1 Mom, don´t let me die: a possible explanation for hatching

2 failure related to laying order in the Pied flycatcher

3
4 Manuel Fuertes-Recuero and Alejandro Cantarero*
5
6 Department of Physiology, Veterinary School, Complutense University of Madrid,
7 Avenida Puerta de Hierro s/n, 28040 Madrid, Spain.
*
9 Corresponding author: alex.cantarero@hotmail.com
10 AC ORCID: 0000-0002-5816-701X; MFR: 0000-0003-0736-8791
11 Abstract
12 Introduction
13 Hatching success is a key parameter that has been associated with fitness in birds
14 (Cooke et al., 1985; Koenig, 1982). Hatching failure, arising from either infertility or
15 embryo mortality, constitutes a significant factor contributing to diminished avian
16 reproductive outcomes (Briskie and Mackintosh, 2004; Jamieson and Ryan, 2000;
17 Spottiswoode and Møller, 2004). Several causes have been related to hatching failure:
18 environmental phenomes such as rainy or cold weather during the incubation and
19 hatching stage (Järvinen and Väisänen, 1984; Stoleson, 1999; Webb, 1987), poor
20 condition of incubating adults which could lead to abandonment of the nest, egg size or
21 quality (Magrath, 1992; Parsons, 1970), clutch size (Blackburn, 1991), parental age or
22 quality (Bolton, 1991; Meathrel et al., 1993; Reid and Boersma, 1990) or laying order
23 (Cabezas-Díaz and Virgós, 2007; Potti and Merino, 1996; Robertson and Cooke, 1993;
24 Williams et al., 1993). While embryo mortality is the primary cause of hatching failure
25 in birds (Birkhead et al., 2008; Hemmings and Birkhead, 2016), the specific reasons
26 behind it remain largely unclarified. Embryo mortality can stem from a combination of
27 internal and external factors, including environmental factors and parental behaviour,
28 factors that may exhibit variations throughout the different stages of development.
29 Consequently, discerning the precise stage at which an embryo dies during development
30 may offer insights into the underlying cause of mortality (Hemmings and Birkhead,
31 2016).

32 The position within the laying order is a factor that has been associated with
33 hatching failure (Robertson and Cooke, 1993; Williams et al., 1993), with higher rates
34 of embryo mortality in the first and last-laid eggs (Potti and Merino, 1996; Williams et
35 al., 1993). Furthermore, studies with artificially incubated eggs have described that
36 embryo death occurs more frequently in last-laid eggs, while infertility was the main
37 cause of hatching failure in first-laid eggs (Cabezas-Díaz and Virgós, 2007).

38 Egg size has been positively associated nestlings growth (Potti and Merino,
39 1996), being an important factor that explain hatching failure in birds. This relationship
40 is often influenced by the quality of the parents and/or the order in which the eggs are
41 laid (Clifford and Anderson, 2002; Meathrel et al., 1993; Murton et al., 1974; Potti and
42 Merino, 1996; Reid and Boersma, 1990; Williams et al., 1993). Almost all bird species
43 take care of their young soon after hatching, neglecting last-laid eggs because of
44 foraging constraints. Hatching asynchrony creates harsh conditions for last-laid eggs
45 and often late-hatched chicks tend to exhibit lower survival and growth rates (Węgrzyn
46 et al., 2023). The timing of the initiation of incubation is the primary determinant of
47 hatching asynchrony. Although synchronized broods generally result in higher fledging
48 success, most altricial birds hatch asynchronously.

