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Abstract Background African swine fever (ASF) is a highly contagious and severe haemorrhagic disease of Suidae, with mortalities that approach 100 percent. Several studies suggested the potential implication of non-biting dipterans in the spread of ASFV in pig farms due to the identification of the ASFV DNA. However, to our knowledge, no study has evaluated the viral DNA load in non-biting dipterans collected in outbreak farms and no risk factors have been analysed. In this context, our study aimed to analyse the risk factors associated with the presence of non-biting dipterans collected from ASF outbreaks in relation to the presence and load of viral DNA. Methods Backyard farms (BF), type A farms (TAF), and commercial farms (CF), were targeted for sampling in 2020. In 2021, no BF were sampled. Each farm was sampled only once. The identification of the collected flies to family, genus, or species level was performed based on morphological characteristics using specific keys and descriptions. Pools were made prior to DNA extraction. All extracted DNA was tested for the presence of the ASFV using a real-time PCR protocol. For this study, we considered every sample with a CT value of 40 as positive. The statistical analysis was performed using Epi Info 7 software (CDC, USA). Results All collected non-biting flies belonged to five families: Calliphoridae, Sarcophagidae, Fanniidae, Drosophilidae, and Muscidae. Of the 361 pools, 201 were positive for the presence of ASFV DNA. The obtained CT values of the positive samples ranged from 21.54 to 39.63, with a median value of 33.59 and a mean value of 33.56. Significantly lower CT values (corresponding to higher viral DNA load) were obtained in Sarcophagidae, with a mean value of 32.56; a significantly higher number of positive pools were noticed in August, mean value = 33.12. Conclusions Our study brings compelling evidence of the presence of the most common synanthropic flies near domestic pig farms carrying ASFV DNA, highlighting the importance of strengthening the biosecurity measures and protocols for prevention of the insect life cycle and distribution. Graphical Abstract

The pressure global food systems exert on the environment has amplified over time, following the growth of the human population, changes in the food consumption patterns and intensification of the production systems. Without changes in food systems, an overpressure on agriculture and food production will be unavoidable, with inevitable consequences for the environment and human health. Animal‐derived protein production like meat and milk is among the most important drivers of this impact. Under this context, the evaluation of the health and sustainability impacts of alternatives for animal‐based food proteins are central aspects of this discussion. Health risk‐benefit assessment (RBA) of foods estimates the overall human health impact following exposure to a particular food or food component, integrating risks and benefits in comparable measures. Methods that simultaneously consider the environmental impact of the food production systems were needed to perform a sustainable holistic assessment. "ALTERNATIVA | Alternative protein sources in the European diets – integrating health risk‐benefit and sustainability" aimed to develop a holistic approach to assess the impact of alternative protein sources, integrating health and sustainability assessment. This project, considering five complementary work packages, developed activities to enhance the capacity‐building among members of the consortium in applying methodologies related to the health RBA and sustainability impact assessment; to develop, in collaboration with different stakeholders, a protocol to integrate the assessment of the impact of foods/diets on health and on sustainability; to apply the derived framework to a case study, to validate it with a real case scenario, estimating the health and sustainability impacts of replacing red meat consumption by pulses in two different countries; and to disseminate and promote the established method of integrated health and sustainability assessment to other potential EU users. This external scientific report summarizes the main outputs and results obtained in the context of the ALTERNATIVA project. [ABSTRACT FROM AUTHOR]

Lysozyme obtained from PC2 hyperimmune egg whites, originating from hens immunized with a complex of bacterial and fungal antigens, was purified by ion exchange chromatography on Amberlite FPC 3500 resin. The purity of lysozyme was analysed by polyacrylamide gel electrophoresis in denaturing system (SDS-PAGE). Based on the migration pattern of the molecular marker, the presence of a single band with a molecular mass of 14.1 kDa was found. The agar gel immunodiffusion test (AGID) showed the presence of lysozyme which was tested in dilutions from 1/2 to 1/32 as compared to standard lysozyme (Sigma) and that obtained from eggs from chickens free of specific germs (SPF) and conventional (CV). The immunological identity between the standard lysozyme and the PC2 lysozyme was established by the AGID test. The PC2 lysozyme showed agglutination reactions with flaky clumps in the presence of Micrococcus lysodeikticus cultures and with granular clumps in the presence of Staphylococcus aureus cultures. The antimicrobial activity of lysozyme was intense against Gram-positive bacteria and less intense against Gram-negatives. The concentration of lysozyme was assessed by the lysoplate method against a Micrococcus lysodeikticus culture. Mean values (x̄ ± ds) of purified lysozyme were obtained, ranging between 12.5 mg/mL and 14,0 mg/mL. Lythic units (mg/mL) were determined in immunologically active products containing lysozyme (gels, emulsions, solutions, powders). The mean values (x̄ ± ds) were between 49.0±0.34 and 60.5±0.84. This study suggests that PC2 lysozyme exhibits an immunologic activity with an important role in the mechanisms of non-specific defense of organisms.

