Effect of egg yolk-based extender and seminal plasma removal on sperm viability of cooled donkey semen / Efeito do diluente à base de gema de ovo e remoção do plasma seminal na viabilidade espermática do sêmen refrigerado de jumento

Developing effective cooled semen protocols is essential to increase pregnancy rates and reproductive efficiency in donkeys. This study aimed to evaluate the effect on sperm kinetic parameters and membrane integrity in cooled donkey semen diluted with defined milk proteins extender with 1% or 2% of egg yolk and the removal of seminal plasma. Twenty-four ejaculates from six jackasses were collected. Each ejaculate was divided into four aliquots that were diluted in extender with 1% (EY1) or 2% (EY2) egg yolk. One sample from each group was centrifuged, seminal plasma was removed (CEY1, CEY2 groups, respectively), and the samples were then refrigerated at 5 °C for 24 h. Fresh and cooled semen samples were assessed for sperm motility, morphology, and plasma membrane integrity. Total motility, progressive motility, sperm kinetic parameters, or live sperm cells were not statistically different when semen was cooled with an extender supplemented with 1% or 2% of egg yolk. Seminal plasma removal does not affect total motility or sperm kinetic parameters. However, progressive motility decreased (P<0.05) when semen was extended with 2% of egg yolk and seminal plasma was removed. Membrane integrity was affected (P<0.05) in centrifuged samples. In conclusion, the obtained results suggest that there is no difference in sperm kinetics and membrane integrity when 1% or 2% of egg yolk was added to the Equiplus extender. Also, the removal of seminal plasma by centrifugation did not have any beneficial effect on cooled donkey semen. Further studies are needed to relate these results with in vivo fertility tests with cooled donkey semen.(AU)

O desenvolvimento de protocolos de sêmen resfriado eficazes é essencial para aumentar as taxas de prenhez e eficiência reprodutiva em jumentos. O objetivo desse estudo foi avaliar o efeito do diluente à base de proteínas do leite com 1 ou 2% de gema de ovo sobre os parâmetros cinéticos do sêmen e integridade da membrana em sêmen resfriado de jumento, com ou sem a remoção do plasma seminal. Vinte e quatro ejaculados de seis jumentos foram coletados. Cada ejaculado foi dividido em quatro alíquotas e diluído em diluente com 1% (EY1) ou 2% (EY2) de gema de ovo. Uma amostra por grupo foi centrifugada e o plasma seminal removido (grupos CEY1 e CEY2, respectivamente). Os pellets foram novamente ressuspendidos nas mesmas concentrações e diluentes. Em seguida, as quatro alíquotas foram refrigeradas a 5°C por 24 horas. Amostras de sêmen fresco e refrigerado foram avaliadas quanto à motilidade espermática e integridade da membrana plasmática. Motilidade total, motilidade progressiva, parâmetros de cinética espermática ou células espermáticas vivas não apresentaram diferença significativa quando o sêmen foi resfriado com diluente suplementado com 1% ou 2% de gema de ovo. A remoção do plasma seminal não afetou a motilidade total ou os parâmetros de cinética espermática; entretanto, a motilidade progressiva diminuiu (P<0,05) quando o sêmen foi diluído com 2% de gema de ovo e o plasma seminal removido. Nas amostras centrifugadas, a integridade da membrana foi afetada (P<0,05). Em conclusão, os resultados sugerem que não há diferença na cinética espermática e na integridade da membrana quando 1% ou 2% de gema de ovo são adicionados ao diluente Equiplus e a remoção do plasma seminal por centrifugação não teve nenhum efeito benéfico no resfriamento de sêmen de jumento. Mais estudos são necessários para relacionar esses resultados com testes de fertilidade in vivo com sêmen resfriado em jumentos.(AU)

Production and evaluation of egg derived hot start antibodies

BACKGROUND: Hot start can greatly improve specificity, sensitivity and yield of PCR. Non-specific amplification can occur in PCR when reaction mixture is prepared at room temperature, because Taq DNA polymerase is active and the primers can hybridize non-specifically. Hot start Taq DNA polymerases remain inactive at room temperature and are activated after heating at 95°C preventing non-specific amplification. Monoclonal antibodies against Taq DNA polymerase is the first line of reagents used for turn on regular Taq DNA polymerase into Hot start one. The goal of this research was to produce and evaluate Hot Start antibodies derived from chicken eggs. RESULTS: We performed affinity purification of yolk immunoglobulin (IgY) and obtained polyclonal Hot Start antibodies. The yield of specific antibodies was 0.36 mg per egg or 0.2% of total yolk antibodies. The protocol for real time measurement and Hot start IgY activity assessment was developed. We found that Hot start IgY can reversibly block Taq DNA polymerase activity at 50°C and have no negative impact neither on the Taq DNA polymerase activity after denaturation nor on the reverse transcriptase. We estimated that 1.0 µg of Hot start IgY effectively blocks 5 U activity of Taq DNA polymerase. CONCLUSIONS: Egg derived Hot Start polyclonal antibodies are the cheapest source of Hot start antibodies, from one immune egg we can isolate 0.36 mg IgY, this quantity is enough for producing 1800 U activity of Hot start Taq DNA Polymerase.