49 Many birds such as the pied flycatcher Ficedula hypoleuca are known to begin
50 incubating before clutch completion, giving rise to an asynchronous hatch, especially of
51 the last chicks (Enemar and Arheimer, 1989; Forbes et al., 1997; Lord et al., 2011; Scott
52 Johnson et al., 2009). This strategy could lead to failure in the hatching of the last egg or
53 to competitive disadvantages if it is hatched (Clotfelter et al., 2000; Jeon, 2008;
54 Ostreiher, 2001, 1997; Pettifor et al., 2001; Soler et al., 2020; Węgrzyn, 2012; Wiebe,
55 1996). Although several hypotheses have been proposed to explain this phenomenon
56 (reviewed in (Węgrzyn et al., 2023)), one of the most plausible is the Brood Reduction
57 Hypothesis (Lack, 1968), which argues that asynchronous hatching is advantageous in
58 unpredictable environments (bad years), producing a reduced brood at the expense of
59 the death of the last chick. However, some studies have shown that asynchronous
60 hatching and the resulting size hierarchy were detrimental to nestling growth of
61 nestlings in both good and bad years in the collared flycatcher Ficedula albicollis
62 (Szöllősi et al., 2007). The failure to hatch the last-egg laid has generally been attributed
63 to the end of the incubation period, , associated with the terminal egg neglect hypothesis
64 (Cabezas-Díaz and Virgós, 2007).

65 Here we analyse hatching failure with laying order in the pied flycatcher,
66 discriminating between hatching failures due to infertility and those due to embryo
67 death and its development stage. To date, the studies that have addressed this topic have
68 not taken into the development stage of the embryo, a fact that may be key to
69 understanding the reproductive strategies of some birds.
 70 Material and methods

71 Field methods
72 The study was conducted during the spring of 2023 in a population of pied flycatchers
73 breeding in nest boxes in a montane forest of Pyrenean oak (Quercus pyrenaica) in
74 Valsaín, central Spain (40°54'N, 04°01'W) where long-term studies on cavity-nesting
75 birds have been carried out since 1991. Of 465 nest-boxes, 112 were occupied by pied
76 flycatchers and the rest by other species, mainly blue tits Cyanistes caeruleus, rock
77 sparrows Petronia petronia, nuthatches Sitta europaea and great tits Parus major.
78 From the beginning of April, nest-boxes were routinely checked to monitor flycatcher
79 reproduction and determine signs of nest building. Comprehensive daily checking was
80 done from the moment the females completed the construction of the nest to detect the
81 laying of the first egg. Once the onset of egg laying was established, every new egg was
82 distinctively marked according to the laying order with an indelible black pencil that did
83 not penetrate the eggshell or damage the embryo.
84 On the fifth day of incubation females were captured in the nest-box during
85 daytime without traps as they are not easily frightened away from the nest at this stage.
86 They were identified by their rings or ringed if necessary and weighed to the nearest
87 0.01 g with a digital balance. We took a digital photograph of the entire clutch by
88 placing the eggs in order into a custom box that allowed us to take pictures reducing the
89 risk of breaking them (Figure X). Digital pictures were analysed with Adobe PhotoShop
90 CS4 version 11.0 (Adobe Systems, San Jose, CA) to estimate the size, the maximum
91 length, and the breadth for each egg to the nearest 0.1 mm. Mean egg size, length and
92 breadth were calculated for each clutch. Moreover, mean egg volume was calculated
93 from egg length and breadth using the formula of Hoyt (1979).
94 On day 3 (hatching day = day 1), we recorded the number of nestlings and
95 calculated hatching success as the proportion of eggs that hatched. Unhatched eggs were
96 collected in sterile tubes and frozen for subsequent laboratory analysis. All adult birds
97 were captured in their nest boxes while feeding 7 days old nestlings. The trap was
98 placed for a maximum of 1h and removed once both adults were captured. All birds
99 were identified by their rings or supplied with individually numbered rings for
100 identification. The following traits associated were recorded: age estimated from ring
101 data, wing length measured with a ruler, body mass recorded with an electronic balance
102 and male forehead patch size (patch breadth × patch height) and dorsal blackness as
103 percentage of black plumage on the mantle. This trait has been associated with male
104 individual quality (Siitari and Huhta, 2002). Exact age of adults older than one year
105 were established if ringed as nestlings in the study area or classified according to Jenni
106 and Winkler (1994) and Svensson (1984) as yearlings or older from the colour of the
107 outer wing coverts. Once they have bred in our nest boxes, surviving adults return each
108 spring to attempt breeding. On day 13, we ringed nestlings and measured in the same
109 way as adults. Fledging success was calculated as the proportion of hatched chicks that
110 fledged.
111
112 Egg examination
113 Unhatched eggs collected were defrost and opened in the laboratory. Eggs content was
114 emptied into a Petri dish and examined under a magnifying glass. Eggs scored as
115 infertile only contained a yellow yolk, while eggs scored as inviable contained a
116 germinal disc or embryo which died before hatching. If this was observed, it was
117 cleaned in a saline solution and examined under a microscope to identify the
118 developmental stage at which death occurred, based on the stages of chick development
119 categorized by Hemmings and Birkhead (2016). Developmental stages were grouped as
120 early (germinal disc corresponding to 0 to 3 days of development), middle (small
121 embryo corresponding to 4 to 7 days of development) or late development stage (clearly
122 visible embryo which died before hatching).
123 For an overall analysis of hatching failure, we have grouped together failures
124 due to infertility and those due to death of the embryo. Thus, nests with hatching failure
125 were those where at least one egg failed to hatch.
126
127 Statistical analyses
128 All the analyses were performed using STATISTICA package. Hatching success
129 exhibited a skewed distribution of data values and attempts to transform the data did not
130 enhance its normal distribution fit. Consequently, hatching success was analyzed with
131 nonparametric tests (Mann – Whitney U tests) or as a categorical variable ("transformed
132 hatching success) by dividing nests into "unsuccessful" (at least one egg failed to hatch)
133 and "successful" (all eggs hatched).
134 Chi-squared (χ2) tests were performed to assess potential relationships between
135 hatching failure in relation to laying order.
136 We analyzed the effects of adults age and their interaction on different traits
137 (hatching success, clutch size and laying date) using generalized linear models. Some
138 potential predictors were included in the models. Backward deletion procedure was
139 applied to reduce the number of predictors in the final model. when the variance
140 explained did not significantly improve the model (α = 0.05). Finally, a Pearson
141 correlation was used for those variables that were normally distributed. In all cases
142 statistical significance was set at P < 0.05.
143 Results

