Training large macaws for artificial insemination procedures

For some endangered parrot species, captive breeding may be the only insurance for their survival. However, many individuals in captivity do not reproduce. Artificial insemination (AI) may help overcome reproductive failures or geographic distance. For semen collection in birds, massage is the most commonly used method. However, this process, which usually requires capture and restraint, involves risk of stress and injuries. The aim of this experiment was to train large macaws to accept the physical manipulations of their body parts needed for the artificial insemination process. Within 15 weeks, a male and a female Buffon's macaw (Ara ambiguus) learned to accept handling without apparent stress. A pair of green-winged macaws (Ara chloropterus) progressed more slowly and displayed some signs of stress. This stress highlights the need to monitor the birds' possible signs of discomfort during the training in order to adapt the working protocol. These results demonstrate that it is possible to train the AI behaviors, thus avoiding the capture, restraint, and anesthesia. An added benefit to this is the potential for the training to provide a form of behavioral enrichment.     

The phylogenetic position of the extinct Cuban Macaw Ara tricolor based on complete mitochondrial genome sequences

The Cuban Macaw Ara tricolor was a species of macaw native to Cuba and Isla de la Juventud in the Caribbean that became extinct in the 1860s. Morphologically, it was similar to, but distinctively smaller than, the large red macaws – Scarlet Macaw Ara macao and Red‐and‐green Macaw Ara chloropterus. A close affinity with the Scarlet Macaw has been suggested based on plumage similarities. In this study we use complete mitochondrial genome sequences to examine the phylogenetic position of the Cuban Macaw. Our results do not indicate a sister‐species relationship with the Scarlet Macaw but place the Cuban Macaw as sister to the two red species and the two large green macaws, the Military Macaw Ara militaris and the Great Green Macaw Ara ambiguus. Divergence estimates suggests that the Cuban Macaw separated from this group approximately 4 million years ago.     

Seed predation by monkeys and macaws in eastern Venezuela: Preliminary findings

Feeding data collected concurrently on bearded sakis (Chiropotes satanas) and red and green macaws (Ara chloroptera) on a large island in Guri, Venezuela provides preliminary evidence that these two seed predators have similar diets. Individuals of both species were equally capable of opening very hard, protected fruits. Of the seven fruit species used by macaws during the study period, four species were also ingested by sakis at the same stage of ripeness, two species were ingested at different stages of ripeness (macaws earlier than sakis), and one species was never observed to have been eaten by sakis. The second finding, that macaws ingest young seeds of the Anacardiaceae and Burseraceae families and the bearded sakis ingest only the ripe mesocarp of these species suggests that the most distinguishable difference in their diets might be a tolerance of toxins by the macaws that act as feeding deterrents for the monkeys. Although we did not document the location of local clay licks in eastern Venezuela, the use of clay licks by macaws in Peru (Munn, 1992) suggests that this activity (that is not practiced by the sakis) may be helpful in detoxifying or ameliorating the effects of ingesting chemically protected fruit. deceased.     

Ecology and Management of Nesting Blue-and-Yellow Macaws (Ara ararauna) in Mauritia Palm Swamps

This paper reports on the nesting ecology of blue-and-yellow macaws (Ara ararauna) and the structure, conservation, and management of the Mauritia flexuosa palm swamps where they nest: clutch size averaged 2.6 and the number of chicks fledged per nesting attempt was 0.5. Macaws nested in tall dead palms in healthy palm swamps and in palms of all heights in open dieing palm stands. All nesting palms rose well above the surrounding vegetation presumably to discourage terrestrial predators. PVC nest boxes failed to attract nesting blue-and-yellow macaws. A small section of palm swamp was managed to encourage macaw nesting by cutting the tops off of M. flexuosa palms and clearing the understory vegetation. The palms remained standing from 4 to 7 years and were occupied by nesting macaws at a rate of 24%. The data presented here suggest that cutting five palms a year in perpetuity would produce a stand of approximately 20 standing dead palms used by 6 or more pairs of macaws annually. However, macaw occupancy rates would depend on the density of macaws and density of naturally occurring nest sites. This management scheme could be conducted on a 100-year rotation in an area of 1–4 ha or more depending on the palm density. Such a colony could be used to increase reproductive success of blue-and-yellow macaw populations, create a valuable ecotourism resource, and concentrate macaw nesting in protected areas.     

