Female Birds Research Articles

Sexual Dimorphism in A Cross River Ecotype Local Chicken – Rose Comb (<em>Gallus gallus domesticus</em>)

Sexual dimorphism in the local chicken (Gallus gallus domesticus)-Rose Comb- in Cross River was investigated. Sixty (60) male and female Gallus gallus domesticus chickens were evaluated for the effect of sex on body weight (BW), body length (BL), body girth (BG), thigh length(TL), shank length (SL) and keel length (KL) which were measured fortnightly for twelve (12) weeks. Sexual Size Dimorphism Index (SSDI) and Sexual Dimorphism Index (SDI) were used to confirm relative contribution of each trait to total dimorphism. The values obtained for SSDI and SDI respectively were 1.61 and 61% for BW; 1.26 and 26.52% for TL; 1.23 and 23.81% for SL; 1.22 and 21.50% for BL; 1.18 and 18.20% for BG; 1.17 and 17.94% for KL. To identify the best trait for predicting sex, stepwise discriminate analysis was employed and the Wilk’s Lamda values obtained were BW(0.471); TL(0.452); SL(0.401); BL(0.283); BG(0.272) and KL(0.271). Results showed that there were significant (p<0.05) differences in all measured parameters at week 6 with mean BW, BL, BG, SL, KL and TL of 300.01 and 253.24; 22 and 18.62; 5.20 and 3.98; 4.60 and 3.50; 5.50 and 3.90; 13.10 and 11.20 for both male and female birds respectively. These results indicate that sexual dimorphism is exhibited in all morphometric traits in favor of the male with BW as the highest dimorphic trait.

Gene explains bird color sex differences

Female birds often sport less colorful plumage than males of the same species. Some birds evolved this sexual dichromatism as an adaptation related to mating and survival, but scientists still don’...

Anti-microbial activity of whole blood and plasma collected from Anna's Hummingbirds (Calypte anna) against three different microbes.

Hummingbirds are essential pollinators in many ecosystems, making their conservation critical. As is the case with many species, hummingbirds are now facing a variety of challenges resulting from anthropogenic changes. As populations shift and species interactions change, disease is likely to pose a significant threat. There is a basic understanding of which pathogens currently affect a variety of hummingbird species, however there is a paucity of information about their immune systems capacity to kill pathogens and what specific factors may affect immunity. The objective of this study was to gain a basic understanding of the effect of age, sex, and molt on the constitutive innate immunity of hummingbirds. An in vitro assay was used to assess the microbiocidal capacity of the whole blood of Anna’s Hummingbirds (Calypte anna) against three different microbes: Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Candida albicans (C. albicans). The effect of age, sex and molt on anti-microbial capacity varied based on the microbe type. After-hatch-year birds tended to have better anti-microbial capacity compared to hatch-year birds. Male birds had higher anti-microbial activity than female birds, although this was not observed against C. albicans. Molting birds had a weaker antimicrobial activity against E. coli and S. aureus than birds that were not molting. These results represent an important first step towards defining the parameters of constitutive innate immunity of Anna’s Hummingbirds as well as providing important knowledge about factors that should be considered when evaluating the health of wild populations.

Trade-off between aerobic performance and egg production in migratory macaroni penguins