Nutritional quality and functional lipids in the free-range egg yolks of Brazilian family farmers / Calidad alimenticia y lípidos funcionales en las yemas de gallina del campo de agricultores de familias Brasileñas

ABSTRACT Nowadays consumers are more concerned about food quality as well as how food is produced. Moreover, consumer demand for more natural, nutritious, safer to eat, and environmentally sustainable food has drawn attention to alternative breeding systems, such as a free-range system. Hens' eggs represent a rich source of important nutrients, including lipids and carotenoids. A detailed characterization of the nutritional and lipid profile of free-range chicken eggs from family farms was carried out. The chemical composition of the free-range egg yolks confirmed the advantages of this farming system, as a relevant source of unsaturated fatty acids and carotenoids, functional compounds in the human diet. These findings can help improve income generation, food supplementation, and consolidation of the family farming system.

RESUMEN Cada vez más los consumidores se preocupan por la calidad de los alimentos, así como tambien de como ellos son producidos. Además, la demanda de los consumidores por alimentos más naturales, nutritivos, más seguros para el consumo y ambientalmente sostenibles ha llamado la atención a sistemas de cría de alternativa, como el sistema de gallina del campo. Los huevos de gallinas representan una fuente rica de sustancias nutritivas importantes, incluyendo lípidos y carotenoids. Fue realizada una caracterización detallada de los nutrientes y del perfil lipídicos de huevos de gallinas del campo de agricultores familiares. La composición química de yemas de gallinas del campo confirmó las ventajas de este sistema de producción, como una fuente importante compuestos funcionales como ácidos grasos insaturados y carotenoides, en la dieta humana. Además, estos hallados pueden ayudar a mejorar la generación de ingreso, la suplementación de alimentos y la consolidación de sistema de agricultura familiar.

Qualidade de ovos brancos comercializados em diferentes estabelecimentos no município de Paragominas – PA / Quality of whit eggs marketed in different establishments in the municipality of Paragominas – PA

Objetivou-se verificar a qualidade de ovos brancos de acordo com os aspectos internos e externos comercializados em diferentes estabelecimentos do município de Paragominas - PA. Os ovos foram coletados de aleatoriamente em diferentes pontos comerciais. As análises consideradas no estudo, foi de: integridade e aparência da casca e peso do ovo, altura da câmara de ar, peso, altura e diâmetro da gema para o cálculo de índice da gema, altura e peso do albúmen e cor da gema. Os dados foram submetidos à análise de variância e ao teste de Tukey. Os ovos dos grandes supermercados apresentaram melhores índices de qualidade interna e os piores resultados quanto a peso, trincas visíveis e sujeiras. Os ovos de pequenos supermercados apresentaram o maior índice de trincas não visíveis e os de médio porte os menores índices de sujeiras e trincas.

A case of anhidrotic ectodermal dysplasia presenting with pyrexia, atopic eczema, and food allergy

Anhidrotic ectodermal dysplasia (AED) is a rare hereditary disorder with a triad of sparse hair, dental hypoplasia, and anhidrosis. Here we report a case of AED with food allergy and atopic eczema. The patient was a 11-month-old boy admitted to our hospital with pyrexia for 2 weeks. He presented with a history of dry skin, eczema, and food allergy to egg. On clinical examination, his body temperature was 38.8°C, with dry skin and eczema almost all over the body, sparse eyebrows, and scalp hair. Laboratory investigations and physical examination did not show any evidence of infection. Radioallergosorbent test was positive to egg yolk, egg white, ovomucoid, milk, house dust, and house dust mite. As the child did not sweat despite the high fever, we performed the sweat test which revealed a total lack of sweat glands. Genetic examination revealed a mutation of the EDA gene and he was diagnosed as AED. His pyrexia improved upon cooling with ice and fan. His mother had lost 8 teeth and her sweat test demonstrated low sweating, suggestive of her being a carrier of AED. Atopy and immune deficiencies have been shown to have a higher prevalence in patients with AED. Disruption of the skin barrier in patients with AED make them more prone to allergic diseases such as atopic eczema, bronchial asthma, allergic rhinitis and food allergy. Careful assessment of the familial history is essential to differentiate AED when examining patients with pyrexia of unknown origin and comorbid allergic diseases.