Parrot behavior at a Rio Manu (Peru) clay lick: temporal patterns,associations, and antipredator responses

Although eating clay at licks (a form of geophagy) has been described, there are few behavioral data on temporal patterns, social interactions, species associations, or reactions to potential predators. We examined the behavior of nine species of macaws, parrots, and parakeets at the Machiguenga Ccolpa, a clay lick on the Rio Manu, Peru in the dry season. Three distinct mixed-species groups used the licks: in the early morning (parrots and small macaws), in mid-morning (large macaws), and in the early afternoon (parakeets), although the latter two groups used the licks at other times of day as well. The first parrots to begin eating at the lick in the early morning were yellow-crowned parrots (Amazona ochrocephala) and dusky-headed parakeets (Aratinga weddellii), followed by blue-headed parrots Pionus sordidus, and then by mealy (Amazona farinosa) and orange-cheeked (Pionopsitta barrabandi) parrots, and chestnut-fronted macaws (Ara severa). Although blue-headed parrots fed in dense groups of over 50, the others rarely exceeded 20 individuals. Scarlet macaws (A. macao) sometimes fed alone or joined the early morning groups, but most associated with a large group of red and green macaws (A. chloroptera) that arrived, often scaring off the smaller birds. On average, about 100 macaws and parrots fed in the early morning, macaw feeding groups averaging just over 40, and the parakeets averaged over 70. Average time at the lick ranged from 28 min for yellow-crowned parrots to 47 min for tui parakeets. Of the early morning group, blue-headed and mealy parrots were the most aggressive and orange-cheeked parrots were the least aggressive. Red and green macaws were more aggressive than scarlet macaws; the parakeets were equally aggressive. All species had more aggressive interactions with conspecifics than with other species. Responses to intruders and predators varied by species of parrot/macaw and type of intruder. In response to intruders or loud calls, responses could be partial (some individuals flew away, circled, and returned), temporary (all individuals flew away but returned within a few minutes), or total (all flew away and abandoned feeding for at least a half hour). The large macaws showed the lowest rate of total abandonment and the parakeets showed the highest. People passing up or down river in boats scared birds from the lick. The local residents (Machiguenga tribespeople in boats) elicited a much greater response than did the researchers. In the recent past, macaws and parrots were hunted for food, feathers, and the pet trade, and the birds response, as well as the presence of parrot and macaw feathers in local villages we visited, suggests some continued exploitation, or a long-term memory in the birds.Communicated by R.F. Oliveira     

The application of non-invasive genetic tagging reveals new insights into the clay lick use by macaws in the Peruvian Amazon

Genetic tagging, the unique identification of individuals by their DNA profile, has proven to be an effective method for research on several animal species. In this study we apply non-invasive genetic tagging from feather samples to reveal the genetic structure and estimate local population size of red-and-green macaws (Ara chloropterus) without the need to capture these animals. The study was centered in the Tambopata region of the Peruvian Amazon. Here macaws frequently visit clay licks and their naturally molted feathers provide a unique source of non-invasively sampled DNA. We analyzed 249 feathers using nine microsatellite loci and identified 221 unique genotypes. The remainder revealed 21 individuals which were ‘recaptured’ one or more times. Using a capture-mark-recapture model the average number of different individuals visiting clay licks within one breeding season was estimated to fall between 84 and 316 individuals per clay lick. Analysis of population genetic structure revealed only small genetic differences among regions and clay licks, suggesting a single red-and-green macaw genetic population. Our study confirms the utility of non-invasive genetic tagging in harsh tropical environment to obtain crucial population parameters about an abundant parrot species that is very difficult to capture in the wild.     

Chromosomal Diversity and Karyotype Evolution in South American Macaws (Psittaciformes,Psittacidae)

Most species of macaws, which represent the largest species of Neotropical Psittacidae, characterized by their long tails and exuberant colours, are endangered, mainly because of hunting, illegal trade and habitat destruction. Long tailed species seem to represent a monophyletic group within Psittacidae, supported by cytogenetic data. Hence, these species show karyotypes with predominance of biarmed macrochromosomes, in contrast to short tailed species, with a predominance of acro/telocentric macrochromosomes. Because of their similar karyotypes, it has been proposed that inversions and translocations may be the main types of rearrangements occurring during the evolution of this group. However, only one species of macaw, Ara macao, that has had its genome sequenced was analyzed by means of molecular cytogenetics. Hence, in order to verify the rearrangements, we analyzed the karyotype of two species of macaws, Ara chloropterus and Anodorhynchus hyacinthinus, using cross-species chromosome painting with two different sets of probes from chicken and white hawk. Both intra- and interchromosomal rearrangements were observed. Chicken probes revealed the occurrence of fusions, fissions and inversions in both species, while the probes from white hawk determined the correct breakpoints or chromosome segments involved in the rearrangements. Some of these rearrangements were common for both species of macaws (fission of GGA1 and fusions of GGA1p/GGA4q, GGA6/GGA7 and GGA8/GGA9), while the fissions of GGA 2 and 4p were found only in A. chloropterus. These results confirm that despite apparent chromosomal similarity, macaws have very diverse karyotypes, which differ from each other not only by inversions and translocations as postulated before, but also by fissions and fusions.     

