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Level 20 Ovo

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Last Updated: 13 October 2020

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General | Latest Info

Otu and ovo require cells autonomously in female germline for germ cell proliferation and differentiation. Xy germ cells do not require otu when developing in testes, but become dependent on otu function for proliferation when placed in ovary. The requirement for ovo is dependent on cell autonomous signal deriving from X: ratio. Differential regulation of otu and ovo genes provides a mechanism for female germline to incorporate both somatic and cell - autonomous inputs required for oogenesis. All ovo alleles are fully penetrant for female sterility; no male function for ovo + is known. Lack of zygotic activity results in complete absence of germ - line cells in adult female; reduction in numbers of germ cells was first evident during early gastrulation. Homozygotes with weaker alleles produce germ cells, but oogenesis defective; egg chambers may degenerate prior to vitellogenesis or proceed through oogenesis and be oviposited, depending on allele; laid eggs are permeable to neutral red and never develop. Heterozygotes for two dominant alleles are phenotypically similar to homozygotes for weak recessive alleles. Ovarian tumors form in females carrying ovo D3 in heterozygous combination with hypomorphic alleles, eg, ovo D1rv20, or with snf. Ovo D2 mutation is partially suppressed by many Sxl alleles. Ovo D1 / + females produce no eggs; extensively used in selection of ovo + germ - line clones. Svb alleles are homozygous lethal. There are many fewer denticles than wild type; remaining denticles are small and with characteristic morphology. Denticles arranged in belts in abdominal segments; absent from thoracic segments. Cell autonomous in mosaics; useful in studying embryonic mosaics.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions

Introduction

For most vertebrate species, steroid signals produced by embryonic gonads during development influence sexual differentiation, but intriguingly, this process appears to be largely unaffected by relatively high levels of steroids in maternal circulation or egg yolk. It appears that maternal steroids are rapidly and efficiently metabolize, effectively buffering embryo from any potential sex - related effects. In placental species, tissues such as placenta and liver possess a suite of enzymes capable of metabolizing steroids as they pass from maternal to fetal circulation. Recently, it has been shown that maternal steroids are also metabolized by embryos of oviparous vertebrates. While the metabolic fate of steroids can vary depending on a number of factors, including steroid of interest, study species and period of embryonic development, general pattern is for steroids to be subject to Phase I metabolism followed by Phase II metabolism which is typically associated with inactivation and clearance. In all vertebrate embryos, Phase II metabolism of maternal steroids is primarily accomplished by sulfonation, replaced with glucuronidation after birth. Much of what we know about sulfonation of maternal steroids in oviparous amniotes comes from work in red - eared slider. Research initially focus on the metabolic fate of maternal oestradiol because exogenous oestradiol can influence gonadal differentiation, and because levels of maternally derived oestradiol in eggs vary seasonally; second clutches contain approximately 10 more oestradiol than first clutches from the same female within nesting season. During the first 10 days of incubation, concentrations of oestradiol in yolk drop to low / undetectable levels regardless of initial concentrations. This decline is driven by conversion of oestradiol to oestrone sulfate, resulting in an increase in oestrone sulfate levels early in development. Exogenous oestrone sulfate can cause gonadal feminization, but only at very high physiological or supraphysiological levels. Consequently, sulfonation has been hypothesized to buffer developing embryos from the effects of maternal oestrogens in similar manner to what is observed in placental amniotes. Many of the same sulfotransferase enzymes that are responsible for sulfonation of steroids also sulfonate exogenous chemicals, including endocrine disrupting chemicals. Thus, maternally derived steroids and EDCs could both be conjugated by the same enzymes during embryonic development. We have recently demonstrated that application of EDC, Bisphenol, to T. Scripta eggs results in increased levels of oestradiol and oestrone and decreased levels of oestrone sulfate present within egg during early development. In this study, we tested the hypothesis that BPA inhibits conversion of oestrone to oestrone sulfate. Because sulfonation of exogenous oestrogens occurs very early in development of T. Scripta, we first characterize sulfonation of oestrone over the first 72 h of incubation, and then examine the effect of BPA on conversion of oestrone to oestrone sulfate.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions

