Plasma Protein Fractions Research Articles

Growth performance, serum metabolites, plasma protein fraction, meat lipid profile and antioxidant activities of breast meat of broiler chicken fed diets containing Cinnamomum ceylon powder, Zingiber officinale powder and Moringa oleifera leafmeal supplem

An experiment was conducted to determine growth performance, serum metabolites, plasma protein fraction, meat lipid profile and antioxidant activities of meat of broilers as affected by Cinnamomium ceylon, Zingiber officinale and Moringa oleifera supplementation. A total of 112 day old chicks were allotted into four treatments replicated four times with seven birds per replicate in a completely randomized design (CRD). Basal diets were formulated for broiler starters (0-28days) and finisher phase (29-56) days. The basal diets were divided into 4 diets : diet 1- control diet without supplementation; diet 2- contained 0.2% Cinnamomum ceylon supplementation (CCS); diet 3- contained 0.2% Zingiber officinale supplementation (ZOS), diet 4-contained 0.2% Moringa oleifera leafmeal (MOLM) supplementation. The experiment lasted for 56 days. Results showed an improvement in final liveweight, weight gain, feed intake and feed conversion ratio of birds on 0.2% MOLM. Birds on phytogenic feed additives have lower values of urea, cholesterol, glucose, high density lipoprotein and low density lipoprotein. Birds administered phytogenic supplements exhibited much greater plasma albumin levels. Low levels of meat cholesterol, triglycerides and low density lipoprotein were found in birds fed phytogenic feed additives while high levels of high density lipoprotein were recorded for birds on phytogenic feed additives. The supplementation of broilers diets with phytogenic feed additives increased the glutathione peroxidase and superoxide dismutase and decreased lipid peroxidation. It was concluded that supplementation of broiler diets with Moringa oleifera leaf meal enhanced body weight gain, antioxidant activities also reduced cholesterol level in both serum metabolites and meat.

Partitioning behavior of short DNA fragments in polymer/salt aqueous two-phase systems.

The development of liquid biopsy as a minimally invasive technique for tumor profiling has created a need for efficient biomarker extraction systems from body fluids. The analysis of circulating cell-free DNA (cfDNA) is especially promising, but the low amounts and high fragmentation of cfDNA found in plasma pose challenges to its isolation. While the potential of aqueous two-phase systems (ATPS) for the extraction and purification of various biomolecules has already been successfully established, there is limited literature on the applicability of these findings to short cfDNA-like fragments. This study presents the partitioning behavior of a 160 bp DNA fragment in polyethylene glycol (PEG)/salt ATPS at pH 7.4. The effect of PEG molecular weight, tie-line length, neutral salt additives, and phase volume ratio is evaluated to maximize DNA recovery. Selected ATPS containing a synthetic plasma solution spiked with human serum albumin and immunoglobulin G are tested to determine the separation of DNA fragments from the main plasma protein fraction. By adding 1.5% (w/w) NaCl to a 17.7% (w/w) PEG 400/17.3% (w/w) phosphate ATPS, 88% DNA recovery was achieved in the salt-rich bottom phase while over 99% of the protein was removed.

A Pilot Study Investigating Plasma Protein Electrophoresis in One Anuran and Six Urodelan Species.

As threats to amphibian health increase, there is a growing need for diagnostic tools to assess and monitor their health status. Plasma protein electrophoresis has proven to be useful in other nonmammalian species. It enables quantification of protein fractions in plasma that may be altered in various disease processes, and is therefore useful in narrowing down differential diagnoses and detecting inflammation, in combination with other modalities such as biochemical and hematologic testing. The amphibian electrophoretogram must be defined before baseline reference intervals are obtained across species. Agarose gel electrophoresis was performed on plasma samples collected from presumed clinically normal individuals of one anuran and six urodelans: Osteopilus septentrionalis (n=2), Gyrinophilus porphyriticus (n=1), Notophthalmus viridescens (n=1), Eurycea guttolineata (n=2), Amphiuma tridactylum (n=2), Cryptobranchus alleganiensis (n=5), and Siren lacertina (n=6). The electrophoretograms varied in number of fractions between each species; however, the number of fractions was consistent within a species. An albumin migrating fraction was consistently observed in all species. A prealbumin migrating fraction was identified in species that primarily use organs other than skin for respiration. This study provides preliminary examples of a normal plasma protein electrophoretogram for seven amphibian species. Further studies quantifying reference intervals and identification of protein fractions will help establish protein electrophoresis as a useful tool in amphibian health investigations.

