Dynamic changes in protein concentrations of keratins in crop milk and related gene expression in pigeon crops during different incubation and chick rearing stages.

1. The objective of this study was to examine the keratin composition of crop milk, the variation of epithelial thickness and keratin (K) gene expression in samples from young pigeon during incubation and chick rearing.2. Crop milk was collected from 1-, 3- and 5-day-old squab crops for keratin content analysis. Results showed that K4 accounted for the highest proportion of all detected keratins.3. In total, 42 pairs of adult pigeons were allocated to seven groups according to different stages to collect crop samples. Gene expression studies showed that the K3 gene expression was maximised at rearing Day 15 (15) and R1 in males and females, respectively. K6a gene level was the greatest at R15 in females, whereas it peaked at incubation Day 4 (I4) in males. The K12, K13, K23 and K80 gene levels were inhibited at the peak period of crop milk formation in comparison with I4. In females, K cochleal expression peaked at I10, whereas it was the greatest at R25 in males. K4 and K14 gene expression was the highest at I10 in females, while K4 and K14 were minimised at I17 and R7 in males, respectively. Gene expressions of K5, K8, K19 and K20 in males and K19 in females were maximised at R1. The K5, K20 and K75 gene levels in females peaked at R7. K75 and K8 expressions in males and females reached a maximum value at R25 and I17, respectively.4. The epithelial thickness of male and female crops reached their greatest levels at R1 and had the highest correlation with K19.5. These results emphasised the importance of keratinisation in crop milk formation, and different keratins probably play various roles during this period. The K19 was probably a marker for pigeon crop epithelium development. The sex of the parent pigeon affected keratin gene expression profiles.

Maternal dietary linoleic acid altered intestinal barrier function in domestic pigeons (Columba livia).

Linoleic acid (LA) is predominantly essential for poultry. Poultry lacking LA show retarded growth and reduced disease resistance. Intestinal barrier function plays an important role in pigeon squab growth, whereas research on the effects of LA on intestinal health in altrices is scant. Considering that squabs are fed by their parents, the study aimed to explore the effects of maternal dietary LA on intestinal morphology, tight junction proteins, immune cytokines and microbial flora in squabs. A completely randomised design with a control group, 1 % LA supplementation group, 2 % LA supplementation group and 4 % LA supplementation group was used. Six squabs from each treatment were randomly sampled at 21 d post-hatching. The results indicated that LA supplementation improved intestinal morphology, as reflected by increased villus height, villus area and the ratio of villi to crypts. Also, 1 % LA supplementation elevated the density of goblet cells in the intestine and strengthened tight junctions by up-regulating claudin-3 and occludin gene expression but down-regulating claudin-2 gene expression. Moreover, 1 % LA supplementation reduced the secretion of proinflammatory cytokines and partly increased anti-inflammatory cytokines. The intestinal microbial diversity in the 1 % LA supplementation group was higher than that in the other groups. As beneficial bacteria, Butyrivibrio was the biomarker of 1 % LA supplementation. However, excessive (4 %) LA supplementation led to adverse impacts on intestinal immunity and microbiota. In conclusion, maternal dietary LA might alter intestinal barrier function in pigeon squabs in a dose-dependent manner. Supplementation with 1 % LA was suggested in parental pigeons.

Enhancing Growth and Gut Health in Squabs: The Impact of Fermented Mixed Feed.