When successive stages of an organism's life-history overlap, conflicts and trade-offs can emerge due to competition among physiological pathways. For example, long periods of sustained locomotion in migrating birds are supported by the androgenic up-regulation of aerobic factors, such as new red blood cell production and hematocrit. However, towards the end of migration, many female birds begin up-regulating 17ß-estradiol (E2) to support vitellogenesis and egg production, but E2 secretion is known to have suppressive effects on red blood cell production (anti-erythropoiesis). We explored potential trade-offs between factors related to aerobic performance (hematocrit, reticulocyte index) and the expression of factors related to E2-mediated vitellogenesis (i.e. yolk precursor production) in female macaroni penguins (Eudyptes chrysolophus), a species in which the physiologies controlling egg production and migratory activity run simultaneously (e.g. females experience a migratory conflict). We collected blood samples from penguins immediately upon their return to the colony, prior to egg laying. Hematocrit was elevated when the penguins returned to the colony (50.05% ± 3.40 SD), which is similar to pre-laying values observed in other migratory bird species. Furthermore, mean reticulocyte levels were elevated (34.87% ± 2.34), which is the highest level yet recorded in birds. Similarly, both plasma vitellogenin and yolk-targeted very low density lipoprotein levels were upregulated (2.30 ± 0.06 μg Zn ml−1, and 9.70 ± 0.19 mmol l−1, respectively), indicating that penguins were reproductively active and producing eggs during migration and upon arrival on land. As predicted, a negative relationship between hematocrit and plasma vitellogenin was found, but we found no evidence to suggest that birds were experiencing reproductive anemia. Alternatively, we attribute the negative relationship to a hemodilution effect of yolk precursor secretion into circulation. It appears that female macaroni penguins are able to preserve hematocrit levels and new red blood cell production when migratory activity overlaps with reproductive processes.

Using a multistate occupancy approach to determine molecular diagnostic accuracy and factors affecting avian haemosporidian infections

The use of a sensitive and accurate parasite detection methodology is crucial in studies exploring prevalence of parasites in host populations or communities, and uncertainty in identifying parasite genera and/or lineages may limit the understanding of host-parasite interactions. Here, we used a multistate occupancy approach that accounts for imperfect detection to assess whether sex and breeding season influenced the prevalence of a specific Haemoproteus lineage (TARUF02) in a white-lined tanager population. Likewise, we explored whether the probability of detecting the target parasite in an infected bird using PCR and sequencing analyses may be influenced by season and host sex. We found little evidence that sex influenced the probability of an individual host being infected by a haemosporidian parasite. Conversely, we found that the probability of infection by Haemoproteus TARUF02 was ~30% higher during the breeding season, reflecting a higher prevalence of this parasite in this season. The probability that PCR detects DNA of haemosporidian parasite was higher for female birds, suggesting that they are more prone to be parasitized with parasitemia levels that are more successfully detected by molecular analysis. Sequencing successfully determined the Haemoproteus TARUF02 lineage in 60% of samples collected during the breeding season and 84% of samples collected during the non-breeding season. Understanding the ecology of hosts and aspects of their physiology that may influence the parasite infection is essential to better understanding of hemoparasite infections and how parasites influence their native hosts, through decreasing reproductive success, lifespan, and/or survival.

Egg investment strategies adopted by a desertic passerine, the Saxaul Sparrow (Passer ammodendri)

BackgroundAs one of the reproductive strategies adopted by bird species, variation in investment in egg production and its influencing factors are important and well-studied subjects. Intraclutch changes in egg size associated with laying order may reflect a strategy of “brood survival” or “brood reduction” adopted by female birds in different situations.MethodsWe conducted field studies on the breeding parameters of the Saxaul Sparrow (Passer ammodendri) in Gansu Province, China from 2010 to 2017, to clarify the factors affecting the egg investment and reproductive performance of this passerine species.ResultsOur results revealed significant differences in clutch size, egg size and the fledging rate between the first and second brood of Saxaul Sparrows and suggested that this typical desert species allocates more breeding resources to the more favourable second brood period, leading to greater reproductive output. Female body size presented a positive relationship with egg size, and male body size presented positive relationships with clutch size and hatchability. The females that started their clutches later laid more eggs, and hatchability and the fledging rate also increased with a later laying date in the first brood period. With successive eggs laid within the 5-egg clutches (the most frequent clutch size), egg size increased for the first three eggs and then significantly decreased.ConclusionsOur results indicate that female Saxaul Sparrows increased egg investment because of good quality of paired males and good environmental conditions. The intraclutch variation of egg size suggests that this species inhabiting an arid environment adopts a “brood reduction” strategy.

Corticosterone and testosterone treatment influence expression of gene pathways linked to meiotic segregation in preovulatory follicles of the domestic hen.