Hyperresponsiveness to Boiled Egg Yolk in Early Life Leads to Prolonged Egg Allergy

Hen's egg is the most common allergen in IgE-mediated food allergy among children in Japan. Although the majority of patients with egg allergy can eat heated egg yolk safely because of its low allergenicity, severely allergic patients show an immediate-type reaction to heated egg yolk. We hypothesized that patients with hyperresponsiveness to boiled egg yolk may have difficulty in acquiring tolerance to egg. The purpose of this study was to examine the prognosis of patients with hyperresponsiveness to boiled egg yolk. Data from 121 patients with egg allergy who underwent oral food challenge (OFC) with boiled egg yolk between January 2012 and December 2013 were analyzed retrospectively. The proportion of patients who could consume heated whole egg 3 years after OFC was 15.4% in the OFC-positive group and 75.8% in the OFC-negative group. Hyperresponsiveness to boiled egg yolk in early life might lead to prolonged egg allergy in children. This finding might aid in the selection of an appropriate population requiring practical immunotherapy.

Clinical and microbiological effects of egg yolk antibody against Porphyromonas gingivalis as an adjunct in the treatment of moderate to severe chronic periodontitis: a randomized placebo-controlled clinical trial

PURPOSE: To evaluate the clinical and microbiological effects of the local use of egg yolk immunoglobulin against Porphyromonas gingivalis (anti-P.g. IgY) as an adjunct to scaling and root planing (SRP) in the treatment of moderate to severe chronic periodontitis. METHODS: This was a randomized, placebo-controlled, double-blind trial involving 60 systematically healthy patients with moderate to severe chronic periodontitis. Subjects (n=20/group) were randomly assigned to receive SRP combined with subgingival irrigation of anti-P.g. IgY and anti-P.g. IgY mouthwash, subgingival irrigation of 0.2% chlorhexidine and 0.2% chlorhexidine mouthwash, or subgingival irrigation of placebo and placebo mouthwash for 4 weeks. Probing pocket depth, clinical attachment level, bleeding on probing, and the plaque index were evaluated at baseline and at 4 weeks. Subgingival plaque, gingival crevicular fluid, and saliva were simultaneously collected for microbiological analysis. RESULTS: Our results showed that anti-P.g. IgY mouthwash was as effective as chlorhexidine at improving clinical parameters over a 4-week period. All the groups showed a significant reduction in levels of P.g. at 4 weeks. No significant difference was observed in the test group when compared to placebo regarding the reduction in the levels of P.g. Anti-P.g. IgY significantly suppressed the numbers of red complex bacteria (RCB) in subgingival plaque and saliva in comparison with placebo. No adverse effects were reported in any of the subjects. CONCLUSIONS: Within the limitations of the study, the present investigation showed that passive immunization with anti-P.g. IgY may prove to be effective in the treatment of chronic periodontitis due to its ability to improve clinical parameters and to reduce RCB. No significant differences were found between the anti-P.g. IgY and placebo groups in the reduction of P.g.

Age-related changes of yolk precursor formation in the liver of laying hens / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

A rapid decline in egg production of laying hens begins after 480 d of age. Such a rapid decrease results predominantly from the ovarian aging, accompanied by endocrine changes, decreased yolk synthesis and accumulation, and the reduction in follicles selected into the preovulatory hierarchy. In this study, hens at 90, 150, 280, and 580 d old (D90, D150, D280, and D580, respectively) were compared for yolk precursor formation in the liver to elucidate effects of aging on laying performance. The results showed that liver lipid synthesis increased remarkably in hens from D90 to D150, but decreased sharply at D580 as indicated by the changes in triglyceride (TG) levels. This result was consistent with the age-related changes of the laying performance. The levels of liver antioxidants and total antioxidant capacity decreased significantly in D580 hens and the methane dicarboxylic aldehyde in D580 hens was much higher than that at other stages. The serum 17β-estradiol level increased from D90 to D280, but decreased at D580 (P<0.05). The expression of estrogen receptor α and β mRNAs in the liver displayed similar changes to the serum 17β-estradiol in D580 hens. Expressions of the genes related to yolk precursor formation and enzymes responsible for fat acid synthesis were all decreased in D580 hens. These results indicated that decreased yolk precursor formation in the liver of the aged hens resulted from concomitant decreases of serum 17β-estradiol level, transcription levels of estrogen receptors and critical genes involved in yolk precursor synthesis, and liver antioxidant status.

