Simultaneous detection of infectious bronchitis virus and avian metapneumovirus genotypes A, B, and C by multiplex RT-qPCR assay in chicken tracheal samples in Ecuador.

Respiratory RNA viruses such as Infectious bronchitis virus (IBV) and Avian metapneumovirus (aMPV), which are characterized by generating both respiratory damage and adverse effects on reproductive organs, affect poultry production economically due to high mortality rate and decrease in egg production and quality. Particularly, aMPV has three genotypes that have been reported with greater frequency in chickens: aMPV-A, aMPV-B, and aMPV-C. The present study proposes the design of a multiplex RT-qPCR assay for the simultaneous diagnosis of the 3 genotypes of interest of aMPV and IBV, followed by testing of 200 tracheal samples of vaccinated chickens with respiratory symptoms and finally a phylogenetic analysis of the sequences found. The assay detected up to 1 copy of each viral genome. The standard curves showed an efficiency between 90 and 100% in the multiplex assay and inter- and intra-assay coefficients of variation of 0.363 and 0.459, respectively and inter- and intra-assay coefficients of variation of 0.363 and 0.459, respectively. 69.5% of samples were found positive alone or in coinfection. 114 samples were positive for IBV, 13 for aMPV-A and 25 for aMPV-B. RNA of aMPV-C was no detected. The most commonly found combination was aMPV-B and IBV within 6 samples, and the least common was aMPV-A and aMPV-B in coinfection in 2 samples. The assay was specific for amplification of the genomes of the studied respiratory viruses (IBV, aMPV-A, aMPV-B, aMPV-C) as no amplification was shown from other viral genomes (ChPV, CAstV, ANV, and FAdV) or from the negative controls. Partial genomic Sanger sequencing enabled to identify circulating vaccine-derived and wild-type strains of IBV and vaccine and vaccine-derived strains of aMPV-B. In conclusion, this newly developed multiplex RT-qPCR was shown to be able to detect individual infections as well as co-infections among the respiratory viruses investigated. It was demonstrated to be a reliable and efficient tool for rapidly and safely diagnosing these infections. Furthermore, this study represents the first report of aMPV strains in Ecuadorian poultry and demonstrates the circulation of aMPV-A, aMPV-B, and GI-13 IBV strains in unvaccinated chicken populations in the country. Thus, it highlights the importance of simultaneously identifying these pathogens in greater detail and on a regular basis in Ecuador.

Effect of Density and Lineage on Dorsal Surface Temperature, Performance, and Carcass Condemnation of Broiler Grillers.

The aims of this study were (i) to evaluate the effect of density, lineage, age, and time of day on dorsal surface temperature and (ii) to evaluate the effect of density and lineage on performance and carcass condemnations in broiler grillers. The evaluations were carried out in barns with the Dark House system, with two densities, 17 and 19 chickens/m2 and two lineages, Cobb and Ross. The dorsal surface temperature of the chickens was measured by infrared thermography at 7, 14, 21, 23, 25 and 27 days of age, four times a day. The average daily weight gain, feed conversion, mortality, partial carcass condemnations, as well as those due to arthritis and dermatosis were also evaluated. The highest dorsal surface temperatures were observed in Cobbs housed at a density of 17 chickens/m2, and in Ross housed at a density of 19 chickens/m2. Cobbs housed at a 17 chickens/m2 density showed the lowest feed conversion compared to Ross at the same density. Ross showed higher dorsal surface temperatures when compared to Cobbs at 14, 21, and 27 days. Cobbs showed higher percentages of partial carcass condemnation and arthritis compared to Ross. The higher density of broiler grillers in the Dark House system does not influence the dorsal surface temperature, performance, dermatosis, arthritis, and partial carcass condemnations.

Nutritional and technological potential of chicken feathers for the food industry.

1. The production of chicken meat has resulted in high volumes of byproducts, such as feathers, bones, skin, viscera, and feet. The structure of feathers is one of the most complex among vertebrates, with a central axis and lateral filamentary structures, providing rigidity, lightness, and flexibility. Chicken feathers are composed of proteins, lipids, and water, with the highest protein content, especially keratin, which is responsible for the material's rigidity.2. Industries still make little use of feathers, which are generally intended for the production of flour or organic fertilisers. These are low added value products, and discarded feathers can harm the environment.3. Keratin extraction techniques and resulting protein hydrolysates have been studied in chicken feathers. Acid, alkaline or enzymatic hydrolysis is the most commonly used method for obtaining molecules with functional properties such as antioxidant, antimicrobial, antihypertensive and antidiabetic activity.4. The development of keratin-based biodegradable films represents an area of interest for reducing the economic and environmental impacts caused by inappropriate disposal of feathers.

