Petronia Research Articles

Heterozygosity, genetic similarity and extra‐pair paternity variation in two populations of rock sparrow Petronia petronia: a within and between populations comparison

Extra‐pair paternity (EPP) has been broadly reported in socially monogamous bird species and it has been hypothesized that females engage in extra‐pair copulations to increase the genetic variability of the offspring and to reduce the risk of inbreeding and genetic incompatibilities. This hypothesis makes two predictions: within populations, females should engage with more dissimilar/heterozygous males and, among populations, females should pursue more frequently EPP in populations characterized by a lower genetic variability and a higher homozygosity. However, support is still unclear throughout literature, and usually involves the study of a single population. We compared a peripheral population of rock sparrow Petronia petronia living at the marginal distribution of the species and characterized by a high EPP level (> 50%) (Italian Alps) with a population located in the centre of the species' distribution (central Spain), to understand if variations in EPP could be linked to differences in mean heterozygosity and genetic similarity both between and within populations. EPP in the Spanish population was 18.1%, three times lower than that observed in the Alpine one (51.2%), and this difference remained fairly constant across different years. Supporting the between populations prediction, we found lower heterozygosity and reduced allelic richness in the Alpine population compared to the Spanish one. In contrast, social and extra‐pair males, as well as within and extra‐pair offspring, did not significantly differ in terms of genetic similarity and heterozygosity within brood in either population. Social and extra‐pair males did not differ in tarsus size, body weight or yellow badge size, suggesting that females were not choosing extra‐pair partners based on heterozygosity, genetic similarity or phenotypic quality. Although based on a limited sample in the within population analysis, our results indicate that EPP may evolve in response to a low level of genetic variability in the population.

The relationships between breeding success and nest site selection for some Passeriformes in Ikh Nart Nature Reserve, Mongolia

Offspring survival is critical to bird conservation and understanding avian life history. We monitored 73 nests belonging to six Passeriformes species in Ikh Nart Nature Reserve, Mongolia in 2017 and 2018. The study focused on nest site selection for breeding visitors and resident avian species in relation to habitat types found in arid environments, such as rocky outcrops and open steppe. Egg laying period varied among the species. Those that nested in the rocky outcrops realized higher reproductive success. The offspring survival decreased in ground-nesting birds after hatching. Resident species, including Rock Sparrow (Petronia petronia), Mongolian Finch (Bucanetes mongolica), and Horned Lark (Eremophila alpestris), showed lower reproductive success during the first half of the season, while breeding visitors, including Northern Wheatear (Eremophila alpestris), Isabelline Wheatear (Oenanthe isabellina), and Pied Wheatear (Oenanthe pleschanka), saw higher reproductive success. Rocky outcrops near water sources in arid environments play an important part in the breeding success of rock-dwelling birds.

A revised multilocus phylogeny of Old World sparrows (Aves: Passeridae)

Abstract The Old World sparrows include some of the best-studied passerine species, such as the cosmopolitan human commensal, the house sparrow (Passer domesticus) as well as poorly studied narrow-range endemics like the Iago sparrow (P. iagoensis) from the Cape Verde Archipelago or specialists from extreme environments like the desert sparrow (P. simplex). It is therefore notable that to date the most complete phylogenetic hypothesis for the Old World sparrows comprised only ten of 43 currently accepted species. With this study we provide an updated phylogeny of Passeridae covering about two third of the family’s species richness. Though still being far from taxon-complete, this new phylogenetic hypothesis provides firm evidence to clarify some open taxonomic questions. All genus-level taxa were reciprocally monophyletic with strong support. Contrary to previous classifications, bush sparrows and rock sparrows were not sister taxa, and therefore their classification in separate genera Gymnoris and Petronia is justified. Plumage color traits like the yellow throat patch of the latter two genera or head color pattern in Passer species do not provide reliable phylogenetic information, except for the large-sized African grey-headed sparrows that resulted as a monophyletic group (P. diffusus, P. griseus, P. gongoensis). Unexpectedly, two small-sized species, P. eminibey and P. luteus that to date are regarded as close relatives were firmly nested in two separate clades of Passer sparrows. Therefore, their separate generic treatment under Sorella eminibey and Auripasser luteus (together with A. euchlorus) does not seem justified.

Climate warming induced a stretch of the breeding season and an increase of second clutches in a passerine breeding at its altitudinal limits.

