Newborn Chicks Research Articles

Lactobacillus casei DBN023 protects against jejunal mucosal injury in chicks infected with Salmonella pullorum CMCC-533

Pullorum disease is one of the most serious poultry diseases. Antibiotic prophylaxis is commonly applied in actual production, but the emergence of drug-resistant strains and drug residues threatens the health of animals and humans. The use of probiotics has become a potential solution for the aforementioned problem. In this study, we investigated the effects of Lactobacillus casei DBN023 (CGMCC-16146) on the intestinal tissue structure and immune functions of the jejunal epithelium in chicks infected with Salmonella pullorum CMCC-533 using histomorphological and immunological methods. A total of 450 newborn chicks were randomly divided into the control group and experimental groups. We used hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), and an enzyme-linked immunosorbent assay (ELISA) to observe the structure of the jejunal mucosa and analyze changes in cytokine and secretory immunoglobulin A (sIgA) levels. The results showed that prophylactic feeding of L. casei DBN023 to chicks significantly increased their jejunum villar height, villar height-to-crypt-depth (V/C) ratio, and muscle thickness; reduced intestinal-crypt depth; and increased the proliferating cell nuclear antigen (PCNA) level compared with S. pullorum infection. L. casei DBN023 enhances intestinal mucosal immunity, regulates cytokine balance, enhances intestinal immune function, and effectively reduces intestinal inflammation. Thus, it protects intestinal tissues and reduces S. pullorum CMCC-533 damage to the intestinal tract.

Automated Study Challenges the Existence of a Foundational Statistical-Learning Ability in Newborn Chicks

What mechanisms underlie learning in newborn brains? Recently, researchers reported that newborn chicks use unsupervised statistical learning to encode the transitional probabilities (TPs) of shapes in a sequence, suggesting that TP-based statistical learning can be present in newborn brains. Using a preregistered design, we attempted to reproduce this finding with an automated method that eliminated experimenter bias and allowed more than 250 times more data to be collected per chick. With precise measurements of each chick’s behavior, we were able to perform individual-level analyses and substantially reduce measurement error for the group-level analyses. We found no evidence that newborn chicks encode the TPs between sequentially presented shapes. None of the chicks showed evidence for this ability. Conversely, we obtained strong evidence that newborn chicks encode the shapes of individual objects, showing that this automated method can produce robust results. These findings challenge the claim that TP-based statistical learning is present in newborn brains.

Effect of vitamin E in ovo feeding to broiler embryos on hatchability, chick quality, oxidative state, and performance.

The objective of the present study was to ascertain the effect of in ovo feeding of vitamin E (VE) on the incubation results, quality, and oxidative state of newborn chicks and on the initial performance results. The design consisted of randomized blocks with treatments at different levels of VE (0.0, 27.5, 38.5, 49.5, and 60.4 IU). On 17.5 d of embryonic development, 780 eggs underwent in ovo injection using a manual needle. VE supplementation of 60.4 IU provided the highest hatching rate (P < 0.05) and shortest hatch window (P < 0.05). Better results regarding chick physical quality were observed in groups supplemented with VE (body weight, length, newborn chick quality score) and higher chick weight/egg weight ratios (P < 0.05). VE inoculation did not have any effect on the chicks' immunological system (P > 0.05). Greater development of the small intestine (intestine weight/yolk free chick weight and higher villi in duodenum) and better feed conversion over all periods studied (1 to 7, 1 to 14, and 1 to 21 d) were observed among chicks that received in ovo VE supplementation (P < 0.05). The total protein concentrations in the liver and striated breast skeletal muscle tissue were highest in chicks that received 60.4 IU of VE (P < 0.05). The highest catalase activity was observed in the livers of newborn chicks supplemented with 60.4 IU of VE (P < 0.05). It was concluded that in ovo VE supplementation improved the chicks' oxidative state, which led to improvements in incubation results, chick quality and performance results.