Effect of West Indian Bay Leaf (Pimenta racemosa) and Turmeric (Curcuma longa) Essential Oils on Preserving Raw Chicken Breasts.

Research background: While the use of chemical preservatives in meat may appear to be tremendously advantageous, they have long been purported to increase the risk of incidence of certain types of cancers. Consequently, many people have opted for minimally processed alternatives. This consumer shift has placed substantial pressure on the food industry to implement more natural alternatives to these synthetic preservatives in the meat industry. Research on plant extracts as potential agents for food additives is increasing. The bioactive components present in West Indian bay leaf and turmeric essential oils have a promising potential for use as novel, green preservatives in the meat industry. Experimental approach: Raw chicken breast samples (28 g) were each treated with different volumes (0.5, 1 and 1.5 mL) of the essential oil of West Indian bay leaf or turmeric or their mixture (1:1 to make up a final volume of 0.5, 1 and 1.5 mL). Physicochemical, microbiological and sensory evaluations were performed on the fresh and treated samples stored for 14 days at 4 °C. Results and conclusions: The West Indian bay leaf oil had a higher extraction yield and total phenolic content, while the turmeric oil had a higher total flavonoid content. The most effective treatments, compared to the control, significantly (p<0.05) minimized the pH increase by 13.9 % (1.5 mL bay leaf oil), reduced texture loss by 44.8 % (1.5 mL oil mixture) and reduced protein loss by 98.9 % (1 mL bay leaf oil). Most treated samples had reduced microbial loads, with the turmeric oil showing the highest efficacy against lactic acid bacteria, yeasts and moulds. Treated samples had significantly higher (p<0.05) sensory scores than the control on the final day of storage, with the 1.5 mL oil mixture proving to be the most effective, as the storage life of the chicken breast sample was extended by 6 days. Novelty and scientific contribution: This study has shown for the first time that the essential oil from turmeric and West Indian bay leaf can extend the shelf life of raw chicken breast and highlights the potential of the oil as natural preservative agents in lieu of synthetic alternatives.

New insights into chicken processing wastewater treatment: the role of the microalgae Parachlorella kessleri on nitrogen removal.

Microalgal Technologies have recently been employed as an alternative treatment for high nitrogen content wastewater. Nitrogen is an essential nutrient for microalgae growth, and its presence in wastewater may be an alternative source to synthetic medium, contributing to a circular economy. This study aimed to investigate the effect of using Parachlorella kessleri cultivated in wastewater from the thermal processing of chicken meat. Experiments were performed to obtain the ideal sampling site, inoculum dosage, and contact time. P. kessleri had better growth in the sample from the settling basin. Nitrogen removal was 95% (0,15 mg TNK/107 cells) in 9 days, and the final nitrogen concentration was lower than 20 mg/L, and the nitrate concentration was lower than 1 mg/L. However, during the third cycle in the kinetic assay, there was a decline in the microalgae growth, occasioned by the accumulation of nitrite (38,4 mg/L) in the inside of the cell. The study demonstrated that nitrogen concentration is directly related to the cell growth of the algae. Parachlorella kessleri efficiently removed nitrogen from chicken meat thermal processing wastewater and is a potential option for tertiary treatment and valorisation of such effluent as a nitrogen source.

Molecular Characterization of the Chicken Parvovirus Based on VP1 Gene Circulating in Brazilian Chicken Flocks.

