Noninvasive Assessment of Chicken Egg Fertility during Incubation Using HSSE-GC-MS VOC Profiling.

Volatile organic compounds (VOCs) carry crucial information about chicken egg fertility. Assessing the fertility before incubation holds immense potential for poultry industry efficiency. Our study used headspace sorptive extraction-gas chromatography-mass spectrometry to analyze egg VOCs before and during the initial 12 incubation days. A total of 162 VOCs were identified. Hexanal was significantly higher in unfertilized eggs, whereas compounds such as propan-2-ol, propan-2-one, and carboxylic acids were higher in fertilized eggs. Furthermore, the obtained multiple logistic regression model outperformed the partial least-squares-discriminant analysis (PLS-DA) model, demonstrating lower complexity and superior performance. Fertile eggs were accurately identified in the validation set in 68-75% of the cases during the initial 4 days, to 85 and 100% on days 6 and 8. Finally, hierarchical cluster analysis in fertilized eggs revealed the clustering of VOCs of the same chemical class, indicative of their shared biochemical origin. This suggests a promising direction for future research aimed at understanding the biological information embedded in VOCs and their relationship to biochemical processes during embryo development.

Non-destructive egg breed separation using advanced VOC analytical techniques HSSE-GC-MS, PTR-TOF-MS, and SIFT-MS: Assessment of performance and systems' complementarity.

Over the past decade, advanced analytical techniques have been utilized to examine volatile organic compounds (VOCs) in eggs. These VOCs offer valuable insights into factors such as freshness, fertility, the presence of cracks, embryo sex, and breed. In our study, we assessed three mass spectrometry-based systems (headspace sorptive extraction gas chromatography-mass spectrometry; HSSE-GC-MS, proton transfer reaction time-of-flight-mass spectrometry; PTR-TOF-MS; and selected ion flow tube mass spectrometry; SIFT-MS) to analyze and identify VOCs present in intact hatching eggs from three distinct breeds (Dekalb white layer, Shaver brown layer, and Ross 308 broiler). The eggs were sampled on incubation days 2 and 8, to identify VOCs that distinguish breeds irrespective of incubation day. VOC measurements were conducted on 15 eggs per breed by placing them together with PDMS-coated stir bars inside inert Teflon® air sampling bags. After an accumulation period of 2 h, the headspace was analyzed using PTR-TOF-MS and SIFT-MS, while the VOCs adsorbed onto the stir bars were analyzed using GC-MS for additional compound identification. Partial least squares discriminant analysis (PLS-DA) models were constructed for breed differentiation, and variable selection was performed. As a result, 111 VOCs were identified using HSSE-GC-MS, with alcohols and esters being the most abundant. The PLS-DA models demonstrated the efficacy of breed discrimination, with the HSSE-GC-MS and the PTR-TOF-MS exhibiting the highest balanced accuracy of 95.5 % using a reduced set of 11 VOCs and 5 product ions, respectively. The SIFT-MS model had a balanced accuracy of 92.8 % with a reduced set of 11 product ions. Furthermore, complementarity was observed between HSSE-GC-MS, which primarily selected higher molecular weight VOCs, and PTR-TOF-MS and SIFT-MS. A higher correlation was found for compound abundances between the HSSE-GC-MS and the PTR-TOF-MS relative to the SIFT-MS, indicating that the PTR-TOF-MS was better suited to quantify specific compounds identified by the HSSE-GC-MS. Finally, the findings support the presence of VOCs originating from both synthetic and natural sources, highlighting the ability of the VOC analysis systems to non-destructively perform quality control and reveal differences in management practices or biological information encoded in eggs.

Bibliographical Mapping of Research into the Relationship between In Ovo Injection Practice and Hatchability in Poultry.

Recent advances in poultry practice have produced new tools enabling the poultry industry to increase productivity. Aiming at increasing production quality, varying protocols of in ovo injection facilitate the introduction of exogenous substances into the egg to complement the nutrients that support embryonic development up to hatching, which are already available in the internal and external compartments. Due to embryonic sensitivity, adding any substance into the egg can be either advantageous or disadvantageous for embryonic survival and can influence hatch rates. Thus, understanding the relationship between poultry practices and production rates is the first step towards successful commercial application. This review aims to assess the influence on hatch rates of injecting different substances in ovo, including effects on embryo and chick health parameters where these are reported. Bibliographic mappings of co-authorship of citations, co-occurrence of keywords, and bibliographic coupling based on the in ovo injection technique and hatchability parameters were also performed. Using the Scopus database, 242 papers were retrieved, reviewed, and submitted for bibliographic mapping using the VOSviewer® software. This review provides a broad overview of just over 38 years' research on the subject, revealing that studies have significantly increased and peaked in 2020, being produced primarily by US researchers and published primarily in the journal Poultry Science. It also reveals that despite negative reports relating to some substances in the embryo, in ovo delivery of substances may possibly change the poultry industry for the better in terms of production rates (hatchability) and/or poultry health.