The purpose of this study was to evaluate the effect of fermented mixed feed (FMF) (soybean meal-rapeseed meal-corn bran (6:3:1, m/m/m)) on the growth performance, intestinal microbial communities, and metabolomes of squabs. One hundred and eighty 1-day-old squabs were randomly allocated to two groups, each containing six replicates of fifteen squabs cared for by 60 pairs of breeding pigeons secreting crop milk. Each pair of breeding pigeons cared for three squabs. The control group was fed a basal diet, while the experimental group was fed the basal diet containing 5% FMF. The results showed that daily weight gain, carcass weight, villus height, and the mRNA level of ZO-1 in the ileum were increased in the birds fed FMF compared to the control squabs (p < 0.05). Greater abundances of beneficial bacteria such as Lactobacillus, Bifidobacteria, and Bacillus as well as fewer harmful bacteria (i.e., Enterococcus, Veillonella, and Corynebacterium) in the ilea of squabs fed FMF. Six differential metabolites were identified in the FMF-treated squabs; one metabolite was increased (ω-salicoyisalicin) and five were decreased (3-benzoyloxy-6-oxo-12-ursen-28-oic acid, estradiol-17-phenylpropionate, aminotriazole, phosphatidyl ethanolamine (22:6/0:0), and 1-arachidonoylglycerophosphoinositol). Positive correlations were observed between the abundance of Lactobacillus and villus height. Overall, FMF treatment improved both growth and intestinal health in pigeons, suggesting potential benefits for pigeon production.

The Identification of Functional Genes Affecting Fat-Related Meat Traits in Meat-Type Pigeons Using Double-Digest Restriction-Associated DNA Sequencing and Molecular Docking Analysis.

The Chinese indigenous Shiqi (SQ) pigeon and the imported White King (WK) pigeon are two meat-type pigeon breeds of economical and nutritional importance in China. They displayed significant differences in such meat quality traits as intramuscular fat (IMF) content and fatty acid (FA) compositions in the breast muscles. In this study, we aimed to screen candidate genes that could affect fat-related meat quality traits in meat-type pigeons. We investigated the polymorphic variations at the genomic level using double-digest restriction-associated DNA (ddRAD) sequencing in 12 squabs of SQ and WK pigeons that exhibited significant inter-breed differences in IMF content as well as FA and amino acid compositions in the breast muscles, and screened candidate genes influencing fat-related traits in squabs through gene ontology analysis and pathway analysis. By focusing on 6019 SNPs, which were located in genes with correct annotations and had the potential to induce changes in the encoded proteins, we identified 19 genes (ACAA1, ACAA2, ACACB, ACADS, ACAT1, ACOX3, ACSBG1, ACSBG2, ACSL1, ACSL4, ELOVL6, FADS1, FADS2, HACD4, HADH, HADHA, HADHB, MECR, OXSM) as candidate genes that could affect fat-related traits in squabs. They were significantly enriched in the pathways of FA metabolism, degradation, and biosynthesis (p < 0.05). Results from molecular docking analysis further revealed that three non-synonymous amino acid alterations, ACAA1(S357N), ACAA2(T234I), and ACACB(H1418N), could alter the non-bonding interactions between the enzymatic proteins and their substrates. Since ACAA1, ACAA2, and ACACB encode rate-limiting enzymes in FA synthesis and degradation, alterations in the enzyme-substrate binding affinity may subsequently affect the catalytic efficiency of enzymes. We suggested that SNPs in these three genes were worthy of further investigation into their roles in explaining the disparities in fat-related traits in squabs.

Effect of tea polyphenols supplement on growth performance, antioxidation, and gut microbiota in squabs.

Early life nutritional supplementation can significantly improve pigeon health. Both the nutritional crops of parental pigeons and the intestinal development of squabs play key roles in the growth rate of squabs. Tea polyphenols (TPs), as natural plant extracts, exhibit potential biological activities. However, the impact of TPs on the intestinal function of squabs is not known. This study evaluated the effects of TPs on growth performance, immunity, antioxidation, and intestinal function in squabs. A total of 432 young pigeons (1 day old) were divided into four groups: a control group (fed a basic diet) and three treatment groups (low, medium, and high dose groups; 100, 200, and 400 mg/kg TPs, respectively). On the 28th day, samples of serum, mucosal tissue, and digests from the ileum of squabs were collected for analysis. The results revealed that TP supplementation significantly reduced the feed-to-meat ratio and improved the feed utilization rate and serum biochemical indices in squabs. Additionally, it enhanced the intestinal barrier function of birds by promoting intestinal development and integrity of tight junctions and regulating digestive enzyme activities and intestinal flora. Mechanistically, TPs activated the Nrf2-ARE signaling pathway, which may be associated with improved antioxidant and immune responses, correlating with an increased abundance of Candida arthritis and Corynebacterium in the ileum.