Parvovirus infection affects several animal species, especially young animals. In birds, parvovirus infection has been described in Muscovy ducks, turkeys, and chickens, all of which had enteric diseases characterized by diarrhea. Chicken parvovirus (ChPV) has been detected in poultry around the world in animals affected by enteric problems, showing dwarfism, cloacal pasting, and diarrhea. In Brazil, ChPV was detected in chickens affected by diarrhea fifteen years ago. However, the genetic characteristics of ChPV circulating in chicken flocks were not determined. Therefore, the aim of the present investigation was to determine the genetic characteristics of the VP1 gene from ChPV detected in chickens affected by enteric diseases in Brazil. For this purpose, a molecular approach was used. Specific primers were designed to flank the complete VP1 gene of ChPV and amplify it using PCR. The amplified products from samples of chickens with enteric diseases were sequenced, and 22 complete CDs of the VP1 gene were obtained. These samples, compared to the ABU-P1 sequence, showed 17 sequences with high nucleotide (NT) similarity of 92.7-97.4% and amino acid (AA) similarity of 94.8-99.5% associated with Runting and Stunting syndrome (RSS); there were also five samples associated with hens with diarrhea with unusual jejunal dilatation (JD) that had less similarity than the RSS sequences (NT of 86.5% and AA of 93-93.1%). The phylogenetic analysis determined four groups. Group I had sequences from Korea. The second group included sequences from Korea, China, and Brazil (not included in this work). The third group had studied RSS sequences grouped with the ABU-P1 strain and sequences from China and the United States. Finally, the sequences from JD were clustered in a separate group with a bootstrap of 100%, a group that was denoted as group IV, and included sequences from China. RDP4 and SimPlot analysis showed one point of recombination with the sequences of group III ChPV in the JD sequences. Herein, we show that circulating strains of ChPV exhibit genetic differences in the VP1 gene in Brazilian chicken flocks. Nevertheless, more studies are needed to determine the probability of a new genetic group of ChPV based on the analysis of the complete genome.

Detection and molecular characterization of chicken parvovirus and chicken megrivirus in layer breeders affected by intestinal dilatation syndrome.

RESEARCH HIGHLIGHTS: IDS presented pathognomonic dilatation of the jejunum up to Meckel's diverticulum.IDS caused weight loss, decreased egg production, and increased culling and mortality.Chicken parvovirus (ChPV) was consistently detected through PCR assays.Chicken megrivirus (ChMV) was consistently detected through viral metagenomics.

miR-137 confers robustness to the territorial restriction of the neural plate border.

The neural plate border (NPB) of vertebrate embryos is segregated from the neural plate (NP) and epidermal regions, and comprises an intermingled group of progenitors with multiple fate potential. Recent studies have shown that, during the gastrula stage, TFAP2A acts as a pioneer factor in remodeling the epigenetic landscape required to activate components of the NPB induction program. Here, we show that chick Tfap2a has two highly conserved binding sites for miR-137, and both display a reciprocal expression pattern at the NPB and NP, respectively. In addition, ectopic miR-137 expression reduced TFAP2A, whereas its functional inhibition expanded their territorial distribution overlapping with PAX7. Furthermore, we demonstrate that loss of the de novo DNA methyltransferase DNMT3A expanded miR-137 expression to the NPB. Bisulfite sequencing revealed a markedly elevated presence of non-canonical CpH methylation within the miR-137 promoter region when comparing NPB and NP samples. Our findings show that miR-137 contributes to the robustness of NPB territorial restriction in vertebrate development.

Insecticidal activity of fluralaner (Exzolt®) administered to Gallus gallus domesticus against triatomines (Hemiptera, Reduviidae, Triatominae).

BACKGROUND: Triatoma infestans, Triatoma brasiliensis, Triatoma pseudomaculata and Rhodnius prolixus are vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. Chickens serve as an important blood food source for triatomines. This study aimed to assess the insecticidal activity of fluralaner (Exzolt®) administered to chickens against triatomines (R. prolixus, T. infestans, T. brasiliensis and T. pseudomaculata). METHODS: Twelve non-breed chickens (Gallus gallus domesticus) were randomized based on weight into three groups: negative control (n = 4); a single dose of 0.5 mg/kg fluralaner (Exzolt®) (n = 4); two doses of 0.5 mg/kg fluralaner (Exzolt®) (n = 4). Nymphs of 3rd, 4th and 5th instars of R. prolixus, T. infestans, T. brasiliensis and T. pseudomaculata (all n = 10) were allowed to feed on chickens before treatment, and at intervals of 1, 7, 14, 21, 28, 35 and 56 days after treatment, with insect mortality determined. RESULTS: Treatment with two doses of fluralaner showed higher insecticidal efficacy against R. prolixus, T. infestans and T. brasiliensis compared to the single-dose treatment. Similar insecticidal efficacy was observed for T. pseudomaculata for one and two doses of fluralaner. Insecticidal activity of fluralaner (Exzolt®) against triatomine bugs was noted up to 21 and 28 days after treatment with one and two doses of fluralaner, respectively. CONCLUSIONS: The results demonstrate that treatment of chickens with fluralaner (Exzolt®) induces insecticidal activity against triatomines for up to 28 days post-treatment, suggesting its potential use as a control strategy for Chagas disease in endemic areas.

Taste Preferences in Broilers: Effect of Age, Delivery Matrix, and Number of Chickens per Pen on Selection and Consumption Behaviour.

