Gentoo penguins are the least numerous of the penguins breeding at Macquarie Island, and the only species to rear two chicks. This project examined the interactions between diving behaviour, diet and reproductive strategy. Data were collected on Time Depth Recorders (TDRs), and stored in hexadecimal format. Hexadecimal files can be read using 'Instrument Helper', a free download from Wildlife Computers (see the url given below).

June 2018 Adélie penguin scats were collected from Signy Island (South Orkney Islands) during crèche (December/January) 2014/15 and 2015/16 and stored in 80% Ethanol. DNA was extracted from ~30 mg of faecal material using a Promega ‘Maxwell 16' instrument and a Maxwell® 16 Tissue DNA kit. A total of 450 samples were analysed: 30 extractions per week for 2015 and 60 per week for samples collected in 2016. Three DNA markers providing different taxonomic information were amplified from penguin faecal DNA. First, ALL faecal DNA samples were characterised using a highly conserved metazoan primer set that amplifies a region of the nuclear 18S gene. In addition, a subset of faecal samples from each year were also characterised with two other primer pairs that amplify a region of the mtDNA 16S gene to allow species-level identification for most fish (16S_Fish) and krill (16S_Krill) species respectively. During amplification of markers, the products were tagged with a unique pair of index primers allowing samples to be pooled and sequenced (2x150bp) on a MiSeq high-throughput DNA sequencer. - See Adelie Pengiun Diet CCAMLR paper for all of the primer/PCR details - See BAS Adelie 18s Krill and Fish subset excel spreadsheet for sample details. - See BAS Adelie 18s ALL samples fastq for 18s fastq files - See BAS Adelie 16s Krill subset fastq for 16s krill fastq files - See BAS Adelie Fish subset fastq for 16s fish fastq files November 2018 In addition we also amplified all 450 samples with the 16S_Fish marker. - See Adelie Experiment Details 16s Fish for sample details, plate layout, first and second round PCR and miseq sheet. - See BAS Adelie Fish ALL Samples fastq for 16s fish fastq files

Metadata record for data from ASAC Project 419 See the link below for public details on this project. From the abstracts of some of the referenced papers: The population size and breeding success of Emperor Penguins (Aptenodytes forsteri) at the Auster and Taylor Glacier colonies were estimated during the 1988 breeding season. At Auster a total of 10963 pairs produced about 6350 fledglings for a breeding success of 58%. At Taylor Glacier about 2900 pairs raised 1774 fledglings for a breeding success of 61%. Fledglings left Taylor Glacier over a period of 33 days at a mean mass of 10.56kg. The accuracy of the tritiated water (HTO) and sodium-22 (22Na) turnover methods as estimators of dietary water and sodium intake was evaluated in emperor penguins fed separate diets of squid and fish. Emperor penguins assimilated 76.2% and 81.8% of available energy in the squid and fish diets, respectively. Both isotopes had equilibrated with body water and exchangeable sodium pools by 2h after intramuscular injection. The tritium method yielded reliable results after blood isotope levels had declined by 35%. On average the tritium method underestimated water intake by 2.9%, with a range of -10.3% to +11.1%. The 22Na method underestimated Na intake on average by 15.9% with the errors among individuals ranging from -37.2% to -1.8%. Discrepancies with 22Na turnover were significantly greater with the squid diet than the fish diet. The results confirm the reliability of the tritium method as an estimator of food consumption by free-living emperor penguins (provided seawater and freshwater ingestion is known) and support the adoption of the 22Na method to derive an approximation of seawater of seawater intake by tritiated emperor penguin chicks and by tritiated adults on foraging trips of short duration. The diet composition of Emperor Penguin Aptenodytes forsteri chicks was examined at Auster and Taylor Glacier colonies, near Australia's Mawson station, Antarctica, between hatching in mid-winter and fledging in mid-summer by 'water-offloading' adults. Chicks at both colonies were fed a similar suite of prey species. Crustaceans occurred in 82% of stomach samples at Auster and 87% of stomachs at Taylor Glacier and were heavily digested; their contribution to food mass could not be quantified. Fish, primarily bentho-pelagic species, accounted for 52% by number and 55% by mass of chick diet at Auster, and squid formed the remainder. At Taylor Glacier the corresponding values were 27% by number and 31% by mass of fish and 73% by number and 69% by mass of squid. of the 33 species or taxa identified, the fish Trematous eulepidotus and the squid Psychroteuthis glacialis and Alluroteuthis antarcticus accounted for 64% and 74% of the diets by mass at Auster and Taylor Glacier, res pectively. The sizes of fish varied temporally but not in a linear manner from winter to summer. Adult penguins captured fish ranging in length from 60 mm (Pleuragramma antarcticum) to 250 mm (T. eulepidotus) and squid (P. glacialis) from 19 to 280 mm in mantle length. The length-frequency distribution of P. glacialis showed seasonal variation, with the size of squid increasing from winter to summer. The energy density of chick diet mix increased significantly prior to 'fledging'.