Results

Total skin RNAs were isolated from frozen and powdered skin using TRIzol Reagent from GIBCO - BRL. Poly + - RNAs were isolated using Oligotex. Ovary poly + - RNAs were purchased from Ambion. Northern blots containing other adult mouse tissues were purchased from Clontech. One of the following probes was used 1. 0 - kb fragment containing 5 - end sequences of mOvo1 cDNA including all four zinc fingers; 350 - bp fragment containing mostly 5 UTR of mOvo1; or 1. The 2 - kb fragment contains 3 UTR and parts of coding sequences including three of four zinc fingers. All three probes give rise to similar results. Hybridizations were performed in ExpressHyb hybridization solution from Clontech at 68C, followed by two washes in 2 SSC, 0. 05% SDS at room temperature, and three washes with 0. 1 SSC, 0. 1% SDS at 65C. In situ hybridizations with digoxygenin - label cRNA probes were performed on frozen sections using the method of Schaeren - Wiemers and Gerfin - Moser with slight modifications: 10% heat - inactivate goat Serum was used as blocking agent prior to incubation with Anti - digoxygenin antibody. Two different probes were used and yielded similar results: 1. 0 - kb cRNA containing four zinc fingers and upstream sequences; 870 - B cRNA containing 3 UTR of mOvo1. Immunofluorescence microscopy was performed as described. The Rabbit polyclonal antibody was generated by Zymed Laboratories to peptide sequence CTSESQEGHVLHLKERHPDS at amino acid position 220 of mOvo1a Protein. Antiserum was affinity purified on peptide - conjugated column using Sulfolink kit according to manufacturers ' suggestions. Rabbit Anti - loricrin was raised in laboratory AE13 was kindly provided by T. - T. Sun. Transglutaminase activity was measured using the following procedure. Frozen skin sections were fixed in methanol for 20 min, followed by 2 5 - min washes in PBS. Sections were then incubated with a solution containing 2. 5 M M DTT, 0. 25% Triton X100 0. 15 M NaCl, 0. 05 M Tris - HCl at pH 8. 0 10 M M CaCl 2, and 20% dansyl cadaverine, wash 3 5 min in PBS, and mount as described. Transglutaminase activity was then detected by fluorescence microscopy. Mouse keratinocyte line UG1 was cultured in media containing < 0. 05 M M CA 2 +. In CA 2 + induction experiments, CaCl 2 was added to subconfluent culture at final concentration of 1. 2 M M, and cells were then maintained at high CA 2 + for the desired period of time before harvesting. Full - length 2. 0 - kb mOvo1a cDNA was released from Bluescript vector by Eco RI - Asp I restriction and clone into Kpn I site of mammalian expression vector pCB6 containing CMV promoter and enhancer. Cos cells were transfected with the resulting plasmid using the calcium - phosphate method. Full - length mOvo1 cDNA was used to screen the 129 / Sv genomic library. One resulting clone was subject to detailed restriction map analysis and used to construct a targeting vector. The targeting vector was electroporated into R1 ES cells at 270 V, 500 F in GenePulser. Es cell culture and drug selection was essentially as described.


Discussion

In Drosophila, ovo is expressed in all germ cells, but its only function appears to be in female germ cells, where it is required for female sex determination and oogenesis. In contrast, movo genes seem to have evolved to play a role in mammalian spermatogenesis. This is perhaps not surprising given that in mammals, default pathway for germ - line sex determination has changed from male to female, and sex determination has evolved to differ quite dramatically from Drosophila. In mice, movo1 does not appear to be required for sex determination, because at least some viable male and female homozygous mutant movo1 germ cells develop, resulting in few successful matings of homozygous movo1 / mice within the population. Defects in movo1 mutant testes occur late, concomitant with late onset of movo1 expression in testis. Interestingly, despite lack of appreciable movo1 expression in spermatogonia, male germ - line stem - cell survival was also affect, process that coincides with major signs of tubule degeneration in adult animals. At present, we surmise that defects in sperm production are primary, whereas those in male germ - cell survival and in testis vascularization are secondary. Regardless of sequence of events, degeneration seen in mutant testes was among the most striking of defects caused by movo1 mutation.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions

