Immuno-protective effect of commercial IBD vaccines against emerging novel variant infectious bursal disease virus in specific-pathogen-free chickens.

IMPORTANCE: Infectious bursal disease (IBD) is an important viral poultry disease that vaccination can control. OBJECTIVE: This study examined the immune protection of immune-complex (Vaccine A) and attenuated live (Vaccine B) IBD vaccines in specific-pathogen-free (SPF) chickens against a novel Malaysian variant IBD virus (vaIBDV) challenge. METHODS: One-day-old (n =75) SPF chickens were divided randomly into the following three groups of 25 chicks each: Control, Vaccine A, and Vaccine B groups. The vaIBDV strain, UPM1432/2019, was used for the challenge at 21 and 28days post-vaccination (dpv). Five birds from unchallenged and challenged groups were sacrificed seven days post-challenge, and blood, bursa, spleen, and cloacal swabs were collected. The IBD antibodies (Abs), lymphoid lesions, and viral load were determined. RESULTS: The UPM1432/2019 virus induced bursal damage in vaccinated SPF chickens despite Ab titers. The mean Ab titers of the Vaccine A challenged group were significantly lower (p < 0.002) than in the unchallenged group at 28 dpv. The bursal indices of the vaccinated unchallenged groups did not differ significantly from those of the vaccinated challenged groups (p = 0.94). Microscopically, the bursae of the challenged groups showed significant atrophy. The bursal lesion score was higher (p < 0.05) in the control and Vaccine B challenged groups than the Vaccine A challenged group. The challenged group had a higher viral load than the vaccinated groups (p < 0.001). CONCLUSIONS AND RELEVANCE: Neither vaccine fully protected against a vaIBDV challenge, highlighting the limitations of current vaccines and the need for further research.

Broadly cross-reactive immune responses in chickens immunized with chimeric virus-like particles of nodavirus displaying the M2e originated from avian and human influenza A viruses.

Avian influenza A viruses (IAVs) pose a persistent threat to poultry industry worldwide, despite the presence of vaccines. Additionally, reverse-zoonosis transmission potentially introduces human-originated IAVs into poultry and complicates the efforts to control the spread of influenza. Current avian influenza vaccines are primarily based upon the rapidly mutating hemagglutinin (HA) and neuraminidase (NA) glycoproteins, which limit their efficacy against diverse strains of IAVs. Hence, the highly conserved ectodomains of matrix 2 protein (M2e) of IAVs are widely studied as alternatives to the HA and NA. However, the differences in the M2e amino acid sequences between avian and human IAVs generate antibodies that do not cross-react reciprocally with IAVs from other origins. To broaden and enhance the immunogenicity of M2e, we fused two copies each of the M2e derived from avian and human IAVs at the C-terminal end of the Macrobrachium rosenbergii nodavirus (MrNV) capsid protein (NvC). Transmission electron microscopic and dynamic light scattering analyses revealed that the chimeric protein self-assembled into virus-like particles (VLPs). Immunization of chickens with the chimeric VLPs demonstrated a robust induction of broadly reactive immune responses against both the M2e of avian and human IAVs. Additionally, the chimeric VLPs elicited the production of cytotoxic T lymphocytes (CTL), macrophages, as well as a well-balanced Th1 and Th2 population, indicating their potential in activating cell-mediated immune responses in chickens. Furthermore, the chimeric VLPs triggered the production of both Th1- and Th2-cytokines, attesting their potential in mounting a robust and balanced immune response in avian species. This study demonstrated the potential of these chimeric VLPs in stimulating and broadening cross-reactive immune responses in chickens against both avian and human IAVs.

Virus-like particles in poultry disease: an approach to effective and safe vaccination.

The poultry industry, a cornerstone of global food security, faces dynamic challenges exacerbated by viral diseases. This review traces the trajectory of poultry vaccination, evolving from traditional methods to the forefront of innovation Virus-Like Particle (VLP) vaccines. Vaccination has been pivotal in disease control, but traditional vaccines exhibit some limitations. This review examines the emergence of VLPs as a game-changer in poultry vaccination. VLPs, mimicking viruses without replication, offer a safer, targeted alternative with enhanced immunogenicity. The narrative encompasses VLP design principles, production methods, immunogenicity, and efficacy against major poultry viruses. Challenges and prospects are explored, presenting VLP vaccines as a transformative technique in poultry disease control. Understanding their potential empowers industry stakeholders to navigate poultry health management with precision, promising improved welfare, reduced economic losses, and heightened food safety.