Due to substantial differences between studies, the understanding of avian taste perception remains incomplete. Also, studies on chicken taste preferences have mainly focused on measuring consumption differences, neglecting consumption behaviour patterns. This study investigated how age, the compound delivery matrix, and the number of birds per pen affect broiler chicken preferences and consumption behaviour, and established their preference values for four taste compounds. Ninety-six one-day-old male broiler chickens (Ross 308) were divided into two age groups (initial: days 7-23; final: days 26-42), with two compound delivery matrices (water or ground wheat) and two numbers of birds (one or two chickens per pen), following a 2 × 2 × 2 factorial design. Four taste compounds (sucrose, monosodium glutamate (MSG), L-lysine, and calcium carbonate) were tested at different concentrations. Preferences were assessed at 2, 4, and 8 h post-test, along with recording various behavioural parameters. Initial-stage birds showed higher (p < 0.001) preference values, time of approach (TA), number of bouts (NB), duration of bouts (DB), and number of pecks (NP) than final-stage birds. Birds exposed to a water matrix also exhibited higher (p < 0.001) preference and NB, while those exposed to a ground wheat matrix showed a higher (p < 0.001) NP. Pens with a pair of birds had a higher (p < 0.003) 2 h preference, TA, NB, DB, and NP, than pens with a single chicken. Chickens showed significant preference values for 100 mM sucrose at 2 h (p = 0.025), 150 mM MSG at 4 h (p = 0.026) and 8 h (p = 0.013), and 300 mM MSG at 2 h (p = 0.013). We concluded that all the variables evaluated influence broilers' taste preferences and consumption behaviour during selection tests. Future studies should prioritize including chickens in the initial stage of the production cycle, testing them in pairs or groups, and delivering compounds via a liquid matrix.

Eggshell translucency: its relationship with specific gravity and eggshell color and its influence on broiler egg weight loss, hatchability, and embryonic mortalities.

Eggshell quality is among the most important factors affecting hatchability in broiler breeders, and therefore several methods for its assessment are available in the poultry industry. Among them, eggshell translucency has received special attention in recent years due to its connection with ultrastructural disorganization of the shell layers. However, there is very limited data on the impact of translucency on hatching eggs and on the possible links between this trait and specific gravity (SG) or shell color. Thus, our study investigated associations and interactions between eggshell translucency, SG, and color on incubation parameters of eggs from the same breeding flock (Ross 308AP, 51 wk of age). To this end, light and dark eggs within 5 different SG categories (≥1.065, 1.070, 1.075, 1.080, and ≤1.085) were selected from 15,976 eggs, graded into 3 translucency scores, and later incubated to evaluate egg weight loss, hatchability and embryonic mortalities. In general, translucency scores were evenly distributed within SG categories (χ2 [8, N = 1,138] = 13.67, P = 0.090) and color (χ2 [2, N = 1,138] = 4.93, P = 0.084). No interactions between eggshell translucency and SG or between translucency and color were found for the analyzed variables. An interaction was observed between SG and eggshell color for the variable egg weight loss, where the light-shelled eggs, in most SG categories lost more weight throughout incubation than dark eggs. Eggshell translucency affected egg weight loss, hatchability, and embryonic mortality on 11 to 18 d of incubation, with highly translucent eggs showing the worst results. At the same time, eggs with SG lower than 1.070 displayed the greatest weight loss, lowest hatchability, and highest contamination. We found no influence of eggshell color on weight loss or hatchability, but light-shelled eggs exhibited higher late embryonic mortality. Together, these data suggest that despite its effects on certain hatching parameters, shell translucency bears no relationship to SG or color.

Prevalence and risk factors of Campylobacter jejuni and Campylobacter coli in fresh chicken carcasses from retail sites in Bogotá, Colombia.

Campylobacter is one of the most common causes of foodborne gastroenteritis. The objective of this study was to estimate the prevalence and risk factors associated with Campylobacter jejuni and Campylobacter coli species in fresh chicken carcasses for human consumption from farmers' markets and small food stores in seven localities of Bogotá, Colombia. Ninety-one samples of fresh chicken carcasses were collected from farmers' markets and small food stores at seven localities in Bogotá. Samples were tested for Campylobacter using the real-time polymerase chain reaction (real time PCR) and isolation by plating. To analyze possible risk factors associated with Campylobacter spp. contamination in retail chicken carcasses, information was collected using a structured questionnaire and a univariate logistic regression analysis (α = 0.05) was used. Forty-two positive samples were obtained for Campylobacter spp., given a prevalence of 46.2%, of which 54.8% were to C. jejuni, 9.52% to C. coli and 35.7% to joint contaminations. C. jejuni was the most prevalent species. Risk factors found included poor cleanliness, in frequency of disinfection, type of establishment, and direct contact of chickens with other food. This study is the first report in the country on the prevalence and risk factors of Campylobacter in retail chicken.