The increase in the average air temperature due to global warming has produced an early onset of the reproduction in many migratory birds of the Paleartic region. According to the “mismatch hypothesis” this response can lead to a decrease in the breeding output when the conditions that trigger the departure from the wintering areas do not match the availability of food resources in the breeding ground. We used 653 brooding events registered during the period 1991–2013 to investigate the link between climatic variables and individual breeding performance of a partially migratory passerine, the Rock Sparrow Petronia petronia, breeding at the altitude limit of its distribution. The laying date (LD) of the earliest first clutch was associated with local spring (minimum) temperatures but did not show a significant trend during the period considered. The LD of the latest first clutch had a positive and statistically significant trend, unrelated to local covariates and resulting in a longer breeding season (∼1.5 days/year). A longer breeding season allowed birds to produce more second clutches, which proportion increased from 0.14 to 0.25. The average breeding success was also positively correlated with the average temperature in July and with the duration of the breeding season. Contrary to expectations, the most important climate-dependent effect was a stretch of the breeding season due to a significant increase of the LD of the latest first-clutches rather than an earlier breeding onset. We show how climate changes act on bird populations through multiple paths and stress the need to assess the link between climatic variables and several aspects of the breeding cycle.

Habitat use during post-pre-breeding period in a Rock Sparrow <em>Petronia petronia</em> population

Most studies focusing on Rock Sparrow populations concern their breeding biology. With this study, we aim to investigate the wintering habitat and social behaviour outside the breeding period. In the study area the species is present all year long, and, outside the breeding period, birds frequently gather together on flocks of different size (3-150 individuals), which are larger in the coldest months. The habitat types most used by the species were cereal fields with crops in their first stage of development (17.71% ± 4.72 Standard Deviation) and arable stubbles, both of them with bushes and trees on their boundaries (82.4%), which likely provide shelters from predators. The bird species most frequently associated with Rock Sparrow flocks were Tree Sparrow, Starling and Crested Lark. Holes in the roof tiles of the same abandoned buildings used for breeding were also used as night shelters. The average hours of leaving and returning to the shelters was strictly related to the photoperiod (rs = -0.9405; 0.8810).

Pair coordination is related to later brood desertion in a provisioning songbird

Raising a family is not free of conflict for parents, as each parent benefits when its partner provides more care for the offspring. Resolving this conflict requires cooperation between the parents. One way to achieve such cooperation might be to coordinate parental provisioning by synchronizing (i.e. returning to the nest at the same time) or alternating (i.e. taking turns) offspring provisioning at the nest. Empirical studies in birds indicate that pair coordination of the nest visits is common; however, it is unknown whether this behaviour is directly related to different outcomes of sexual conflict, such as brood desertion. We used the rock sparrow, Petronia petronia, a species with high levels of sexual conflict, to explore whether alternation and synchrony of the nest visits were related to later brood desertion. Pairs with no desertion alternated and synchronized their nest visits more than pairs in which one sex deserted. This difference in coordination was not simply a by-product of differences in provisioning by the parents. Synchrony of the visits also increased with offspring age in the pairs with no desertion. We provide evidence, for the first time to our knowledge, that the degree of parental coordination is strongly associated with the ultimate consequence of sexual conflict, brood desertion, supporting the idea that coordination in parental behaviour might promote conflict resolution.

Parental feeding responses to experimental short-term partner removal in a species with male and female brood desertion

As parental care is costly, it can be expected that there will be a sexual conflict between parents over the individual levels of parental investment because each parent has limited resources to invest in a reproductive event. Theoretical models of parental investment predict that when one parent reduces its parental effort, the other parent should adjust its effort facultatively to compensate for the decrease in the mate’s contribution. Here, we tested for facultative adjustments in care in the rock sparrow, Petronia petronia. In this species, both sexes can desert the brood, creating the potential for strong sexual conflict over parental care to occur. To that end, we examined how rock sparrow parents adjust their level of care in response to an experimental mate removal for a limited time period, mimicking the starting phase of the desertion process. We compared male and female provisioning rates before and after an experimental mate removal. Males behaved according to the prediction of compensatory adjustment, as they allocated more care to the offspring and returned faster to the nest after female removal, whereas females showed no response. Our study shows sexual differences in response to mate removal in a species with biparental care and strong sexual conflict over parental care, and suggests that males may use the female absence to determine their actual parental effort.Significance statementStrong differences in parental care, with one sex providing more care than the other, are widespread across the animal kingdom. Theoretical models of parental investment predict that when one parent reduces its care, the mate should adjust its care facultatively to compensate. Here, we tested for facultative adjustments in care in the rock sparrow when their mates were experimentally removed for a limited time period. We show that only male behavior was consistent with theoretical predictions as they increased their frequency of provisioning significantly after mate removal, whereas females showed no response. Our study suggests that members of a pair do not make independent decisions, but negotiate to modify their effort in direct response to the prior effort of their mate.