Betacyanins from red pitahaya (Hylocereus polyrhizus) exhibit antiviral response against influenza A virus.

Seasonal influenza affects millions of lives worldwide, with the influenza A virus (IAV) responsible for pandemics and annual epidemics, causing the most severe illnesses resulting in patient hospitalizations or death. With IAV threatening the next global influenza pandemic, it is a race against time to search for antiviral drugs. Betacyanins are unique nitrogen-containing and water-soluble reddish-violet pigments that have been reported to possess antiviral properties against the dengue virus. This study aimed to examine the antiviral effect of betacyanins from red pitahaya (Hylocereus polyrhizus) on IAV-infected lung epithelial A549 cells. HPLC and LC-MS analysis of extracted betacyanin showed four betacyanins in the betacyanin fraction: phyllocactin, hylocerenin, betanin, and isobetanin. Cytotoxicity assay showed that betacyanin fractions were not cytotoxic to A549 cells at concentrations below 100 µg/mL. Betacyanin fraction concentrations of 12.5, 25.0, and 50.0 µg/mL prevented the formation of viral cytopathic effect and reduced virus titer in IAV-infected cells up to 72 h. A downregulation of protein and mRNA nucleoprotein expression levels was observed after treatment with 25.0 and 50.0 µg/mL of betacyanin fraction after 24 h, thereby providing evidence for the antiviral activity of betacyanin from red pitahaya against IAV in vitro.

Campylobacteriosis and Control Strategies against Campylobacters in Poultry Farms.

Campylobacteriosis is a significant foodborne illness caused by Campylobacter bacteria. It is one of the most common bacterial causes of gastroenteritis worldwide, with poultry being a major reservoir and source of infection in humans. In poultry farms, Campylobacters colonize the intestinal tract of chickens and contaminate meat during processing. Vaccines under development against Campylobacters in poultry showed partial or no protection against their cecal colonization. Therefore, this review will elaborate on campylobacteriosis and emphasize the control strategies and recent vaccine trials against Campylobacters in poultry farms. The epidemiology, diagnosis, and treatment of Campylobacter infection, along with specific mention of poultry Campylobacter contamination events in Malaysia, will also be discussed.

Sequence Analysis of the Malaysian Low Pathogenic Avian Influenza Virus Strain H5N2 from Duck.

The avian influenza viruses (AIV) of the H5 subtype have the ability to mutate from low pathogenic (LPAI) to highly pathogenic (HPAI), which can cause high mortality in poultry. Little is known about the pathogenic switching apart from the mutations at the haemagglutinin cleavage site, which significantly contributes to the virus virulence switching phenomenon. Therefore, this study aimed to compare the molecular markers in the haemagglutinin (HA), neuraminidase (NA), and matrix (M) genes of a locally isolated LPAI AIV strain H5N2 from Malaysia with the reference HPAI strains using bioinformatics approaches, emphasising the pathogenic properties of the viral genes. First, the H5N2 strain A/Duck/Malaysia/8443/2004 was propagated in SPF eggs. The viral presence was verified by haemagglutination assay, RT-PCR, and sequencing. Results showed successful amplifications of HA (1695 bp), NA (1410 bp), and M (1019 bp) genes. The genes were sequenced and the deduced amino acid sequences were analysed computationally using MEGA 11 and NetNGlyc software. Analysis of the HA protein showed the absence of the polybasic cleavage motif, but presence of two amino acid residues that are known to affect pathogenicity. There were also two glycosylation sites (glycosites) compared to the reference HPAI viruses, which had three or more at the HA globular head domain. No NA stalk deletion was detected but the haemadsorbing and active centres of the studied NA protein were relatively similar to the reference HPAI H5N2 isolates of duck but not chicken origins. Six NA glycosites were also identified. Finally, we observed a consistent M1 and M2 amino acid sequences between our LPAI isolate with the other HPAI H5N1 or H5N2 reference proteins. These data demonstrate distinct characteristics of the Malaysian LPAI H5N2, compared to HPAI H5N2 or H5N1 from ducks or chickens, potentially aiding the epidemiological research on genetic dynamics of circulating AIV in poultry.

Translating conventional wisdom on chicken comb color into automated monitoring of disease-infected chicken using chromaticity-based machine learning models.