Lysozyme is a biologically active protein present in egg whites, having an essential role in the immune defense of the embryo. Due to its antimicrobial properties, it is extracted and used in the composition of various products as a preservative and/or a bioactive ingredient. Its qualitative and quantitative analysis can be performed by various test methods widely presented in the literature. The present paper presents the results obtained in assessing the lyzozyme content of aqueous solutions derived from hyperimmune egg white and/or from biologically active products in which it was included by using the sandwich ELISA technique.

In recent decades, avian antibody technology has become more popular in the medical field, successfully replacing the immunoglobulins obtained from mammals. Y antibodies (IgY) are extracted from the yolk of hyperimmune eggs harvested from hens (Gallus domesticus) immunized with different antigens and have an already proven applicability in various medical fields such as the manufacture of diagnostic kits and the development of passive antibody therapy protocols for various infectious diseases. It is well known that avian immunoglobulin therapy is effective in the prevention and treatment of bacterial diseases in humans and animals, especially with gastrointestinal localization. In the present study, the production of avian immunoglobulins with antiviral activity, their extraction, and purification, as well as the formulation of therapeutic products for administration to human patients exhibiting clinically symptomatic infections with rotavirus, Herpes simplex virus type 1, human papillomavirus or Varicella zoster virus. These are highly pathogenic viruses, leading to significant human morbidity and mortality, for which the therapeutic options are currently quite limited.

The research carried out describes one preparation procedure of the monovalent and polyvalent hyperimmune eggs, obtained from hens immunized with a bacterial and fungal antigen complex, pathogenic for humans, resistant or sensitive to antibiotics. It was prepared a monovalent antigen of Staphylococcus aureus MRSA strain, isolated from a patient with urinary infection. Lots of laying hens have been formed, at the beginning of the laying period, which was immunized on days, 0, 14 and 28. From the hyperimmune eggs obtained the specific immunoglobulin Y (IgY) was extracted and purified by polyethylene glycol precipitation (PEG 6000). Purity analysis was performed by polyacrylamide gel electrophoresis in a denatured system (SDS-PAGE) obtaining a single fraction with highlighting of heavy chains (molecular weight=75KDa) and light chains (molecular weight=25KDa). By the agar gel immune-diffusion assay (IDGA), the identity between analyzed IgY and standard IgY (MyBiosurse) was highlighted. The specificity of IgY antibodies obtained from hyperimmune egg yolk was proved by the ELISA immune enzymatic assay, obtaining high titers for each antigen present in the inoculums. The hyperimmune eggs were used for the preparation of biologically active products from the range IMUNOINSTANT, recommended in the therapy of diseases caused by multi-resistant germs.

Immunoglobulins Y (IgY) are the main antibody synthesized by the body of birds, reptiles, amphibians and some fish species. In the last decades, IgY has been receiving growing attention from medical specialists. It presents a number of structural differences from mammalian IgG and multiple benefits compared to mammalian antibodies. IgY can be applicable in the diagnosis, prevention, and therapy of infectious diseases of pathogenic bacteria, fungi or viruses. This study aimed to evaluate the specific antibody titer against 18 antigens for a period of 13 months. Throughout the experiment, ELISA assays were performed to determine the total immunoglobulin concentration in the obtained hyperimmune eggs and the specificity of the extracted IgG to each antigen.

Infections caused by multidrug-resistant microbial agents often complicate the treatment of patients with severe primary conditions such as neoplasms, organ failure or chronic diseases. The methicillin-resistantStaphylococcus aureus (MRSA) is one of the main global-spread nosocomial bacteria for which significant efforts are currently being made to develop new therapeutic options. Avian immunoglobulin (IgY) is used in various fields of human and veterinary medicine, with proven action against a wide range of microbial infections (bacterial, fungal and viral). In this study, IgY was successfully administered to a patient with MRSA-overloaded neoplasm, being incorporated into therapeutic products for general administration and local applications.