144 Laying order and hatching failure

145 In our study, 8.97 % (56 of 624) of pied flycatcher eggs failed to hatch. From those 56
146 unhatched eggs, 23.21% (n=13) and 76,79% (n=43) were classified as infertile and as
147 inviable respectively. Of the inviable eggs, 37.3% (n=16) were classified as early
148 development stage, 20.93% (n=9) as middle stage and 41.86% (n=18) as late stage. We
149 detected that laying order significantly affected the viability of the eggs. Thus, first and
150 last positions in the laying sequence were more prone to fail (χ 26= 13.99, P = 0.029; Fig.
151 X). Neither infertile eggs nor early and mid-stage embryos were affected by laying
152 order (infertile: χ26= 4.61, P = 0.595; early development stage; χ 26= 5.30, P = 0.506;
153 middle development stage; χ26= 2.73, P = 0.842).

18
16
14
Number of eggs unhatched

12
10
8 Inviable eggs
Infertile eggs
6
4
2
0
1 2 3 4 5 6 7
Laying order
154
155
156 However, death of late development embryos depended on laying order (χ 26= 14.32, P =
157 0.027, Fig. X), especially, in the last positions of the laying sequency.
158
 18
16
14
Number of eggs unhatched

12
10
8 Rest of unhatched eggs
6 Late development stage

4
2
0
1 2 3 4 5 6 7
Laying order
159
160
161 Hatching success
162 Female age on her own did not affect hatching success (yearling: 85.63 ± 24.14 vs old:
163 88.04 ± 18.25; Mann-Whitney tests, z = -0.16, P = 0.87). However, taking into
164 consideration the age of the male, old females showed increased hatching success when
165 paired with yearling males (z = -1.42, P = 0.042; Fig. X). Transformed hatching success
166 was significantly affected by the interaction between the age of the male and the dorsal
167 blackness, with old males being more black and presenting less hatching success than
168 yearling (interaction F82 = 7.05, P = 0.009; Fig. X).
169

100.00
95.00 94.17
90.00 89.07
85.00 84.12
82.16
Hatching success (%)

80.00
75.00
70.00
65.00
60.00
55.00
50.00
Yearling Female vs Yearling Female vs Older Female vs Older Female vs
Yearling Male Older Male Yearling Male Older Male
170

91.50 96.00

93.75 94.00
91.00
92.00
Hatching success (%)

Dorsal blackness (%)

90.50
90.00

90.00 88.00
87.25
86.00
89.50
84.00
89.00
82.00

88.50 80.00
Yearling Male Older Male
171

172 Hatching success was not related to laying date, female condition or eggs measurements
173 (all P >0.05).

174
175
176
177
178
179
180
181 Eeggs volume and hatching success
182 The volume of the eggs was significant in relation to the laying order (interaction F547 =
183 21.84, P = <0.001; Fig. X).