Bacteria- or virus-infected chicken is conventionally detected by manual observation and confirmed by a laboratory test, which may lead to late detection, significant economic loss, and threaten human health. This paper reports on the development of an innovative technique to detect bacteria- or virus-infected chickens based on the optical chromaticity of the chicken comb. The chromaticity of the infected and healthy chicken comb was extracted and analyzed with International Commission on Illumination (CIE) XYZ color space. Logistic Regression, Support Vector Machines (SVMs), K-Nearest Neighbors (KNN), and Decision Trees have been developed to detect infected chickens using the chromaticity data. Based on the X and Z chromaticity data from the chromaticity analysis, the color of the infected chicken's comb converged from red to green and yellow to blue. The development of the algorithms shows that Logistic Regression, SVM with Linear and Polynomial kernels performed the best with 95% accuracy, followed by SVM-RBF kernel, and KNN with 93% accuracy, Decision Tree with 90% accuracy, and lastly, SVM-Sigmoidal kernel with 83% accuracy. The iteration of the probability threshold parameter for Logistic Regression models has shown that the model can detect all infected chickens with 100% sensitivity and 95% accuracy at the probability threshold of 0.54. These works have shown that, despite using only the optical chromaticity of the chicken comb as the input data, the developed models (95% accuracy) have performed exceptionally well, compared to other reported results (99.469% accuracy) which utilize more sophisticated input data such as morphological and mobility features. This work has demonstrated a new feature for bacteria- or virus-infected chicken detection and contributes to the development of modern technology in agriculture applications.

Effects of Phage Cocktail, Probiotics, and Their Combination on Growth Performance and Gut Microbiota of Broiler Chickens.

Phages, which are often used therapeutically, have begun to receive interest as alternatives to antibiotic growth promoters (AGPs) for enhancing chicken growth. Another option that has been extensively studied as a growth promoter in chickens is probiotics. To the best of our knowledge, there is currently no study available on the use of phages and probiotics in combination as potential feed additives for broiler chickens. Therefore, this study demonstrated the effects of a phage cocktail, probiotics, and their combination on the growth performance and gut microbiota of broiler chickens. A total of 288 one-day-old male Cobb 500 broilers were randomly allotted to one of six treatments in a completely randomised design. The treatments were (i) C (basal diet (BD) only), (ii) 1ϕ (BD + 0.1% phage cocktail), (iii) 2ϕ (BD + 0.2% phage cocktail), (iv) P (BD + 0.1% probiotic), (v) 1ϕP (BD + 0.1% phage cocktail + 0.1% probiotic), and (vi) 2ϕP (BD + 0.2% phage cocktail + 0.1% probiotic). The 1ϕP treatment had significantly (p < 0.05) better BW (35 days), BWG (22-35 days, 1-35 days), and FCR (1-21 days, 22-35 days, 1-35 days) compared to C. Unique gut microbiota diversity was also found between the ϕP (1ϕP and 2ϕP) and non-ϕP groups (C, 1ϕ, 2ϕ, and P) in ilea, particularly in the 35-day-old chickens. Microorganisms associated with short-chain fatty acid (SCFA) producers were significantly (p < 0.05) more present in the ϕP group than in the non-ϕP group. The predicted genes related to carbohydrate and amino acid metabolism were significantly upregulated in ϕP groups compared to non-ϕP groups. These genes were involved in the digestion and absorption of nutrients, as well as the production of energy. Our findings showed that the 1ϕP treatment could be a potential alternative to AGPs for poultry, as growth performance was enhanced, and gut microbiota was positively modulated.

Expression Profiles of Immune-Related Genes and Apoptosis Study of Avian Intraepithelial-Natural Killer Cells in Chickens Inoculated with Vaccine Strain of Newcastle Disease Virus (NDV) and Challenged with Virulent NDV.