Abstract Novel foods could represent a sustainable alternative to traditional farming and conventional foodstuffs. Starting in 2018, Regulation (EU) 2283/2015 entered into force, laying down provisions for the approval of novel foods in Europe, including insects. This Approved Regulation establishes the requirements that enable Food Business Operators to bring new foods into the EU market, while ensuring high levels of food safety for European consumers. The present risk profile tackles the hazards for one of the most promising novel food insects, the house cricket (Acheta domesticus). The risk profile envisages a closed A. domesticus crickets rearing system, under Hazard Analysis and Critical Control Points (HACCP) and good farming practices (GFP), in contrast with open cricket farms. The methodology used involves screening the literature and identifying possible hazards, followed by adding relevant inclusion criteria for the evidence obtained. These criteria include animal health and food safety aspects, for the entire lifespan of crickets, based on the farm to fork One Health principle. When data were scarce, comparative evidence from close relatives of the Orthoptera genus was used (e.g. grasshoppers, locusts and other cricket species). Nevertheless, significant data gaps in animal health and food safety are present. Even if HACCP‐type systems are implemented, the risk profile identifies the following considerable concerns: (1) high total aerobic bacterial counts; (2) survival of spore‐forming bacteria following thermal processing; (3) allergenicity of insects and insect‐derived products; and (4) the bioaccumulation of heavy metals (e.g. cadmium). Other hazards like parasites, fungi, viruses, prions, antimicrobial resistance and toxins are ranked as low risk. For some hazards, a need for additional evidence is highlighted.

Feline Immunodeficiency Virus (FIV) infection represents a viral immunosuppressive condition that predominantly affects the immune system, having no curative therapeutic options. The present research focused on the biological and physical effect of the use of poly-specific avian immunoglobulin Y (IgY) on some immune molecules of FIV infected cats, especially on gamma-globulins and white blood cells, as well as on how this therapy can amend or improve the quality of life of infected cats. The polyspecific avian immunoglobulin Y was used to treat two groups of feline patients: 10 patients infected with FIV and 10 patients free of the disease. The patients were monitored via blood samples collected on days 0, 20 and 40. The samples were used for testing the biochemical profile, CBC, WBC and the plasma concentration of 10 cytokines: Fas, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12p40 and RANTES. The study also attempted to establish the eventual side effects associated with the suppressed immune system of FIV infected cats. [ABSTRACT FROM AUTHOR]

In this study, the effect of ejaculate sampling frequency on the morphological properties of sperm in Plymouth Rock roosters was determined. Four experimental groups were tested: of five, four, three, and two weekly ejaculates. The age of the roosters at the beginning of the experiment was 28 weeks and the duration of the experiment was 30 weeks. Increasing the frequency of semen sampling from one weekly sampling to five samplings per week had the following effects: 1. increase in ejaculate volume, but decrease in sperm density; 2. no significant changes in sperm motility; 3. increase of spermatozoa viability and 3. significant influences on the percentage of anomalies but no significant influences on the types of anomalies (the most common being the sperm head abnormalities). [ABSTRACT FROM AUTHOR]

Among skin pathologies, psoriasis is one of the most challenging disorders in terms of therapy and clinical monitoring of the patient.Psoriasis is a chronic inflammatory skin condition that has an autoimmune mechanism mediated by T cells, with involvement of innate immune cells.An important role is played by different subsets ofT-helper lymphocytes, as well as a number of proinflammatory cytokines that maintain the chronic inflammatory state. Although some evidence suggests that NK cells play animportant role in psoriasis, their contribution to disease's development and progression remains incompletely understood. Experimental models are an indispensable tool in psoriasis research for both pathogenesis evaluation and for possible therapeutic targets identification. Due to its advantages, experimental models such as the Imiquimod-based mice model is one of the most popular animalmodel to study psoriasiform dermatitis in mice. In order to highlight the role of NK cells in the development of psoriasis, we performed the Imiquimod-based mice model and analysed NK cells distribution and phenotype by flow cytometry. Skin inflammation induced by Imiquimodwas assessed based on in vivo parameters (erythema, desquamation, thickening), PASI score, evaluation of splenomegaly and pathological examination. NK cells distribution was evaluated on three immune levels: peripheral blood spleen and skin There were quantified NK cells subsets based on the expression of CD27 and CDUb, and NK1.1+CD11c+CD45R+subset. The expression of CD43, KLRG1 and CD69, markers with an important role in NK cells maturation and activation, was analysed.Our data show that, although a somewhat neglected cell in psoriasis, NKs correlate with the gravity of the auto-immune disease. [ABSTRACT FROM AUTHOR]