These spreadsheets provide the proportions of prey DNA sequences in the scats of Adelie penguins at Bechervaise Island and Whitney Point in East Antarctica. Samples were collected during two stages of the breeding season: mid brood guard (Bechervaise Island-January 4-6th 2013, Whitney Point 23- 28th December 2012) and mid creche (23-26th January 2013). Scat samples were collected from breeding birds, chicks and non-breeders at Bechervaise Island and breeding birds and chicks at Whitney Point. 'Breeders' were identified as individuals brooding or provisioning a chick, whereas 'non-breeders' were usually pairs that had reoccupied the colony and were building new practice nests with no chick present. Non-breeders in the colony include immature birds that have not yet bred and mature birds of breeding age that did not breed in a particular season (e.g. no partner or insufficient body condition) DNA from each sample was extracted and sequenced as per the protocols in the following paper: Jarman, S.N., McInnes, J.C., Faux, C., Polanowski, A.M., Marthick, J., Deagle, B.E., Southwell, C. and Emmerson, L. 2013 Adelie penguin population diet monitoring by analysis of food DNA in scats. PLoS One 8, e82227. (doi:10.1371/journal.pone.0082227). The Raw Data spreadsheet contains the proportion of each prey group of each individual sample, plus the total sequence count of prey items. Only samples with greater than 100 prey sequences are included in the dataset. The summary datasheet contains only prey taxa which contained greater than 2% of the proportion of sequences. Analysis of these data have been published in: McInnes JC, Emmerson L, Southwell C, Faux C, Jarman SN. (2016) Simultaneous DNA-based diet analysis of breeding, non-breeding and chick Adelie Penguins http://dx.doi.org/10.1098/rsos.150443

Adélie penguin and snow petrel scats were collected at Béchervaise Island (67°35’S, 62°49’E) in the austral summers 2014/2015, 2017/2018 and 2019/2020 and stored in 80% Ethanol. DNA was extracted using the Maxwell RSC48 instrument with the Maxwell RSC 48 Tissue DNA Kit (Promega). ~30 mg of the scat was added to 250 μL of S.T.A.R buffer (Roche Diagnostics). All remaining steps followed the manufacturer’s instructions. Reagent blank controls (n=5) were added to the extraction process. DNA was plated out and diluted 1:5. In total, there were 465 scat samples; 302 collected from Adélie penguins and 163 from snow petrels. Three DNA markers providing different taxonomic information were amplified. 18s - All samples (n=480 includes positives and negatives) were amplified with a primer set that amplifies ~170bp of the nuclear 18S gene (McInnes et al., (2017b) DNA Metabarcoding as a Marine Conservation and Management Tool: A Circumpolar Examination of Fishery Discards in the Diet of Threatened Albatrosses. Frontiers in Marine Science). A PNA clamp was also added to suppress bird and mammal DNA . Krill - We characterised the taxonomic identity of krill by amplifying ~250bp of the 16S rDNA gene (Ratcliffe et al, (2021), Changes in prey fields increase the potential for spatial overlap between gentoo penguins and a krill fishery within a marine protected area. Diversity and Distributions). Samples were considered positive for krill if the Ct value was less than 35 (n=120). PCR amplifications were performed in two rounds, the first to amplify the target gene and add sample-specific 6 or 7 bp multiplex-identifier (MID) tags (forward and reverse primer) and Illumina sequencing primers, the second to add sequencing adapters and additional 8 bp MIDs. PCR products from all samples including the blanks, positive and negative controls (n=600) were pooled and purified using Agencourt Ampure (Beckman Coulter, USA) magnetic beads. The pool was diluted to 2 nM and paired-end reads generated on a MiSeq (Illumina, San Diego, CA, USA) with a MiSeq Reagent Kit V2 (1 x 150 bp). The 480 18S_SSU and 120 positive 16S_Krill were sequenced on one chip (n=600). -See 18s and Krill PCR excel sheet for samples, primers, 1st round PCR with MID tags, second round PCR with MID tags and miseq sheet. -See 18s and Krill folder for Fastq files In addition we also amplified 500 samples (465 scats, 17 repeats, extraction blanks, positives and negative controls) with the 16S_Fish marker (Deagle, et al (2007) Studying Seabird Diet through Genetic Analysis of Faeces: A Case Study on Macaroni Penguins (Eudyptes chrysolophus). PLOS ONE 2:e831). PCR amplifications were performed in two rounds, the first to amplify the target gene and add sample-specific 6 bp multiplex-identifier (MID) tags (forward and reverse primer) and Illumina sequencing primers, the second to add sequencing adapters and additional 8 bp MIDs. PCR products from all samples including the blanks, positive and negative controls (n=500) were pooled and purified using Agencourt Ampure (Beckman Coulter, USA) magnetic beads. The pool was diluted to 2 nM and paired-end reads generated on a MiSeq (Illumina, San Diego, CA, USA) with a MiSeq Reagent Kit V2 (1 x 150 bp). -See Fish PCR excel sheet for samples, primers, 1st round PCR with MID tags, second round PCR with MID tags and miseq sheet. -See Fish Fastq folder for Fastq files The sex of each sample was determined with a real-time melt curve analysis (Faux et al, (2014) High-throughput real-time PCR and melt curve analysis for sexing Southern Ocean seabirds using fecal samples. Theriogenology 81:870-874). Known male and female Adélie penguin, snow petrel samples and Gentoo penguin samples were included on each run. Sexing reaction mix contained 1 μM for each forward and reverse primer, 2 μg BSA, 1 x LightCycler 480 Probes Master (Roche), 1 x EvaGreen (Biotium). Thermal cycling conditions were 95 degrees for 5 min; followed by 40 cycles of 95 degrees for 10s, 55