Using automation to combat the replication crisis: A case study from controlled-rearing studies of newborn chicks

The accuracy of science depends on the precision of its methods. When fields produce precise measurements, the scientific method can generate remarkable gains in knowledge. When fields produce noisy measurements, however, the scientific method is not guaranteed to work – in fact, noisy measurements are now regarded as a leading cause of the replication crisis in psychology. Scientists should therefore strive to improve the precision of their methods, especially in fields with noisy measurements. Here, we show that automation can reduce measurement error by ∼60% in one domain of developmental psychology: controlled-rearing studies of newborn chicks. Automated studies produce measurements that are 3–4 times more precise than non-automated studies and produce effect sizes that are 3–4 times larger than non-automated studies. Automation also eliminates experimenter bias and allows replications to be performed quickly and easily. We suggest that automation can be a powerful tool for improving measurement precision, producing high powered experiments, and combating the replication crisis.

Characteristics of the intestinal flora of specific pathogen free chickens with age

Specific pathogen-free (SPF) experimental animals are recognized as standard laboratory animals in the fields of biomedical, animal husbandry and veterinary research and production. Intestinal flora plays a critical role in nutrient absorption, improving health and protecting the host from pathogens. We therefore explored the variation and maintenance of intestinal flora in SPF chicks in order to better understand the composition of intestinal microflorain SPF chickens, and provide reference for the study of intestinal flora of SPF experimental animals. Five chicks were randomly selected at each of 14, 28, and 42 days, and ceca were removed for DNA extraction. The Illumina Miseq platform was used for microbiome analysis of the V3–V4 region of the 16S rRNA gene. During the course of chick gut microbiome development, we observed major changes in diversity, especially between day 14 and day 28. Firmicutes, Proteobacteria, and Bacteroidetes were the main bacterial taxa, and Firmicutes increased significantly with age. The genus with the highest relative abundance was Lactobacillus, followed by Faecalibacterium. In addition, while abundance of Ruminococcaceae spp., Ruminococcus, and Blautia increased with age, Lactobacillus, Enterobacteriaceae spp., and Oscillospira decreased with age. Interestingly, the abundance of Faecalibacterium first increased and then decreased over time. The characteristics of SPF chicken gut flora at different ages establish a basis for the regulation of intestinal flora in the early stage of brooding, and also provide a theoretical foundation for controlling and preventing infections and poultry diseases in newborn chickens.

Mapping the Neural Substrates of Recent and Remote Visual Imprinting Memory in the Chick Brain.

Social attachment formed by filial imprinting in newborn chicks undergoes a process of memory consolidation that involves rearrangement of its neural storage substrates. In the first 3 h after imprinting it depends on the integrity of the intermediate medial mesopallium (IMM) and beyond that time on unidentified memory storage structures dubbed S’. To search for the S’ memory system in the chick brain, we mapped and compared patterns of activity induced by retrieval of filial attachment memory before and after this critical transition. Chicks were trained in the visual imprinting task, and their memory was reactivated by imprinting stimulus either 1 h (recent memory retrieval) or 24 h (remote memory retrieval) after the completion of training. Patterns of brain activity were mapped by in situ hybridization to mRNA of an immediate early gene c-fos. We also mapped c-fos expression induced by the first presentation of the imprinting stimulus. Memory retrieval triggered massive c-fos expression in the chick brain both 1 and 24 h after the end of training. These activity patterns mostly coincided with the c-fos expression induced by the first presentation of imprinting stimulus. However, in the hippocampus c-fos induction was observed only after the first exposure to imprinting stimulus but not after memory retrieval. In the IMM, medio-rostral nidopallium/mesopallium, and hyperpallium densocellulare c-fos activation was induced by retrieval of only the remote but not of the recent memory. These c-fos mapping data point to the candidate brain structures for systems reorganization of imprinting memory in chicks.