A Case Study Employing Operant Conditioning to Reduce Stress of Capture for Red-Bellied Tamarins (Saguinus labiatus)

Traditional techniques used to capture New World monkeys, such as net capture, can induce high levels of acute stress detrimental to welfare. Alternatively, training nonhuman animals via operant conditioning to voluntarily participate in husbandry and/or veterinary practices is accepted as a humane process that can reduce stress and improve welfare. This study details the use of operant conditioning using positive reinforcement training (PRT) and target training to train a family of 5 captive red-bellied tamarins (Saguinus labiatus) in a wildlife park to voluntarily enter a transportation box and remain calm for 1 min after 54 training sessions. Observations of 2 unrelated net-capture processes provided measures of locomotion and vocalizations as indicators of stress behavior that were compared with those of the trained tamarins. Net-captured monkeys exhibited rapid erratic locomotion and emitted long, high-frequency vocalizations during capture whereas the trained tamarins exhibited minimal locomotion and emitted only 4 brief vocalizations (root mean square 35 dB) during capture. This indicates that the use of PRT considerably reduced potential for stress and improved welfare during the capture and containment of the tamarins.     

Genetic patterns and conservation of the Scarlet Macaw (<Emphasis Type="Italic">Ara macao</Emphasis>) in Costa Rica

Once widely distributed throughout the lowland forests of Costa Rica, scarlet macaws (Ara macao) have been reduced to two major, geographically separated, populations along the Pacific slope. Past demographic declines raise conservation concerns regarding the detrimental effects of population fragmentation. This investigation aimed to evaluate the current status of scarlet macaws along the Pacific slope by examining levels of genetic variation and patterns of genetic structure within and among remnant populations. Statistical analyses using multilocus genotypes revealed strong differentiation between Central and South Pacific populations, suggesting local geographic barriers have historically restricted gene flow between these localities. High genetic diversity suggests neither population suffers from genetic erosion, likely resulting from relatively large population sizes and high dispersal capacity and longevity. However, evidence of disequilibrium within the Central Pacific population infers anthropogenic threats have disrupted natural population dynamics. These results advocate on focusing available resources on habitat restoration and nest protection, as a means to assist in reestablishing demographic stability and maintain the genetic health of wild scarlet macaws in Costa Rica.     

Forensic species identification of large macaws using DNA barcodes and microsatellite profiles

Using mitochondrial and nuclear markers species identification was conducted in the case of seized feathers. Earlier, we had sequenced cytochrome c oxidase subunit I (COI) both from 10 seized specimens and 43 validation specimens from captive macaws belonging to 4 Ara species (A. macao, A. chloropterus, A. ararauna, and A. ambiguus) and identified 19 haplotypes based on COI sequences. Species-level identification using Barcode of Life Data Systems showed that seized feathers shared the highest similarity with scarlet macaws (A. macao), and this result was supported by the tree-base identification with high bootstrap values. Moreover, microsatellite profiles in AgGT17 locus showed that patterns of allelic distribution in the seized feathers were apparently distinct from those of red-and-green macaw (A. chloropterus), but were overlapped with those of A. macao, suggesting that all of seized feathers were derived from several individuals of A. macao. We also determined the parentage of hybrid macaws by the combination of COI barcodes and microsatellite profiles. The technique presented here will contribute to forensic identification and future conservation of large macaws that have been lost due to deforestation.     

Scarlet Macaw, Ara macao, (Psittaciformes: Psittacidae) diet in Central Pacific Costa Rica

From 1993 to 1997, we observed Scarlet Macaw (Ara macao) feeding behavior in Central Pacific Costa Rica. Feeding data acquired in this study were not collected systematically, but opportunistically whenever macaws were observed feeding. To supplement feeding observations, we conducted interviews with local residents. Scarlet Macaws fed on seeds, fruits, leaves, flowers and/or bark of 43 plant species. Various plant parts eaten by macaws from several tree species contain secondary compounds toxic to humans, and additional species included in their diet are nonnative, introduced for agricultural purposes. Important macaw feeding tree species are Ceiba pentandra, Schizolobium parahybum, and Hura crepitans; these species are also crucial to this macaw population because of nest cavities they provide. The results of this study contribute to the conservation of Scarlet Macaws in Central Pacific Costa Rica through promoting protection of individual trees, and through local elementary school reforestation programs focusing on tree species that macaws use for feeding and/or nesting. Scarlet Macaw conservation is extremely important, as numerous population pressures have caused significant declines in macaw numbers in Costa Rica.     