Intestinal Immune Development Is Accompanied by Temporal Deviation in Microbiota Composition of Newly Hatched Pigeon Squabs.

Identifying the interaction between intestinal mucosal immune system development and commensal microbiota colonization in neonates is of paramount importance for understanding how early life events affect resistance to disease later in life. However, knowledge about this interaction during the early posthatch development period in altrices is limited. To fill this gap, samples of intestinal content and tissue were collected from newly hatched pigeon squabs at four time points (days 0, 7, 14, and 21) for microbial community analysis and genome-wide transcriptome profiling, respectively. We show that the first week after hatching seems to be the critical window for ileal microbiota colonization and that a potentially stable microbiota has not yet been well established at 21 days of age. Regional transcriptome differences revealed that the jejunum rather than the ileum plays a crucial role in immunity at both the innate and adaptive levels. In the ileum, temporal deviation in innate immune-related genes mainly occurs in the first week of life and is accompanied by a temporal change in microbiota composition, indicating that the ileal innate mucosal immune system development regulated by microbial colonization occurs mainly in this period. Furthermore, we provide evidence that colonization by Escherichia and Lactobacillus within the first week of life is likely one of the causative factors for the induction of proinflammatory cytokine expression in the ileum. We also demonstrate that cellular adaptive immune responses mediated by Th17 cells following commensal-induced proinflammatory cytokine production in the ileum begin as early as the first week posthatch, but this cellular immunity seems to be less effective in terms of maintaining the inflammatory response balance. Because the induction of high levels of mucosal secretory IgA (SIgA) seems to take approximately 3 weeks, we favor the idea that humoral adaptive immunity might be less active, at least, during the first 2 weeks of life. Our data may help to explain the phenomenon of the occurrence of intestinal infections mainly in the ileum of pigeon squabs during the early posthatch period. IMPORTANCE The pigeon (Columba livia), an altricial bird, is one of the most economically important farmed poultry for table purposes. Identifying the interaction between intestinal mucosal immune system development and commensal microbiota colonization in neonates is of paramount importance for understanding how early life events affect resistance to disease and potential productivity later in life. However, knowledge about this interaction during the early posthatch development period in altricial birds is limited. The study described herein is the first to try to provide insights into this interaction. Our data provide evidence on the mutual relationship between intestinal mucosal immune system development and commensal microbiota colonization in pigeon squabs and may help to explain the phenomenon of the occurrence of intestinal infections mainly in the ileum of pigeon squabs during the early posthatch period.

Effects of early weaning on intestinal morphology, digestive enzyme activity, antioxidant status, and cytokine status in domestic pigeon squabs (Columba livia).

The aim of this study was to explore the effects of early weaning on growth performance, intestinal morphology, digestive enzyme activity, antioxidant status, and cytokine status in domestic pigeon squabs (Columba livia). The conclusion is based on body weight (BW) and average daily gain (ADG), length index and weight index of small intestine, small intestinal morphology, activity of digestive enzymes in duodenum content, the concentrations of jejunal antioxidant status and cytokines. A completely randomized design with 2 treatments, the control group (CON) and early weaning (EW) group, was utilized. Eight squabs per treatment were sampled at the age of 25 d. The results showed that early weaning reduced BW (P < 0.05), ADG (P < 0.05), ileac length index (P < 0.05), and weight index (P < 0.01). Compared with the CON group, small intestinal morphology was altered in the EW group. Ileac crypt depth (CD) increased significantly (P < 0.01). The villus area was decreased in the duodenum (P < 0.05), jejunum (P < 0.01), and ileum (P < 0.05). The ileac ratio of villus height to crypt depth (VCR) in the EW group was lower than the ileac ratio of villus height to VCR in the CON group (P < 0.01). The activity of trypsin (P < 0.05), sucrase (P < 0.01) and aminopeptidase-N (APN) (P < 0.01) in the duodenum was reduced. Jejunal malondialdehyde (MDA) (P < 0.01) was increased and total superoxide dismutase (T-SOD) (P < 0.01) was reduced significantly. Early weaning decreased the concentrations of interferon-γ (IFN-γ) (P < 0.01), interleukin-4 (IL-4) (P < 0.05) and interleukin-10 (IL-10) (P < 0.01) but induced significant upregulation of interleukin-2 (IL-2) (P < 0.05). In conclusion, our results suggested that early weaning did harm the BW and ADG, intestinal length index and weight index, intestinal morphology, activity of digestive enzymes, and antioxidant and cytokine status.