Effect of Seminal Plasma Protein Fractions on Cooled Dog Semen Kinetics

Abstract Semen cooling and cryopreservation have revolutionized the field of reproductive biotechnologies. However, challenges persist in maintaining sperm quality and viability during these processes. The unsatisfactory results and the main problems are associated with low quality, viability, morphological, structural, and DNA integrity, changes in plasma membrane, ability to interact with female tract and decrease in sperm fertilization potential. The objective of this study was to determine the effect of seminal plasma proteins with different molecular weights on the kinetic parameters of dog spermatozoa during cold storage at 4 °C. The proteins were isolated from the dogs’ seminal plasma ranging between 6-200 kDa, separated in four fractions. The ejaculates (n=15) were collected from 6 heathy dogs which were pooled. Spermatozoa were extended with Tris medium alone (control) or with addition of one of the isolated protein fractions, and were then incubated at 4 °C for 2 hours. Sperm incubated with seminal plasma proteins ranging between 10-15 kDa had significantly higher total motility (37.01±2.98%), sperm progressiveness (15.97±1.91%), curvilinear line velocity (37.46±3.75 μm/s), linearity (26.18±1.00%), and straightness (45.94±2.03%) compared to the other groups incubated with higher molecular weight proteins and the control group. The findings of this study indicated that the use of certain proteins in seminal plasma can be beneficial for reducing the detrimental effect of cooling at 4 °C and preserving the viability of dog spermatozoa. It seemed that the presence of 10-15 kDa proteins from canine seminal plasma rendered the spermatozoa less amenable to the negative influence during cooling.

Phytoplankton Biological Pump Controls the Spatiotemporal Bioaccumulation and Trophic Transfer of Antibiotics in a Large Subtropical River.

The spatiotemporal bioaccumulation, trophic transfer of antibiotics, and regulation of the phytoplankton biological pump were quantitatively evaluated in the Pearl River, South China. The occurrence of antibiotics in organisms indicated a significant spatiotemporal trend associated with the life cycle of phytoplankton. Higher temporal bioaccumulation factors (BAFs) were found in phytoplankton at the bloom site, while lower BAFs of antibiotics in organisms could not be explained by phytoplankton biomass dilution but were attributed to the low bioavailability of antibiotics, which was highly associated with distribution coefficients (R2 = 0.480-0.595, p < 0.05). Such lower BAFs of antibiotics in phytoplankton at higher biomass sites hampered the entry of antibiotics into food webs, and trophic dilutions were subsequently observed for antibiotics except for ciprofloxacin (CFX) and sulfamerazine (SMZ) at sites with blooms in all seasons. Distribution of CFX, norfloxacin (NFX), and sulfapyridine (SPD) showed further significant positive relationships with the plasma protein fraction (R2 = 0.275-0.216, p < 0.05). Both mean BAFs and trophic magnification factors (TMFs) were significantly negatively correlated with phytoplankton biomass (R2 = 0.661-0.741, p < 0.05). This study highlights the importance of the biological pump in the regulation of spatiotemporal variations in bioaccumulation and trophic transfer of antibiotics in anthropogenic-impacted eutrophic rivers in subtropical regions.