Detection of multidrug resistant Escherichia coli in the ovaries of healthy broiler breeders with emphasis on extended-spectrum β-lactamases producers

In the last few years, antimicrobial resistant (AMR) Escherichia coli have been detected in newborn chickens suggesting their vertical transmission from breeding birds to their offspring. However, little is known about the presence of AMR E. coli in the reproductive organs of broiler breeders. The aim of this study was to investigate the presence of E. coli in the ovaries of healthy broiler breeders and to study their antimicrobial resistance. Samples from broiler breeders (n = 80) collected from 80 different broiler breeder flocks were included in this study. Antibiotic susceptibility testing was performed using disk diffusion method according to Clinical and Laboratory Standards Institute guidelines. Minimal inhibitory concentrations (MICs) of five antimicrobial agents were determined by Etest. PCR and sequencing were used to detect the blaESBL genes. E. coli were detected in the ovaries of thirty seven out of 80 (46.25%) sampled flocks. High levels of resistance to various first-line antimicrobial agents were recorded in E. coli isolates. This study showed that 89.18% of E. coli isolates were multidrug resistant (MDR). Furthermore, MDR extended-spectrum β-lactamases (ESBL)-producing E. coli were detected in the ovaries of four different broiler breeder flocks. Molecular characterization revealed that three isolates harboured blaCTX-M-1 gene and one isolate expressed blaSHV-12 gene. In addition, one blaCTX-M-1 -producing E. coli co-harboured the blaTEM-1 gene. These findings would contribute to a better epidemiological understanding of MDR E. coli for improve existing preventive strategies in order to reduce the dissemination of antimicrobial resistance in the broiler production system.

Using automated controlled rearing to explore the origins of object permanence.

What are the origins of object permanence? Despite widespread interest in this question, methodological barriers have prevented detailed analysis of how experience shapes the development of object permanence in newborn organisms. Here, we introduce an automated controlled-rearing method for studying the emergence of object permanence in strictly controlled virtual environments. We used newborn chicks as an animal model and recorded their behavior continuously (24/7) from the onset of vision. Across four experiments, we found that object permanence can develop rapidly, within the first few days of life. This ability developed even when chicks were reared in impoverished visual environments containing no object occlusion events. Object permanence failed to develop, however, when chicks were reared in environments containing temporally non-smooth objects (objects moving on discontinuous spatiotemporal paths). These results suggest that experience with temporally smooth objects facilitates the development of object permanence, confirming a key prediction of temporal learning models in computational neuroscience.

Enhancement of bone mineral density and body mass in newborn chickens by in ovo injection of ionic-hydroxyapatite nanoparticles of bacterial origin

Using non-drug, non-surgical treatments for improving bone mineral diseases in newborn babies is an important topic for neonatologists. The present study introduces bacterial synthesized ionic nano-hydroxyapatite (Bio-HA) for the development of bone mineral density in the chicken embryo model. In vitro cytotoxicity analyses were demonstrated the optimal concentrations of Bio-HA compared to a chemically-synthesized hydroxyapatite (Ch-HA). Toxicity of Bio-HA on MCF-7 human cell lines was negligible at the concentrations less than 500 μg/mL whereas Ch-HA showed similar results at the concentrations less than 100 μg/mL. Therefore, concentrations at 50 μg/mL and 100 μg/mL were selected for in ovo injection of both materials into the fertilized eggs. The newly hatched chickens were sacrificed in order to monitor their serological factors, total body mass, bone mineral contents and bone mineral density. The results confirmed that Bio-HA increased the average body weight and bone mineral indices of chickens in comparison to the Ch-HA and negative controls (normal saline and intact groups). In view of the intact group, no liver or kidney damage occurred in the groups receiving Bio-HA which promises the effectiveness of these nanoparticles for the treatment of afterbirth bone mineral deficiency.

Effect of in ovo feeding of amino acids and dextrose solutions on hatchability, body weight, intestinal development and liver glycogen reserves in newborn chicks.

Early development of the digestive tract is crucial for achieving maximal growth and development of chickens. This study examined the effects of in ovo (IO) feeding of 0.70 mL of dextrose (10.00% and 20.00%) or amino acids solutions into the yolk sac at day 14 of incubation on small intestine histomorphometry and histomorphology, intestinal development, hatchability, body weight, and liver glycogen reserves in newborn chicks. Results showed body weight in amino acid fed hatchlings was higher than control and dextrose groups non-significantly, but hatchability was lower in amino acid group than others. Also, diameter of glycogen vacuoles in all IO treatment groups was more than control. Administration of exogenous dextrose and amino acids solutions into the yolk sac enhanced intestinal development by increasing the size and surface area of the villi and changed villi shape as well. It seems that dextrose or amino acids solutions could improve the intestinal villi development, while they did not affect finger-like villi in jejunum.