In spite of the available information on the role of natural killer (NK) cells in several viral infections, the interactions between chicken intraepithelial-NK (IEL-NK) cells and Newcastle disease virus (NDV) are poorly understood. In this study, we investigated these interactions following the inoculation of chickens with NDV vaccine strain LaSota and subsequent challenge with velogenic NDV (vNDV) genotype VII (GVII) and VIII (GVIII), through quantification of IEL-NK cell's apoptosis and expression profiling of its surface receptors. Specific-pathogen-free chickens were randomly divided into six groups, as follows: one group of an uninfected control, one group infected with NDV LaSota, two groups each infected with either GVII or GVIII, and two groups inoculated with NDV LaSota and challenged with either GVII (LaSota-genotype VII [LSGVII]) or GVIII (LaSota-genotype VIII [LSGVIII]). Avian intraepithelial lymphocytes (IEL) were isolated from the duodenal loops, and CD3- cells were characterized. Immunophenotyping and apoptosis analysis of CD3-/CD25+/CD45+IEL NK cells were conducted using a flow cytometer. In addition, a gene expression study was conducted using real-time quantitative PCR. Data were analyzed using two-way analysis of variance. The results showed that vNDV GVII or GVIII caused apoptosis of IEL-NK cells; however, following inoculation of LSGVII or LSGVIII, the effect of vNDV GVII and GVIII to cause a reduction in the population of viable IEL-NK cells was significantly reduced. Furthermore, the expression profiles of activating receptors CD69, NK-lysin, and IFN-γ, were generally upregulated in chickens inoculated with LSGVII or LSGVIII. In contrast, B-NK, an inhibitory receptor, was downregulated in these treatment groups. In NDV GVII- and GVIII-challenged groups, however, B-NK was upregulated, whereas the other receptors were generally downregulated. The findings of this study showed that NDV vaccine strain LaSota may prevent apoptosis and cause upregulation of activating receptors of chicken IEL-NK cells in velogenic virus-challenged settings.

Perfiles de expresión de genes relacionados con la inmunidad y estudio de apoptosis de células asesinas naturales intraepiteliales aviares en pollos inoculados con la cepa vacunal del virus de la enfermedad de Newcastle (NDV) y desafiados con de Newcastle virulento. A pesar de la información disponible sobre el papel de las células asesinas naturales (NK) en varias infecciones virales, se conoce poco acerca de las interacciones entre las células NK intraepiteliales de pollo (IEL-NK) y el virus de la enfermedad de Newcastle (NDV). En este estudio, investigamos estas interacciones luego de la inoculación de pollos con la cepa vacunal LaSota y con el desafío posterior con los genotipo VII (GVII) y VIII (GVIII) velogénico de NDV (vNDV), mediante la cuantificación de la apoptosis de las células IEL-NK y los perfiles de expresión de sus receptores de superficie. Los pollos libres de patógenos específicos se dividieron aleatoriamente en seis grupos, de la siguiente manera: un grupo de control no infectado, un grupo infectado con LaSota, dos grupos cada uno infectado con GVII o GVIII, y dos grupos inoculados con LaSota y desafiados con ya sea el genotipo GVII (LaSota-genotipo VII [LSGVII]) o con el genotipo GVIII (LaSota-genotipo VIII [LSGVIII]). Se aislaron células NK intraepiteliales de pollo de las asas duodenales y se caracterizaron las células CD3-. La inmunofenotipificación y el análisis de apoptosis de las células NK CD3-/CD25+/CD45+IEL se realizaron utilizando citometría de flujo. Además, se realizó un estudio de expresión de genes mediante PCR cuantitativa en tiempo real. Los datos se analizaron utilizando un análisis de varianza de dos vías. Los resultados mostraron que el virus de Newcastle genotipos GVII o GVIII causaron apoptosis de células NK intraepiteliales; sin embargo, después de por los tratamientos LaSota-genotipo VII o LaSota-genotipo VIII, el efecto de del virus virulento de Newcastle GVII y GVIII para provocar una reducción en la población de células NK intraepiteliales viables se redujo significativamente. Además, los perfiles de expresión de los receptores activadores CD69, NK-lisina e IFN-γ generalmente aumentaron en pollos inoculados con los tratamientos LaSota-genotipo VII o LaSota-genotipo VIII. Por el contrario, B-NK, que es un receptor inhibidor, se reguló a la baja en estos grupos de tratamiento. Sin embargo, en los grupos expuestos a los virus de Newcastle genotipos GVII y GVIII, el gene B-NK estaba regulado al alza, mientras que los otros receptores generalmente estaban regulados a la baja. Los hallazgos de este estudio mostraron que la cepa vacunal LaSota puede prevenir la apoptosis y causar una regulación al alza de los receptores activadores de las células NK intraepiteliales de pollo en entornos expuestos al virus velogénico.

Fucoxanthin Production of Microalgae under Different Culture Factors: A Systematic Review.