184

2
1.95
1.9
1.85
Eggs Volume (mm3)

1.8
1.75
1.7
1.65
1.6
1.55
1.5
0 1 2 3 4 5 6 7
Laying order
185
186 Discussion
187 -Overall, our results suggest that laying order had a significant effect on egg viability,
188 showing that the first and last positions in the laying sequence were more predisposed to
189 failure. In addition, the death of late-stage embryos was dependent on laying order,
190 particularly in the last positions of the laying sequence.
191
192 -We described that 8.97 % of eggs fail to hatch, a proportion consistent with rates
193 documented in prior studies involving Pied Flycatcher and other bird species with
194 cavity nest (Di Giovanni et al., 2023; Morrow et al., 2002; Spottiswoode and Møller,
195 2004; Stewart and Westneat, 2013).
196

197 - The result shows a discernible pattern in the overall hatching failure, demonstrating a
198 concave-upward trajectory along the laying sequence. Higher failure rates were
199 observed in both the initial and final positions within the clutches, the majority of which
200 typically contained 5 or 6 eggs. This pattern suggests a distinct optimal hatchability at
201 intermediate positions within the clutch. These findings are in line with those reported
202 by Potti and Merino (1996), and Ylimaunu and Jarvinen (1987) in their studies of the
203 Pied Flycatcher which also observed reduced hatching or fledgling success associated
204 with both the first and last eggs in the laying sequence.
205 --The diminished hatchability of eggs laid early in the sequence may be attributed to
206 enhanced physiological effectiveness in egg production as the laying sequence
207 progresses (Leblanc, 1987). Alternatively, the inceased temporal exposure of early-laid
208 eggs to environmental factors, could also contribute to this phenomenon (Arnold et al.,
209 1987; Veiga and Viñuela, 1993; Veiga, 1992).
210 --The lower hatchability of eggs laid in the last position in the sequence may be due to
211 the asynchronous hatching of passerines such as the Pied Flycatcher (Enemar and
212 Arheimer, 1989; Forbes et al., 1997; Lord et al., 2011; Scott Johnson et al., 2009;
213 Slagsvold, 1985). Several hypotheses have been put forward to explain this strategy,
214 which could lead to the failure of the last eggs to hatch. The predominant hypothesis
215 that may elucidate the elevated failure rate observed in the last eggs is the "Terminal
216 Egg Neglect" (Cabezas-Díaz and Virgós, 2007). According to this hypothesis, females
217 may exhibit a tendency to neglect the incubation of the final eggs, potentially influenced
218 by the feeding demands of the recently hatched nestlings. Alternatively, the embryonic
219 development and hatching of last eggs might be impeded by the presence of already
220 hatched siblings, resulting in increased mortality of the last-laid eggs. An alternative
221 hypothesis could be the “Brood Reduction Hypothesis” (Pijanowski, 1992). This
222 hypothesis posits that asynchronous hatching is advantageous in unpredictable
223 environments, leading to a decreased brood size at the cost of mortality in the later-eggs
224 or hatching individuals.
225

226

227

228
229

230 - There is a significant increase of mortality of late development embryos in comparison

231 with the rest of unhatched eggs, related with the later positions of the laying sequence,
232 in contrast with Potti and Merino (1996) study were there was not statistically
233 significant. A possible plausible explanation of this event could be related as well with
234 the hypothesis of terminal egg neglected (Slagsvold, 1985), increasing the mortality of
235 embryos in later positions especially of those in the late development close to hatching,
236 due to the asynchronous hatching characteristics of species such as Pied Flycatcher.
237 Although the increase of infertile eggs in the first and last eggs of the clutch could be
238 explained by the increase of phylogenetic efficacy with laying order (Leblanc, 1987)
239 and the decrease of sperm efficacy, sperm depletion or ova viability (Graves, 1992),
240 respectively, no relationship with laying order was found in comparison with the rest of
241 the unattached eggs. Furthermore, no difference was found between early and mid-stage
242 embryos compared to the rest of the unattached eggs in relation to laying order. This
243 may be due to a different distribution of egg deaths in the clutch, with different causes
244 leading to infertility or embryo death.
245