Decades of work indicate that female birds can control their offspring sex ratios in response to environmental and social cues. In laying hens, hormones administered immediately prior to sex chromosome segregation can exert sex ratio skews, indicating that these hormones may act directly on the germinal disc to influence which sex chromosome is retained in the oocyte and which is discarded into an unfertilizable polar body. We aimed to uncover the gene pathways involved in this process by testing whether treatments with testosterone or corticosterone that were previously shown to influence sex ratios elicit changes in the expression of genes and/or gene pathways involved in the process of meiotic segregation. We injected laying hens with testosterone, corticosterone, or control oil 5h prior to ovulation and collected germinal discs from the F1 preovulatory follicle in each hen 1.5h after injection. We used RNA-sequencing (RNA-seq) followed by DESeq2 and gene set enrichment analyses to identify genes and gene pathways that were differentially expressed between germinal discs of control and hormone-treated hens. Corticosterone treatment triggered downregulation of 13 individual genes, as well as enrichment of gene sets related to meiotic spindle organization and chromosome segregation, and additional gene sets that function in ion transport. Testosterone treatment triggered upregulation of one gene, and enrichment of one gene set that functions in nuclear chromosome segregation. This work indicates that corticosterone can be a potent regulator of meiotic processes and provides potential gene targets on which corticosterone and/or testosterone may act to influence offspring sex ratios in birds.

Female song in eastern bluebirds varies in acoustic structure according to social context

Most of our knowledge about the function, evolution, neurobiology and ontogeny of complex vocalizations in birds comes from previous work on male song. However, the recent increase in focus on female bird song over the last few decades has helped bridge that gap in our knowledge and has vastly changed our understanding of the function, phylogeny and evolution of bird song. Here we present evidence that female song may be modifiable based on signal function. Eastern bluebirds primarily use female song for within-pair communication with their social mate. We examined the female songs of eastern bluebirds sung during within-pair communication and compared the acoustic structure to female songs used in other contexts. Within-pair communication songs differed significantly from other female songs that we measured by six of the eight acoustic variables we measured. Additionally, we examined the song structure of within-pair communication songs at variable distances between mates and between neighbour females. Song structure did not vary over distance. However, we found significant differences between neighbouring females and the number of syllables and frequency modulations each female included in her within-pair communication songs. We propose that female songs have the potential to indicate individual identity, through acoustic structure modification. Our findings further highlight the need for additional study of female song and functional acoustic plasticity in bird song. Animal signals may be stereotyped to convey honest information, they may be functionally plastic to convey additional information, or elements of a signal may fall in both categories. Historically, bird song has been primarily studied as a stereotyped male trait. Whereas male bird song may have variable acoustic structure in some instances, such as extreme aggression, the functional plasticity of female song is largely unknown. We examined female song plasticity for within-pair communication songs of female eastern bluebirds. We present some of the first evidence that female song structure may be variable between functional contexts and that females may alter their song structure to encode additional information, such as individual identity. Our results inform future work on the function and ontogeny of female bird song. Additionally, our findings support the need for further study on female bird song and the functional plasticity of complex acoustic signals.

Relationships between mercury burden, sex, and sexually selected feather ornaments in crested auklet (Aethia cristatella).

Individuals with higher contaminant burdens are expected to be in poorer physical health and be of lower individual body condition and energetic status, potentially resulting in reduced ornamentation or increased asymmetry in bilateral features. The degree and magnitude of this effect also would be expected to vary by sex, as female birds depurate contaminants into eggs. We tested for relationships among mercury in feathers, sex, and elaborate feather ornaments that relate to individual quality in crested auklets (Aethia cristatella), small planktivorous seabirds in the North Pacific Ocean. We found no relationships between mercury and the size of individuals' forehead crest or degree of measurement asymmetry in auricular plumes, both of which are favoured by intersexual selection. Females had significantly greater mercury concentrations than males (females. 1.02 ± 0.39μg/g; males, 0.75 ± 0.32μg/g); but concentrations were below that known to have physiological effects, as expected for a secondary consumer. Sex differences in overwintering area for this long-distance migrant species (more females in the Kuroshio Current Large Marine Ecosystem than males) could be the reason for this seemingly counterintuitive result between sexes. Further research relating mercury burden to overwintering ecology and diet contents would build on our results and further elucidate interrelationships between sex, sexually selected feather ornaments and contaminant burden.