Discussion

Genomic organization of Ovo locus, which encodes three transcripts: Ovo -, Ovo - B, and svb. Exons, introns, Protein coding regions, and zinc - finger DNA - binding domains are show. Regions corresponding TO probes for in situ hybridization and Ovo - and Ovo - B - specific region amplified by qRT - PCR analysis are also indicate. Sites where EGFP was inserted are indicated by green triangles. Ovoa - Nterm - EGFP and ovoB - Nterm - EGFP knock - in alleles contain genes in the N - terminal regions of Ovo - and Ovo - B proteins, respectively. Expression of Ovo transcripts during embryogenesis. Probes used for in situ hybridization are indicated in. Embryos and gonads were hybridized with common probe and svb - specific probe. Gonads were double - stain for Ovo or svb transcript and Vasa Protein, marker for germ plasm and PGCs. The developmental stage of each embryo is shown in lower right. Arrowheads show germ plasm and PGCs. Scale bars: 20 m. Levels of Ovo - and Ovo - B transcript relative TO expression level of Ovo - B in. 4 PGCs were determined by qRT - PCR, and plotted against developmental stages of embryogenesis. Mean SEM of three biological replicates are show. Expression of maternal and / or zygotic Ovo Protein during embryogenesis. Ovo - EGFP fusion Protein encoded by ovoB - Nterm - EGFP was detected using anti - GFP antibody. Developmental stages are shown in lower right. Insets show close - up images of germ plasm and PGCs. Maternal and zygotic Ovo - EGFP Protein was detected in embryos derived from female homozygous for ovoB - Nterm - EGFP mat with male hemizygous for ovoB - Nterm - EGFP. Maternal Ovo - EGFP was detected in embryos with YFP Staining, derived from ovoB - Nterm - EGFP / FM7c, Dfd - GMR - nvYFP females mat with y w males. Zygotic expression of Ovo - EGFP was detected in only female embryos produced from y w females mat with ovoB - Nterm - EGFP hemizygous males, since the Ovo gene is on the X chromosome. Scale bars: 20 m. Distribution of number of Vasa - positive germline cells per gonad of Ovo - Aexpressing and control females and males at embryonic stage 15 - 16, first, second, and third instar larval stages. Ovo - was expressed under control of mzNG4 in progeny derived from nos - Gal4 homozygous females mat with UASp - Ovo - homozygous males. Progeny derived from nos - Gal4 homozygous females mat with y w males were used as controls. Furthermore, Ovo - was overexpressed under control of mNG4 in YFP - positive progeny derived from nos - Gal4 / TM6B, Dfd - GMR - nvYFP females mat with UASp - Ovo - homozygous males. Yfp - positive progeny derived from nos - Gal4 / TM6B, Dfd - GMR - nvYFP females mat with y w males were used as controls. N: number of gonads observe. The significance of differences relative TO controls was calculated using Fishers exact test. Ovaries and testes of adult progeny derive from nos - Gal4 homozygous females mat with y w males, nos - Gal4 homozygous females mat with UASp - Ovo - homozygous males, or Ovo amk and nos - Gal4 homozygous females mat with UASp - Ovo - homozygous males; and YFP - positive adult progenies derive from nos - Gal4 / TM6B, Dfd - GMR - nvYFP females mat with y w males or nos - Gal4 / TM6B, Dfd - GMR - nvYFP females mat with UASp - Ovo - homozygous males. Ovo amk transgene expresses only Ovo - B under control of Ovo - B promoter 14.