Fucoxanthin is one of the light-harvesting pigments in brown microalgae, which is increasingly gaining attention due to its numerous health-promoting properties. Currently, the production of microalgal fucoxanthin is not yet feasible from an economic perspective. However, the cultivation of microalgae at favourable conditions holds great potential to increase the viability of this fucoxanthin source. Hence, this study aimed to review the fucoxanthin production of microalgae under different conditions systematically. A literature search was performed using the Web of Science, Scopus and PubMed databases. A total of 188 articles were downloaded and 28 articles were selected for the current review by two independent authors. Microalgae appeared to be a more reliable fucoxanthin source compared to macroalgae. Overall, a consensus fucoxanthin production condition was obtained and proposed: light intensity ranging from 10 to 100 µmol/m2/s could achieve a higher fucoxanthin content. However, the optimal light condition in producing fucoxanthin is species-specific. The current review serves as an antecedent by offering insights into the fucoxanthin-producing microalgae response to different culture factors via a systematic analysis. With the current findings and recommendations, the feasibility of producing fucoxanthin commercially could be enhanced and possibly achieve practical and sustainable fucoxanthin production.

Efficacy of genotype-matched Newcastle disease virus vaccine formulated in carboxymethyl sago starch acid hydrogel in chickens vaccinated via different routes.

BACKGROUND: The commercially available Newcastle disease (ND) vaccines were developed based on Newcastle disease virus (NDV) isolates genetically divergent from field strains that can only prevent clinical disease, not shedding of virulent heterologous virus, highlighting the need to develop genotype-matched vaccines. OBJECTIVES: This study examined the efficacy of the NDV genotype-matched vaccine, mIBS025 strain formulated in standard vaccine stabilizer, and in carboxymethyl sago starch-acid hydrogel (CMSS-AH) following vaccination via an eye drop (ED) and drinking water (DW). METHODS: A challenge virus was prepared from a recent NDV isolated from ND vaccinated flock. Groups of specific-pathogen-free chickens were vaccinated with mIBS025 vaccine strain prepared in a standard vaccine stabilizer and CMSS-AH via ED and DW and then challenged with the UPM/NDV/IBS362/2016 strain. RESULTS: Chickens vaccinated with CMSS-AH mIBS025 ED (group 2) developed the earliest and highest Hemagglutination Inhibition (HI) NDV antibody titer (8log2) followed by standard mIBS025 ED (group 3) (7log2) both conferred complete protection and drastically reduced virus shedding. By contrast, chickens vaccinated with standard mIBS025 DW (group 5) and CMSS-AH mIBS025 DW (group 4) developed low HI NDV antibody titers of 4log2 and 3log2, respectively, which correspondingly conferred only 50% and 60% protection and continuously shed the virulent virus via the oropharyngeal and cloacal routes until the end of the study at 14 dpc. CONCLUSIONS: The efficacy of mIBS025 vaccines prepared in a standard vaccine stabilizer or CMSS-AH was affected by the vaccination routes. The groups vaccinated via ED had better protective immunity than those vaccinated via DW.

Pathogenesis of Chicken Astrovirus Related Illnesses.

Of the several known viruses, chicken astrovirus (CAstV) has been associated with diarrhea, runting-stunting syndrome, severe kidney disease, and gout, and white chick syndrome (WCS) in young broiler chicks. Discovered in 2004, CAstV consists of two genogroups with an expanding subgroup because of the diversity exhibited in its viral capsid sequence. Despite these findings, there exists a dearth of knowledge on its pathogenesis. This review highlights the pathogenesis and development of in vivo and in vitro models.

Metagenomics Insights Into the Microbial Diversity and Microbiome Network Analysis on the Heterogeneity of Influent to Effluent Water.