Research Note: Effects of the rearing method and stocking density on carcass traits and proximate composition of meat in small-sized meat ducks

The present study was conducted to evaluate the effects of different rearing methods and stocking densities on carcass yield and proximate composition of meat in small-sized meat ducks. A total of 555 one-day-old birds were randomly allocated to six treatment groups (three replicates per treatment, sex ratio 1/1) with a 2 × 3 factorial arrangement of two rearing methods (reared in cage or net) and three stocking densities (5 [low], 7 [medium], or 9 [high] birds/m2) until day 70. Five male and five female birds from each replicate were randomly selected and processed to determine the carcass yield. Proximate composition was determined by proximate analysis using the breast and thigh muscles. There was no interaction effect between the rearing method and stocking density on carcass yield. The rearing method affected the thigh muscle rate, which was higher in the cage groups (P < 0.05). The final BW and abdominal fat rate decreased with increasing density (P < 0.05), whereas the thigh muscle rate increased (P < 0.05). There were significant interaction effects (P < 0.05) between the rearing method and stocking density on the content of protein, fat, and collagen. The content of fat and moisture was greater and lower, respectively, in the cage groups (P < 0.05). The content of moisture, fat, and collagen with a medium density was higher (P < 0.05). In addition, the content of protein and fat was lower in the ducks fed in nets at low and high densities (P < 0.05), respectively; the collagen content of breast and thigh muscle was lower in the ducks fed in cages and nets, respectively, at a low density (P < 0.05). Our findings provide valuable insights into the single and interactive effects of the rearing method and stocking density on duck slaughter performance and proximate composition of meat. The results indicate that a rearing system with a cage pattern and a medium density is better than other arrangements for small-sized meat ducks.

Food access modifies GnIH, but not CRH, cell number in the hypothalamus in a female songbird

Food deprivation or restriction causes animals to mount a stereotypical behavioral and physiological response that involves overall increases in activity, elevated glucocorticoid production, and (often) inhibition of the reproductive system. Although there is increasing evidence that these responses can differ in their degree or covariation between the sexes, most studies to-date on food restriction/deprivation have focused on male songbirds. We therefore aimed to characterize the behavioral, physiological, and neuroendocrine response to acute food deprivation in a female songbird using a nomadic species, the zebra finch. We quantified behavior during a 6.5 h food deprivation and then measured physiological and neuroendocrine responses of female birds at the 6.5 h timepoint. Within 1 h of acute food deprivation, female zebra finches increased foraging behaviors, and after 6.5 h of food deprivation, females lost 5% of their body mass, on average. Change in body mass was positively associated with elevated corticosterone and (contrary to findings in male zebra finches) negatively related to the number of gonadotropin inhibitory hormone-immunoreactive cells in the hypothalamus. However, there was no effect of food deprivation on corticotropin releasing hormone-immunoreactive cells in the hypothalamus. There was also no relationship between corticotropin releasing hormone-immunoreactive cell number and circulating corticosterone. Our results are consistent with the hypothesis that neuroendocrine responses to food deprivation differ between male and female songbirds. Future studies should work to incorporate sex comparisons to evaluate sex-specific neuroendocrine responses to acute stress.

Cobb MV × Cobb 700 broiler responses to eight varying levels of amino acid density with emphasis on digestible lysine

Summary The percentage of broiler complex processing plants slaughtering big birds (e.g., those greater than 3.5 kg) has steadily increased in the United States the past 15 yr. Precision amino acids in dietary formulation is key as the growing cycle for these birds is extended, as well as total feed consumed. Furthermore, broilers reared for big-bird slaughter plants are destined for specialty breast meat yield portioning, making digestible Lys one of the key nutrients in feed formulation. This work assesses dietary Lys levels in graduations across feed phases to 53 D in Cobb MV × Cobb 700 female and male broilers. Optimizing digestible Lys in the starter phase (to 14 D) occurred between 1.26 and 1.31%. On average, dietary Lys intake per bird needed to optimize live performance, and white meat yield was higher in male birds (64 g) than in female birds (58 g). Optimization of Lys amino acid density in the Cobb MV × Cobb 700 broilers resulted in increased white meat yields in larger broilers, resulting in increased fillet weights per bird with white striping and woody breast being unaffected.