The fatty acid profile in the yolk and yolk sac from incubated goose eggs depends on the breeder age and laying period.

1. The study analysed the content of fatty acids in the lipids of the yolk and yolk sac of hatching eggs obtained from geese in four reproductive flocks and three laying periods at different incubation dates.2. A total of 1080 hatching eggs were used in the study (90 eggs from each age group in three laying periods). The geese were kept on one farm under the same conditions.3. On days 0, 16, 22, and 28 of incubation, the yolk/yolk sac was sampled. Saturated and unsaturated (mono- and poly-) fatty acids were determined, including myristic acid, palmitic acid, palmitoleic acid, margaric acid, stearic acid, oleic acid, linoleic acid, α-linolenic acid, behenic acid, eicosapentaenoic acid. The ratio of unsaturated to saturated fatty acids was calculated.4. Embryo fatty acid utilisation in eggs from different age groups of geese was similar. The fatty acid profile depended mostly on the laying period. The different proportions of fatty acids in the yolk during incubation indicated changes in the activity of various enzymatic processes in the membrane of the yolk sac of embryos from the beginning and at the end of the laying period.5. When analysing the interactions between the age of the parent flock and the laying period, the most significant effect on the fatty acid composition was found in fresh eggs. On d 16 of lay the myristic, stearic, linoleic, and behenic acids and PUFA; on d 22 of lay linolenic acid, and on day 28th palmitoleic and margaric acids were involved in this interaction.

Effects of Sanitizers on Microbiological Control of Hatching Eggshells and Poultry Health during Embryogenesis and Early Stages after Hatching in the Last Decade.

The sanitization of hatching eggs is the backbone of the hygienic-sanitary management of eggs on farms and extends to the hatchery. Poultry production gains depend on the benefits of sanitizers. Obtaining the maximum yield from incubation free of toxic sanitizers is a trend in poultry farming, closely following the concerns imposed through scientific research. The toxic characteristics of formaldehyde, the primary sanitizer for hatching eggs, are disappointing, but it is a cheap, practical and widely used antimicrobial. To overcome this shortcoming, multiple synthetic and natural chemical sanitizers have been, and continue to be, tested on hatching eggs. This review aims to evaluate the effects of different sanitizers on the microbiological quality of hatching eggshells and poultry health during embryogenesis and early stages after hatching.

Occurrence, Risk Factors, Serotypes, and Antimicrobial Resistance of Salmonella Strains Isolated from Imported Fertile Hatching Eggs, Hatcheries, and Broiler Farms in Trinidad and Tobago.

ABSTRACT: This cross-sectional study was conducted to determine the occurrence, risk factors, and characteristics of Salmonella isolates recovered from imported fertile broiler hatching eggs, hatcheries, and broiler farms in Trinidad and Tobago. Standard methods were used to isolate and characterize Salmonella isolates from two broiler hatcheries and 27 broiler farms in the country. The frequency of isolation of Salmonella was 0.0% for imported fertile hatching eggs (0 of 45 pools of 10 eggs each, i.e., 450 eggs), 7.6% for hatcheries (12 of 158 samples), and 2.8% for broiler farms (24 of 866 samples) (P = 0.006). Stillborn chicks at hatcheries had the highest prevalence of Salmonella (7 of 28 samples, 28.0%), whereas on broiler farms the cloacal swabs had the highest prevalence of Salmonella (15 of 675 samples, 2.2%). None of the 15 farm management and production practices investigated were significantly associated (P > 0.05) with the isolation of Salmonella. The predominant Salmonella serotypes were Kentucky (83.3%) and Infantis (62.5%) among hatchery and farm isolates, respectively. The disk diffusion method revealed frequencies of antimicrobial resistance (i.e., resistance to one or more agents) of 44.0% (11 of 25 isolates) and 87.5% (35 of 40 isolates) at hatcheries and broiler farms, respectively (P = 0.0002). Antimicrobial resistance among hatchery isolates was highest (28.0%) to doxycycline and kanamycin and was very high (>65%) among farm isolates to sulfamethoxazole-trimethoprim, gentamicin, ceftriaxone, kanamycin, and doxycycline. Multidrug resistance (MDR; i.e., resistance to antimicrobial agents from three or more classes) was exhibited by 4.0 and 85.7% of Salmonella isolates recovered from several environmental and animal sources at the hatcheries and farms, respectively (P < 0.0001). The high level of antimicrobial resistance and the presence of MDR among Salmonella isolates from broiler farms highlight the therapeutic implications and the potential for MDR strains to enter the food chain.