Monitoring the variations in physicochemical characteristics of squab meat during the braising cooking process.

Braised squabs are traditional Chinese foods. However, the processing is highly experience dependent and lacks a theoretical basis. Hence, a comparative study of the physicochemical properties in different processing stages of braised squabs was necessary. We observed the physicochemical changes in the processing stages of braised squabs (raw meat, braised meat, and fried meat). The color parameters, moisture content, and drip loss rate gradually decreased during the processing. On the contrary, crude protein content and pH value were upregulated in the processing stages of braised squabs. Furthermore, the diameter of muscle fiber significantly increased in the braised meat and further decreased in the fried meat compared with the raw muscle fiber. Similarly, hardness, springiness, and chewiness were also increased in the braised step and decreased in the fried step. Additionally, the contents of essential amino acids remain unchanged. Hence, our results provided a certain reference value on the production of braised squabs.

Dietary Supplementation of Chitosan Oligosaccharide-Clostridium butyricum Synbiotic Relieved Early-Weaned Stress by Improving Intestinal Health on Pigeon Squabs (Columba livia).

According to a previous study, we had found that early weaning causes harm to growth performance, intestinal morphology, activity of digestive enzymes, and antioxidant status in pigeon squabs (Columba livia). Chitosan oligosaccharides (COS) and Clostridium butyricum have been reported to have great potential to improve the growth performance and intestinal health of early-weaned animals. Therefore, the aim of this study is to explore whether dietary supplementation with COS-C. butyricum synbiotic could relieve early-weaned stress by evaluating its effects on growth performance and intestinal health in pigeon squabs. A total of 160 squabs (weaned at 7 days of age) were randomly divided into 5 groups: the control group, fed with artificial crop milk; the COS supplementation group, fed with artificial crop milk + 150 mg/kg COS; and three synbiotic supplementation groups, fed with artificial crop milk + 150 mg/kg COS + 200, 300, and 400 mg/kg C. butyricum. The results showed that a diet supplemented with COS-C. butyricum synbiotic benefitted the growth performance of early-weaned squabs; even so the differences were not significant among the five groups (p > 0.05). In addition, dietary supplementation of 150 mg/kg COS + 300~400 mg/kg C. butyricum significantly improved the intestinal morphology (especially villus surface area and the ratio of villus height to crypt depth), the activity of digestive enzymes (lipase, trypsin, and leucine aminopeptidase) in duodenum contents, and the production of total short-chain fatty acids and acetic acid in ileum content (p < 0.05). Additionally, dietary supplementation of 150 mg/kg COS + 400 mg/kg C. butyricum benefitted gut health by improving the antioxidant capacity (glutathione peroxidase and total antioxidant capacity) and cytokine status (IL-4 and IL-10) (p < 0.05), as well as by improving the intestinal microbiota diversity. In conclusion, our results revealed that dietary supplementation with synbiotic (150 mg/kg COS + 300~400 mg/kg C. butyricum) could relieve early-weaned stress by maintaining intestinal health in pigeon squabs.

Corrigendum: The Composition and Function of Pigeon Milk Microbiota Transmitted From Parent Pigeons to Squabs.

[This corrects the article DOI: 10.3389/fmicb.2020.01789.].