A study on risk factors for macroscopic gut abnormalities in intensively reared broiler chickens

ABSTRACT Intensification of broiler production has coincided with an increase in enteric disorders. Enteric syndromes of unknown aetiology are often associated with an increased feed conversion ratio and are given the general term “dysbiosis”. Despite the importance of dysbiosis, information on factors contributing to this condition are scarce. Therefore, the aim of this study was to describe dysbiosis in broilers (Ross 308) during one production round and to identify risk factors. Fifteen farms in Flanders (Belgium) were followed up, with visits at days 10, 17, 20, 24 and 28 of production. At every visit, 10 random birds were inspected for footpad lesions, hock burns and breast blisters. Also, coccidiosis and enteric abnormalities were scored after necropsy. A gut appearance score (GAS) was given based on 10 macroscopically visible parameters, where a higher GAS equalled more enteric abnormalities. Footpad lesions were seen in 14 farms and increased in prevalence with the age of the birds. Hock burns were seen less frequently, and no breast blisters were detected. Eimeria acervulina lesions were most frequently observed, followed by E. maxima and E. tenella lesions. The average GAS increased from day 10 until day 20. The strong correlations between the GAS at days 10, 17 and 20 indicate that prevalence of gut abnormalities at day 10 can be predictive for scores later on. A higher amount of intestinal defects was seen in older female birds, in the presence of a conceivable E. tenella infection and at farms with a higher productivity and sanitary status. RESEARCH HIGHLIGHTS Gut lesions found in young broilers can predict further development of defects. Caecal lesions showed the strongest correlation with GAS in a multivariate model.

Why female birds of prey are larger than males

AbstractThe causes of the reversed sexual size dimorphism (RSD; females larger than males) in birds of prey are subject to a centuries-old, passionate debate. A crucial difficulty is to distinguish whether the postulated benefits derive from the proposed causal process(es) or are incidental. After reviewing the existing literature, we present a methodology that overcomes this difficulty and renders unnecessary any speculative a priori distinctions between evolved function and incidental effects. We can thus justify the following novel version of the well-known nest defence hypothesis as the most likely to explain the phenomenon in all birds of prey that show RSD: if the female predominates in actively defending the eggs and young against predators, then she is the heavier sex, and her relatively greater body mass is adaptive. That is, heavier females are favoured (independently of males) by natural selection. The attractiveness of this hypothesis is that it has the potential to explain the phenomenon in all raptors exhibiting RSD, can deal with the exceptional cases in this group, explains the direction of the dimorphism, focuses on a key factor in the reproductive success of most raptors, is parsimonious, i.e. does not require supporting hypotheses, and is supported by a substantial body of evidence.

Identifying medullary bone in extinct avemetatarsalians: challenges, implications and perspectives.

Medullary bone (MB) is a sex-specific tissue produced by female birds during the laying cycle, and it is hypothesized to have arisen within Avemetatarsalia, possibly outside Avialae. Over the years, researchers have attempted to define a set of criteria from which to evaluate the nature of purported MB-like tissues recovered from fossil specimens. However, we argue that the prevalence, microstructural and chemical variability of MB in Neornithes is, as of yet, incompletely known and thus current diagnoses of MB do not capture the extent of variability that exists in modern birds. Based on recently published data and our own observations of MB distribution and structure using computed tomography and histochemistry, we attempt to advance the discourse on identifying MB in fossil specimens. We propose: (i) new insights into the phylogenetic breadth and structural diversity of MB within extant birds; (ii) a reevaluation and refinement of the most recently published list of criteria suggested for confidently identifying MB in the fossil record; (iii) reconsideration of some prior identifications of MB-like tissues in fossil specimens by taking into account the newly acquired data; and (iv) discussions on the challenges of characterizing MB in Neornithes with the goal of improving its diagnosis in extinct avemetatarsalians. This article is part of the theme issue 'Vertebrate palaeophysiology'.