246 - In our study, we found no relationship between egg size and hatching success, in
247 contrast to previous study in Tree Sparrows (Pinowska et al., 2002) or even in the Pied
248 Flycatcher observed by Potti and Merino (1996). The different results between our
249 study and the one carried out by Potti and Merino (1996) may be due to differences
250 between the two studies in the way the eggs were measured (dobe PhotoShop versus
251 dial calipers), the different study areas with distinct Pied Flycatcher populations, the fact
252 that the study was conducted in a population with more than 30 years of monitoring
253 versus a newly established population, or the fact that our study used data from a single
254 season versus using data from several seasons in which repeat females could be
255 included.
256 -We have shown that young males have a higher hatching success rate than older males,
257 which tend to have more dorsal blackness. Although the blackness has been described as
258 an individual quality factor (Siitari and Huhta, 2002), and as a factor that makes males
259 more likely to have extra pair mating (Canal et al., 2011), the fact that young males have
260 more effective semen (Pizzari et al., 2008; Vega-Trejo et al., 2019) may be one of the
261 reasons for this finding. In addition, older males are more prone to polygyny (Canal et
262 al., 2011), which would result in them not being able to fully take care of feeding the
263 both females during the incubation time, leading to a reducing hatching success of those
264 clutches (Garamszegi et al., 2004).
265 When comparing female and male age in relation to hatching success, our results show
266 that older females mating with younger males have a higher hatching success rate. This
267 could be explained by the fact that adult females, with previous experience in nest
268 establishment, incubation and chick care, have a higher hatching success rate and
269 fledging success than inexperienced young females (Garamszegi et al., 2004; Lv et al.,
270 2016; Woodard and Murphy, 1999).
271
272
273
274
275 - While hatching failure has been extensively investigated, there has been limited focus
276 on the precise causes of embryo mortality, distinct from issues of infertility.
277 Additionally, scant attention has been given to identifying the critical periods of
278 vulnerability during embryo development in wild birds. The ability to identify the
279 specific stages at which embryos perish during incubation would allow embryo
280 mortality to be correlated with local environmental or other factors. This study could in
281 turn help to identify potential factors contributing to embryo mortality.
282
283 In conclusion, the position of the eggs in the laying sequence is related to hatching
284 failure, with the first and last positions being more susceptible. Further studies on
285 embryonic deaths should be carried out to investigate the possible causes.
286
287
288 References
289 Arnold, T.W., Rohwer, F.C., Armstrong, T., 1987. Egg Viability, Nest Predation, and the Adaptive
290 Significance of Clutch Size in Prairie Ducks. The American Naturalist 130, 643–653.
291 https://doi.org/10.1086/284736
292 Birkhead, T.R., Hall, J., Schut, E., Hemmings, N., 2008. Unhatched eggs: methods for
293 discriminating between infertility and early embryo mortality. Ibis 150, 508–517.
294 https://doi.org/10.1111/j.1474-919X.2008.00813.x
295 Blackburn, T.M., 1991. An Interspecific Relationship Between Egg Size and Clutch Size in Birds.