Spraying Hatching Eggs with Clove Essential Oil Does Not Compromise the Quality of Embryos and One-Day-Old Chicks or Broiler Performance.

The objective of this study was to evaluate whether sanitizing hatching eggs with clove essential oil in the preincubation phase affects broiler performance and influences the hatch window and quality of embryos and one-day-old chicks. Hatching eggs (n = 1280; mean weight = 58.64 ± 0.49 g) from a batch of 37-week-old broiler breeder hens of the CPK (Pesadão Vermelho) lineage were randomly distributed into four treatments in the preincubation phase. The treatments consisted of three different sanitization procedures (spraying with grain alcohol, spraying with clove essential oil, and fumigation with paraformaldehyde) and a control treatment (nonsanitized). The lengths of the embryos and one-day-old chicks (one of the parameters used to assess bird quality) were not significantly different among the treatments, with means of 15.30 ± 1.41 and 18.37 ± 0.76 mm, respectively. Body weight, body weight gain, feed consumption, and feed conversion rate in different rearing periods did not differ significantly among the treatments. However, there was a significant difference in the percentage of survivability during the initial period (1 to 28 days) among the treatments. In conclusion, clove essential oil treatment did not negatively affect the quality of embryos and one-day-old chicks or the performance of broilers.

Effects of coated hatching eggs obtained from old broiler breeders with chitosan on embryonic growth, hatching results and chick quality.

The aim of this research was to determine the effects of coating broiler hatching eggs with chitosan on egg quality, embryonic growth, hatching results and chick quality. Eggs obtained from old broiler breeder, aged 59 weeks, were used. Eggs were divided into two groups (coated with chitosan and uncoated) groups. Each group was divided into three groups according to the storage period. Eggs were stored for periods of 1, 4 and 7 days at 15°C and 80% humidity. Then, they were incubated. Storage period of eggs up to 7 days did not affect the egg quality, egg weight loss, embryo growth, chick properties and hatchability of fertile eggs. However, embryo development, relative residual yolk sac weight and relative chick weight were affected by coating eggs with chitosan. Interaction was not found between storage lengths and coating of eggs with chitosan for all examined parameters. The results of the present study suggested that hatching eggs obtained from old broiler breeder flocks were immersed into chitosan solution once and quickly have negative effect on the embryo weight, yolk sac absorption and relative chick weight. It has been observed that covering the hatching eggs with chitosan is not suitable in this respect even if immersion takes place once and in a short time.

Effects of a dry hydrogen peroxide disinfection system used in an egg cooler on hatchability and chick quality.

A sanitation method that could continually clean and disinfect the air and surfaces in a hatchery could provide a second layer of microbial reduction on top of routine cleaning and disinfection. A gaseous dry hydrogen peroxide (DHP) system has been used in other facilities for this purpose and could have potential for use in chicken hatcheries. Because the DHP is a true gas and can permeate through the entire hatchery space, contact with eggs during storage and incubation could potentially interfere with normal hatching processes. Therefore, the aim of this study was to evaluate the effects of the DHP system on hatching parameters and chick quality. A total of 3,960 hatching eggs were collected from an ∼40-week-old Ross 308 broiler breeder flock and distributed in 2 treatments: treated and nontreated. For the treated group, the egg cooler was cleaned, and 1 DHP generator was placed inside. Two other DHP generators were placed in the common area outside as well. Both areas were treated for 7 D before placement of eggs, and then eggs were collected and placed inside the cooler over a 4-day period. Eggs were then stored for an additional 3 D after the last collection. Dry hydrogen peroxide levels were recorded each day during storage. For the nontreated group, all DHP machines were removed from the cooler and external room, and the egg cooler was cleaned. Eggs were collected in the same way for the control group as the treated group. After storage, eggs were placed into a single stage Natureform incubator. The eggs exposed to DHP showed higher (P < 0.05) hatchability of fertile eggs and lower (P < 0.05) early embryonic dead than eggs from the nontreated group. No other parameters evaluated were different between groups. Based on this work, the DHP treatment of fertile eggs had no detrimental effect on any performance parameter, with potential positive effects seen on hatch of fertile eggs and early embryonic dead embryos.

Impact of egg disinfection of hatching eggs on the eggshell microbiome and bacterial load.

