipectus, Ictinaetus malayensis and Falco sp. In addition, there was one type of potential maleo predator, Macaca tongkeana, with as many as seven individuals. Two potential factors could disturb the maleo population: the laying location factor and the dense undergrowth factor.

Phylogeography of the Maleo Senkawor (Macrocephalon maleo Sal. Muller 1846) Based on Cytochrome B Gene in Sulawesi And Their Sex Determination using Molecular Sexing

The Maleo Senkawor bird (Macrocephalon maleo) is endemic and widely distributed in Sulawesi. Maleo Senkawor bird is monomorphic so it is difficult to distinguish between male and female individuals. This study aimed to determine the kinship relationship among M. maleo in various geographic areas in Sulawesi through the use of the gene marker Cytochrome-b (Cyt-b), determine the sex ratio, and examine the reliability of the 2550F/2718R primer. Fifteen (feather and eggshell) samples from North Sulawesi, Southeast Sulawesi, and Central Sulawesi were collected. Total DNA isolation was performed using the Dneasy® Blood and Tissue kit (Cat. No. 69504) following a Qiagen protocol with modification. PCR amplification (35 cycles) used a forward primer MMCytb_F (5'- GAAAATCCCACCCCCTACTA-3'), a reverse primer MMCytb_R (5'- GTTGGCTACGAGGAGTCAGA-3') and a primer for sexing used 2550F/2718R. Analysis was performed on the length of 903 bp Cyt-b mtDNA gene sequences of M. maleo along with their amino acid sequences (301 AA) and Chromo Helicase DNA-Binding (CHD) gene sequences. Based on the genetic distance of the Kimura 2-Parameter model and p-distance, the interpopulation of M. maleo in Sulawesi (the population of Southeast Sulawesi was separated from the population of Central Sulawesi and North Sulawesi) was 0.002 (0.2%) – 0.003 (0.3%). Each population has formed a different haplotype. Primer 2550F/2718R was able to amplify the CHD gene and could distinguish the sex identification. Male individuals are more dominant than female individuals with a sex ratio of 6,5:1

Degree of egg-taking by humans determines the fate of maleo (Macrocephalon maleo) nesting grounds across Sulawesi

The maleo (Macrocephalon maleo) of Sulawesi, Indonesia, is culturally iconic and Critically Endangered, but the causes of its decline have never been systematically analyzed nor its nesting grounds comprehensively surveyed. We visited 122 previously known and 58 previously unrecorded sites, collecting data and interviewing local people at each site. We used ordinal logistic regression to fit models with combinations of 18 different predation, habitat, and nesting ground variables to determine the strongest predictors of nesting ground success, as represented by maleo numbers. At least 56% of known nesting grounds are now inactive (abandoned), and 63% of remaining active sites host ≤ 2 pairs/day at peak season. Egg-taking by humans is the single biggest driver of maleo decline. Protecting eggs in situ predicts higher maleo numbers than protecting eggs through hatchery methods. After egg-taking, quality (not length) of the travel corridor connecting nesting ground to primary forest best predicts nesting ground success. Being inside a federally protected area is not a primary driver of success, and does not ensure persistence: 28% of federally protected nesting grounds have become inactive. Local conservation efforts protected nesting grounds 2‒3 times better than federal protection. We update the methodology for assessing nesting ground status, and recommend five measures for maleo conservation, the foremost being to protect nesting grounds from egg-taking by humans at all remaining active sites.

Nesting Site Nesting Site Characteristics and Egg Hatching Success of Maleo (Macrocephalon maleo) in Hungayono Sanctuary, Gorontalo

Maleo population is decreasing due to the decline in suitable breeding site. The establishment of a maleo sanctuary in Hungayono aims to increase maleo population by protecting its eggs and increasing the success rate of eggs hatched. This study aims to analyze the characteristics of maleo nesting habitats, to describe the semi-natural maleo egg hatching management in the sanctuary, and analyze the success rate of maleo egg hatching in Hungayono. The results showed that the Maleo nesting sites are clean of bushes and shrubs, but there was still shades from trees or bamboo. The nesting sites are close to rivers and geothermal sources with mixed soil structure. The natural egg burrows has an average depth of 92.8 cm, width of 82.7 cm, temperature of 32.5 oC, and humidity of 90%. Hungayono has seven hatcheries that utilized geothermal heat to hatch Maleo egg semi-naturally. The burrows size in the hatcheries were made according to the size of Maleo eggs, with an average depth of 32 cm and width of 10 cm, the average temperature is 32.5 oC and humidity is 90.8%. The incubation period for the semi-natural hatcheries is 60-80 days. Maleo egg hatching success rate reached 60.18% in which is medium category.