Nestling sex ratio is associated with both male and female attractiveness in rock sparrows

According to theory, in species in which male variance in reproductive success exceeds that of the females, sons are more costly to produce; females mated with high quality males or those in better condition should produce more sons. In monogamous species, however, the variance in the reproductive success of the two sexes is often similar and mate choice is often mutual, making predictions regarding sex allocation more difficult. In the rock sparrow Petronia petronia, both males and females have a sexually selected yellow patch on the breast, whose size correlates with individual body condition. We investigated whether the brood sex ratio co‐varies with the size of the yellow patch of the father and the mother in a sample of 173 broods (818 chicks) over 8 breeding seasons. While the size of the yellow patch of the mother and the father did not predict per se a deviation from the expected 1:1 sex ratio, brood sex ratios were predicted by the interaction of male and female yellow patch size. This result is surprising, as the ornament is sexually selected by both males and females as an indicator of quality in both sexes and should therefore be inherited by all offspring irrespective of their sex. It indirectly suggests that other sex‐specific traits associated with patch size (e.g. polygyny in males and fecundity in females) may explain the sex allocation bias observed in rock sparrows. Thus, female individual quality alone, as expressed through the size of the yellow patch, was not associated with the biases in sex ratios reported in this study. Our results rather suggest that sex allocation occurs in response to male attractiveness in interaction with female attractiveness. In other words, females tend to preferentially allocate towards the sex of the parent with more developed ornament within the pair.

A novel gene organization of the rock sparrow Petronia petronia (Aves: Passeriformes) revealed by complete mitochondrial genome

The whole sequence of the Rock Sparrow (Petronia petronia) mitochondrial genome was determined using La-PCR and conserved primer walking approaches. The entire mitogenome was 17,426 bp in length and harboured 13 protein-coding genes, 2 rRNA genes, 23 tRNA genes, and 1 noncoding control region. The mitogenome of this species resembled other avian species in gene arrangement and composition, except that tRNA-Glu had two copies and the control region contained two parts. One part of the control region is between tRNA-Glu and tRNA-Phe and a second between the two tRNA-Glu copies. The overall nucleotide composition are A (30.7%), T (23.8%), G (14.4%), and C (31.1%) with 54.5% A + T content. These mitochondrial data are potentially important for the study of molecular evolution and conservation genetics.

Oxidative status in nestlings shows different associations with parental carotenoid-based plumage ornaments depending on parental sex and year: a study of rock sparrows Petronia petronia

Carotenoid-based colouration in birds has been suggested to be a sexually selected signal, allowing individuals to obtain reliable information about the quality of antioxidant resources. Moreover, oxidative stress levels during early development may have medium- and long-term fitness consequences. Rock sparrows (Petronia petronia) display a carotenoid-based breast patch in both sexes which has been shown to function as a signal of phenotypic quality in sexual and social interactions. We studied a rock sparrow population in central Spain aiming to test whether the extent of the carotenoid-based breast patch is associated with offspring growth and oxidative status. We have used plasma malondialdehyde (MDA) level in plasma as a measure of oxidative damage, and total antioxidant status of plasma (TAS) and total glutathione (tGSH) levels in red blood cells as measures of antioxidant defences. The study included the extremely late breeding season of 2013 and the phenologically more normal season of 2014. Unfavourable conditions for breeding resulted in smaller nestlings in 2013. Male breast patch size showed a negative association with nestling tarsus length and, strikingly, a positive association with nestling MDA. It also showed a positive relationship with nestling wing length in interaction with year. Nestlings in late and larger broods showed higher tGSH levels. Female breast patch size showed positive associations with nestling tGSH levels, indicating positive links of maternal ornamentation with offspring condition. Moreover, TAS values were only positively associated with female patch size under favourable conditions, as shown by the interaction with year. Associations of carotenoid-based parental plumage ornaments with offspring growth and oxidative status depends on parental sex and environmental conditions. Carotenoid-based plumage traits signal parental quality in female rock sparrows, while paternal ornaments seemingly do not reflect parental quality. To our knowledge, this is the first study to investigate the association between female plumage ornaments and offspring antioxidant capacity. The same signal may reflect different parental attributes in the two sexes in sexually monomorphic species.