Disinfection of hatching eggs is essential to ensure high quality production of broilers. Different protocols are followed in different hatcheries; however, only limited scientific evidence on how the disinfection procedures impact the microbiome is available. The aim of the present study was to characterize the microbiome and aerobic bacterial load of hatching eggs before disinfection and during the subsequent disinfection steps. The study included a group of visibly clean and a group of visibly dirty eggs. For dirty eggs, an initial wash in chlorine was performed, hereafter all eggs were submitted to two times fumigation and finally spray disinfection. The eggshell microbiome was characterized by sequencing of the total amount of 16S rRNA extracted from each sample, consisting of shell surface swabs of five eggs from the same group. In addition, the number of colony forming units (cfu) under aerobic conditions was established for each disinfection step. The disinfection procedure reduced the bacterial load from more than 104 cfu (initially visibly clean eggs) and 105 cfu (initially visibly dirty eggs) to less than 10 cfu per sample after disinfection for both groups of eggs. The microbiome of both initially visibly clean and initially visibly dirty eggs had the highest abundances of the phyla Firmicutes, Proteobacteria and Bacteroidetes. Within the phyla Firmicutes the relative abundances of Clostridiales decreased while Lactobacillus increased from before to after final disinfection. In conclusion, the investigated disinfection procedure is effective in reducing the bacterial load, and by adding a chlorine wash for initially visibly dirty eggs, the microbiome of initially visibly clean and initially visibly dirty eggs had a highly similar microflora after the final disinfection step.

Evaluation of chlorine dioxide based product as a hatchery sanitizer.

Formaldehyde is commonly used to overcome contaminants introduced by hatching eggs or water supply in the hatcher cabinets. However, health risks associated with its use make economical alternatives important. This project evaluated a chlorine dioxide based product (CDBP) (0.3% concentrate) as a hatchery sanitizer in decontaminating microbial populations on the shell surface of hatching eggs (>18 d old), as well as its impact on hatchability and chick performance. Hatchers (0.20 m2) designed to hold approximately 50 eggs and equipped with circulation fans, heaters, and thermostats were used for the evaluation. For each of the 2 trials conducted, 450 hatching eggs were obtained and incubated in a common setter. Eggs used in trial 1 were floor eggs whereas in trial 2 nest eggs were used. On d 18 of incubation, eggs were removed from the setter, and viable eggs were randomly allocated to 9 hatchers. Pre-treatment egg rinse samples (10 eggs per hatcher) were collected for initial microbial analysis. Three hatchers were treated with CDBP and 3 hatchers with a formaldehyde based product (FBP). Three untreated hatchers served as control (C). Prior to hatch, 10 eggs/incubator, not previously rinsed, were used for post treatment microbial counts. The hatched chicks were reared until d 21 in floor pens with a common starter diet. For the CDBP treated eggs, hatchability and chick performance (weight gains, mortality, and FCR on d 7 and d 21) were similar to the other treatments. The application rate of CDBP evaluated in this study was not an effective antimicrobial alternative to formaldehyde for sanitizing hatching eggs in hatcher cabinets prior to hatch.

Presence of antimicrobial resistance in coliform bacteria from hatching broiler eggs with emphasis on ESBL/AmpC-producing bacteria.

Antimicrobial resistance is recognized as one of the most important global health challenges. Broilers are an important reservoir of antimicrobial resistant bacteria in general and, more particularly, extended-spectrum ß-lactamases (ESBL)/AmpC-producing Enterobacteriaceae. Since contamination of 1-day-old chicks is a potential risk factor for the introduction of antimicrobial resistant Enterobacteriaceae in the broiler production chain, the presence of antimicrobial resistant coliform bacteria in broiler hatching eggs was explored in the present study. Samples from 186 hatching eggs, collected from 11 broiler breeder farms, were inoculated on MacConkey agar with or without ceftiofur and investigated for the presence of antimicrobial resistant lactose-positive Enterobacteriaceae, particularly, ESBL/AmpC-producers. Escherichia coli and Enterobacter cloacae were obtained from the eggshells in 10 out of 11 (10/11) sampled farms. The majority of the isolates were recovered from crushed eggshells after external decontamination suggesting that these bacteria are concealed from the disinfectants in the egg shell pores. Antimicrobial resistance testing revealed that approximately 30% of the isolates showed resistance to ampicillin, tetracycline, trimethoprim and sulphonamides, while the majority of isolates were susceptible to amoxicillin-clavulanic acid, nitrofurantoin, aminoglycosides, florfenicol, neomycin and apramycin. Resistance to extended-spectrum cephalosporins was detected in eight Enterobacteriaceae isolates from five different broiler breeder farms. The ESBL phenotype was confirmed by the double disk synergy test and blaSHV-12, blaTEM-52 and blaACT-39 resistance genes were detected by PCR. This report is the first to present broiler hatching eggs as carriers and a potential source of ESBL/AmpC-producing Enterobacteriaceae for broiler chicks.