In this paper, we determined the evolution of ejaculate volume, sperm concentration, motility and viability along with the evolution of the activity of some enzymes (TGO, alkaline phosphatase, ATPase and lactic dehydrogenase) in seminal plasma of Cornish roosters at 30, 42, 57 and 63 weeks of age. The monitored parameters showed a specific evolution that characterizes the physiological ageing process in Cornish roosters. Thus, the volume of ejaculate showed a significant decrease (P = 0.011) from 0.39 to 0.32 microliters. The number of sperm per ejaculate decreased from 0.86 to 0.29x109 (P = 0.002). Motility decreased from 69% to 41% (P = 0.001) and sperm viability decreased from 89 to 74% (P = 0.00). The percentage of abnormal spermatozoa decreased not significantly, from 96 to 94%, some types of abnormalities increasing and others decreasing in frequency during the monitoring period. The activity of the monitored enzymes characterized the decrease of the anaerobic metabolism of spermatozoa and the increase of the membrane permeability of the spermatozoa in the physiological ageing process. [ABSTRACT FROM AUTHOR]

Lysozyme obtained from PC2 hyperimmune egg whites, originating from hens immunized with a complex of bacterial and fungal antigens, was purified by ion exchange chromatography on Amberlite FPC 3500 resin. The purity of lysozyme was analysed by polyacrylamide gel electrophoresis in denaturing system (SDS-PAGE). Based on the migration pattern of the molecular marker, the presence of a single band with a molecular mass of 14.1 kDa was found. The agar gel immunodiffusion test (AGID) showed the presence of lysozyme which was tested in dilutions from 1/2 to 1/32 as compared to standard lysozyme (Sigma) and that obtained from eggs from chickens free of specific germs (SPF) and conventional (CV). The immunological identity between the standard lysozyme and the PC2 lysozyme was established by the AGID test. The PC2 lysozyme showed agglutination reactions with flaky clumps in the presence of Micrococcus lysodeikticus cultures and with granular clumps in the presence of Staphylococcus aureus cultures. The antimicrobial activity of lysozyme was intense against Gram-positive bacteria and less intense against Gram-negatives. The concentration of lysozyme was assessed by the lysoplate method against a Micrococcus lysodeikticus culture. Mean values (x ± ds) of purified lysozyme were obtained, ranging between 12.5 mg/mL and 14,0 mg/mL. Lythic units (mg/mL) were determined in immunologically active products containing lysozyme (gels, emulsions, solutions, powders). The mean values (x ± ds) were between 49.0±0.34 and 60.5±0.84. This study suggests that PC2 lysozyme exhibits an immunologic activity with an important role in the mechanisms of non-specific defence of organisms. [ABSTRACT FROM AUTHOR]

Copyright of Veterinary Drug / Medicamentul Veterinar is the property of Romanian National Association of the Veterinary Products Manufacturers and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The research carried out describes one preparation procedure of the monovalent and polyvalent hyper immune eggs, obtained from hens immunized with a bacterial and fungal antigen complex, pathogenic for humans, resistant or sensitive to antibiotics. It was prepared a monovalent antigen of Staphylococcus aureus MRSA strain, isolated from a patient with urinary infection. Lots of laying hens have been formed, at the beginning of the laying period, which were immunized on days, 0, 14 and 28. From the hyper immune eggs obtained the specific immunoglobulin Y (IgY) was extracted and purified by polyethylene glycol precipitation (PEG 6000). Purity analysis was performed by polyacrylamide gel electrophoresis in a denatured system (SDS-PAGE) obtaining a single fraction with highlighting of heavy chains (molecular weight=75KDa) and light chains (molecular weight=25KDa). By the agar gel immune-diffusion assay (IDGA), the identity between analyzed IgY and standard IgY (MyBiosurse) was highlighted. The specificity of IgY antibodies obtained from hyper immune egg yolk was proved by the ELISA immune enzymatic assay, obtaining high titers for each antigen present in the inoculum. The hyper immune eggs were used for preparation of biologically active products from the range IMUNOINSTANT, recommended in the therapy of diseases caused by multi resistant germs. [ABSTRACT FROM AUTHOR]