Effect of superoxide dismutase, catalase, and glutathione peroxidase supplementation in the extender on chilled semen of fertile and hypofertile dogs

This study investigated the correlation between oxidative stress status and key canine sperm parameters and the effect of addition of a superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) combination in egg yolk tris-citrate glucose (EYT-G) extender on semen during 10 days of storage at 4℃. Ten Boxer dogs were divided into two groups, fertile (F) and hypofertile (H), depending on pregnancy and live birth rate status in the previous year. Semen evaluation was performed on the day of collection (D0) and after 5 (D5) and 10 (D10) days of cooled storage. Sperm motility, kinetic parameters, and DNA integrity were assessed. A correlation between oxidative status and key semen parameters in both F and H groups was observed. Total and progressive motilities were significantly higher in the treated (SOD, CAT, and GPx addition) versus control groups at D10 in both F and H groups, and at D5 in the H group. DNA integrity was significantly higher in both treated groups (H and F) at D5 and D10. In conclusion, the addition of SOD, CAT, and GPx in the extender allows preservation of semen quality for up to 10 days of storage at 4℃ in both fertile and hypofertile dogs.

Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs / Teste de ligação espermática à membrana perivitelínica da gema de ovo de galinha como avaliação funcional do sêmen de cães

During fertilization, spermatozoa interact with the zona pellucida (ZP) through the binding between the acrosome and proteins 2 and 3 (ZP2 and ZP3). The perivitelline membrane of chicken egg yolk is homologous to the mammalian ZP3, which allows the binding of sperm of several species. The aim of this study was to standardize and evaluate the efficiency of sperm-binding to the perivitelline membrane of chicken eggs as a functional method for canine semen evaluation. For this purpose, nine post-thaw sperm samples were used, which were divided into two aliquots: the first was kept in water bath at 37ºC (live sample) and the second was submitted to cold shock to induce cellular damage (dead sample). The two aliquots were mixed on five proportions, corresponding to 0%, 25%, 50%, 75%, and 100% of viable cells, and the binding test was performed by analyzing the number of spermatozoa bonded to the perivitelline membrane by means of computerized assessment of sperm motility (CASA) or conventional microscopy. Additionally, samples were submitted to sperm motility analysis, evaluation of plasmatic and acrosomal membrane integrity, and sperm mitochondrial activity. The sperm-binding test to the perivitelline membrane of chicken egg yolk was considered a feasible sperm analysis test for both fertilizing capacity and overall sperm attributes evaluation, mainly when the analysis is performed by a conventional microscope, which expands its practicality to the majority of canine reproduction laboratories.(AU)

Durante a fecundação, os espermatozoides interagem com a zona pelúcida (ZP) por meio da ligação entre o acrossomo e as proteínas 2 e 3 (ZP2 e ZP3). A membrana perivitelínica da gema de ovo de galinhas é homóloga à ZP3 de mamíferos, possibilitando a ligação espermática de diversas espécies. Este trabalho padronizou e avaliou a eficiência do teste de ligação espermática à membrana perivitelínica da gema de ovo de galinhas como avaliação funcional do sêmen de cães. Para tal, foram utilizadas nove amostras seminais previamente criopreservadas. Cada amostra foi dividida em duas alíquotas: a primeira foi mantida em banho-maria à 37ºC (vivos) e a segunda submetida a choque térmico com o intuito de induzir dano celular (mortos). As duas alíquotas foram misturadas, correspondendo a 0, 25, 50, 75 e 100% de células viáveis. As amostras foram avaliadas quanto ao número de espermatozoides ligados à membrana perivitelínica por meio da análise computadorizada da motilidade (CASA) ou microscopia convencional. Ademais, as amostras foram avaliadas quanto à motilidade espermática, integridade das membranas acrossomal e plasmática e atividade mitocondrial espermática. O teste de ligação espermática à membrana perivitelínica de ovos de galinha foi considerado um teste de análise seminal exequível tanto para avaliar a capacidade fecundante dos espermatozoides como atributos seminais gerais, especialmente quando realizado em microscopia convencional, expandindo sua praticidade para a maioria dos laboratórios de análise de sêmen canino.(AU)

Effect of Syzygium cumini leaves on laying hens performance and egg quality

ABSTRACT The aim of this study was to evaluate the effect of Syzygium cumini leaves (SCL) in laying hens diets on productive performance, egg quality and lipid oxidation of yolk. A total of 108 Hysex White laying hens were distributed in a completely randomized design with three treatments and six replicates of six birds each. The treatments consisted of SCL inclusion at dietary levels of 0, 5 and 10 g/kg. There was no significant effect of SCL inclusion on feed intake, laying percentage, weight and egg mass, feed conversion ratio, Haugh units, specific gravity, percentage of yolk, albumen and egg shells and shell thickness. However, the inclusion of SCL significantly influenced the yolk color and yolk lipid oxidation measured by TBARS values. Yolk color increased and TBARS values ​​decreased with the inclusion of SCL. The inclusion of SCL in laying hens diets improves pigmentation and lipid stability of yolk.