Folic Acid Reduced Triglycerides Deposition in Primary Chicken Hepatocytes.

Abdominal fat or fatty liver cause huge economic losses in the poultry industry, and nonalcoholic fatty liver disease (NAFLD) is also a global health issue in humans. More than 90% of de novo lipogenesis in humans and chickens is undertaken by the liver, which is proved to be full of lipids in new-born chickens. Folic acid was thought to have correlation with lipid metabolism. Primary hepatocytes from new-born chickens were employed as a natural model of early stage fatty liver in vitro and further to explore whether folic acid could prevent fatty liver in the current study. We found that folic acid addition reduced triglyceride deposition by suppressing de novo fatty acid synthesis and coordinately promoting triglyceride hydrolysis and exportation in primary chicken hepatocytes from new-born chickens. In addition, lipogenesis suppression was through the PI3K/AKT/SREBP pathway mediated by weakening insulin/IGF signal. Our data suggested that folic acid may be considered as a precautionary strategy for abdominal fat deposition in broilers or fatty liver in laying hens and humans. In addition, mechanism regulation also implied that an IGF2 inhibitor and PI3K inhibitor may be used for the NAFLD precautionary measure to reduce TG deposition.

Prenatal Sensory Stimulation Induces BDNF Gene Expression in the Brain and Potentiates the Development of Species-Specific Predisposition in Newborn Chicks.

We studied the effects of light and non-specific sound stimulation of domestic chick embryos on their filial preference as well as on the expression of two transcriptional factors c-Fos and Egr-1 and neurotrophin BDNF in the embryo brain. Prenatal light stimulation increased preference of the "natural" object, thus producing a priming effect. In the brain of E19 embryos, c-Fos and Egr-1 were expressed at a high basal level and neither light nor sound stimulation affected the number of cells expressing these factors. BDNF mRNA was also present in a number of brain areas of non-stimulated embryos, but light and sound stimulation enhanced the expression of BDNF mRNA in brain structures associated with filial imprinting. These findings suggest that BDNF is probably involved in the effects of prenatal priming on the development of species-specific behavior.

Short-term memory in habituation and dishabituation of newborn chicks' freezing response.

Cognitive models of habituation and dishabituation postulate that the latter is attributable to the perturbation of the model of the repeated stimulation stored in short-term memory (STM) by the occurrence of a new stimulus, called dishabituator. However, although both behavioral phenomena depend on STM, previous studies in Aplysia have found that dishabituation seems to require further steps of development of the STM system to emerge. Here, we addressed whether this is a universal condition for the appearance of the 2 forms of learning, namely whether dishabituation must necessarily follow habituation. To this aim, we studied habituation and dishabituation of the freezing response to a sudden acoustic stimulation in newly hatched chicks (1 day old vs. 3 days old). The results showed that in chicks, dishabituation was fully present a few hours after hatching, a pattern of results indicating that, in this precocial avian species, habituation and dishabituation share the same developmental trajectory and the underlying STM mechanisms are simultaneously operative soon after birth. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

In Ovo Administration of Innate Immune Stimulants and Protection from Early Chick Mortalities due to Yolk Sac Infection.

Omphalitis or yolk sac infection (YSI) and colibacillosis are the most common infectious diseases that lead to high rates of early chick mortalities (ECMs) in young chicks. Out of numerous microbial causes, avian pathogenic Escherichia coli (APEC) or extraintestinal pathogenic E. coli infections are considered the most common cause of these conditions. YSI causes deterioration and decomposition of yolk, leading to deficiency of necessary nutrients and maternal antibodies, retarded growth, poor carcass quality, and increased susceptibility to other infections, including omphalitis, colibacillosis, and respiratory tract infection. Presently, in ovo injection of antibiotics, heavy culling, or after hatch use of antibiotics is practiced to manage ECM. However, increased antibiotic resistance and emergence of "super bugs" associated with use or misuse of antibiotics in the animal industry have raised serious concerns. These concerns urgently require a focus on host-driven nonantibiotic approaches for stimulation of protective antimicrobial immunity. Using an experimental YSI model in newborn chicks, we evaluated the prophylactic potential of three in ovo-administered innate immune stimulants and immune adjuvants for protection from ECM due to YSI. Our data have shown >80%, 65%, and 60% survival with in ovo use of cytosine-phosphodiester-guanine (CpG) oligodeoxynucleotides (ODN), polyinosinic:polycytidylic acid, and polyphosphazene, respectively. In conclusion, data from these studies suggest that in ovo administration of CpG ODN may serve as a potential candidate for replacement of antibiotics for the prevention and control of ECM due to YSI in young chicks.