296 Bolton, M., 1991. Determinants of Chick Survival in the Lesser Black-Backed Gull: Relative
297 Contributions of Egg Size and Parental Quality. Journal of Animal Ecology 60, 949–960.
298 https://doi.org/10.2307/5424
299 Briskie, J.V., Mackintosh, M., 2004. Hatching failure increases with severity of population
300 bottlenecks in birds. Proc Natl Acad Sci U S A 101, 558–561.
301 https://doi.org/10.1073/pnas.0305103101
302 Cabezas-Díaz, S., Virgós, E., 2007. Adaptive and Non-Adaptive Explanations for Hatching Failure
303 in Eggs of the Red-Legged Partridge Alectoris rufa. arde 95, 55–63.
304 https://doi.org/10.5253/078.095.0106
305 Canal, D., Potti, J., Dávila, J.A., 2011. Male phenotype predicts extra-pair paternity in pied
306 flycatchers. Behaviour 148, 691–712.
307 Clifford, L.D., Anderson, D.J., 2002. Clutch size variation in the Nazca booby: a test of the egg
308 quality hypothesis. Behavioral Ecology 13, 274–279.
309 https://doi.org/10.1093/beheco/13.2.274
310 Clotfelter, E.D., Whittingham, L.A., Dunn, P.O., 2000. Laying order, hatching asynchrony and
311 nestling body mass in Tree Swallows Tachycineta bicolor. Journal of Avian Biology 31,
312 329–334. https://doi.org/10.1034/j.1600-048X.2000.310308.x
313 Cooke, F., Findlay, C.S., Rockwell, R.F., Smith, J.A., 1985. Life History Studies of the Lesser Snow
314 Goose (anser Caerulescens Caerulescens). Iii. the Selective Value of Plumage
315 Polymorphism: Net Fecundity. Evolution 39, 165–177. https://doi.org/10.1111/j.1558-
316 5646.1985.tb04089.x
317 Di Giovanni, A.J., Miller, M.J., Jones, T.M., Benson, T.J., Ward, M.P., 2023. Hatching failure is
318 greater in altricial bird species with cavity nests and large clutches. Ornithology 140,
319 ukac048. https://doi.org/10.1093/ornithology/ukac048
320 Enemar, A., Arheimer, O., 1989. Developmental asynchrony and onset of incubation among
321 passerine birds in a mountain birch forest of Swedish Lapland.
322 Forbes, S., Thornton, S., Glassey, B., Forbes, M., Buckley, N.J., 1997. Why parent birds play
323 favourites. Nature 390, 351–352. https://doi.org/10.1038/37025
324 Garamszegi, L., Török, J., Michl, G., Moller, A., 2004. Female survival, lifetime reproductive
325 success and mating status in a passerine bird. Oecologia 138, 48–56.
326 https://doi.org/10.1007/s00442-003-1408-z
327 Graves, J., 1992. Sperm competition in birds. Evolutionary causes and consequences. Animal
328 Behaviour 44, 389–390. https://doi.org/10.1016/0003-3472(92)90047-D
329 Hemmings, N., Birkhead, T.R., 2016. Consistency of passerine embryo development and the use
330 of embryonic staging in studies of hatching failure. Ibis 158, 43–50.
331 https://doi.org/10.1111/ibi.12336
332 Jamieson, I.G., Ryan, C.J., 2000. Increased egg infertility associated with translocating inbred
333 takahe (Porphyrio hochstetteri) to island refuges in New Zealand. Biological
334 Conservation 94, 107–114. https://doi.org/10.1016/S0006-3207(99)00158-5
335 Järvinen, A., Väisänen, R.A., 1984. Reproduction of Pied Flycatchers (Ficedula hypoleuca) in
336 Good and Bad Breeding Seasons in a Northern Marginal Area. The Auk 101, 439–450.
337 https://doi.org/10.1093/auk/101.3.439
338 Jeon, J., 2008. Evolution of parental favoritism among different-aged offspring. Behavioral
339 Ecology 19, 344–352. https://doi.org/10.1093/beheco/arm136
340 Koenig, W.D., 1982. Ecological and Social Factors Affecting Hatchability of Eggs. The Auk 99,
341 526–536. https://doi.org/10.1093/auk/99.3.526
342 Lack, D., 1968. Ecological adaptations for breeding in birds. Methuen, London.