Evaluation of Neck Lymph Node Metastasis on Contrast-Enhanced Ultrasound: An Animal Study

OBJECTIVES: To assess the performance of contrast-enhanced ultrasound (CEUS) intended to differentiate hyperplastic from malignant neck lymph nodes in an animal model. METHODS: Twenty-four New Zealand rabbits were randomly divided into two groups: neck lymph node metastasis group (12 rabbits) and reactive hyperplastic lymph node group (12 rabbits). Tongue VX2 carcinoma with cervical lymph node metastasis was induced in 12 rabbits by injecting VX2 carcinoma suspension into the left tongue submucosa. Hyperplastic neck lymph nodes were induced by injecting egg yolk in the submandibular region of the rabbits in hyperplastic group. CEUS were performed in both groups before and after intravenous administration of SonoVue. The site, number, echogenicity, longitudinal and transverse nodal dimensions, patterns of enhancement of the neck lymph nodes were observed and recorded. RESULTS: In both groups only one lymph node was found in the left (tumor) side of the neck. CEUS found 12 of 12 metastatic lymph nodes in metastasis group, and diagnosed 11 of 12 lymph nodes as metastatic. Histopathologic analysis revealed metastatic lesions in all 12 rabbits, each with one lymph node, and all 12 lymph nodes in hyperplastic group is inflammation lymph nodes. All 12 cases in the hyperplastic group showed centripetal homogeneous enhancement while in the metastasis group one case showed centripetal homogeneous enhancement, three cases showed centrifugal heterogeneous enhancement, and eight cases showed diffused heterogeneous enhancement. Only one lymph node was dissected on the left side of the neck in each rabbit in both groups. CONCLUSION: CEUS can play a role in discriminating metastatic from hyperplastic lymph nodes in head and neck carcinoma.

Synthetic pigments for Japanese quail fed diets with sorghum / Pigmentantes sintéticos para codornas japonesas alimentadas com rações à base de sorgo

Corn is the major energy ingredient in diets, and many ingredients have been tested aiming to replace it. In this regard, sorghum stands out for its chemical profile similar to that of corn. However, because it is low in carotenoids, its inclusion in diets reduces the egg yolk color pigmentation, which can be corrected by the addition of synthetic pigments. This study aimed to evaluate the performance and egg quality of Japanese quail (Coturnix japonica) supplemented with red (canthaxanthin) and yellow (apo-ester 10%) synthetic pigments. A total of 150 quail at 70 days of age were distributed according to the experimental diet [R1: corn-based control diet (DC); R2: sorghum-based diet (DS); R3: DS + yellow; R4: DS + yellow + red; and R5: DS + red] with six replications and five birds per experimental unit, for 28 days. Performance, egg quality, yolk color, and feed cost characteristics were evaluated. Regression equations were estimated for the effects of color as a function of periods, and treatment means were compared by Tukey's test at 0.05 probability. There was no significant effect (P>0.05) of additives on the quail productive traits. However, addition of synthetic pigments significantly improved the chromatic profile of the yolks (P<0.05). Inclusion of synthetic pigments improves yolk color, but should be evaluated according to market demands.(AU)

O milho é o principal ingrediente energético nas rações. A fim de substituí-lo, vários ingredientes foram testados. Nesse sentido, destaca-se o sorgo, pois apresenta perfil bromatológico semelhante ao do milho. No entanto, por ser deficiente em carotenoides, sua inclusão na ração reduz a pigmentação da gema do ovo, o que pode ser corrigido por meio da adição de pigmentos sintéticos. Objetivou-se, com este estudo, avaliar o desempenho zootécnico e a qualidade dos ovos de codornas japonesas (Coturnix japonica) suplementadas com os pigmentantes sintéticos vermelho (cantaxantina) e amarelo (apoéster 10%). Foram utilizadas 150 codornas japonesas com 70 dias de idade, distribuídas de acordo com a ração experimental (R1: ração referência à base de milho; R2: ração à base de sorgo (RS); R3: RS + amarelo; R4: RS + amarelo + vermelho; R5: RS + vermelho), com seis repetições e cinco aves por unidade experimental, durante 28 dias. Foram avaliadas as características de desempenho, qualidade dos ovos, cor da gema e custo das rações. Equações de regressão dos efeitos da cor em função dos períodos foram estimadas, e as médias de tratamento foram comparadas pelo teste de Tukey com 0,05 de probabilidade. Não houve efeito significativo (P>0,05) dos aditivos sobre as características produtivas das codornas. Entretanto, a adição de pigmentantes sintéticos melhorou significativamente o perfil cromático das gemas dos ovos de codornas (P<0,05). A inclusão de pigmentantes sintéticos melhora a cor das gemas, porém deve ser avaliada de acordo com as exigências de mercado.(AU)