Selective enrichment of plasma cell-free messenger RNA in cancer-associated extracellular vesicles

Extracellular vesicles (EVs) have been shown as key mediators of extracellular small RNA transport. However, carriers of cell-free messenger RNA (cf-mRNA) in human biofluids and their association with cancer remain poorly understood. Here, we performed a transcriptomic analysis of size-fractionated plasma from lung cancer, liver cancer, multiple myeloma, and healthy donors. Morphology and size distribution analysis showed the successful separation of large and medium particles from other soluble plasma protein fractions. We developed a strategy to purify and sequence ultra-low amounts of cf-mRNA from particle and protein enriched subpopulations with the implementation of RNA spike-ins to control for technical variability and to normalize for intrinsic drastic differences in cf-mRNA amount carried in each plasma fraction. We found that the majority of cf-mRNA was enriched and protected in EVs with remarkable stability in RNase-rich environments. We observed specific enrichment patterns of cancer-associated cf-mRNA in each particle and protein enriched subpopulation. The EV-enriched differentiating genes were associated with specific biological pathways, such as immune systems, liver function, and toxic substance regulation in lung cancer, liver cancer, and multiple myeloma, respectively. Our results suggest that dissecting the complexity of EV subpopulations illuminates their biological significance and offers a promising liquid biopsy approach.

Serum-Derived Bovine Immunoglobulin Stimulates SCFA Production by Specific Microbes in the Ex Vivo SIFR® Technology.

Serum-derived bovine immunoglobulins (SBI) exert health benefits mediated by their ability to bind microbial components, thereby preventing translocation and subsequent inflammation. While in vivo studies have shown that a fraction of SBI also reaches the colon, little is known about the impact of SBI on the dense colonic microbiota that has great potential to impact human health. This study, therefore, investigated the impact of three bovine plasma protein fractions (SBI, bovine plasma (BP) and albumin-enriched bovine plasma (ABP)) on the gut microbiota of six human adults using the novel ex vivo SIFR® technology, recently demonstrated to generate predictive findings for clinical studies. When dosed at an equivalent of 5 g/day, all protein fractions significantly increased health-related metabolites-acetate, propionate, and butyrate. Upon simulating small intestinal absorption, SBI still markedly increased acetate and propionate, demonstrating that SBI is more resistant to small intestinal digestion and absorption compared to the other protein sources. Despite noticeable interindividual differences in microbiota composition among human adults, SBI consistently stimulated a narrow spectrum of gut microbes, which largely differed from the ones that are typically involved in carbohydrate fermentation. The SBI-fermenting consortium included B. vulgatus and L. edouardi (correlating with acetate and propionate) along with Dorea longicatena, Coprococcus comes and the butyrate-producing bacterium SS3/4 (correlating with butyrate). Overall, this study revealed that protein bovine fractions can contribute to health benefits by specifically modulating the human gut microbiota. While health benefits could follow from the production of SCFA, a broader range of protein-derived metabolites could also be produced. This study also confirms that the concept of prebiotics (substrates selectively utilized by host microorganisms conferring a health benefit) could go beyond the use of ingestible carbohydrates and extend to partially indigestible proteins.

Low-dose and repeated exposure to nickel leads to bioaccumulation and cellular and metabolic alterations in quails

Nickel (Ni) is a widespread environmental pollutant commonly released into effluent due to industrial activities, the use of fuels, or wastewater disposal. Many studies confirm the toxic effects of this heavy metal. However, there is a lack of knowledge and data on bioaccumulation patterns in tissues as well as cellular and molecular responses following the exposure of living organisms to Ni. In this study, Japanese quails were exposed to low (10 μg/L) and high (2000 μg/L) Ni concentrations in the form of nickel(II) chloride via drinking water. Sub-chronic exposure lasted 30 days while nominal concentrations represented average Ni content in drinking water (low dose) and average Ni levels in highly polluted aquatic environments (high dose). It was revealed that a high dose of Ni was correlated with increased water intake and decreased body weight. Overall, Ni exposure induced the development of microcytic anemia and alterations in measured blood indices. Moreover, Ni exposure impaired immunological activation as seen through the increased number of the white blood cells, increased heterophile/lymphocyte (H/L) ratio, and pronounced thrombocytosis. Ni elicited changes in the albumin, glucose, cholesterol, and triglyceride serum levels in a concentration-dependent manner. Alterations of plasma protein fractions suggested liver functional impairment while high levels of urea and creatinine indicated potential kidney injury. Granulation of heterophiles and an increase in erythroblasts in the bone marrow showed that the hematopoietic tissue was also impacted by Ni toxicity. On average each quail bioaccumulated 5.87 μg of Ni per gram of tissue. Moreover, the distribution and bioaccumulation of Ni in terms of relative concentration were as follows: feathers > kidneys > heart > liver > pectoral muscles. Assessed bioaccumulation levels and associated cellular and metabolic alterations have revealed new multilayer toxicological data that will help in the extrapolation of Ni toxicity in other vertebrates, including humans.