Molecular Bird Sexing on Fischeri Lovebird (Agapornis fischeri) by Using Polymerase Chain Reaction

Fischeri Lovebird (Agapornis fischeri) found originally in Africa which has spread to many countries. In Indonesia, Fischeri Lovebird is popular as a pet animal. This lovebird is a monomorphic bird, so it is difficult to differentiate morphologically between male and female birds. In general, a male lovebird has ZZ homozygotes, whereas females' lovebird has ZW heterozygous of their sex chromosome. These sex chromosomes set used as study targets for molecular bird sexing of many species of birds because this method is effective and simple to perform. This method targeted to amplify the Chromodomain Helicase DNA-binding (CHD) gene, which found into the sex chromosome of male and female birds. The objective of this study was to rapid molecular bird sexing of Fischeri Lovebird by using PCR methods. Research samples were collected from feather calamus of A. fischeri. The total sample was 11 feathers from A. fischeri. which were collected three to six feathers for each lovebird. Then the research was followed by DNA extraction from calamus feathers, DNA amplification by PCR and agarose gel electrophoresis of PCR products and visualization of PCR predicts by UV-Transilluminator in darkroom. It concluded that PCR amplification using NP, MP and P2 primers produced double DNA bands in size of 400 bp on Z chromosome and bp on W chromosome for female Fischeri Lovebird, whereas for male Fischeri Lovebird only produced a single DNA band in size of 400 bp on Z chromosome. From eleven samples of Fischeri Lovebird showed a total of five females and six male Fischeri Lovebirds.

Long-term Devocalization of Zebra Finches.

Songbirds, such as the zebra finch, are a popular animal model for studying the neural basis of vocal and complex skill learning. Adult male zebra finches produce courtship song toward females (referred to as 'directed song') and recording and analyzing sounds of directed song along with underlying neural activity is important for investigating behavioral and neural mechanisms of song production and learning. However, recording of directed song is easily contaminated by calls that are often as loud as directed songs and frequently produced by a female bird is presented in the same sound-recording chamber to elicit directed song. We developed a new surgical procedure to relatively easily and almost completely devocalize female zebra finches semi-permanently, without affecting other behaviors. This procedure enables researchers to record directed songs with almost no contamination by female calls. The procedure can also be used to devocalize male birds as well and, thus, has great potential for a variety of experimental purposes, such as long-term elimination of auditory feedback during singing in male birds.

INVESTIGATION OF ALKALINE PHOSPHATASE EXPRESSION IN THE SMALL INTESTINES OF THE BRONZE TURKEY (MELEAGRIS GALLOPAVO)

The aim of our investigation was to study the expression of the enzyme Alkaline phosphatase in the small intestines of the bronze turkey in the age aspect. Forty clinically healthy bronze turkeys (twenty males and twenty females) were studied. The groups of the birds were at age 1, 7, 14, 28, 35, 49, 56, 90, 120, and 240 days. Each group consisted of five males and five female birds. The enzyme’s expression was investigated by Gomori staining. The most significant expression of tissue alkaline phosphatase was observed in the epithelial cells of the duodenum. It was weaker in the jejunum and weakest in the ileum. In the three intestinal segments, high enzyme activity was observed during the first weeks of hatching (from the 1st to the 14th day in the duodenum and ileum, and from the 1st to the 28th day in the jejunum).