Breeding records of the birds of south-east Sulawesi, Indonesia: a collation of observations encompassing nearly 20 years of research in Wallacea

The islands of the Wallacean biodiversity hotspot support diverse and highly endemic bird communities, yet remain ornithologically poorly studied. There is a particular paucity of data regarding breeding biology for the region's birds. Here, we help to address this research gap by collating observations of breeding evidence made during nearly 20 years of field work between 1999 and 2018 in south-east Sulawesi and its offshore islands. Data were collected between April and September, with a special focus on July and August, which is the dry season across the southern half of Sulawesi. In total, we summarise 1,064 observations of potential breeding in 66 species, of which 27 are Wallacean endemics (including the Critically Endangered Maleo Macrocephalon maleo), and 39 species of wider range (including the Endangered Milky Stork Mycteria cinerea). Records include species with little or no previously published information on their breeding biology, such as Pygmy Hanging Parrot Loriculus exilis, Sulawesi Pitta Erythropitta celebensis and an undescribed Zosterops species, provisionally referred to as Wangi-wangi White-eye.

DNA barcoding of Macrocephalon maleo originating from Sulawesi, Indonesia

Abstract. Samad A, Solihin DD, Sumantri C, Purwantara B. 2022. DNA barcoding of Macrocephalon maleo originating from Sulawesi, Indonesia. Biodiversitas 23: 4400-4408. Conservation of Maleo Senkawor (Macrocephalon maleo Sal. Muller, 1846) continues to be carried out to sustain this bird species. The genetic information from this study can be used to support future conservation activities. This study aimed to provide information on the genetic characteristics of the endangered Maleo Senkawor based on the partial sequence of the Cytochrome Oxidase 1 (COI) gene in mitochondrial DNA. Samples were collected from Central Sulawesi, Southeast Sulawesi, and North Sulawesi. Total DNA isolation was performed using the Dneasy® Blood and Tissue kit through a modified Qiagen protocol. PCR amplification used a forward primer MMCOI_F (5'-ccgatgattattctcaacca-3') and a reverse primer MMCOI_R (5'- gtagccgaatggttcttttt-3'). Analysis was performed on the partial sequences of the M. maleo COI mtDNA gene and its amino acid sequences. The genetic distance between the M. maleo population from North Sulawesi, Southeast and Central Sulawesi is 0.001 (0.1%). Macrocephalon maleo population of Southeast and Central Sulawesi have 0% genetic distance. Analysis of the COI M. maleo gene from 3 population locations showed that they came from the same species. Based on the genetic distance of the Kimura model with 2 parameters in the 3 populations, the M. maleo formed the same group (ingroup species) but had a different haplotype. These differences can be used as characterizing markers in each population. Furthermore, the conservation of M. maleo from these 3 populations was better treated as a separate group based on the mitochondrial DNA COI gene to avoid the negative impact of interpopulation breeding.

Vegetation Characteristics of The Maleo Bird (Macrochepalon maleo) Habitat at Natural Tourism Park of Towuti Lake, South Sulawesi

Maleo birds (Macrocephalon maleo) generally have habitats in forests near hot springs, but in East Luwu Regency their habitat is found on the shores of Lake Towuti. It is suspected that the habitat of maleo birds at the site has different characteristics. This study aims to identify the vegetation characteristics of maleo bird habitat in the sleeping area and tread where to lay eggs. Vegetation analysis was carried out by purposive sampling with a plot size of 20 m x 20 m in the sleeping area, and 5 m x 5 m in the laying site where the egg-laying was located. The data collected are the name of the type, diameter, the individual number of trees, poles, stakes, and seedlings. The results showed that in sleeping areas, macadamia hildebrandii trees have the highest INP values. Macadamia hildebrandii fruit is thought to be one of [1]the feeds for Maleo birds. In egg-laying habitats, Imperata cylindrica has the highest INP compared to other types. On the tread of the egg-laying nests can be seen maleo birds eating grass grains. There are differences in ecological index [2]values found in sleeping habitats, namely H'= 2.24 (medium), Dmg= 3.02 (low), and E= 0.81 (high). However, in egg-laying habitats, the Ecological Index values found were H'= 0.62 (low), Dmg= 0.93 (low) and E= 0.30 (low). The discovery of maleo birds in Lake Towuti TWA makes the area one of the remaining in situ habitats in South Sulawesi and can be a consideration for the government to maintain and preserve it.