Simvastatin reduces plasma membrane caveolae and caveolin-1 in uterine leiomyomas

AimsUterine leiomyomas, or fibroids, are estrogen dependent benign tumor in women, however, they have limited treatment options. Simvastatin, a drug commonly used to treat high cholesterol. Recently we demonstrated that simvastatin alters estrogen signaling by reducing the expression and trafficking of the estrogen receptor-α (ER-α) in human uterine leiomyoma cells. Caveolae are invaginations of the plasma membrane where ER-α is known to localize and directly interacts with the caveolar protein caveolin-1 (CAV1). This study examines the effects of simvastatin on plasma membrane caveolae and the expression and palmitoylation of CAV1 in human leiomyomas which may influence ER-α signaling. Main methodsWe performed in vitro experiments using primary and immortalized human uterine leiomyoma cells. The caveolae were quantified using transmission electron microscopy. Additionally, we examined the impact of simvastatin treatment (40 mg orally per day for 12 weeks) on human leiomyoma tissue obtained from a randomized controlled trial. The CAV1 protein and mRNA levels were determined using quantitative real-time polymerase chain reactions, western blotting, and immunofluorescence analyses. Key findingsSimvastatin decreased the number of caveolae in primary leiomyoma cells and reduced CAV1 abundance in whole cells and remarkably the plasma protein fraction. It also decreased CAV1 palmitoylation, a post-translational modification associated with CAV1 activation. The effects of simvastatin on CAV1 were recapitulated in human leiomyoma tissue samples. SignificanceOur results identify caveolae and CAV1 as novel targets of simvastatin which may contribute to the recently described effects of simvastatin on ER-α signaling and plasma membrane trafficking.

Addition ofseminal plasma proteins effecting theinvitro kinetic properties ofcanine spermatozoa.

The objective ofthis study isto evaluate the changes inthe motility and kinetic patterns ofcanine spermatozoa, capacitated and decapacitated, after the addition ofseminal plasma protein fractions with different molecular weight. Ithas been proposed that proteins inseminal plasma support the survival ofthe spermatozoa and exert adual effect: capacitation and decapacitation. The seminal plasma from fresh ejaculates was subjected tochromatographic separation and four protein fractions were obtained. Computer-assisted sperm analysis was used todetermine the sperm subpopulations with specific motion and kinetic characteristics after incubation with each ofthe four protein fractions. Two-dimensional electrophoresis ofthe fractions that exhibit asignificant effect onthecapacitation and decapacitation was performed. Bysperm class analyser, capacitation changes were observed inthe sperm subpopulation with ahigh curvilinear velocity and amplitude oflateral head displacement incubatedwith the seminal plasma protein fraction with ahigh molecular weight, which was also reflected inthedecreased linearity, straightness, and progressive motility. The sperm subpopulation incubated with theseminal plasma protein fraction with alow molecular weight seemed toundergo aprocess ofdecapacitation (decreasing ofthe curvilinear velocity, increasing ofthe linearity, straightness and showing progressive motility). Despite their ample panorama ofactions, the role ofseminal plasma proteins regarding capacitation and decapacitation isstill undetermined.

The amniotic fluid proteome changes with gestational age in normal pregnancy: a cross-sectional study