Interactive effects of dietary vitamin K3 and Bacillus subtilis PB6 on the growth performance and tibia quality of broiler chickens with sex separate rearing

Both vitamin K and probiotics can promote the bone health of poultry and mammals. The present study was conducted to investigate the interactive effects between vitamin K3 (VK3) and Bacillus subtilis PB6 on the growth performance and tibia quality of broiler chickens with sex separate rearing. In a 3 × 2 × 2 factorial arrangement, 720 one-day-old broiler chicks (Arbor Acres) were assigned to 12 groups with three levels of dietary VK3 (0, 0.5 and 4.0 mg/kg), with or without probiotic supplementation (500 g/t) and with sex separation (male and female). Each group included 3 replicates with 20 birds per replicate. During day 1 to 21, 0.5 and 4.0 mg/kg of VK3 increased average daily gain (ADG) of all birds and average daily feed intake of male birds (P < 0.05). During day 22 to 42, probiotic supplementation increased the ADG of birds (P < 0.05). Probiotic addition increased the weight, length, diameter and strength of tibia in all birds, and 0.5 and 4.0 mg/kg of VK3 increased the tibial breaking strength of male birds at day 21 (P < 0.05). Vitamin K3 and probiotic synergistically increased tibial breaking strength at day 42 and ash content at day 21 (P < 0.05). Three factors exhibited interactive effects on the chemical composition of tibia at day 42, and female birds fed 4 mg/kg of VK3 and probiotic had the highest contents of ash, calcium and phosphorus (P < 0.05). Bacillus subtilis PB6 increased the serum phosphorus level of male birds at day 21 and serum calcium level of female ones at day 42 (P < 0.05). At day 21, in the probiotic-supplemented birds, serum osteocalcin (OCN) and bone-specific alkaline phosphatase (BALP) were increased by 0 and 4.0 mg/kg of VK3, respectively (P < 0.05). Probiotic increased serum OCN and cooperated with VK3 to increase the serum BALP at day 42 (P < 0.05). Vitamin K3 and probiotic synergistically down-regulated the mRNA expression of Runt-related transcription factor 2 and OCN at day 21 (P < 0.05). Vitamin K3 down-regulated the alkaline phosphatase (liver/bone/kidney) expression in male birds at day 21 and 42, but probiotic up-regulated the expression of these genes at day 42 (P < 0.05). In conclusion, VK3 and B. subtilis PB6 promoted the growth performance of broilers during starter and grower phases, respectively. They synergistically improved the physical and chemical traits of tibias, especially in grower phase, by modulating calcium and phosphorus metabolism as well as osteogenic gene expression.

Transcriptome analysis and identification of genes associated with chicken sperm storage duration

The sperm storage tubules located in the mucosal folds of the uterovaginal junction (UVJ) are the primary site of sperm storage in chicken hens after natural mating or artificial insemination (AI). The short-term sperm storage (24 h after mating or AI) in hens was highly associated with immunity and pH-related pathway genes. However, the underlying mechanism of longer duration of sperm storage in female birds remains largely unclear. In the present study, transcriptome analysis was applied to uncover the dynamic gene expression changes in chicken UVJ tissues at two time points (day 3 and day 9) after AI. A total of 574 differentially expressed genes (DEG) were enriched, including 266 upregulated and 308 downregulated DEG. The validation of 5 DEG using quantitative PCR showed a similar expression tendency with RNA sequencing results. The gene ontology terms of DEG were highly enriched in heparin binding (9 genes including COMP, CTGF, and IMPG2), glycosaminoglycan binding (10 genes including PCOLCE, POSTN, and RSPO3), and response to estradiol and ion transport (AREG, RAMP3, SFRP1, and SSTR1). Kyoto encyclopedia of genes and genomes pathway-enrichment analyses of DEG revealed 10 significant pathways (P < 0.05) represented by calcium signaling pathway (7 genes including CACNA1G, PDE1C, PDGFRB, and SLC8A1) and glycosaminoglycan biosynthesis (B3GNT7, CSGALNACT1, GLCE, and ST3GAL1). Protein-protein interaction network of DEG established the connection-regulating epithelial cell or cell-matrix adhesion and migration. The enriched pathways and genes were highly correlated with temporary sperm storage in and possibly sequential sperm release from chicken UVJ overtime after AI. Of these, HIP1, PDE1C, and calcium-related genes were the most interesting candidates associated with sperm storage duration. This report provided a global gene expression profile of the chicken UVJ regarding the capacity of sperm storage overtime after AI. The outcome of this study will contribute to further understanding of the long-term sperm maintenance in avian females and eventually improving the duration of fertile egg performance by selected chicken breeding.