343 Leblanc, Y., 1987. Intraclutch variation in egg size of Canada geese. Can. J. Zool. 65, 3044–3047.
344 https://doi.org/10.1139/z87-461
345 Lord, A.M., McCleery, R., Cresswell, W., 2011. Incubation prior to clutch completion accelerates
346 embryonic development and so hatch date for eggs laid earlier in a clutch in the great
347 tit Parus major. Journal of Avian Biology 42, 187–191. https://doi.org/10.1111/j.1600-
348 048X.2010.05256.x
349 Lv, L., Komdeur, J., Li, J., Scheiber, I.B.R., Zhang, Z., 2016. Breeding experience, but not mate
350 retention, determines the breeding performance in a passerine bird. Behavioral
351 Ecology 27, 1255–1262. https://doi.org/10.1093/beheco/arw046
352 Magrath, R.D., 1992. The effect of egg mass on the growth and survival of blackbirds: a field
353 experiment. Journal of Zoology 227, 639–654. https://doi.org/10.1111/j.1469-
354 7998.1992.tb04420.x
355 Meathrel, C.E., Bradley, J.S., Wooller, R.D., Skira, I.J., 1993. The effect of parental condition on
356 egg-size and reproductive success in short-tailed shearwaters Puffinus tenuirostris.
357 Oecologia 93, 162–164. https://doi.org/10.1007/BF00317665
358 Morrow, E.H., Arnqvist, G., Pitcher, T.E., 2002. The evolution of infertility: does hatching rate in
359 birds coevolve with female polyandry? Journal of Evolutionary Biology 15, 702–709.
360 https://doi.org/10.1046/j.1420-9101.2002.00445.x
361 Murton, R.K., Westwood, N.J., Isaacson, A.J., 1974. Factors Affecting Egg-Weight, Body-Weight
362 and Moult of the Woodpigeon Columba Palumbus. Ibis 116, 52–73.
363 https://doi.org/10.1111/j.1474-919X.1974.tb00223.x
364 Ostreiher, R., 2001. The importance of nestling location for obtaining food in open cup-nests.
365 Behav Ecol Sociobiol 49, 340–347. https://doi.org/10.1007/s002650000308
366 Ostreiher, R., 1997. Food division in the Arabian babbler nest: adult choice or nestling
367 competition? Behavioral Ecology 8, 233–238. https://doi.org/10.1093/beheco/8.2.233
368 Parsons, J., 1970. Relationship between Egg Size and Post-hatching Chick Mortality in the
369 Herring Gull (Larus argentatus). Nature 228, 1221–1222.
370 https://doi.org/10.1038/2281221a0
371 Pettifor, R.A., Perrins, C.M., McCleery, R.H., 2001. The individual optimization of fitness:
372 variation in reproductive output, including clutch size, mean nestling mass and
373 offspring recruitment, in manipulated broods of great tits Parus major. Journal of
374 Animal Ecology 70, 62–79. https://doi.org/10.1111/j.1365-2656.2001.00465.x
375 Pijanowski, B.C., 1992. A Revision of Lack’s Brood Reduction Hypothesis. The American
376 Naturalist 139, 1270–1292.
377 Pinowska, B., Barkowska, M., Pinowski, J., Hahm, K.-H., Lebedeva, N., 2002. The Effect of Egg
378 Size on Hatching Rate in the Tree Sparrow Passer montanus (Study in Central Poland).
379 aorn 37, 7–14. https://doi.org/10.3161/068.037.0102
380 Pizzari, T., Dean, R., Pacey, A., Moore, H., Bonsall, M.B., 2008. The evolutionary ecology of pre-
381 and post-meiotic sperm senescence. Trends Ecol Evol 23, 131–140.
382 https://doi.org/10.1016/j.tree.2007.12.003
383 Potti, J., Merino, S., 1996. Causes of Hatching Failure in the Pied Flycatcher. The Condor 98,
384 328–336. https://doi.org/10.2307/1369151
385 Reid, W.V., Boersma, P.D., 1990. PARENTAL QUALITY AND SELECTION ON EGG SIZE IN THE
386 MAGELLANIC PENGUIN. Evolution 44, 1780–1786. https://doi.org/10.1111/j.1558-
387 5646.1990.tb05248.x
388 Robertson, G.J., Cooke, F., 1993. Intraclutch egg-size variation and hatching success in the