The factors that control the number of animals in a population are often difficult to understand. However, this basic understanding is central to managing those populations and assessing how they might respond to human induced pressures. For animals living in the Antarctic, like penguins, the marine environment that they depend on for food can vary due to natural events such as El Nino, and potentially due to human induced changes such as global warming. This study uses modern computer technology to track Royal penguins at sea and to monitor their time on land. By relating where the birds go to feed, what they feed on, and how successfully they catch their food to the survival rates of their chicks, this study will describe how fluctuations in a major Antarctic oceanographic feature (the Antarctic Polar Front) can influence the size of the Royal penguin population at Macquarie Island. Information on breeding success, diet and foraging success were collected each year between 1997-2001. Diving behaviour and at-sea movements were also quantified between 1997 and 1999. These data will also be available in the ARGOS satellite tracking database. Attached to this metadata record are ARGOS tracking data collected by Cindy Hull between 1994 and 2000. The tracking data have been collected from 19 different royal penguins. The download file contains a csv file with tracking data.

Metadata record for data from ASAC Project 1252 See the link below for public details on this project. Currently three datasets are attached to this metadata record. Dive data collected in 1988, track data from adult birds collected in 1994 and track data from fledglings collected in 1995. Dive data are available in Microsoft Word format, while the track data are available in Microsoft Excel format. A readme file (txt) is included in each download file to explain column headings, etc. ---- Public Summary from Project ---- To breed successfully the winter-breeding emperor penguins must fatten on two occasions: once before the onset of moult in January, and again prior to the commencement of the new breeding season in March. Interference with the capacity of the penguins to fatten in summer might be detrimental to the their breeding performance and survival later on in winter. This study seeks to determine the likely impact of commercial fishing operations on emperor penguin colonies at the Mawson Coast. More specifically, the data pertains to the locations of emperor penguins when fattening prior to the moult, and prior to the new breeding season. Project objectives: 1. To determine the extent and location of foraging areas of post-breeding adult Emperor penguins in summer. 3. To determine the extent and locations of foraging areas of fledgling Emperor penguins on their first trip to sea. 4. To identify interseasonal and interannual variations in foraging areas in conjunction with changes in seaice conditions and compare these with results from different colonies. 5. To survey the coastline of the AAT to verify the existence (or non-existence) of Emperor penguin colonies. Emperor penguins are icons of Antarctic wildlife and their conservation is of paramount interest to the wider community. They are also key consumers of marine resources in several areas and consequently there is great potential for interactions between feeding penguins and harvesting of fish and krill. Emperor penguins are one of the few species to breed on the fast ice (although there are three known land-based colonies, one of which has all but ceased to exist in recent years). Thus, the breeding habitat of Emperor penguins is subject to direct alteration as a result of climate change. Colonies of Emperors are found across a wide latitudinal range, from deep in the Ross Sea to the tip of the Antarctic Peninsula. This range includes breeding areas where significant changes in seaice are not (yet?) thought to be occurring to areas where seaice is changing rapidly. Accordingly, studies at multiple locations will provide valuable clues on how this species will be affected by a warming Antarctic. Additionally, Emperor penguins are large animals that live in a relatively small number of discrete locations. It is therefore more than feasible, using an international effort, to study an entire species and to make some predictions about their response to a warming world and to current and future fishing practices. This project aims to make the first steps towards an overall conservation assessment of Emperor penguins through studies in several locations around the Antarctic continent. Should these attempts be successful, then a more ambitious international project will be launched to take a species-wide perspective.