Introduction

Campylobacteriosis is the most frequent bacterial zoonosis with an estimated 9 million human cases and an economic burden of around 2. 4 billion EUR each year in the European Union alone. The main etiologic agent of human campylobacteriosis is Campylobacter jejuni. Symptomatic C. Jejuni infection usually manifests as enterocolitis with watery or bloody diarrhea, mostly accompanied by fever and abdominal pain. Infections may be followed by serious sequelae like reactive arthritis and Guillain - Barre syndrome. The majority of human infections can be attributed to consumption of C. Jejuni - contaminated poultry meat products. Reduction of Campylobacter in chicken reservoirs is therefore considered an effective strategy to reduce the public health burden. It is estimated that a 2 - log reduction of C. Jejuni on chicken carcasses is sufficient to reduce the incidence of human campylobacteriosis by 30%. One of potential strategies to reduce C. Jejuni colonization in broiler chickens is vaccination. C. Jejuni flagellin, major subunit of bacterial flagellum is an attractive candidate vaccine antigen. Bacterial flagellins are highly immunogenic antigens in chickens and their immunostimulatory properties, including activation of chicken Toll - like receptor 5, make them potent vaccine adjuvants. Natural flagellin - specific antibodies likely contribute to maternal immunity in chickens. Yet, vaccination of chickens with flagellin - base vaccines has yielded variable success. Intraperitoneal immunization of 16 day - old chickens with heat - kill C. Jejuni enriched with native flagellin followed by booster two weeks later resulted in 1 - 2 log reduction in cecal colonization. Similarly, recombinant Campylobacter flagellin fuse to Escherichia coli heat - labile toxin administered orally at 14 days of age lower number of colonized animals. Administration of heat - kill C. Jejuni cells or flagellin to 16 day - old chicken embryos with increased levels of flagellin - specific IgY, IgM and IgA antibodies in chicken sera, levels of sIgA in bile and intestinal scrapings, and numbers of immunoglobulin - containing cells in spleen and intestine. Yet, in Ovo immunization combined with oral or intraperitoneal booster given 7 days after hatch does not confer protection upon C. Jejuni challenge. In attempt to achieve protection against C. Jejuni, we decided to improve immune response to Campylobacter flagellin by introducing intrinsic adjuvant activity to protein. In its natural form, C. Jejuni flagellin is unable to activate TLR5 in contrast to flagella subunits of most other bacterial species. Engineering of chimeric C. Jejuni flagellin that has TLR5 activating domains of Salmonella enterica serovar Enteritidis results in recombinant NHC flagellin that is able to activate TLR5 and thus potentially has intrinsic adjuvant activity. In the present study, we determined immunogenicity and protective efficacy of recombinant NHC flagellin after Ovo delivery. Our results indicate that in Ovo immunization with flagellin - base subunit vaccine is an effective way to generate specific systemic antibody response against C. Jejuni but that this strategy is still not sufficient to provide protection against C. Jejuni challenge.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions

2. Material and methods

Box - Behnken design in response surface methodology was used to investigate the response of broiler chicks to in ovo feeding of betahydroxy betamethylbutyrate, dextrin and timing of first water and feed deprivation. On day 18 of incubation, 1 500 eggs were randomly assigned to 15 experimental runs of BBD, each with 4 replicates, as 3 levels of IOF of HMB and dextrin, and 3 levels of first water and feed deprivation. Dayold chicks from each replicate were then used to assess the effect of IOF and time first water and feed access on the chick's responses. Iof of dextrin leads to respectively 9. 7% - 15. 5% lower hatchability for 20% and 40% inclusion, whereas HMB inclusion appears with no effect on hatchability. Administration of dextrin or HMB into amnion of embryos elevates length, width and surface area of the villus, and increases the glycogen content of the liver and breast. In all parameter models, linear terms show the highest contribution to explaining existing variation in chick's responses. First, water and feed deprivation had the largest effect on BW2 and glycogen content of liver and breast. It is concluded that if possible, place chicks before 7 hr of hatch to preserve BW loss and have maximum response from IOF. If not possible, use IOF with 40% dextrin + 0. 5% HMB to preserve gut integrity and energy status up to 48 hr. This should give advantage to chicks to recover fast after feeding, but that would have to be confirmed by trials growing birds to slaughter age.