The cell-free transcriptome in amniotic fluid (AF) has been shown to be informative of physiologic and pathologic processes in pregnancy; however, the change in AF proteome with gestational age has mostly been studied by targeted approaches. The objective of this study was to describe the gestational age-dependent changes in the AF proteome during normal pregnancy by using an omics platform. The abundance of 1310 proteins was measured on a high-throughput aptamer-based proteomics platform in AF samples collected from women during midtrimester (16–24 weeks of gestation, n = 15) and at term without labor (37–42 weeks of gestation, n = 13). Only pregnancies without obstetrical complications were included in the study. Almost 25% (320) of AF proteins significantly changed in abundance between the midtrimester and term gestation. Of these, 154 (48.1%) proteins increased, and 166 (51.9%) decreased in abundance at term compared to midtrimester. Tissue-specific signatures of the trachea, salivary glands, brain regions, and immune system were increased while those of the gestational tissues (uterus, placenta, and ovary), cardiac myocytes, and fetal liver were decreased at term compared to midtrimester. The changes in AF protein abundance were correlated with those previously reported in the cell-free AF transcriptome. Intersecting gestational age-modulated AF proteins and their corresponding mRNAs previously reported in the maternal blood identified neutrophil-related protein/mRNA pairs that were modulated in the same direction. The first study to utilize an aptamer-based assay to profile the AF proteome modulation with gestational age, it reveals that almost one-quarter of the proteins are modulated as gestation advances, which is more than twice the fraction of altered plasma proteins (~ 10%). The results reported herein have implications for future studies focused on discovering biomarkers to predict, monitor, and diagnose obstetrical diseases.

НОВЕ СЛОВО В МЕТОДИЦІ ВИГОТОВЛЕННЯ І ВИКОРИСТАННЯ ЕКСПРЕСНОГО ДІАБЕТИЧНОГО МЕДУ З ОЗДОРОВЧОЮ МЕТОЮ

The article is devoted to the problem of making express diabetic honey to prevent and improve health, as well as to treat diabetic individuals. The disease has become extremely widespread both globally and in individual countries. It was registered that over 425 million people have diabetes for the current year. According to this situation, the medical expenses reach up to 727 billion US dollars, which could be spent to the improvement of the quality of living standards. The number of cases of diabetes is about 1.3 million adults against the backdrop of difficult demographic situation in Ukraine. Unfortunately, there is a negative trend in our country towards the spread of the disease among young people and children, even preschoolers. In accordance with a prognostic point of view, the prospect of today's young generation seems highly undesirable – that is, to be among people with this pathology all their lives. Therefore, solving the problem through prevention and healthy lifestyle is important. Due to shortages of medicines and financial support for certain population category, the solution in this case is seen in the expansion of folk remedies for prevention and treatment, the leading place among which are bee products. The aim of the study was to develop a method of making diabetic express honey from blueberries (Vaccinium myrtillus L.), Jerusalem artichoke (Helianthus tuberosus L.), common beans (Phaseolus vulgaris L.) and to study its effect on changes in glucose, total protein and plasma protein fractions (PPF) of diabetics. According to studies of two groups of males (control and experimental) aged 50-70 years with a total of 24 people, it was found that the use of developed by the author's methodology of express diabetic honey from blueberries, Jerusalem artichokes and common beans helps to stabilize blood glucose levels; it has a positive effect on protein metabolism and immunity of diabetics, as well as enhances the effect of oral antidiabetic drugs.

Plasma electrophoresis profiles of Blanding's turtles (Emydoidea blandingii) and influences of month, age, sex, health status, and location.

Baseline plasma electrophoresis profiles (EPH) are important components of overall health and may aid in the conservation and captive management of species. The aim of this study was to establish plasma protein fractions for free-ranging Blanding's turtles (Emydoidea blandingii) and evaluate differences due to age class (adult vs. sub-adult vs. juvenile), sex (male, female, or unknown), year (2018 vs. 2019), month (May vs. June vs. July), health status, and geographical location (managed vs. unmanaged sites). Blood samples were obtained from 156 Blanding's turtles in the summer of 2018 and 129 in 2019 at two adjacent sites in Illinois. Results of the multivariate analysis demonstrated that age class, sex, year, month, health status, and geographical location all contributed to the variation observed in free-ranging populations. Adult females had the highest concentration of many protein fractions, likely associated with reproductive activity. Juveniles had lower protein concentrations. Temperature and rainfall differences between years impacted concentrations between 2018 and 2019, while May and June of both years saw higher levels in some protein fractions likely due to peak breeding and nesting season. Individuals with evidence of trauma or disease also showed increased plasma protein fractions when compared to those that were considered healthy. The two sites showed a wide/large variation over the two years. All of these factors emphasize the importance of considering multiple demographic or environmental factors when interpreting the EPH fractions. Establishing ranges for these analytes will allow investigation into disease prevalence and other environmental factors impacting this endangered species.