The Composition and Function of Pigeon Milk Microbiota Transmitted From Parent Pigeons to Squabs.

Mammalian neonates obtain antibodies, nutrients, and microbiota from breast milk that help them resist the complex growth environment. Similar to mammals' lactation behavior for their offspring, parent pigeons regurgitate pigeon milk (PM) from their crops to feed the squabs. Whether pigeon milk is as valuable as mammalian milk is not clear, especially in terms of microbiota. This study adopted 16S rRNA gene sequencing to investigate the microbial composition and function in pigeon milk. We found abundant microbiota in pigeon milk. The dominant genera in parent pigeons' milk were Lactobacillus, Enterococcus, Veillonella, and Bifidobacterium. An analysis of squab milk (SM) showed that Lactobacillus also accounted for a considerable proportion, followed by Bifidobacterium. Most of the squab milk microbial genera were also detected in parent pigeons. Microbial functional analysis showed that the squab milk microbes were more involved in the pathways of carbohydrate metabolism, amino acid metabolism, and energy metabolism. These findings indicated that microbiota play an important role in squabs and can be transmitted from parent pigeons to squabs by pigeon milk. The presence of plentiful probiotics in squabs also suggests that adding probiotics in artificial pigeon milk may promote the growth and development of squabs and improve the production performance of pigeons.

Prevalence of intestinal helminth parasites of pigeons (Columba livia domestica Gmelin 1789) in Kano State, North-Western Nigeria.

In Nigeria, helminths have over the last decades been established as important parasite of poultry including pigeons. However, the prevalence of these parasites of pigeons in Kano State is yet to be established. The prevalence of helminth parasites of domestic pigeon (Columba livia domestica) in Kano State, Nigeria was investigated in this study. The differences in the prevalence of infection between the pigeons examined on the basis of sex of the pigeon and season of the year were analyzed statistically using the students' "t" -test. The intestinal contents of 144 pigeons were examined for gastrointestinal helminths between February and April representing the dry season and between June and August for the wet season. Pigeons were grouped according to Squabs (chicks) (0-4 weeks), Squeakers (Juveniles) (5-8 weeks) and Youngsters (Adults) (9 weeks onwards). Intestinal parasites found include Cestodes: Raillietina tetragona, 20 (13.80%), R. echinobothrida, 11 (7.64%), Amoebotaenia cuneata, 5 (3.47%), Hymenolopis contaniana, 10 (6.95%), Davainea proglottina, 1(0.69%) and Ornithostrongylus quadriatus 1(0.69%). Nematodes: Capillaria obsignata, 10(6.95%) and Ascaridia columbae, 9(6.25%). Forty-two (29.16%) of the birds had mild infection, 17 (11.81%) had moderate infection, 9 (6.25%) and 2(1.39%) had heavy and severe infection, respectively. The prevalence was higher during the wet season (36.65%) than during the dry season (27.97%). Of all the age groups, only Youngsters, 34 (23.61%) were infected with intestinal helminths. However, there were statistically significant differences between the age, sex and seasons in this study (P < 0.005). Pigeons raised on semi-intensive had the highest rate of infection (37.50%) and those raised on intensive management had the lowest rate of infection (9.03%). A number of measures are recommended for the control of these helminthic parasites.

Effects of Dietary Supplementation with dl-Methionine and dl-Methionyl-dl-Methionine in Breeding Pigeons on the Carcass Characteristics, Meat Quality and Antioxidant Activity of Squabs.