Vegetation Characteristics of The Maleo Bird (Macrochepalon Maleo) Habitat at Natural Tourism Park of Towuti Lake, South Sulawesi

Maleo birds (Macrocephalon maleo) generally have habitats in forests near hot springs, but in East Luwu Regency their habitat is found on the shores of Lake Towuti. It is suspected that the habitat of maleo birds at the site has different characteristics. This study aims to identify the vegetation characteristics of maleo bird habitat in the sleeping area and tread where to lay eggs. Vegetation analysis was carried out by purposive sampling with a plot size of 20 m x 20 m in the sleeping area, and 5 m x 5 m in the laying site where the egg-laying was located. The data collected are the name of the type, diameter, the individual number of trees, poles, stakes, and seedlings. The results showed that in sleeping areas, macadamia hildebrandii trees have the highest INP values. Macadamia hildebrandii fruit is thought to be one of [1]the feeds for Maleo birds. In egg-laying habitats, Imperata cylindrica has the highest INP compared to other types. On the tread of the egg-laying nests can be seen maleo birds eating grass grains. There are differences in ecological index [2]values found in sleeping habitats, namely H'= 2.24 (medium), Dmg= 3.02 (low), and E= 0.81 (high). However, in egg-laying habitats, the Ecological Index values found were H'= 0.62 (low), Dmg= 0.93 (low) and E= 0.30 (low). The discovery of maleo birds in Lake Towuti TWA makes the area one of the remaining in situ habitats in South Sulawesi and can be a consideration for the government to maintain and preserve it.

Short Communication: A new primer set in CHD1 gene for bird sex identification

Abstract. Vera F, Wajjwalku W, Yuda P, Daryono BS. 2021. Short Communication: A new primer set in CHD1 gene for bird sex identification. Biodiversitas 22: 4977-4982. Determine sex is difficult for many bird species that are sexually monomorphic or only dimorphic in the adult stage. Many molecular markers have been developed for DNA sexing, which were mostly based on identifying Z and W chromosomes of avians. The sex determination of birds was mostly applied universal primer set such as P2/P8 or 2550F/2718R. However, those universal primers that were designed sometimes could not good result consistency in non-ratite birds or ratite birds. Therefore, we improved a specific primer design with deletion site in W chromosome to amplify the female-specific segments on Chromodomain-helicase DNA binding protein-1 (CHD1) gene from alignment sequences CHD1-Z and CHD1-W of Macrocephalon maleo S. Müller, 1846. This study aims to design a new pair of primer targeting a section of the CHD1 gene that can be amplified in non-ratite birds, the name of a new primer is In-Sex F/In-Sex R. A new primer set amplified DNA fragments in around 650 or 550 base pairs of CHD1-Z and also, 350 base pairs of CHD1-W. The result has successfully amplified the sex of multiple species on orders Galliformes, Passeriformes, Accipitriformes, and Strigiformes. This analysis can be helpful to the effort of in-situ or ex-situ management and conservation programs e.g the mating system in birds. And also, the analysis can be helpful for sexing data in the case of captive birds before releasing in natural habitat or reintroduction programs.

“Someday we will all learn anew through hands that love us”: Protecting a key insular population of the Sulawesi Fruit Bat Acerodon celebensis

“Someday we will all learn anew through hands that love us”: Protecting a key insular population of the Sulawesi Fruit Bat <i>Acerodon celebensis</i>

Maleo Macrocephalon maleo population recovery at two Sulawesi nesting grounds after community engagement to prevent egg poaching