Results and Discussion

Several investigators have shown significance of Ovo administration in composition and diversity of intestinal microbiota of neonate broiler chickens. Additionally, Bacteroides, Clostridium cluster XIVa, and Clostridium cluster IV have been described to have a profound role in intestinal homeostasis and reduction of inflammation. In the present study, bacterial composition at family level revealed that Enterobacteriaceae was numerically higher in the PBS control group, whereas Ruminococcaceae was significantly higher in the LAM treat group. Ruminococcaceae is a family in class Clostridia, which includes Clostridium and other similar genera. Ruminococci spp. They are among the most abundant cellulose - degrading bacteria in rumen and may also make a significant contribution to plant cell wall breakdown in large intestine in other mammals. They belong to clostridial cluster IV, contributing to up to 20% of bacteria present in humans and are important short - chain fatty acid producers. Scfas induce profound physiological responses in gut integrity and reduce inflammation. Interestingly, Ruminococcus spp., Faecalibacterium spp., And Lachnospiraceae spp. They are essential butyric acid contributors. Gastrointestinal inflammation has been associated with significant reduction of Clostridium clusters XIVa and IV, such as Lachnospiraceae, Ruminococcus, and Roseburia, hence, importance of differentiating beneficial clostridial strains from pathogenic strains such as Clostridium perfringens and Clostridium difficile. In the present study, SCFA - producing bacteria of the family Ruminococcaceae and genus Butyricicoccus were not detected in PBS control, whereas they were found in LAM. Moreover, unidentified genera that belong to Lachnospiraceae were significantly higher in embryos inoculated with LAM, whereas Enterobacteriaceae family was significantly higher in embryos inoculated with PBS. Lachnospiraceae is abundant in digestive tracts of many mammals and is a crucial bacteria because of its role in production of SCFA. In mice, probiotics have been shown to induce significant changes in SCFA. This, in turn, has profound impact on intestinal physiology as well as pathogen control for enteropathogens such as enterohemorrhagic E. Coli O157: H7. In chickens, in Ovo application of probiotics suggests that they can improve performance and immune functions and provide resistance against enteropathogens without affecting the hatchability of chickens. Likewise, in the present study, hatchability was not affected by Ovo treatment in both trials. Moreover, embryos inoculated with LAM show significant reduction in total number of gram - negative bacteria at DOH and d7. This reduction was also accompanied by a significant increase in total LAB at DOH in the LAM - treat group when compared with that in PBS control. These results were associated with significant differences in both beta diversity and alpha diversity, suggesting that LAM treatment may drive large - scale changes in microbial community structure and composition as has been published previously. In summary, in Ovo administration of defined LAM isolate from adult hens does not affect hatchability, improved BWG and reduced mortality at d7, induced variations in cecae microbiota structure, and reduced Enterobacteriaceae colonization on virulent E. Coli horizontal infection model in broiler chickens.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions

SIZE MATTERS

The advent of in ovo devices has been one of the driving forces behind the increasing size and efficiency of hatcheries in developed and developing poultry markets over the last two decades. But it is not only larger hatcheries that can benefit from in - ovo devices. Technology also has potential to allow smaller hatcheries to improve the quality of chicks they supply and avoid labor, efficacy and safety issues associated with subcutaneous vaccination. In 2012, Zoetis introduced the world's first semi - automate, compact in ovo device: Inovoject M BioDevice. This device has the same quality and unique design features that have made Inovoject System world most widely used in ovo device - but in compact form for hatcheries that do not have throughput or floor space suitable for standard Inovoject System. The new device can handle 12 000 to 20 000 eggs per hour, depending on configuration, and can be operated by two or three people. Whatever size of your operation, we will send a survey team to assess your needs and make sure you get the most appropriate setup. Our technicians will be on hand to help every step of the way, from installation to initial run - up and operation.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions

WHY COMPROMISE?

The Superior design and quality of Inovoject System has helped to make the device industry standard for in ovo Technology. No other device can deliver the same level of performance and support after 20 years of experience. And with the range of new and additional technologies available from Zoetis, if you are thinking of adopting in ovo Technology in your hatchery, why compromise? 1 Avakian AP, Wakenell PS, Bryan T, Schaeffer JL, Williams CJ, Whitfill C. In ovo administration of Mareks Disease vaccine: Importance of vaccine deposition site in fertile egg, Proceedings. 51 Western Poultry Disease Conference 2002; 119 - 121. 2 Barbosa T, Williams C, Villalobos T. Efficacy and Mareks Disease protection comparison between different vaccination methods, in Proceedings. 18 Congress World Veterinary Poultry Association 2013; 217.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions

Sources

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions.

* Please keep in mind that all text is machine-generated, we do not bear any responsibility, and you should always get advice from professionals before taking any actions

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