This study aimed to investigate the effects of dietary supplementation with dl-methionine (dl-Met) and dl-methionyl-dl-methionine (dl-Met-Met) in breeding pigeons on the carcass characteristics, meat quality and antioxidant activity of squabs. A total of 324 pairs of breeding pigeons were selected and allotted to 9 treatments in a completely randomized design, and the birds were fed dietary treatments for 45 d, including a Met-deficient basal diet (BD, crude protein = 15%, Met = 0.25%) and BD + 0.15%, 0.30%, 0.45%, or 0.60% dl-Met or dl-Met-Met diets. Compared with the diet fed to the BD group, dietary dl-Met or dl-Met-Met supplementation effectively increased the carcass yield, semieviscerated yield, eviscerated yield, breast muscle yield, thigh muscle yield, a* value, catalase activity, total superoxide dismutase activity and glutathione peroxidase activity, but decreased the L* value, malonaldehyde concentration, drip loss and cooking loss of squabs (p < 0.05). The relative bioavailability values of dl-Met-Met relative to those of dl-Met were 467% and 376% based on carcass yield and breast muscle yield, respectively (p < 0.001). Moreover, dl-Met-Met was more effective than dl-Met in decreasing the drip loss and improving the antioxidant activity of the breast and thigh muscles of squabs (p < 0.05). As a source of Met, dl-Met-Met, rather than dl-Met, was more beneficial to squabs.

Effects of in ovo feeding of L-lysine on hatchability, hatching time, and early post-hatch development in domestic pigeons (Columba livia)1.

This study was conducted to evaluate the effects of in ovo feeding (IOF) of L-lysine (Lys) on hatchability and development in pigeon neonates. At day 13 of incubation, fertile eggs were injected with 200 µL of sterilized saline (0.75%, SC group), 1% Conc. Lys solution (Lys group, 2.11 mg Lys dissolved in 200 µL of sterilized saline), with controls not injected (NC group). The results showed that IOF of Lys decreased the hatchability (P < 0.05) compared with other groups, whereas the hatching time was not affected among groups. On day 14 of post-hatch (D14), the body weight (BW) of squabs received Lys IOF was increased relative to the NC group (P < 0.05). Squabs in Lys group exhibited higher (P < 0.05) body weight gain (BWG) than other groups from D14 to day of hatch (DOH). Meanwhile, IOF of Lys increased the brain relative weight on DOH (P < 0.05), and organ index of heart, legs, and gizzard on day 7 of post-hatch (D7) compared with other groups. In addition, the length index of duodenum, jejunum, or ileum had no difference between groups, as well as the weight index except the weight index of duodenum in Lys group was higher than that of SC group on DOH. However, squabs received Lys IOF showed lower villus height, crypt depth, and villus surface area of jejunum than NC group (P < 0.05) on DOH, and lower crypt depth than NC and SC groups on D7. The situation was improved by D14, although there was no significant difference in morphometric trait of jejunum between Lys group and NC group, squabs received Lys IOF showed higher villus height, crypt depth, and villus surface area of jejunum than NC group. In conclusion, the present study demonstrates that IOF of Lys has negative effects on hatchability and development of early post-hatch squabs, but the situation will be improved with the growth of age.

Comparing the Effect of Different Management and Rearing Systems on Pigeon Squab Welfare and Performance after the Loss of One or Both Parents.

Pigeon squabs completely depend on their parents for care and nourishment. The loss of one or both parents affects squabs' successful fledging. This study was carried out on young squabs to compare the effect of pigeon parent sex and different fostering methods on squab welfare (behavior and growth performance). Two experiments were carried out. In the first experiment, the squabs were divided into three groups. Group 1 (control) consisted of 10 parent pairs with 20 brooding squabs; group 2 consisted of 10 male parents with 20 brooding squabs; and group 3 consisted of 10 female parents with 20 brooding squabs. In the second experiment, the squabs were also divided into three groups. Group 1 (control) consisted of 10 parent pairs with 20 brooding squabs; group 2 consisted of 20 brooding squabs fostered by 10 foster parent pigeons (either male or female); and group 3 consisted of 20 brooding squabs fostered by the hand-rearing method. A significant improvement in growth performance, behavioral welfare (head waggle, squab note and squab wing shake); increased repetition of these behaviors indicates stress and discomfort), and survival rate was observed to be higher in the group brooded by both parents compared to the group brooded by either a male or a female parent. In addition, the group fostered by hand-rearing showed a significant improvement in growth performance, behavioral welfare, and survival rate compared to the group brooded by foster pigeon parents.