A technique for transcervical intrauterine insemination of ewes

In commercial artificial insemination (AI) of sheep, fresh extended semen is deposited into the vagina or cervical os, or fresh extended or frozen semen is placed laparoscopically into the uterus. Transcervical intrauterine insemination of the ewe is not used commercially. In this study, methods of restraint and instrumentation for AI were evaluated and modified to produce a transcervical intrauterine technique suitable for commercial application. Four methods of restraint, four vaginal specula, three forceps and four instruments suitable for transcervical passage were compared. From these comparisons a technique was developed in which the ewes were positioned in dorsal recumbency with their hindquarters elevated. The vagina was dilated using a duck-billed speculum, the cervix was grasped and retracted using forceps, and an inseminating instrument was introduced into the cervical opening and manipulated through the cervical canal. The technique was repeated on 89 mature, multiparous ewes: the difficulty in locating the cervical opening, the force required to retract the cervix and the time required to penetrate into the uterus were recorded. Uterine penetration was achieved in 82% of the ewes. This technique has the potential to be applied in commercial artificial insemination programs of sheep.     

Short-duration insemination with frozen semen increases fertility rate in nulliparous dairy goats

Standard artificial insemination (AI) using a speculum in dairy goats does not result in acceptable fertility rates in nulliparous does. An explanation might be the difficulties to pass the cervical canal in nulliparous females with the insemination gun, increasing the time needed for semen deposition. Nulliparous Alpine dairy goats were used to evaluate whether time interval from insertion to withdrawal of the speculum is a factor influencing pregnancy rates to first AI with frozenthawed semen. Oestrus was synchronized using fluorogestone acetate intravaginal sponges (FGA, 40 mg) for 11 days, associated with 50 mg i.m. of cloprostenol and 250 IU i.m. eCG 48 ± 2 h before sponge removal. In the first experiment (n = 52; 3 herds), the average duration of the AI procedure was 42 ± 10 s, with a median of 39 s. AI performed in less than 39 s resulted in higher pregnancy rates (75%, n = 28) than AI lasting for more than 39 s (46%, n = 24). In the second experiment, does (n = 325; 5 herds) were randomly assigned into two treatment groups according to a short (20 s) or long (60 s) AI procedure. We showed that the duration of AI affected fertility after a first insemination, and that pregnancy rate was significantly improved using a short-duration AI (61.2%; n = 169) compared with a long-duration AI (44.2%; n = 156). We have previously shown in the ewe that genital stimulation during AI enhanced uterine motility. Other authors reported a negative correlation between increased uterine motility at the time of AI and fertility rates in small ruminants. The results of this study suggest that rapid semen deposition may limit the reflex activation of uterine contractions provoked by the speculum and the movement of the insemination gun, and thus ameliorates reproductive performance to first AI in nulliparous goats.     

Acute restraint stress produces behavioral despair in weanling rats in the forced swim test

Stressful experiences in the rat during early life increase the vulnerability to later signs of behavioral despair in adulthood, reflected in increased immobility in the forced swim test (FST). However, the possible immediate effects of stress in weanling rats have only been partially described. The present study tested whether a single session of mild restraint stress modifies immobility in the FST in 21-day-old Wistar rats. After evaluating any possible changes in locomotion using the open field test (OFT), the latency and total duration of immobility were assessed in a single FST session. Regardless of gender, mild restraint stress significantly reduced crossings in the OFT, shortened the latency to the first period of immobility, and increased immobility in the FST compared with a control group devoid of stress. We conclude that a single mild physical stress session, as early as postnatal day 21, produces signs of behavioral despair.     