Predicting Risk of Ammonia Exposure in Broiler Housing: Correlation with Incidence of Health Issues.

The study aimed to forecast ammonia exposure risk in broiler chicken production, correlating it with health injuries using machine learning. Two chicken breeds, fast-growing (Ross®) and slow-growing (Hubbard®), were compared at different densities. Slow-growing birds had a constant density of 32 kg m-2, while fast-growing birds had low (16 kg m-2) and high (32 kg m-2) densities. Initial feeding was uniform, but nutritional demands led to varied diets later. Environmental data underwent selection, pre-processing, transformation, mining, analysis, and interpretation. Classification algorithms (decision tree, SMO, Naive Bayes, and Multilayer Perceptron) were employed for predicting ammonia risk (10-14 pmm, Moderate risk). Cross-validation was used for model parameterization. The Spearman correlation coefficient assessed the link between predicted ammonia risk and health injuries, such as pododermatitis, vision/affected, and mucosal injuries. These injuries encompassed trachea, bronchi, lungs, eyes, paws, and other issues. The Multilayer Perceptron model emerged as the best predictor, exceeding 98% accuracy in forecasting injuries caused by ammonia. The correlation coefficient demonstrated a strong association between elevated ammonia risks and chicken injuries. Birds exposed to higher ammonia concentrations exhibited a more robust correlation. In conclusion, the study effectively used machine learning to predict ammonia exposure risk and correlated it with health injuries in broiler chickens. The Multilayer Perceptron model demonstrated superior accuracy in forecasting injuries related to ammonia (10-14 pmm, Moderate risk). The findings underscored the significant association between increased ammonia exposure risks and the incidence of health injuries in broiler chicken production, shedding light on the importance of managing ammonia levels for bird welfare.

Phenotypic divergence between broiler and layer chicken lines is regulated at the molecular level during development.

BACKGROUND: Understanding the molecular underpinnings of phenotypic variations is critical for enhancing poultry breeding programs. The Brazilian broiler (TT) and laying hen (CC) lines exhibit striking differences in body weight, growth potential, and muscle mass. Our work aimed to compare the global transcriptome of wing and pectoral tissues during the early development (days 2.5 to 3.5) of these chicken lines, unveiling disparities in gene expression and regulation. RESULTS: Different and bona-fide transcriptomic profiles were identified for the compared lines. A similar number of up- and downregulated differentially expressed genes (DEGs) were identified, considering the broiler line as a reference. Upregulated DEGs displayed an enrichment of protease-encoding genes, whereas downregulated DEGs exhibited a prevalence of receptors and ligands. Gene Ontology analysis revealed that upregulated DEGs were mainly associated with hormone response, mitotic cell cycle, and different metabolic and biosynthetic processes. In contrast, downregulated DEGs were primarily linked to communication, signal transduction, cell differentiation, and nervous system development. Regulatory networks were constructed for the mitotic cell cycle and cell differentiation biological processes, as their contrasting roles may impact the development of distinct postnatal traits. Within the mitotic cell cycle network, key upregulated DEGs included CCND1 and HSP90, with central regulators being NF-κB subunits (RELA and REL) and NFATC2. The cell differentiation network comprises numerous DEGs encoding transcription factors (e.g., HOX genes), receptors, ligands, and histones, while the main regulatory hubs are CREB, AR and epigenetic modifiers. Clustering analyses highlighted PIK3CD as a central player within the differentiation network. CONCLUSIONS: Our study revealed distinct developmental transcriptomes between Brazilian broiler and layer lines. The gene expression profile of broiler embryos seems to favour increased cell proliferation and delayed differentiation, which may contribute to the subsequent enlargement of pectoral tissues during foetal and postnatal development. Our findings pave the way for future functional studies and improvement of targeted traits of economic interest in poultry.

Effects of cold stress on physiologic metabolism in the initial phase and performance of broiler rearing.