Effect of Stocking Density, Dietary Vitamin D3 and Probiotic Supplementation on Carcass Traits and Blood Parameters of Broiler Chickens

The present study was conducted to evaluate the effect of stocking density, dietary supplementation of probiotic and Vitamin D3 on carcass yield and blood biochemistry of broiler chickens. A total number of 360 unsexed broiler chicks (Evian), one-day old, nearly similar in live body weight were used in this study. Chicks were randomly divided into equal 12 treatments in a 2X3X2 factorial arrangement experiment. Birds were allocated in two main experimental groups (each of 180 birds), the first main group kept under normal stocking density (LSD) with 10 birds /m2 and the second main group kept under high stocking density (HSD) with16 birds/m2. Each main group was divided into three groups (each of 60 birds) fed on diets supplemented with probiotic at a levels of 0, 1 and 2 g/kg diet, respectively. Each group divided into two sub-group (each of 30 birds) and supplemented with fed vit.-D3 at a level of 250 and 500 IU/kg diet, respectively. Results obtained showed that raised broiler chickens under LSD had significantly improved absolute and relative carcass, giblets and total edible weight Moreover, it has significantly increased plasma total protein, albumin (A), globulin (G), cholesterol, triglyceride and it has decreased plasma transaminase enzymes of AST and ALT, compared with those rose under HSD. Broiler chicks fed on diet supplemented with probiotic at level of 2 g/ kg diet showed higher plasma total protein, albumin and A/G ratio, cholesterol and triglycerides than those fed on the other levels of probiotic supplementations. While, it has recorded lowest plasma AST and ALT levels, compared with the other experimental groups. Feeding broiler chickens on high level of Vit.-D3 (500 IU/ Kg diet) has significantly increased plasma protein fractions and decreased plasma AST and ALT levels, compared with those fed diet supplemented with low level of Vit.-D3 (250 IU/Kg) that caused significant decreased plasma cholesterol and triglycerides. Finally, it could be recommended to apply LSD (10 birds/ m2), high level of Vit.-D3 (500 IU/ Kg) and probiotic at a level of 2 g/ kg diet to improve carcass yield and blood biochemistry of broiler chickens.

Sample Displacement Batch Chromatography of Proteins.

In downstream processing, large-scale chromatography plays an important role. For its development, screening experiments followed by pilot-plant chromatography are mandatory steps. Here we describe fast, simple, and inexpensive methods for establishing a preparative chromatography for the separation of complex protein mixtures, based on sample displacement batch chromatography. The methods are demonstrated by anion-exchange chromatography of a human plasma protein fraction (Cohn IV-4), including the screening step and upscaling of the chromatography by a factor of one hundred. The results of the screening experiments and the preparative chromatography are monitored by SDS-PAGE electrophoresis. In summary, we provide a protocol, which should be easily adaptable for the chromatographic large-scale purification of other proteins, in the laboratory as well as in the manufacturing of biopharmaceuticals. These protocols cover the initial piloting steps for establishing a large-scale sample batch chromatography. The results from the piloting steps may also be applied for packed columns for performing simulated-moving-bed (SMB) chromatography rather than batch chromatography.

PROTEIN ELECTROPHORESIS OF PLASMA SAMPLES FROM BOA CONSTRICTORS WITH AND WITHOUT REPTARENAVIRUS INFECTION.