An experimental test on time constraint and sexual conflict over parental care.

Because parental care is costly, a sexual conflict between parents over parental investment is expected to arise. Parental care behavior is an adaptive decision, involving trade-offs between remating, and consequently desertion of the brood, and continuing parental effort. If the main advantage of desertion is remating, then this will be a time constraint, because the deserting individual will require a certain minimum period of time to breed again in the same breeding season. So, a short breeding season should force certain individuals to desert the first brood to have enough time to successfully complete their second breeding attempt. The rock sparrow, Petronia petronia, is an unusual species in which brood desertion can occur in both sexes and the breeding season is quite short so it is a good species to investigate the role of time constraint on brood desertion. For 3 years, I investigated the brood desertion modality of the rock sparrow. Then, for 2 years, I removed a group of experimental nest boxes during the autumn. Later, I re-installed the experimental nest boxes after the start of the breeding season (2 weeks after the first egg was laid), mimicking a shortening of the breeding season for the (experimental) pairs that used experimental nest boxes. I found that in the experimental pairs, the percentage of deserting individuals was significantly higher than in the control groups, and the deserting individuals were older females. This experiment adds to our knowledge of timing of reproduction effects on individual decisions to desert by showing that a short and delayed breeding season may have different effects on males and females. To my knowledge, this is the first experimental study that demonstrates a direct link between time constraint and brood desertion.

Nest decorations: an ‘extended’ female badge of status?

Extended phenotypes as signals are widely distributed among animal taxa. For example, many bird species build eye-catching nests or structures, which can potentially mirror the quality or ability of the builder. Rock sparrow, Petronia petronia, nests are usually overly decorated with feathers belonging to different species. Feather carrying in this and other species seems to play a role beyond their supposed thermoregulatory function, that is, to provide insulation to eggs and developing chicks. In this study, we documented for the first time this intriguing pattern of behaviour in the rock sparrow and experimentally tested its potential role as a sexually selected or status signal by means of a feather supplementation experiment carried out in two distinct populations from Italy and Spain. We found that females were responsible for feather carrying, laid larger clutches and provisioned their young at a lower rate in those nests with experimentally added feathers. Decorated nests sustained fewer intrusions by floater individuals and were defended with greater intensity by both parents than control nests, which supports the role of nest ornamentation as a status signal to conspecifics. Presence of experimental feathers did not significantly increase the frequency with which males provisioned their young but males tended to desert their brood less often and spent more time guarding the brood in experimental nests, indicating that feather presence may also play a role in an intersexual context. Overall, our results allow us to exclude the thermoregulation hypothesis as a likely explanation for the occurrence of these decorations and provide partial evidence for the idea that feather carrying conveys information to the partner and potential competitors. Our study thus supports the notion that nonbodily traits serving a direct (naturally selected) function can also evolve a signalling component.

Mating strategies, parental investment and mutual ornamentation in Iberian Rock Sparrows (Petronia petronia)

Relatively few bird species show complex social mating systems whose preponderance in a population is likely to affect the patterns of parental care observed there. In turn, parental investment is likely to be related to the expression of certain ornaments, which may reveal information on the bearer’s individual quality. Here we address both issues in a species characterised by several forms of parental care (both biparental and uniparental care) and in which both sexes possess a yellow breast patch, the rock sparrow (Petronia petronia). In our population, males contributed more to the care of the young in comparison with other populations. Social monogamy was the most frequent mating pattern and the percentage of cases of female (or male) brood desertion was lower with respect to that reported in previous studies, suggesting a flexible behaviour of this species to deal with different social environments. Birds did not pair assortatively with respect to the size of the yellow breast patch and we found no significant relationship between this trait and the frequency with which parents provisioned their chicks. However, we observed a positive relationship between male yellow patch size and nestling tarsus length, which suggests that more ornamented males are better parents. Males, but not females, differentially allocated parental investment in response to female ornamentation, although the benefits that males may gain from choosing more attractive females remain unidentified. Our results on paternal care investment along with previous studies on this species, reinforcing the view that the rock sparrow constitutes a good model to study sexual conflict over parental care under different social environments.