Bacterial communication between Lactobacillus spp. isolated from poultry in the inhibition of Salmonella Heidelberg-proof of concept.

Bacterial communication has become an increasingly studied topic aiming at the discovery of new products to aid the treatment of diseases for which conventional options do not work. The production of safe foods, free of pathogens, has been receiving increasing attention due to market demands for food products of high quality and free of residues. This study assessed the communication between Lactobacillus spp. during the in vitro inhibition of Salmonella Heidelberg (SH) and the impact an autoinducer produced by a strain of Lactobacillus plantarum has on communication in the normal microbiota and inhibiting SH in newborn chicks. For this purpose, the isolates of Lactobacillus spp. were isolated cloacal swabs of broilers and identified through biochemical and molecular assays and were obtained from broiler farms. They later had their inhibitory potential against SH stimulated after contact with the autoinducer. For assessing bacterial communication (quorum sensing) during the inhibition of SH by Lactobacillus spp., a spot on the lawn assay was conducted. For the in vivo, 75 one-day-old chicks were divided in 5 experimental groups: control with no treatment; treatment with a Lactobacillus spp. pool on the first day; treatment with autoinducer on the second day; treatment with Lactobacillus spp. on the first day; and autoinducer on the second day and treatment without autoinducer. The autoinducer was assessed through an SH count in the ceca of the birds. The autoinducer produced by the strain of L. plantarum proved to be efficient for communicating with the other Lactobacillus spp. isolates as previous contact with SH induced the production of an autoinducer capable of increasing inhibition of SH both in vitro (in average 132.73%) and in vivo, acting similarly to the Lactobacillus spp. pool (probiotic) by decreasing the SH count in the ceca (64%-24h, 42%-96h, and 46%-168h).

Regulation of filial imprinting and structural plasticity by mTORC1 in newborn chickens

Dysregulation of the mechanistic target of rapamycin complex 1 (mTORC1) signaling leads to memory deficits and abnormal social behaviors in adults. However, whether mTORC1 is involved in critical periods of early learning remains largely unexplored. Our study addressed this question by investigating imprinting, a form of learning constrained to a sensitive period that supports filial attachment, in newborn chickens. Imprinting to virtual objects and sounds was assessed after acute manipulations of mTORC1. To further understand the role of mTORC1 during the critical period, structural plasticity was analyzed using DiOlistic labeling of dendritic spines. We found that mTORC1 is required for the emergence of experience-dependent preferences and structural plasticity within brain regions controlling behavior. Furthermore, upon critical period closure, pharmacological activation of the AKT/mTORC1 pathway was sufficient to rescue imprinting across sensory modalities. Thus, our results uncover a novel role of mTORC1 in the formation of imprinted memories and experience-dependent reorganization of neural circuits during a critical period.

Harina De Ají De Ratón (Capsicum Mínimum) Como Anticoccidial Natural En Pollos De Engorde, Manabí-Ecuador

The use of natural plants for the control and treatment of diseases is an alternative to drugs and antibiotics. In the present test the hot pepper (Capsicum Minimum) was used as an anticoccidial in broiler chickens, it was taken as a reference to salinomycin (control) including 350g / ton, faced with three increasing levels of hot pepper flour (350, 500 and 1000g / ton). For the effect, 160 newborn chicks of the Cobb 500 line were used, distributed in four treatments and four repetitions under a completely randomized design, for the comparison of means the discrete Tukey test (0,05) was used, the data were analysed in the free version of InfoStat 2014 program. The variables analysed were weight, feed consumption, feed conversion, mortality and counting of Eimeria oocysts in faeces. Regarding the weight, the treatments presented statistical equality (p&gt; 0.05), numerically T3 registered the highest (2405.13 ± 43.33); and food conversion T1 and T2 (1.69), T3 and T4 (1.80) did not present differences between their means. Regarding mortality, T3 did not present deaths, not so, T4 (2.5%); T2 (5%) and T1 (10%); for oocyte count of E. Tenella and Acervulina T3 and T4 do not register eggs, T1 (3000 and 4500) and T2 (2500 and 1500, respectively). The hot pepper flour has anticoccidial activity in broiler chickens, its inclusion did not alter the productive parameters of the flock, which is recommended to be used during the fattening stage of the chickens or fowls.