Avaliação da qualidade interna de ovos de consumo pela pesquisa do teor de aminas bioativas / Evaluation of the internal quality of commercial eggs by searching of the bioactive levels

Com o objetivo de avaliar a qualidade dos ovos de consumo pela pesquisa dos níveis de aminas bioativas, foram coletados, pelos serviços de inspeção oficiais, 224 amostras de ovos provenientes de cinco regiões distintas do estado de Minas Gerais, durante o período de um ano. As aminas biogênicas (putrescina, cadaverina, feniletilamina, histamina e tiramina) e as poliaminas (espermidina e espermina) foram pesquisadas por cromatografia líquida de alta eficiência e detecção ultravioleta (CLAE/UV) após derivação pré-coluna com cloreto de dansila. Os resultados demonstraram que a putrescina estava presente, em baixas concentrações, em todas as amostras de gema e de albúmen. As demais aminas também foram detectadas, porém em menor frequência, e a espermina somente foi encontrada em uma amostra de albúmen. Foi concluído que os ovos de consumo produzidos no estado de Minas Gerais não são uma fonte considerável de poliaminas, importantes para o crescimento e a proliferação celular, e os baixos teores de aminas biogênicas, formadas pela descarboxilação de aminoácidos por enzimas bacterianas, não representam riscos à saúde do consumidor, o que indica que o ovo apresenta boa qualidade, tomando por base o critério de aminas bioativas.

In order to evaluate the quality of commercial eggs by searching the bioactive amine levels, 224 samples of eggs from the five regions of Minas Gerais State were collected during one year by the official inspection service. The biogenic amines (putrescine, cadaverine, phenylethylamine, histamine and tyramine) and the polyamines (spermidine and spermine) were determined by high performance liquid chromatography with ultraviolet detection (HPLC/UV) and pre-column derivatization with dansyl chloride. The results demonstrated the presence of putrescine in all samples of yolk and albumen, but in low concentrations. The other amines were also detected, however, with a lower frequency, and spermine was found only in one sample of albumen. It was concluded that the commercial eggs produced in Minas Gerais State are not a considerable source of polyamines, important for growth and cell proliferation; and low levels of biogenic amine, formed by decarboxylation of amino acids by bacterial enzymes, do not represent risks to consumer health, indicating that it has good quality, based on the bioactive amine criterion.

Hens immunized with live attenuated Salmonella strains expressing virulence-associated genes in avian pathogenic Escherichia coli passively transfer maternal antibodies to chicks

We investigated whether maternal antibodies (mAbs) elicited by dams immunized with recombinant vaccine candidates against avian pathogenic Escherichia coli (APEC) can passively confer protective immunity to chicks. In the present study, pBP244 plasmids carrying selected antigens of APEC were transformed into Salmonella Typhimurium JOL912, which was used as a vaccine candidate against APEC. The hens were immunized with the vaccine candidates using prime or booster doses. The levels of IgG and sIgA specific to the selected antigens increased significantly following prime immunization. To evaluate the persistence of passively transferred mAbs, the levels of IgY and IgA were determined in egg yolks and whites, respectively. The eggs from the immunized group showed consistently increased levels of IgY and IgA until week 16 post-laying (PL) and week 8 PL, respectively, relative to the control group. The presence of mAbs was observed in chicks that hatched from the hens, and titers of plasma IgY were consistently raised in those from the immunized hens by day 14 post-hatching. Further, chicks from the immunized hens were protected from challenge with a virulent APEC strain, whereas those from non-immunized hens showed acute mortality.

Qualidade interna de diferentes tipos de ovos comercializados durante o inverno e o verão / Internal quality of different types of eggs sold during the winter and summer

Objetivou-se no presente estudo avaliar a qualidade interna de ovos brancos e vermelhos, comercializados durante o inverno e o verão, no estado de São Paulo. Para tal análise, utilizaram-se ovos classificados como tamanho grande, e a avaliação da qualidade interna realizada por meio das seguintes variáveis: massa média do ovo, gravidade específica, unidade Haugh, índice de gema, coloração da gema e incidência de partículas estranhas nos ovos. O delineamento experimental foi inteiramente ao acaso, em fatorial 2 x 2 (cor da casca x épocas do ano), sendo as cores da casca: branca e vermelha, e as épocas do ano: inverno e verão. Verificou-se que, no verão, foram obtidos os menores valores de massa média do ovo, de unidade Haugh e de índice de gema, o que evidencia maior perda da qualidade do produto, provavelmente devido às elevadas temperaturas, comparativamente ao inverno. Entretanto, durante o inverno, foram observados ovos contendo discos germinativos fecundados. Conclui-se que os ovos comercializados no Vale do Ribeira, SP, apresentaram baixa qualidade e, portanto, estavam impróprios ao consumo e à industrialização.