This study aimed to investigate the effects of 8 h of cold stress (18 °C) every day in broiler chicks during the first 7 days of rearing on crop filling analysis, yolk sac consumption, digestive and immune organs weights, and physiological metabolism at seven days and performance between 1 and 35 days. Cobb500 male broiler chickens (n = 274) were randomly assigned to two treatments. The treatments consisted of varying environmental temperatures during the first week post-housing. Chicks were reared at a thermoneutral temperature (32 °C) or under cold stress (18 °C) for 8 h/day during the first week, and both groups were subsequently reared at a thermoneutral temperature for 8-35 days. The thermoneutral group reached 90% full crop after 48 h of housing (P < 0.05), while the cold-stressed group had more empty crops at 2 h and 48 h after housing (P < 0.05). The chick cloacal temperature was not affected by the treatments (P > 0.05). Additionally, the treatment did not affect serum amylase and corticosterone levels, feed intake, body weight gain, or feed conversion ratio (P > 0.05, while the cold-stressed group had elevated heterophil/lymphocyte count at day 7 (P < 0.05). The thermoneutral group showed higher viability (%) at 7 and 35 days and a higher production factor at 35 days (P < 0.05). Broiler chickens under cyclic cold stress experienced decreased yolk sac absorption during the first week and increased feed intake and feed conversion ratio after 35 days of rearing. Viability was also lower in the cold-stressed group. An appropriate strategy to minimize these adverse effects is to rear the chicks in a thermoneutral environment during the first week.

Characterization of the genetic diversity, structure, and admixture of 7 Chilean chicken breeds.

The Mapuche fowl is an autochthonous breed raised in Chile and represents an important zoogenetic resource for the local economy. This study aimed at investigating the genetic diversity, relationship and population structure of 96 local Chilean chickens derived from 3 ecotype of Mapuche fowl (Kollonka, Ketro, and Kollonka de aretes), 2 ecotype Chilean (Trintre, Cogote pelado) and 2 breeds (Light Brahma and Barred Plymouth Rock) using 12 microsatellite markers. In total, 113 alleles were detected in all populations, with a mean of 7.6 alleles per population. In all population chicken breeds, the observed and expected heterozygosity ranged from 0.91 to 0.98 and from 0.69 to 0.79. Furthermore, all populations showed significant deviations from Hardy-Weinberg expectations. Across each population, the global heterozygosity deficit (FIT) was -0.174, population differentiation index (FST) was 0.073, and the global inbreeding of individuals within breed (FIS) was -0.267. The phylogenetic relationships of chickens were examined using neighbor-joining trees constructed at the level of population. The highest Nei's standard genetic distance value of 0.559 was observed between Barred Plymouth Rock and Light Brahma, whereas the minimum value (0.099) was found between Kollonka and Trintre. The neighbor-joining tree constructed at population level revealed 2 main clusters, with Light Brahma, Barred Plymouth Rock, Ketro and Kollonka de aretes in 1 cluster, and Kollonka, Trintre and Cogote pelado breeds in the second cluster. Based on the results of the STRUCTURE analysis, the most likely number of clustering of the population evaluated was at K = 3, with Light Brahma and Barred Plymouth Rock breeds forming their own distinct clusters, while Kollonka, Ketro, Kollonka de aretes, Trintre and Cogote pelado breeds clustered together. This study represents the first report of genetic diversity in these populations in Chile. These results can be used as baseline genetic information for genetic conservation program, for instance, to control inbreeding and to implement further genetic studies in local Chilean chickens.

Antivenom production in chicken against Sind krait (Bungarus sindanus) venom and its efficacy assessment using different immunoassays / Investigação e desenvolvimento de imunoensaios comparativos para produção de imunoglobulinas contra veneno da cobra (Bungarus sindanus) em aves

Present study aimed for detection, purification, quantification of Sind Krait (Bungarus sindanus) antivenom from chicken eggs and to determine extracted antivenom efficacy in mice. Hens' three groups were immunized by sub-lethal doses of Sind Krait venom with adjuvant paraffin oil+lecithin. Booster doses were injected subcutaneously on pectorals muscles at multiple sites after every two weeks upto eight weeks. Antibodies-IgY produced against Sind Krait venom was purified form eggs' yolk by precipitation method with PEG-6000. Purified antivenom-IgY protein contents were quantified by Nanodrop-photometer, purity accessed by SDS-PAGE, specificity checked by Ochterloneys method and titer estimated by indirect ELISA. Antivenom efficacy was assessed in albino mice. Purified antivnom-IgY exhibited single protein band 180-190 kDa on SDS-PAGE under non-reduced condition and two-bands 63 - 65 kDa and 22 - 25 kDa correspondingly under-reduced condition. Immunodiffusion exhibited sharp precipitation lines of immune-complex (venom and extracted-IgY). In all groups (G1, G2 and G3) antivenom level sharply increase from 3rd to 4th week and maintained thereafter. G2 and G3 presented high titer upto 1:2048 dilutions, while G1 showed upto 1:1024 dilutions, as tested by indirect ELISA. In neutralization assay ED50 dose of G2 and G3 obtained antivenom was 400.23 µg/mice for more than twofold LD50 dose of venom and 100% protection was at 508.84 µg/mice that completely neutralized highly lethal dose of venom. But G1 ED50 was 405.66 µg/mice and provides 100% protection at 554.21 µg/mice. Extracted antivenom, against Sind Krait venom were highly pure, and with high neutralization capacity were produced successfully from eggs yolk first time in Pakistan.