Sanitizing the water sources of local communities is important to control the spread of microbial resistance genes, especially those for water-borne illnesses. The activities of antibiotic resistance gene (ARG)-host pathogens pose a threat to public health, and it has been estimated that the infection will lead up to 10 million deaths globally by the year 2050. Hence, in this study, we aim to analyze the efficiency of our municipal wastewater treatment plant (WWTP) process in producing pathogen-free water by investigating the microbial composition between influent and effluent water sites. Shotgun metagenomics sequencing using the Illumina platform was performed on the influent and effluent samples of six different WWTP sites located in Johore, Malaysia. After raw data pre-processing, the non-redundant contigs library was then aligned against BLASTP for taxonomy profiling and the Comprehensive Antibiotic Resistance Database for ARG annotation. Interestingly, the alpha-diversity result reported that effluent site samples showed higher abundance and diverse heterogeneity compared to the influent site. The principal component analysis (PCA) and non-metric multidimensional scaling (NMDS) plots also suggested that effluent sites showed high variation in the genetic material due to loosely clustered sample plots, as compared to the tightly clustered influent samples. This study has successfully identified the top three abundant phyla in influent-Proteobacteria, Firmicutes, and Bacteroidetes-and effluent-Proteobacteria, Actinobacteria, and Bacteroidetes-water. Despite the overlap within the top three abundant phyla in influent and effluent sites (Proteobacteria and Bacteroidetes), the ARG composition heat map and drug class phenotype plot bar exhibits a general trend of a downward shift, showing the efficiency of WWTP in reducing opportunistic pathogens. Overall, it was demonstrated that our municipal WWTP efficiently eliminated pathogenic microbes from the influent water before its total discharge to the environment, though not with the total elimination of microorganisms. This metagenomics study allowed for an examination of our water source and showed the potential interaction of species and ARGs residing in the influent and effluent environment. Both microbial profile structure and co-occurrence network analysis provide integrated understanding regarding the diversity of microorganisms and interactions for future advanced water sanitation treatments.

Clinical and Preclinical Studies of Fermented Foods and Their Effects on Alzheimer's Disease.

The focus on managing Alzheimer's disease (AD) is shifting towards prevention through lifestyle modification instead of treatments since the currently available treatment options are only capable of providing symptomatic relief marginally and result in various side effects. Numerous studies have reported that the intake of fermented foods resulted in the successful management of AD. Food fermentation is a biochemical process where the microorganisms metabolize the constituents of raw food materials, giving vastly different organoleptic properties and additional nutritional value, and improved biosafety effects in the final products. The consumption of fermented foods is associated with a wide array of nutraceutical benefits, including anti-oxidative, anti-inflammatory, neuroprotective, anti-apoptotic, anti-cancer, anti-fungal, anti-bacterial, immunomodulatory, and hypocholesterolemic properties. Due to their promising health benefits, fermented food products have a great prospect for commercialization in the food industry. This paper reviews the memory and cognitive enhancement and neuroprotective potential of fermented food products on AD, the recently commercialized fermented food products in the health and food industries, and their limitations. The literature reviewed here demonstrates a growing demand for fermented food products as alternative therapeutic options for the prevention and management of AD.

An Insight into the Molecular Characteristics and Associated Pathology of Chicken Astroviruses.

The chicken astrovirus (CAstV) is a ubiquitous enteric RNA virus that has been associated mainly with conditions, such as the runting-stunting syndrome, severe kidney disease, visceral gout, and white chick syndrome, in broiler-type chickens worldwide. Sequence analysis of the capsid genes' amino acids of the strains involved in these conditions reveals a genetic relationship and diversity between and within the CAstV genogroups and subgroups based on phylogenetic analysis, genetic distance (p-dist), and pathogenicity. While the two genogroups (A and B) are demarcated phylogenetically, their pairwise amino acid sequence identity is 39% to 42% at a p-dist of 0.59 to 0.62. Group-A consists of three subgroups (Ai, Aii, and Aiii) with an inter- and intra-subgroup amino acid identity of 78% to 82% and 92% to 100%, respectively, and a p-dist of 0.18 to 0.22. On the other hand, the six subgroups (Bi, Bii, Biii, Biv, Bv, and Bvi) in Group-B, with a p-dist of 0.07 to 0.18, have an inter- and intra-subgroup amino acid identity of 82% to 93% and 93% to 100%, respectively. However, these groupings have little to no effect on determining the type of CAstV-associated pathology in chickens.

Expression of Toll-like receptors 3, 7, 9 and cytokines in feline infectious peritonitis virus-infected CRFK cells and feline peripheral monocytes.