Reptarenaviruses infect a variety of boid and pythonid snake species worldwide and have been shown to be the cause of inclusion body disease (IBD). Little is known about the correlations between virus infection and clinical disease, as well as the effects of viral infection on the immune system and the blood protein fractions. The goal of this study was to examine the differences in the plasma protein fractions in reptarenavirus reverse transcription polymerase chain reaction (RT-PCR)-negative and -positive tested snakes with and without clinical signs of disease. Blood from a total of 111 boa constrictors (Boa constrictor) was evaluated. Reverse transcription PCRs and H&E staining for inclusion bodies were carried out on each sample for the detection of reptarenavirus, and the plasma protein fractions were evaluated by capillary zone electrophoresis (CZE). Thirty four of the 111 evaluated snakes were positive by RT-PCR and 19 of the 34 showed clinical signs of disease. In comparison with IBD-negative healthy boa constrictors, the positive snakes with clinical signs had significantly lower albumin levels (P = 0.0052), lower A: G ratios (P = 0.0037), and lower α-globulin levels (P = 0.0073), while their γ-globulin levels were significantly higher (P = 0.0004). In the same comparison, clinically healthy arenavirus-positive boas showed only significantly lower α-globulin (P = 0.0124) and higher γ-globulin levels (P = 0.0394). The results of the present study indicate that reptarenavirus infection may influence plasma protein fractions in boa constrictors.

Physiological and biochemical response of broiler chickens to amino acids administration

The main objective of this study was to evaluate the effect of amino acids supplementation to drinking water of meat-type broiler chickens on their blood parameters and productive traits. A total of sixty one day old, unsexed broiler chicks were obtained from a local hatchery. They were divided into three treatment groups: control group, and two treatment groups supplemented (via drinking water) with amino acids preparation (Amino-2500 USA Premium Nutrition) at 0.5 and 1.0 g/L, respectively. All chicks were fed a commercial low protein diet containing 19% crude protein and 2900 kcal metabolizable energy per kilogram of the diet. Chicks of the control group was fed the recommended (standard) diet containing 23%crude protein and 3000 kcal of ME/Kg. The duration of the experiment was five weeks. The criteria of productive and physiological responses were live body weight, weight gain, blood plasma protein fractions, plasma lipids (cholesterol, triglycerides, high density lipoprotein (HDL) and low density lipoprotein (LDL). Transaminase (ALT and AST) activity indicative of liver function was measured. Kidney function test in terms of plasma creatinine, urea nitrogen and uric acid were also determined. Results showed that live body weight and weight gain of broiler chickens were significantly increased by AA administration either at three or five weeks of age. At marketing age (5 weeks), birds from the 3rd and 2nd groups achieved heavier body weigh by 32.3 and 26.8% more than the control chickens. Blood plasma total protein, albumin and globulin levels were significantly increased in the AA treatment groups for T3 and T2, respectively as compared to the control chicks. Addition of AA resulted in an obvious increase in the activity of ALT and AST indicative of negative influence of excess AA on livers function-related enzymes, especially with the higher dose of AA (T3).This was also observed for plasma uric acids and creatinine concentrations as end products of protein metabolism. The histological sections from lymphoid organs revealed better histological appearance indicative of better immunity and bird's health. It is concluded that excessive AA administration for broilers fed on low protein diet, could be used to increase live body weight, enhance protein metabolism without negative impacts on blood biochemistry, and improve their immune response. However, further study is needed to assess the cost and the net revenue from AA supplementation.

Vitamin E Supplementation and Early Age Heat Conditioning to Alleviate the Negative Effects of Heat Stress in Quail Chicks.

Inclusion of vitamin E in poultry diets and manipulation of temperature at early age has been known to help birds cope with heat stress at later age of their life. This study was conducted to investigate the effects of early age heat conditioning (EHC) and vitamin E addition on the performance and blood parameters to alleviate deleterious impact of heat stress in quail chicks. Three hundred one-day-old quail chicks were randomly divided into 4 groups of 5 replicates with 15 birds of each. Treatments were: Control, vitamin E: Chicks were fed the basal diet supplemented with 200 IU kg-1 diet vitamin E, EHC: Chicks were exposed to 40±1°C for 2 h at days 7th and 13th of embryogenesis and EHC+vitamin E: Chicks were exposed to 40±1°C for 2 h at days 7th and 13th of embryogenesis and fed the basal diet supplemented with 200 IU kg-1 vitamin E. The experiment lasted from 1-40 days of age. The results indicated that using early age heat conditioning and/or supplementation of vitamin E significantly (p<0.05) improved body weight, feed intake and feed conversion ratio at 40 days of age. Significantly (p<0.05) improvement was observed in blood pH, H/L ratio and Hb concentration, thyroid hormones (T3 and T4), plasma protein fractions and antioxidant status (Total antioxidant capacity, catalase and superoxide dismutase). Exposed quail chicks to early age heat conditioning and addition of vitamin E at 200 IU kg-1 diet can effectively alleviate the adverse effects of heat stress.