389 common eider. Can. J. Zool. 71, 544–549. https://doi.org/10.1139/z93-075
390 Scott Johnson, L., Brubaker, J.L., Johnson, B.G.P., Masters, B.S., 2009. Evidence for a maternal
391 effect benefiting extra-pair offspring in a songbird, the house wren Troglodytes aedon.
392 Journal of Avian Biology 40, 248–253. https://doi.org/10.1111/j.1600-
393 048X.2009.04777.x
394 Siitari, H., Huhta, E., 2002. Individual color variation and male quality in pied flycatchers
395 (Ficedula hypoleuca): a role of ultraviolet reflectance. Behavioral Ecology 13, 737–741.
396 https://doi.org/10.1093/beheco/13.6.737
397 Slagsvold, T., 1985. Asynchronous Hatching in Passerine Birds: Influence of Hatching Failure and
398 Brood Reduction. Ornis Scandinavica (Scandinavian Journal of Ornithology) 16, 81–87.
399 https://doi.org/10.2307/3676471
400 Soler, M., Ruiz-Raya, F., Sánchez-Pérez, L., Ibáñez-Álamo, J.D., 2020. Parental feeding
401 preferences rather than sibling competition determine the death of smaller nestlings in
402 asynchronous broods (preprint). Animal Behavior and Cognition.
403 https://doi.org/10.1101/2020.12.01.404590
404 Spottiswoode, C., Møller, A.P., 2004. Genetic similarity and hatching success in birds. Proc Biol
405 Sci 271, 267–272.
406 Stewart, I.R.K., Westneat, D.F., 2013. Patterns of hatching failure in the house sparrow Passer
407 domesticus. Journal of Avian Biology 44, 069–079. https://doi.org/10.1111/j.1600-
408 048X.2012.05795.x
409 Stoleson, S., 1999. The importance of the early onset of incubation for the maintenance of egg
410 viability. pp. 600–613.
411 Szöllősi, E., Rosivall, B., Török, J., 2007. Is hatching asynchrony beneficial for the brood?
412 Behavioral Ecology 18, 420–426. https://doi.org/10.1093/beheco/arl100
413 Vega-Trejo, R., Fox, R.J., Iglesias-Carrasco, M., Head, M.L., Jennions, M.D., 2019. The effects of
414 male age, sperm age and mating history on ejaculate senescence. Functional Ecology
415 33, 1267–1279. https://doi.org/10.1111/1365-2435.13305
416 Veiga, J., Viñuela, J., 1993. Hatching Asynchrony and Hatching Success in the House Sparrow:
417 Evidence for the Egg Viability Hypothesis. Ornis Scandinavica 24, 237–242.
418 https://doi.org/10.2307/3676739
419 Veiga, J.P., 1992. Hatching Asynchrony in the House Sparrow: A Test of the Egg-Viability
420 Hypothesis. The American Naturalist 139, 669–675. https://doi.org/10.1086/285351
421 Webb, D.R., 1987. Thermal Tolerance of Avian Embryos: A Review. The Condor 89, 874–898.
422 https://doi.org/10.2307/1368537
423 Węgrzyn, E., 2012. In the Blackcap Sylvia atricapilla Last-Hatched Nestlings Can Catch up with
424 Older Siblings. arde 100, 179–186. https://doi.org/10.5253/078.100.0209
425 Węgrzyn, E., Węgrzyn, W., Leniowski, K., 2023. Hatching asynchrony as a parental reproductive
426 strategy in birds: a review of causes and consequences. J Ornithol 164, 477–497.
427 https://doi.org/10.1007/s10336-023-02066-8
428 Wiebe, K.L., 1996. The Insurance-Egg Hypothesis and Extra Reproductive Value of Last-Laid
429 Eggs in Clutches of American Kestrels. The Auk 113, 258–261.
430 https://doi.org/10.2307/4088961
431 Williams, T.D., Lank, D.B., Cooke, F., 1993. Is Intraclutch Egg-Size Variation Adaptive in the
432 Lesser Snow Goose? Oikos 67, 250–256. https://doi.org/10.2307/3545469
433 Woodard, J.D., Murphy, M.T., 1999. Sex roles, parental experience and reproductive success of
434 eastern kingbirds,Tyrannus tyrannus. Animal Behaviour 57, 105–115.
435 https://doi.org/10.1006/anbe.1998.0998
436