Rapid plasticity attenuation soon after birth revealed by habituation in newborn chicks.

Habituation reflects a form of experience-dependent plasticity whereby the organism progressively learns to ignore the irrelevant information repeatedly encountered. Here, we measured the freezing response to a repeated loud noise in three groups of newborn chicks (Gallus gallus) of different ages (1-2 Day old, 2-3 Day old, and 3-4 Day old) to investigate the ontogeny of habituation in this avian species. Habituation was already present 1 Day after hatching, revealing that the neural mechanisms underlying this form of plasticity are immediately active. Unexpectedly, however, we also found that in the second Day of stimulation the amount of learning was significantly attenuated in chicks of 3-4 days of age as compared to the younger animals, thus showing that 24-48 hr of maturation were sufficient to reduce plasticity. While previous findings in other animal species have proved the existence of a precocious critical period of plasticity in early cortical areas by means of sensory deprivation, our results demonstrate that during the initial development of an intact avian brain also the degree of plasticity underlying a learning process like habituation is maximal soon after birth, and then is subject to a rapid attenuation.

The Development of Invariant Object Recognition Requires Visual Experience With Temporally Smooth Objects.

How do newborns learn to recognize objects? According to temporal learning models in computational neuroscience, the brain constructs object representations by extracting smoothly changing features from the environment. To date, however, it is unknown whether newborns depend on smoothly changing features to build invariant object representations. Here, we used an automated controlled-rearing method to examine whether visual experience with smoothly changing features facilitates the development of view-invariant object recognition in a newborn animal model-the domestic chick (Gallus gallus). When newborn chicks were reared with a virtual object that moved smoothly over time, the chicks created view-invariant representations that were selective for object identity and tolerant to viewpoint changes. Conversely, when newborn chicks were reared with a temporally non-smooth object, the chicks developed less selectivity for identity features and less tolerance to viewpoint changes. These results provide evidence for a "smoothness constraint" on the development of invariant object recognition and indicate that newborns leverage the temporal smoothness of natural visual environments to build abstract mental models of objects.

Maternal betaine administration modulates hepatic type 1 iodothyronine deiodinase (Dio1) expression in chicken offspring through epigenetic modifications

As a feed additive, betaine is widely used in livestock production for its ability to promote growth. Our previous studies had reported that maternal betaine supplementation altered hepatic metabolism in offspring. But it remains unknown whether and how maternal betaine modulates metabolism of thyroid hormones in the offspring chickens by epigenetic modification. In this study, one hundred and twenty Rugao yellow-feathered laying hens were randomly divided into two groups, and were fed basal diet with or without 0.5% betaine supplementation for 28 days. After that, all the hens were artificially inseminated and then four hundreds fertilized eggs were selected. After hatching, the newborn chicks were raised until 56 days old. Betaine fed female chicks showed significantly lower body weight and lower level of biologically active thyroid hormone in plasma compared to control group, which was associated with significantly decrease in expression of type 1 iodothyronine deiodinase (Dio1). Moreover, betaine also changed hepatic expression of betaine-homocysteine -S-methyltransferase (BHMT) and DNA methyltransferase 1 (DNMT1), which may contribute to hypermethylation of the Dio1 promoter. Interestingly, betaine treatments of hens caused none of these effects in male chicks except Dio1 expression. These results indicate that maternal betaine administration effects growth of offspring through differential modification of Dio1 gene methylation and expression in liver and this model of transgenerational effects may help elucidate the mechanisms of maternal effects arise in natural systems.