At Hop Island in the Rauer Group during the 2012/13 field season combinations of data loggers were deployed on different adelie penguins. The data loggers were GPS (two types), time-depth recorders and accelerometers. The accelerometer records head movement to identify when the bird captures prey. The units were later retrieved and the data downloaded. A document included with the data has further information about the data. The data were collected following protocols approved by the Australian Antarctic Animal Ethics Committee and supported through the Australian Antarctic program through Australian Antarctic Science project 4087. Data from GPS units deployed at Hop Island in 2011/12 is described by the metadata record with ID AAS_4087_adelie_penguin_tracking_hop_island_2011_12.

This dataset contains data on the habitats, distribution and numbers of Adelie Penguins (Pygoscellis adeliae) in the Mawson area, Antarctica during 1981 and 1988. The data are obtained from aerial photographs obtained at various times, during the 1981-82 and 1988-89 seasons. The results are listed in the documentation. Comparisons are made with census data collected in the 1971-72 summer. Data from this record has been incorporated into a larger Adelie penguin dataset described by the metadata record - Annual population counts at selected Adelie Penguin colonies within the AAT (SOE_seabird_candidate_sp_AP). It also falls under ASAC project 1219 (ASAC_1219).

Metadata record for data from ASAC Project 465 See the link below for public details on this project. From the abstracts of the referenced papers: The diet composition of King penguins Aptenodytes patagonicus at Heard Island (53deg 05S; 73 deg 30E) was determined from stomach contents of 98 adults captured as they returned to the island throughout 1992. During the two growth seasons, the diet was dominated by the myctophid fish Krefftichthys anderssoni (94 % by number, 48 % by mass). The paralepidid fish Magnisudis prionosa contributed less than 1 % by numbers but 17 % by mass. Mackerel icefish Champsocephalus gunnari accounted for 17 % by mass of chick diet in late winter, when chicks were malnourished and prone to starvation, although its annual contribution to the penguins diet was only 3 %. Squid was consumed only between April and August; Martialia hyadesi was the commonest squid taken, comprising 40 to 48 % of the winter diet. The remainder of the diet consisted of the squid Moroteuthis ingens and fish other than K. anderssoni. The energy content of the diet mix fed to the chicks varied seasonally being highest during the growth seasons (7.83 plus or minus 0.25 kJ.g-1) and lowest in winter (6.58 plus or minus 0.19 kJ.g-1). From energetic experiments we estimated that an adult penguin consumed 300 kg of food each of which its chick received 55 kg during the 1992 season. The chicks received large meals at the beginning of winter (1.2 plus or minus 0.3 kg) and during the middle of the second growth season (1.2 plus or minus 0.3 kg), and their smallest meals in late winter (0.4 plus or minus 0.1 kg). The gross energy required to rear a King penguin chick was estimated to be 724 MJ. The potential impact of commercial fisheries on the breeding activities of King penguins is discussed. 23 king penguins (Aptenodytes patagonicus) from Macquarie Island were tracked by satellite during the late incubation period in 1998-1999 to determine the overlap in the foraging zone of king penguins with an area to be declared a marine protected area (MPA) near the island. While all penguins left the colony in an easterly direction and travelled clockwise back to the island, three penguins foraged in the northern parts of the general foraging area and stayed north of 56 south. The remaining 20 penguins ventured south and most crossed 59 south before returning to the island. The total foraging area was estimated to be 156,000 square kilometres with 36,500 square kilometres being most important (where penguins spend greater than 150 hours in total). North-foraging penguins reached on average 331 plus or minus 24 kilometres from the colony compared to 530 plus or minus 76 kilometres for the south-foraging penguins. The latter travelled an average total distance of 1313 p lus or minus 176 kilometres, while the northern foragers averaged 963 plus or minus 166 kilometres. Not only did the penguins spend the majority of their foraging time within the boundaries of the proposed MPA, they also foraged chiefly within the boundaries of a highly protected zone. Thus, the MPA is likely to encompass the foraging zone of king penguins, at least during incubation. The foraging strategies of king penguins from Heard and Macquarie islands were compared using satellite telemetry, time-depth recorders and diet samples. Trip durations were 16.8 plus or minus 3.6 days and 14.8 plus or minus 4.1 days at Macquarie and Heard islands, respectively. At Macquarie Island, total distances travelled were 1281 plus or minus 203 km compared to 1425 plus or minus 516 km at Heard Island. The total time the penguins spent at sea was 393 plus or minus 66 h at Macquarie Island and 369 plus or minus 108 h at Heard Island. The penguins from Macquarie Island performed more deep dives than those from Heard Island. King penguins from Macquarie Island travelled 1.5 plus or minus 0.2 km h-1 day-1 compared to 1.3 plus or minus 0.1 km h-1 day-1. At Macquarie Island, 19% of dives were up to