EFFECT OF IN-OVO INJECTION WITH IRON–METHIONINE CHELATES OR IRON NANO-PARTICLES AND POST HATCH DIETARY FOLIC ACID ON GROWTH PERFORMANCE AND PHYSIOLOGICAL RESPONSES OF BROILER CHICKENS

A study was carried out to evaluate the effect of in-ovo injection of iron–methionine chelates (Fe-Meth) or iron nano-particles (FeNPs) and post hatch feeding with diets fortified with folic acid on productive performance and some physiological responses of Ross broiler chickens. A total of 450 fertile eggs were allocated to 5 treatment groups in the incubator, each of which included 3 replicates (30 eggs each). Experimental treatments consisted of a 5 × 2 factorial arrangement design with 5 levels of in-ovo injection solutions at day seven of incubation and two dietary levels of folic acid (FA) (0.0 or 0.5g/kg, diet) after hatching. In-ovo injection treatments were divided into 5 groups: control non- injected (T1), injected with 0.1 ml physiological saline (T2), sham-injected control (T3),injected with 0.1 ml of 75 ppb of iron nano-particles (T4, FeNPs), while the last group (T5) was injected with 0.1 ml of 75 ppb organic iron (Fe- meth). Results showed that in-ovo injection of different iron sources significantly improved live body weight and weight gain of broiler chicks, and enhanced FCR at 3 and 5 weeks of age compared to other groups. Broilers fed folic acid fortified diets displayed significantly heavier Live body weight (LBW) and body weight gain (BWG) than the those fed the control un-supplemented diet. The combined interaction effect of in-ovo iron injection and dietary folic acid supplementation showed the best productive performance figures during the whole experimental period. Different iron sources had no significant effect on all plasma protein fraction, Lipids profile except a higher concentration of high density lipoprotein (HDL) in plasma recorded in-ovo injection of different iron sources compared with different controls. Plasma hemoglobin, globulin fractions(α, β, and γ-globulin), transferrin and Tri-iodothyronine concentration were significantly higher in both injected groups compared to the different control groups. Birds fed FA treatment recorded the lower significant values of plasma cholesterol, triglyceride and LDL, but, the birds fed FA recorded the higher significant value of plasma HDL, plasma hemoglobin, globulin fractions(α, β, and γ-globulin), transferrin and tri-iodothyronine concentration levels compared with birds fed with or without FA. In conclusion, in-ovo injection of Fe-Meth. or FeNPs and fed the diet fortified with FA were sufficient to improve broiler performance or physiological response in broiler chickens.

Blood plasma high abundant protein depletion unintentionally carries over 100 proteins

AbstractThere is a constant interest in blood‐based protein biomarkers, which can help to improve diagnosis and treatment outcomes of multifactorial human pathologies. In this regard, proteomic studies usually employ plasma immunoaffinity fractionation to deplete the most abundant plasma proteins, due to the high dynamic concentration range of proteins. The depletion of high abundant proteins allows to obtain less abundant and, oftentimes, more interesting proteins. However, the removal of the fraction of the high abundant plasma proteins ‐ the depletome ‐ may co‐elute many unintended proteins due to protein‐protein interactions. Little data is available about the depletome and potential protein biomarkers may be lost during this process. To visualize and characterize these proteins, we analyzed the depletome of 20 plasma samples by shotgun mass spectrometry‐based proteomics. Thus, using immunoaffinity depletion followed by 2‐D liquid chromatography coupled to an ion mobility‐enhanced mass spectrometer, our analysis identified that over 100 proteins are co‐eluting with the high abundant fraction. These proteins play roles in several biological processes, such as receptor‐mediated endocytosis, complement activation, and regulation of immune response. This study supports that investigating the depletome is important in the quest for biomarkers.