O presente estudo teve como objetivo a detecção, purificação, quantificação do soro antiofídico em ovos de galinha e determinar a eficácia do soro antiofídico extraído em camundongos. Os três grupos de galinhas foram imunizados por doses subletais de veneno de cobra Sindhi krait com óleo de parafina adjuvante + lecitina. As doses de reforço foram injetadas por via subcutânea nos músculos do peito em vários locais após cada duas semanas até oito semanas. Anticorpos-IgY produzidos contra o veneno de Krait foram purificados da gema de ovo pelo método de precipitação com PEG-6000. Os conteúdos de proteína antiveneno-IgY purificada foram quantificados por fotômetro nanodrop enquanto a pureza foi acessada por SDS-PAGE. A especificidade do antiveneno-IgY foi verificada pelo método de Ochterloneys e o título foi estimado por ELISA indireto. A eficácia do antiveneno foi avaliada em camundongos albinos. A IgY purificada exibiu uma única banda de proteína 180-190 KDa em SDS-PAGE sob condição não reduzida e duas bandas 63 - 65 KDa e 22 - 25 KDa correspondentemente condição sub-reduzida. A imunodifusão exibiu linhas de precipitação nítidas de imunocomplexo (veneno e IgY extraída). Em todos os grupos (G1, G2 e G3) o nível do soro aumentou acentuadamente da 3ª para a 4ª semana e manteve-se a partir daí. G2 e G3 apresentaram títulos elevados até diluições de 1:2048, enquanto G1 apresentou diluições de até 1:1024, testado por ELISA indireto. No ensaio de neutralização, a dose ED50 de antiveneno G2 e G3 obtida foi de 400,23ug/camundongos para mais de duas vezes a LD50 e 100% de proteção foi @ 508,84ug/camundongos que neutralizam completamente a dose altamente letal de veneno. Mas G1 ED50 foi de 405,66ug/camundongos e forneceu 100% de proteção @ 554,21ug/camundongos. Antivenenos extraídos, contra veneno de Bungarus sindanus eram altamente puros, seu título de anticorpos e capacidade de neutralização foram produzidos com sucesso a partir de gemas de ovos de galinhas imunizadas pela primeira vez no Paquistão.

Extraction and quantification of antivenomous antibodies in chicken eggs against scorpion venom / Extração e quantificação de anticorpos entivenenos em ovos de galinha contra veneno de escorpião

Avian-derived IgY is thought to be the best therapy for scorpion bites concerning low-level side effects. The present study analyzed a hypothesis about the neutralization of scorpion venom Androcotonus australis through antibodies produced in the egg yolks of chickens. The venom used for inoculation was obtained from Androctonus australis (yellow fat-tailed scorpion) from southern Punjab, Pakistan. The lethal dose of LD50 against scorpion venom was calculated in chickens and mice. Safe doses were given to egg-laying chickens to produce IgY antibodies. The antivenom IgY antibodies were extracted from the egg yolks of immunized chicken using the polyethylene glycol (PEG) method. Moreover, IgY was confirmed through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the Ouchterlony double immunodiffusion assay test. The antibody titers were evaluated by the enzyme-linked immunosorbent assay (ELISA). The neutralisation capacity of extracted anti-scorpion antibodies was tested on mice. The calculated LD50 of scorpion venom for chicken and mice was 4 mg/kg and 2.5 mg/kg, respectively. SDS-PAGE and Ouchterlony double immunodiffusion confirmed the presence of IgY against scorpion venom. The maximum titer value of specific IgY produced against scorpion venom was 3.5 ug/ml. A concentration of 220 ul/LD50 was effective to neutralize 1 mg of scorpion venom. It is suggested that IgY obtained from egg yolks is safe against targeted venom and can be used as an effective alternative to equine IgG antibodies against scorpion envenoming.