Rock sparrow song reflects male age and reproductive success.

The evolution of mating signals is closely linked to sexual selection. Acoustic ornaments are often used as secondary sexual traits that signal the quality of the signaller. Here we show that song performance reflects age and reproductive success in the rock sparrow (Petronia petronia). In an Alpine population in south-east France, we recorded the songs of males and assessed their genetic breeding success by microsatellite analysis. In addition to temporal and spectral song features, we also analysed for the first time whether the sound pressure level of bird song reflects reproductive success. Males with higher breeding success sang at a lower rate and with a higher maximum frequency. We found also that older males gained more extra-pair young and had a higher overall breeding success, although they also differed almost significantly by having a higher loss of paternity in their own nests. Older males could be distinguished from yearlings by singing at lower rate and higher amplitudes. Our findings suggest that song rate may be used as a signal of age and together with song pitch as a signal of reproductive success in this species. Alternatively, younger and less successful males might try to compensate their inferior status by increased song rates and lower pitch. Independent of age and quality, high-amplitude songs correlated with paternity loss in the own nest, suggesting that in this species song amplitude is not an indicator of male quality but high-intensity songs may be rather a response to unfaithful social mates.

Breeding Biology of Rock SparrowsPetronia petroniain the Tibetan Plateau, with Special Reference to Life History Variation Across Altitudes

Abstract. The Rock Sparrow Petronia petronia has been well studied with respect to reproductive biology at low-altitude in Europe, but not at high altitudes. This study presents the information on a Rock Sparrow population breeding in an alpine meadow at 3400 m altitude and compares the life history traits with their lower-altitude counterparts studied in Europe. The birds are resident all year round in this area. Nests of Rock Sparrows tended to cluster and were mainly located in abandoned burrows of the Ground Tit Parus humilis. Fairs were monogamous and territorial behaviors were absent, which differed from European populations, where Rock Sparrows show a series of mating systems and display strong territoriality around the nest site. Egg-laying took place between late May and late June, with every pair making a single nesting attempt. Clutch size averaged 5.1 ± 0.9, incubation undertaken by female only lasted 12.7 ± 0.8 days, and young in the nest were fed by both parents for 19.9 ± 0.7 days. Breeding...

Leaders Are More Attractive: Birds with Bigger Yellow Breast Patches Are Followed by More Group-Mates in Foraging Groups

Social network theory provides a perfect tool to better understand the population-level consequences of how individuals interact and make their decisions; however, this approach is generally overlooked among evolutionary biologists interested in social relationships. Here, we used social network analysis to examine the patterns of leader-follower interactions in relation to individual characteristics in foraging groups of free-living rock sparrows (Petronia petronia). We found that yellow feather ornamentation, a carotenoid-based trait, was the best predictor of leadership: birds with bigger ornaments exerted greater influence in the foraging groups and were followed by more group-mates than less elaborate individuals. An individual's tendency for eliciting followings was not influenced by sex, condition or the level of parental investment. None of the above individual characteristics had significant effect on the tendency of individuals to follow others. Our results indicate that a sexually selected trait can also play a significant role in group coordination and social organization of a species.

White tail markings are an indicator of quality and affect mate preference in rock sparrows

In birds, colourful and elaborate feathers are important traits in mate choice. Distinct tail white patches are present in many species of birds, but they remain little studied. Tail markings may indeed have a signal function because in many species males spread the tail offering a good view of these markings to females during courtship behaviour. Here, we investigated whether white tail spots in male rock sparrow, Petronia petronia, play a role in mate choice. In a free-living population of rock sparrows, we found a reduction in white tail spots size as the breeding season progressed due to abrasion, which was expected if tail spots act as a reliable quality indicator (i.e. a handicap). The same reduction was found under captive conditions, and males in worse condition (individuals that lost more weight) abraded a bigger part of white. This suggests that white tail markings are an indicator of male quality. In captivity, we measured female preference for males differing in white patch size in a mate choice experiment. The experimental reduction of the size of the males’ white spots resulted in a lower sexual interest by females. During courtship display, male rock sparrow shows a yellow breast patch (a carotenoid-based, sexually selected ornament) together with the white spots in the tail. The sizes of these two traits are positively correlated, but only the abraded white area in the tail correlates with a surrogate of individual quality (lost of weight). In conclusion, we can assert that the size of the white spots is preferred by female rock sparrows and it is a part of a multiple signal system.