The aim of this study was to evaluate the internal quality of white and red eggs marketed during the winter and summer in the state of São Paulo. For this analysis, eggs classified as big were used, with the evaluation of the internal quality carried out according the following variables: Average egg mass, specific gravity, Haugh unit, yolk index, yolk color and strange particles in eggs. The experimental design was completely randomized in a 2 x 2 factorial (shell color x seasons), with shell color: white and red; and seasons: winter and summer. Through the results we found that summer had the lowest values of average egg weight, Haugh unit and yolk index, showing a greater loss of product quality, probably due to the high temperatures compared to the winter. However, during the winter we observed fertilized eggs containing germ discs. We conclude that eggs sold in the Ribeira Valley - SP are of low quality and therefore unfit for consumption and industrialization.

Detection of anticancer and apoptotic effect of the produced IgYs against the three extracellular domain of human DR5 protein

TNF alpha cytokine family in the body plays divers' roles in the cellular events such as cell proliferation, differentiation, necrosis, septic shock and apoptosis. In response to TNF therapy, several cell signaling pathways activated in cells which in different manners can lead to apoptosis or necrosis. However induction of apoptosis is depended on one of its important members, TRAIL and its receptors that will be followed by apoptosis activity. Tumor necrosis factor-related apoptosis-inducing ligand [TRAIL] and especially the DR5, is generating considerable interests as a possible anticancer therapeutic agent because of its selective activation in apoptosis of this receptor as a superior affinity to ligands. The study was performed in invitro assay and the anticancer effects of the produced antibodies were assumed by MTT and flowcytometric methods. In the first step for immunization of the hens, three selective small peptides from extracellular domain of DR5 which were chemically synthesized, injected to hens and after the proper immunization of them, IgYs were extracted from the egg yolk. After assumption of specificity of the purified IgYs against the whole DR5 protein, they were performed in MTT assay and flowcytometric colorimeter. After confirmation of synthesized peptides they were injected to hens with Fround`s complete adjuvant. With completing the immunization procedure the specificity of purified IgYs were confirmed by ELISA. The antibodies were significantly killed the MCF7 breast cancer cells, but had divers affect [proliferative] on normal hepatocyte cells. Additionally, significantly they induced apoptosis on the cancerous cells in contrast to control cells. The results clearly demonstrated that the produced IgYs with reduced cost and time managing could remarkably use as an effective anticancer drug

Adição de óleos de copaíba (Copaifera langisdorffii) e sucupira (Pterodon emarginatus) na alimentação de poedeiras: estabilidade lipídica de gema de ovos armazenados em diferentes temperaturas / Effect of dietary supplementation with oils of copaíba (Copaifera langsdorffii) and sucupira (Pterodon emarginatus): lipid oxidation of egg yolks stored in different temperatures

The aim of this study was to evaluate the antioxidant effect of dietary supplementation of plant oil resins in laying hens on the oxidative stability of cooked egg yolk kept at 4ºC for 30 days, and fresh eggs stored under refrigeration (R) at the same temperature for 60 days or kept in room temperature (RT) for 30 days. Hens were fed corn- and soybean-based diets (15% CP and 2,900 kcal kg-1) and supplemented with two levels of Copaifera langsdorffii oil resin (CP-0.03; 0.06 and 0.09%) or Pterodon emarginatus oil resin (SC-0.03 and 0.06%), plus a negative control (CN). At 37 weeks of age, 667 eggs were collected and randomly distributed in different storage conditions, in natura or cooked. The progression of lipid oxidation of egg yolk in natura was quantified in quadruplicate and cooked egg yolks in duplicate, using pool of 3 egg yolks/treatment to analyze TBARS (thiobarbituric acid reactive substances) concentration in quadruplicate. Data analysis was performed using a mixed model and Tukey test, at a 5% significance level. The storage period was considered a longitudinal factor, which varied from five times, for R cooked yolk and TA fresh yolk (0-30 days), to nine times, for R fresh yolk (0-60 days). For fresh eggs stored at RT or R, the supplementation of plant oils did not protect egg yolks from lipid oxidation, compared to NC. However, for cooked egg yolks, the addition of 0.03 and 0.06% of CP oil resin showed antioxidant activity since it reduced lipid oxidation up to day 21 of storage, but had a prooxidant effect for 0.09%. Therefore, it can be concluded that the supplementation of copaíba oil resin had an antioxidant protection of cooked egg lipids...