In recent decades, avian antibody technology has become more popular in the medical field, successfully replacing the immunoglobulins obtained from mammals. Y antibodies (IgY) are extracted from the yolk of hyperimmune eggs harvested from hens (Gallus domesticus) immunized with different antigens and have an already proven applicability in various medical fields such as the manufacture of diagnostic kits and the development of passive antibody therapy protocols for various infectious diseases. It is well known that avian immunoglobulin therapy is effective in the prevention and treatment of bacterial diseases in humans and animals, especially with gastrointestinal localization. In the present study, the production of avian immunoglobulins with antiviral activity, their extraction and purification, as well as the formulation of therapeutic products for administration to human patients exhibiting clinically symptomatic infections with rotavirus, Herpes simplex virus type 1, human papillomavirus or Varicella zoster virus. These are highly pathogenic viruses, leading to significant human morbidity and mortality, for which the therapeutic options are currently quite limited. [ABSTRACT FROM AUTHOR]

Infections caused by multidrug-resistant microbial agents often complicate the treatment of patients with severe primary conditions such as neoplasms, organ failure or chronic diseases. The methicillin-resistant Staphylococcus aureus (MRSA) is one of the main global-spread nosocomial bacteria for which significant efforts are currently being made to develop new therapeutic options. Avian immunoglobulin (IgY) is used in various fields of human and veterinary medicine, with proven action against a wide range of microbial infections (bacterial, fungal and viral). In this study, IgY was successfully administered to a patient with MRSA-overloaded neoplasm, being incorporated into therapeutic products for general administration and local applications. [ABSTRACT FROM AUTHOR]

Copyright of Veterinary Drug / Medicamentul Veterinar is the property of Romanian National Association of the Veterinary Products Manufacturers and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The present research focuses on the determination of the immunoglobulin Y (IgY) levels derived from egg yolks obtained from hens previously inoculated with a combination of bacterial and fungal antigens (multiple antigens). The purpose of the work consists of establishing the frequency of the inoculations, in order to obtain a high level of antibodies throughout the duration of the experiment. The antigens were obtained from inactivated bacterial and fungal humans pathogenic strains. Egg-laying hen lots were formed out of individuals at the beginning of the egg-laying period. The hens were inoculated with multiple antigens, three times, at the first, 14th and the 28th day of the experiment. The control for the immune response was performed by sampling eggs on the 14th day from the third inoculation process. The IgY was extracted from the egg yolk in order to obtain the aqueous phase. The characterization of the IgY titers was performed every 30 days, for a period of 9 months, by using qualitative and quantitative ELISA assays. Following the end of the 9 months period since the 3rd inoculation process, the specific IgY titers maintained at high levels, another (4th) inoculation shouldn't be necessary. [ABSTRACT FROM AUTHOR]

Novel foods could represent a sustainable alternative to traditional farming and conventional foodstuffs. Starting in 2018, Regulation (EU) 2283/2015 entered into force, laying down provisions for the approval of novel foods in Europe, including insects. This Approved Regulation establishes the requirements that enable Food Business Operators to bring new foods into the EU market, while ensuring high levels of food safety for European consumers. The present risk profile tackles the hazards for one of the most promising novel food insects, the house cricket ( Acheta domesticus ). The risk profile envisages a closed A. domesticus crickets rearing system, under Hazard Analysis and Critical Control Points (HACCP) and good farming practices (GFP), in contrast with open cricket farms. The methodology used involves screening the literature and identifying possible hazards, followed by adding relevant inclusion criteria for the evidence obtained. These criteria include animal health and food safety aspects, for the entire lifespan of crickets, based on the farm to fork One Health principle. When data were scarce, comparative evidence from close relatives of the Orthoptera genus was used (e.g. grasshoppers, locusts and other cricket species). Nevertheless, significant data gaps in animal health and food safety are present. Even if HACCP-type systems are implemented, the risk profile identifies the following considerable concerns: (1) high total aerobic bacterial counts; (2) survival of spore-forming bacteria following thermal processing; (3) allergenicity of insects and insect-derived products; and (4) the bioaccumulation of heavy metals (e.g. cadmium). Other hazards like parasites, fungi, viruses, prions, antimicrobial resistance and toxins are ranked as low risk. For some hazards, a need for additional evidence is highlighted., (© 2018 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority.)