Artificial insemination in marsupials

Assisted breeding technology (ART), including artificial insemination (AI), has the potential to advance the conservation and welfare of marsupials. Many of the challenges facing AI and ART for marsupials are shared with other wild species. However, the marsupial mode of reproduction and development also poses unique challenges and opportunities. For the vast majority of marsupials, there is a dearth of knowledge regarding basic reproductive biology to guide an AI strategy. For threatened or endangered species, only the most basic reproductive information is available in most cases, if at all. Artificial insemination has been used to produce viable young in two marsupial species, the koala and tammar wallaby. However, in these species the timing of ovulation can be predicted with considerably more confidence than in any other marsupial. In a limited number of other marsupials, such precise timing of ovulation has only been achieved using hormonal treatment leading to conception but not live young. A unique marsupial ART strategy which has been shown to have promise is cross-fostering; the transfer of pouch young of a threatened species to the pouches of foster mothers of a common related species as a means to increase productivity. For the foreseeable future, except for a few highly iconic or well studied species, there is unlikely to be sufficient reproductive information on which to base AI. However, if more generic approaches can be developed; such as ICSI (to generate embryos) and female synchronization (to provide oocyte donors or embryo recipients), then the prospects for broader application of AI/ART to marsupials are promising.     

Trucking stress at breeding does not lower conception rate of beef heifers

The goal of this study was to determine whether 1 h of trucking stress before or after artificial insemination (AI) altered the conception rate of beef heifers. Estrus was synchronized in heifers with prostaglandin F(2alpha).The 3 treatment groups consisted of 1) AI (control heifers, n = 93); 2) Truck + AI (trucked for 1 h immediately before AI, n = 81); and 3) AI + Truck (trucked for 1 h immediately after AI, n = 82). All heifers were artificially inseminated by a single technician with semen from a single ejaculate. Blood samples were collected for cortisol measurement 1 h before AI, immediately before and after AI, and 1 h after AI in the AI (n = 6), Truck + AI (n = 9), and AI + Truck (n = 8) groups Pregnancy in heifers was confirmed either at slaughter or by palpation per rectum. Trucking before AI elevated (P < 0.01) serum cortisol concentrations. Artificial insemination alone increased (P < 0.01) serum cortisol concentrations in AI heifers. The increase in serum cortisol concentrations caused by trucking after AI was not significant (P > 0.05). Areas under the cortisol curves in Truck + AI heifers are greater (P < 0.05) than in AI heifers. The conception rates of AI heifers (50.5%), Truck + AI heifers (51.9%) and AI + Truck heifers (58.5%) are not different (P > 0.05). This study demonstrates that 1 h of trucking stress either before or after AI did not lower the conception rate of heifers.     

Taking the sting out of darting: Risks,restraint drugs and procedures for the chemical restraint of Southern Hemisphere otariids

The need to manage otariid populations has necessitated the development of a wide range of capture methods. Chemical restraint by remote drug delivery (i.e., darting) is a highly selective method that can be used to facilitate otariid capture in a range of scenarios, when other methods may be impracticable. However, the risks associated with darting otariids are not widely known and guidelines necessary to promote and refine best practice do not exist. We review the risks associated with darting and in light of our findings, develop darting guidelines to help practitioners assess and minimize risks during capture, anesthesia and recovery. Published studies reveal that mortalities associated with darting predominantly result from complications during anesthetic maintenance (e.g., prolonged respiratory depression, apnea, or hyperthermia), rather than from complications during capture or recovery. In addition to monitoring vital signs and proper intervention, the risk of irreversible complications during anesthesia can be reduced by administering drug doses that are sufficient to enable the capture and masking of animals, after which anesthetic depth can be regulated using gas anesthesia.     

Comparative analysis of microsatellite variability in five macaw species (Psittaciformes, Psittacidae): Application for conservation

Cross-amplification was tested and variability in microsatellite primers (designed for Neotropical parrots) compared, in five macaw species, viz., three endangered blue macaws (Cyanopsitta spixii [extinct in the wild], Anodorhynchus leari [endangered] and Anodorhynchus hyacinthinus [vulnerable]), and two unthreatened red macaws (Ara chloropterus and Ara macao). Among the primers tested, 84.6% successfully amplified products in C. spixii, 83.3% in A. leari, 76.4% in A. hyacinthinus, 78.6% in A. chloropterus and 71.4% in A. macao. The mean expected heterozygosity estimated for each species, and based on loci analyzed in all the five, ranged from 0.33 (A. hyacinthinus) to 0.85 (A. macao). As expected, the results revealed lower levels of genetic variability in threatened macaw species than in unthreatened. The low combined probability of genetic identity and the moderate to high potential for paternity exclusion, indicate the utility of the microsatellite loci set selected for each macaw species in kinship and population studies, thus constituting an aid in planning in-situ and ex-situ conservation.