Sulawesi's endemic maleo bird Macrocephalon maleo, a megapode that lays its eggs in communal nesting areas of warm sand or soil, is classified as Endangered on the IUCN Red List, with its decline generally attributed to habitat loss and poaching of its large eggs. Despite decades of protection under Indonesian law and numerous field conservation efforts, no conservation projects have yet documented a recovery in local numbers, and the overall maleo population continues to decline. We aimed to test whether the maleo's decline could be reversed in two community-based conservation programs in the Tompotika region of Central Sulawesi, in which maleo eggs were left undisturbed in situ and egg-taking (poaching) by humans ceased by community agreement. Subsequently, maleo numbers have increased four-fold after 14 years and three-fold after 5 years at two different locations. Our results suggest that: a) at these sites, egg poaching by humans is the primary threat to maleos, above habitat loss and natural predation; b) maleo populations can recover if egg poaching is prevented and eggs are left undisturbed; c) interventions such as translocating and/or artificially incubating eggs may be unnecessary if poaching is stopped; d) communities may voluntarily choose to end poaching with appropriate outreach work, which may or may not include economic incentives.

PREDICTING THE GROWTH CURVE OF BODY WEIGHT IN MALEO BIRDS (MACROCEPHALON MALEO)

Maleo bird (Macrocephalon maleo) is the one of protected bird’s species and originated from Sulawesi island of Indonesia. This study was carried out to obtain the growth curve of body weight (BW) in mixed-sex Maleo birds from hatching to yearling ages. The growth curve in this study was calculated with non-linear regression of Logistic (L) and Gompertz (G) models using CurveExprt 1.4. computer program. The primary data in this study was cited from previous study through a literature study. Research showed that the asymptotic weight (A) in birds was 1825.34 g (L) and 3429.23 g (G). The weight of inflection (Wi) in studied birds were 912.67 g (L) and 1260.75 g (G). The time of inflection (ti) in birds was 8.51 months (L) and 11.00 months (G). The maximum growth rate in birds was 159.72 g/month (L) and 151.29 g/month (G). The coefficient of determination (R2) in both models included of very high category (0.80&lt;R2&lt;1.00) but the lower of standard error (SE) value showed in L model. It can be concluded that the non-linear regression of Logistic model can be used as BW predictors in mixed-sex Maleo birds.

A preliminary study of Maleo (Macrocephalon maleo) nesting in Kambunong Cape, West Sulawesi

Maleo (Macrocephalon maleo) is an endemic species inhabiting Sulawesi island and is listed as Endangered under IUCN Red Data List. However, number of adult individuals of the species found in the wild was predicted to be decreasing. Most data available for the species are scattered and or not reported. Therefore, more published research is needed as regard to population size, distribution, and trends of maleo. One site identified as habitat of maleo (Macrocephalon maleo) was located in the coastal area of Kambunong Cape, West Sulawesi Province. This study attempts to examine the nesting site, population trends of the species and to identify nthropogenic and non-anthropogenic threats towards the population in the area. Index point of abundance (IPA) combined with line transect techniques were applied to estimate population of the species. Seven line transects were deployed and combined with 30 circular plots distributed across the lines covering 100 ha of the coastal area in Kambunong Cape. Meanwhile, types of threats posed to the population were explored by using semi-structure interviews to the locals. The results have shown that the total of relative abundance estimated for the species in the area was 5.091 individuals/ha for 30 observation plots or 16.97 (=17) individuals found in 100 ha. There was only one site utilized by the species located in a poor environment. Decreasing trends of the population was mostly due to habitat modification and fragmentation combined with illegal egg hunting/collection as well as high intensity of human activities nearby their home range.

Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo)

Background: Noninvasive DNA sampling has been applied across many avian genetic studies for a variety of purposes including conservation and management of endangered birds. However, its application in megapodes is still lacking. The previous genetic studies on megapodes used either blood or fresh tissue. Here we present the first demonstration of the use of eggshell membrane for research on endangered Maleo ( Macrocephalon maleo). Methods: We used 24 post-hatched eggshell membranes collected from two different sites, Tambun and Tanjung Binerean, in North Sulawesi, 12 samples in each. Two different DNA extraction methods: alkaline lysis method and gSYNC TM DNA Extraction Kit were applied. To determine the sex of Maleo, we utilized PCR-based DNA sexing using CHD genes, with the primer set 2550F/2718R. Results: We successfully extracted all samples; the mean sample concentration was 267.5 ng/µl (range 47-510.5 ng/µl) and samples were of high purity (A260/280 ratio 1.85±0.03). All samples were used to successfully identified sexes, 9 females and 15 males. Conclusions: Our research clearly illustrates that eggshell membranes can be used for DNA sexing and open the possibility to build noninvasive DNA collections over large spatial scales for population study of endangered birds.