Sex‐dependent response of primary moult to simulated time constraints in the rock sparrow Petronia petronia

There is growing evidence that moult speed affects plumage quality. In many bird species, males and females differ in terms of breeding effort, survival expectation and the relationship between fitness and plumage quality. Consequently, differences in moult strategies between the sexes can be expected. The aim of this study was to assess whether, under simulated time constraints and with no parental investment in the previous breeding season, males and females differed in: a) timing and duration of primary moult, b) growth rates of individual primary feathers, and c) number of concurrently growing feathers. We investigated the effect of time constraints generated by a treatment consisting of two decreasing photoperiods (slow changing photoperiod, SCP=2 min day−1 and fast changing photoperiod, FCP=8 min day−1) on the primary post‐nuptial moult of captive rock sparrows Petronia petronia. Females started to moult on average 14 and 15 days later than males in both experimental groups. Primary moult duration was 10 (FCP) and 24 (SCP) days longer in males than in females, and, within sex, 34 (females) and 48 (males) days longer in SCP birds than in FCP ones. Females renewed a larger number of primaries simultaneously (5.7% in FCP and 12.8% in SCP) and had a higher total daily feather mass grown (9.9% in FCP and 22.4% in SCP), even though daily growth rates of individual primaries did not differ between sexes. As a result, males and females completed their primary moult at the same time within treatment. The observed differences in timing, duration and energy allocation for primary moult between the sexes probably have a genetic basis, as birds did not engage in reproduction during the preceding breeding season.

Female ornamentation, parental quality, and competitive ability in the rock sparrow

The evolution of female ornaments is poorly understood. Recent evidence suggests not only that female ornaments may be genetic correlates of selection on males but may also have evolved through male mate choice and/or through female–female aggressive interactions. In the rock sparrow, Petronia petronia, both sexes have a carotenoid-based yellow patch that is sexually selected by both sexes. The benefits that male may gain from choosing an attractive female remain unidentified. Both parents participate in caring for the young, so there should be mutual mate choice because males and females should both benefit from choosing a good parent (good parent hypothesis; GPH). Moreover, it has already been demonstrated that the yellow patch in males is also a badge of status (armament). Therefore, the yellow patch could also serve as both ornament and armament in females (dual utility hypothesis; DUH). We investigated the hypothesis that male and female yellow patch size signals parental quality in the field. We tested by an experiment in captivity the signal function of the yellow patch in female–female aggressive interactions for access to food. Yellow patch size correlated with paternal, but not maternal, feeding rates. Thus, this study supports the hypothesis that yellow patch dimension signals male parental quality, but there is no evidence for the GPH to explain female ornamentation. In the experiment females with relatively large yellow patches had earlier access to food than those with small patches. These results seem to suggest that a sexually selected carotenoid-feather signal may be used in female–female competition, in agreement with the DUH. Males may benefit from choosing well ornamented females because these may be superior competitors.

Nestlings’ carotenoid feather ornament affects parental allocation strategy and reduces maternal survival

In some birds, feather ornaments are expressed in nestlings well before sexual maturation, possibly in response to parental favouritism towards high-quality offspring. In species with synchronous hatching, in which nestling ornaments may vary more among than within broods, parents may use this information to adjust their parental allocation to the current brood accordingly. We tested this hypothesis in the rock sparrow, in which a sexually selected yellow feather ornament is also expressed in nestlings. We experimentally enlarged nestlings' breast patch in a group of broods and sham-manipulated another group of control broods. Nestlings with enlarged ornament were fed more frequently and defended more actively from a dummy predator than their control counterparts. Mothers from the enlarged group were more likely to lay a second clutch and showed a reduced survival to the next breeding season. These results provide one of the first evidences of differential parental allocation among different broods based directly on nestlings' ornamentation, and the first, to our knowledge, to show a reduction in maternal survival.