BACKGROUND: The role of Toll-like receptors (TLRs) in a feline infectious peritonitis virus (FIPV) infection is not completely understood. OBJECTIVES: This study examined the expression of TLR3, TLR7, TLR9, tumor necrosis factor-alpha (TNF-α), interferon (IFN)-ß, and interleukin (IL)-10 upon an FIPV infection in Crandell-Reese feline kidney (CRFK) cells and feline monocytes. METHODS: CRFK cells and monocytes from feline coronavirus (FCoV)-seronegative cats and FCoV-seropositive cats were infected with type II FIPV-79-1146. At four, 12, and 24 hours post-infection (hpi), the expression of TLR3, TLR7, TLR9, TNF-α, IFN-ß, and IL-10, and the viral load were measured using reverse transcription quantitative polymerase chain reaction. Viral protein production was confirmed using immunofluorescence. RESULTS: FIPV-infected CRFK showed the upregulation of TLR9, TNF-α, and IFN-ß expression between 4 and 24 hpi. Uninfected monocytes from FCoV-seropositive cats showed lower TLR3 and TLR9 expression but higher TLR7 expression compared to uninfected monocytes from FCoV-seronegative cats. FIPV-infected monocytes from FCoV-seropositive cats downregulated TLR7 and TNF-α expression between 4 and 24 hpi, and 4 and 12 hpi, respectively. IFN-ß was upregulated early in FIPV-infected monocytes from FCoV-seropositive cats, with a significant difference observed at 12 hpi compared to FCoV-seronegative cats. The viral load in the CRFK and FIPV-infected monocytes in both cohorts of cats was similar over time. CONCLUSION: TLR7 may be the key TLR involved in evading the innate response against inhibiting TNF-α production. Distinct TLR expression profiles between FCoV-seronegative and FCoV-seropositive cats were observed. The associated TLR that plays a role in the induction of IFN-ß needs to be explored further.

Alteration in the Population of Intraepithelial Lymphocytes and Virus Shedding in Specific-Pathogen-Free Chickens Following Inoculation with Lentogenic and Velogenic Newcastle Disease Virus Strains.

Intraepithelial lymphocytes (IELs) provide the first line of immunological defense after the invasion of the intestine by a pathogen. To understand the changes of IEL response in chickens, we measured the population of different subsets of avian IELs at different time points after primary inoculation of Newcastle disease virus (NDV) lentogenic strain (LaSota) and subsequent challenge with NDV velogenic strain- genotypes VII and VIII. Furthermore, NDV shed after each treatment was quantified. Specific-pathogen-free chickens were randomly divided into six groups of chickens, one to six, inoculated with phosphate buffered saline; NDV lentogenic strain (LaSota); genotype VII (GVII); LaSota and challenged with GVII (LSGVII); genotype VIII (GVIII); and group of LaSota and challenged with GVIII (LSGVIII). The chickens were euthanized at 12, 36, and 60 h postchallenge. Immunophenotyping of CD25+ IEL, CD3+ cells, CD4+ cells, and CD8+ cells was conducted using flow cytometer. Furthermore, virus shedding was measured using reverse transcriptase-quantitative polymerase chain reaction. Data were analyzed using a two-way analysis of variance (ANOVA). The results showed that the percentage population of IEL subsets was generally lower in the chickens inoculated with GVII or GVIII when compared with LaSota, LSGVII and LSGVIII inoculated groups. The NDV copy number was significantly higher in chickens challenged with NDV GVII or GVIII when compared with chickens inoculated with LaSota, LSGVII or LSGVIII. Taking together, NDV velogenic strain caused decrease in the population of subsets of chickens' IEL. However, inoculation of NDV LaSota may increase the population of avian IEL subsets and decrease shedding of virulent NDV.

Efficacy, humoral, and cell-mediated immune response of inactivated fowl adenovirus 8b propagated in chicken embryo liver cells using bioreactor in broiler chickens.

Background and Aim: Fowl adenovirus (FAdV) 8b causes inclusion body hepatitis, resulting in major economic losses globally among chickens. The objectives were to inactivate FAdV 8b isolate propagated in chicken embryo liver (CEL) cells using a stirred tank bioreactor (UPM08136P5B1) and determine the humoral and cell-mediated immune response, efficacy, and virus shedding in broiler chickens. Materials and Methods: The FAdV 8b isolate UPM08136P5B1 was inactivated using binary ethyleneimine, adjuvanted with Montanide 71VG, inoculated into day-old broiler chickens in a booster group (BG) and non-booster group (NBG), and challenged with a pathogenic FAdV 8b strain. Clinical signs, gross lesions, body weight (BW), liver: body weight ratio, FAdV antibody titer using enzyme-linked immunosorbent assay, and histopathological changes were recorded. The CD3+, CD4+, and CD8+ T-lymphocyte profiles of the liver, spleen, and thymus using flow cytometry, and viral load in liver and cloacal shedding using quantitative polymerase chain reaction were evaluated. Results: Chickens in the challenged control group (CCG) exhibited mild clinical signs, gross lesions, and histopathological changes, which were absent in the inoculated groups, and had lower BW and higher liver BW ratio than chickens in the unchallenged control group (UCG); BG and NBG on 35- and 42-days post-inoculation (DPI). Chickens in NBG and BG had higher antibodies than UCG on 7, 21, 35, and 42 DPI. The challenged BG and NBG produced higher antibodies than the CCG on 35 DPI. T-lymphocytes were higher among the inoculated groups than UCG in the liver, spleen, and thymus. Inoculated challenged groups recorded higher CD3+, CD4+, and CD8+ T-lymphocytes on 35 and 42 DPI than CCG. The challenged control group had a significantly higher viral load in the liver than challenged that in BG on 35 DPI and BG and NBG on 42 DPI. The challenged control group had significantly higher challenge FAdV shedding than challenged inoculated groups on 35 and NBG on 42 DPI. Conclusion: UPM08136P5B1 was successfully inactivated and mixed with Montanide 71VG. The inactivated vaccine candidate that induced humoral and cellular immunity was effective, reduced FAdV load in the liver, and shedding in the cloaca, and could be useful against FAdV 8b infections in chickens.