Acredita-se que a IgY derivada de aves seja a melhor terapia para picadas de escorpião em relação aos efeitos colaterais. O presente estudo teve como objetivo analisar uma hipótese sobre a neutralização do veneno do escorpião Androcotonus australis através de anticorpos produzidos na gema de ovos de galinhas. O veneno usado para inoculação foi obtido de Androctonus australis (escorpião amarelo de cauda gorda) do sul de Punjab, Paquistão. A dose letal de LD50 contra veneno de escorpião foi calculada em galinhas e camundongos. Doses seguras foram dadas a galinhas poedeiras para produzir anticorpos IgY. Os anticorpos antiveneno IgY foram extraídos das gemas de ovos de galinhas imunizadas pelo método do polietilenoglicol (PEG). Além disso, a IgY foi confirmada por eletroforese em gel de poliacrilamida e dodecil sulfato de sódio (SDS-PAGE) e pelo teste de imunodifusão dupla de Ouchterlony. Os títulos de anticorpos foram avaliados pelo ensaio imunoenzimático (ELISA). A capacidade de neutralização dos anticorpos anti-escorpião extraídos foi testada em camundongos. A LD50 calculada do veneno de escorpião para galinhas e camundongos foi de 4 mg/kg e 2,5 mg/kg, respectivamente. SDS-PAGE e imunodifusão dupla Ouchterlony confirmaram a presença de IgY contra veneno de escorpião. O valor máximo do título de IgY específico produzido contra veneno de escorpião foi de 3,5 ug/ml. Uma concentração de 220 ul/LD50 foi considerada eficaz para neutralizar 1 mg de veneno de escorpião. Sugere-se que a IgY obtida da gema do ovo seja segura contra o veneno direcionado e possa ser usada como uma alternativa eficaz aos anticorpos IgG equinos contra o envenenamento por escorpiões.

Unraveling the biological potential of chicken viscera proteins: a study based on their enzymatic hydrolysis to obtain hydrolysates with antioxidant properties.

Chicken meat production has increased over the years, leading to a proportional increase in waste generation, which often contains high levels of proteins, such as viscera. Therefore, this study aimed to investigate the enzymatic hydrolysis of chicken viscera proteins as a strategy to value solid waste from the poultry industry. The hydrolysates were characterized for their antioxidant properties and molecular weight distribution. Additionally, the enzymatic hydrolysis process was scaled up from 125 mL flasks with 50 mL of protein solution to 3 L using a 6 L bioreactor. The enzymatic hydrolysis of chicken viscera proteins using a binary mixture of proteases (85.25 U/mL of each enzyme, Alcalase and Flavourzyme, totaling 170.5 U/mL) resulted in an increase of up to 245% in 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, 353% 2,2-diphenyl-1-picryl-hydrazyl (DPPH) in radical scavenging, 69% in Ferric Reducing Antioxidant Power Assay (FRAP) and 146% in total reducing capacity (TRC). The antioxidant properties of the protein hydrolysates are preserved during the scale-up of enzymatic hydrolysis. Protein fractions smaller than 5 kDa showed the highest ABTS and DPPH radical scavenging activities, while fractions greater than 30 kDa showed the best results for the FRAP method.

Subclinical circulation of Chicken infectious anaemia virus- A Sero-molecular study.

Chicken infectious anaemia-an important immunosuppressive viral disease of chicken-gained much attention in the recent past. Based on huge mortality and production loss observed in the fast-growing poultry sector, the present study aimed to find out the current status of the chicken infectious anaemia virus (CIAV), among chicken flocks in the Punjab state of India by sero-molecular study. The sera from the blood samples were tested for anti-CIAV antibodies by indirect ELISA and also compared with haematological parameters. DNA from sero-positive samples underwent PCR amplification, sequencing and phylogenetic analysis of the most conserved genomic region (VP3 gene) to detect viraemia in asymptomatic birds. The serological study using indirect ELISA showed a high sero-positivity of 77.27% in chicken flocks. Additionally, the present study also revealed the high molecular evidence (72.54%) of CIAV in apparently healthy birds. Genetic analysis showed that all CIAVs have conserved VP3 genes without any nucleotide substitutions, indicating presence of CIAV and its subclinical circulation among apparently healthy flocks. The wide distribution of CIAV among birds may be the reason for huge mortality and production loss. Further, it is suggested that studies be conducted to find out the co-involvement of CIAV with other immunosuppressive microbial agents and the immunosuppressive effect of CIAV in apparently healthy birds. Also, its role in vaccine failure and outbreaks of various other avian diseases needs to be explored.