O objetivo deste trabalho foi avaliar a atividade antioxidante da adição de óleos de copaíba (CP) e sucupira (SC) na alimentação de poedeiras sobre a oxidação lipídica de ovos in natura armazenados em temperatura ambiente (TA) por 30 dias e sob refrigeração (R) a 4ºC por 60 dias, e de gemas cozidas mantidas sob R por 30 dias. As poedeiras foram alimentadas com ração isoproteica (15% PB) e isoenergética (2900 kcal kg-1) à base de milho e farelo de soja, com inclusão de óleo de Copaifera langsdorffii (CP) nas proporções de 0,03; 0,06 e 0,09% ou de Pterodon emarginatus (SC) nas proporções de 0,03 e 0,06%, mais um controle negativo (CN). Foram coletados 667 ovos às 37 semanas de idade e distribuídos aleatoriamente nas diferentes condições de armazenamento (TA ou R). A oxidação dos lipídios de ovos in natura foi quantificada em quadruplicata e das gemas cozidas em duplicata, utilizando-se pool de 3 gemas/tratamento para as análises de TBARS (thiobarbituric acid reactive substances). Os dados foram avaliados adotando um modelo misto e as médias foram comparadas pelo teste de Tukey em 5% de nível de significância e o período de armazenamento foi considerado como um fator longitudinal, variando de cinco tempos no experimento com gemas cozidas, e nos ovos in natura sob R e em TA (0 a 30 dias), até nove tempos sob R (0 a 60 dias). Foi observado que a adição de óleo de CP e SC não reduziu os valores de TBARS em ovos in natura armazenados em TA e sob R em relação ao CN. No armazenamento de gemas cozidas, a inclusão CP (0,03 e 0,06%) protegeu os lipídios até os 21 dias, mas apresentou efeito pró-oxidante quando suplementado a 0,09%. Concluiu-se que a inclusão de até 0,06% de óleorresina de CP nas rações de poedeiras pode proteger os lipídios da gema cozida contra a oxidação durante o armazenamento refrigerado por até 21 dias...

Production of specific IgY Helicobacter pylori recombinant OipA protein and assessment of its inhibitory effects towards attachment of H. pylori to AGS cell line

PURPOSE: The common triple therapy for Helicobacter pylori is challenged by the increasing cases of antibiotic resistant infections, raising the need to explore alternative therapies. Oral administration of egg yolk immunoglobulin Y (IgY) has been previously reported as a means of passive immunization therapy for H. pylori infections. In this work, we investigated the inhibitory effect of IgY on the attachment of H. pylori to AGS cell line. MATERIALS AND METHODS: Recombinant OipA was prepared. Hens were immunized with recombinant protein three times. IgY was purified from egg yolks of immunized hens using polyethylene glycol precipitation method. The inhibitory effect of the specific immunoglobulin was evaluated in AGS cell line infected with H. pylori. RESULTS: The presence of recombinant OipA (30 kD) was confirmed via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunization of hens was confirmed using enzyme-linked immunosorbent assay. The purified IgY from egg yolks were assessed using SDS-PAGE and confirmed by western blot. CONCLUSION: The results showed that IgY-OipA had inhibitory effect on attachment of H. pylori to AGS cell line and may be utilized as a therapeutic or prophylaxis material.

Diagnostic utility of skin prick test to cooked egg in children with egg allergy

PURPOSE: Reliable predictors of tolerance to cooked egg in an egg allergic population are not established. We investigated the usefulness of the skin prick test to cooked egg in children with egg allergy. METHODS: We studied 36 children with egg allergy. Skin prick tests (SPTs) for the uncooked or cooked form of egg white and egg yolk, whole egg, ovomucoid (OVM), and ovalbumin (OVA) were performed at diagnosis. The reagents of cooked egg for SPT were prepared by baking for 25 minutes in 200 degree oven. We also examined specific IgE levels to whole egg, egg white, egg yolk, OVM, and OVA. RESULTS: Patients with history of allergic reaction to extensively heated egg showed significantly increased wheal size for cooked egg white (median [interquartile range]), 10.5 [7.0-14.6] vs. 4.2 [0.0-5.6], P<0.001) and OVM (9.6 [7.3-13.8] vs. 5.6 [0.0-7.8], P=0.001) than those without the history. The strongest positive correlation was found between wheal size for cooked egg white and OVM (r=0.788, P<0.001). SPT wheal size for cooked egg white were positively correlated with serum OVM-specific IgE levels (r=0.691, P<0.001). Cutoff value was 7.0 mm in SPT wheal size for cooked egg white, the sensitivity was 73.1% and specificity was 99.0%. SPT for cooked egg white showed significantly higher area under curve than serum egg white specific IgE. CONCLUSION: Our results suggest that SPT to cooked egg white may be useful predictor of allergic reaction to cooked egg. Further investigations will be needed.