Molecular characterization and pathogenicity of novel Malaysian chicken astrovirus isolates.

ABSTRACTChicken astrovirus (CAstV) has for over a decade been associated with runting stunting syndrome, severe kidney disease and visceral gout, and white chick syndrome. However, knowledge of the molecular characteristics and pathogenicity of the virus in day-old specific pathogen-free (SPF) chicks is scarce. This study focused on the characterization of near-complete genome of three Malaysian CAstV isolates following virus propagation in SPF embryonated chicken eggs and pathogenicity in day-old SPF chicks. The three isolates demonstrated unique features including a point mutation in their intergenic regions and an additional stem-loop II-like motif (s2m) in ORF-2. Pairwise sequence comparison and phylogenetic analysis of the ORF-2 amino acid sequence of the three isolates revealed an identity share of 86-91% with group B CAstVs while forming a new subgroup in addition to the known four subgroups (Bi, Bii, Biii and Biv) that exhibit high identity of between 95% and 100% within the subgroups. In the pathogenicity study, birds in the infected and exposed sentinel groups exhibited lethargy and diarrhoea 3 days post-inoculation (dpi) that declined by 6 dpi, and 20% growth retardation by 9 dpi. Mild lymphocytic aggregates in the duodenum, tubular degeneration and interstitial nephritis were observed in the intestines and kidneys, respectively, in both groups. In addition, the mean virus copy numbers of the cloacal swabs were log10 13.23 at 3 dpi and log10 9.04 at 6 dpi for the infected and exposed sentinels, respectively. The study suggests that the Malaysian isolates should be assigned to a new subgroup, Bv within group B CAstV. RESEARCH HIGHLIGHTSA single run of NGS protocol is capable of generating a near-complete genome sequence of CAstV.The Malaysian CAstV isolates cluster together and exhibit 86-91% identity with published group B CAstVs.The Malaysian CAstVs encode an additional stem-loop II-like motif (s2m) in ORF-2.The isolates are pathogenic to day-old SPF chicks with lesions mainly in the intestine and kidneys.

The Critical Studies of Fucoxanthin Research Trends from 1928 to June 2021: A Bibliometric Review.

Fucoxanthin is a major carotenoid in brown macroalgae and diatoms that possesses a broad spectrum of health benefits. This review evaluated the research trends of the fucoxanthin field from 1928 to June 2021 using the bibliometric method. The present findings unraveled that the fucoxanthin field has grown quickly in recent years with a total of 2080 publications. Japan was the most active country in producing fucoxanthin publications. Three Japan institutes were listed in the top ten productive institutions, with Hokkaido University being the most prominent institutional contributor in publishing fucoxanthin articles. The most relevant subject area on fucoxanthin was the agricultural and biological sciences category, while most fucoxanthin articles were published in Marine Drugs. A total of four research concepts emerged based on the bibliometric keywords analysis: "bioactivities", "photosynthesis", "optimization of process'', and "environment". The "bioactivities" of fucoxanthin was identified as the priority in future research. The current analysis highlighted the importance of collaboration and suggested that global collaboration could be the key to valorizing and efficiently boosting the consumer acceptability of fucoxanthin. The present bibliometric analysis offers valuable insights into the research trends of fucoxanthin to construct a better future development of this treasurable carotenoid.