Search

The purpose of this study was to determine the effect of mulberry (Morus alba L.) leaf (ML) supplementation on the productivity, blood metabolism, rumen fermentation characteristics and rumen microbial population in lactating dairy cows. Twelve healthy multiparous Holstein dairy cows (initial characteristics: body weight = 442.83 ± 86.87 kg; milk yield = 12.55 ± 1.91 kg/d; days in milk = 150.0 ± 47.0 d) were randomly allocated to two treatment groups of six cows each. The cows were fed two different diets: a control diet (basal diet without ML; CTRL group) and a ML-supplemented diet (basal diet + 300 g ML; ML group). The trial lasted for a total of 28 d after an adjustment period of 14 d. The total rumen volatile fatty acid (VFA) levels (p = .063) and the propionate level (p

Background and Aim: Canine transmissible venereal tumor (CTVT), a unique transmissible cancer in dogs, affects the external genitalia and potentially spreads to other parts of the body. While somatic mutations in oncogenic and tumor-suppressing genes are linked to CTVT development, the impact of DNA methylation, which affects gene expression, remains unclear. This study explored whether DNA methylation in the promoter regions of the MYC oncogene and CDKN2B tumor suppressor genes in CTVTs is associated with their expression, both at the gene and protein levels. Materials and Methods: To investigate promoter DNA methylation of MYC and CDKN2B in CTVTs, we analyzed frozen tissue samples from genital CTVT (GTVTs) and extragenital CTVT (ETVTs). Genomic DNA was extracted, bisulfite-treated, and analyzed using bisulfite polymerase chain reaction (PCR) and sequencing. The messenger RNA and protein of MYC and CDKN2B were also extracted and assessed by real-time PCR and Western blotting. Matching formalin-fixed, paraffin-embedded blocks were used for immunohistochemical staining to visualize protein distribution in GTVT and ETVT tissues. Results: Although both GTVT and ETVT samples showed MYC promoter methylation, the extent of methylation differed significantly. GTVTs displayed a much higher degree of methylation, potentially explaining the more pronounced downregulation of MYC gene expression and reduction in c-MYC protein levels observed in GTVTs compared with ETVTs. Our data revealed a prevalent hypermethylation pattern in the CDKN2B promoter across both sample types. However, DNA methylation, which was expected to have a suppressive effect, did not correlate with gene/protein expression. GTVTs displayed high protein levels despite significantly reduced CDKN2B expression. Conversely, ETVTs maintained regular CDKN2B expression but exhibited reduced protein production, suggesting a complex interplay between methylation and expression in these tumors. Conclusion: MYC demonstrated a clear association between its promoter methylation status, gene expression, and protein levels; however, CDKN2B lacked this correlation, implying the involvement of methylation-independent regulatory mechanisms and highlighting the need for further investigation.

Abstract Background At present, porcine reproductive and respiratory syndrome (PRRS) caused by the PRRS virus (PRRSV) is one of the most severe epidemics impacting pig farming globally. Despite the fact that a number of studies have been conducted on potential solutions to this problem, none have proven effective. The focus of problem solving is the use of natural ingredients such as plant extracts. Popular throughout Asia, Caesalpinia sappan (CS) is a therapeutic plant that inhibits PRRSV in vitro. Therefore, this study was performed to determine the efficacy of CS extract dietary supplementation on the productive performance, antibody levels, immunological indicators, and lung pathology of PRRSV-challenged weaned pigs. A total of 32 weaned piglets (28 days old) were randomized into 4 groups and kept separately for 14 days. The treatments were organized in a 2 × 2 factorial design involving two factors: PRRSV challenge and supplementation with 1 mg/kg CS extract. The pigs in the PRRSV-challenged groups were intranasally inoculated with 2 mL of PRRSV (VR2332) containing 104 TCID50/mL, while those in the groups not challenged with PRRSV were inoculated with 2 mL of normal saline. Results In the PRRSV-challenged group (CS + PRRSV), supplementation with CS extract led to an increase in white blood cells (WBCs) on Day 7 post infection (p

ABSTRACTBovine mastitis is primarily caused by a group of bacteria known as Staphylococcus and Streptococcus. However, additional types of bacteria, such as bovine non-aureus staphylococci and mammaliicocci (NASM) as well as lactic acid bacteria (LAB), are considered minor pathogens and have less impact on cows. Modulating bovine neutrophil activities and gene expressions in response to bacterial stimuli prompted the cells to execute effector functions to combat udder infections. Although neutrophils can manage major mastitis-causing bacteria, this strategy has not been tested against minor pathogens, i.e. NASM, Weissella spp. Our main objective was to investigate how neutrophils interacted with major and minor pathogens during in vitro bacterial stimulation. The results reveal that neutrophils performed offensive duties regardless of the type of bacteria encountered. Neutrophils generated high levels of reactive oxygen species, efficiently phagocytosed both types of bacteria, and facilitated extracellular killing by releasing NET structures against all bacteria. In addition, neutrophils migrated preferentially towards the majors rather than the minors, although myeloperoxidase (MPO) degranulation did not differ substantially across bacteria. Furthermore, the killing capacity of neutrophils was not dependent on any particular bacterium. The correlation of effector functions is intimately linked to the up-regulation of genes associated with the above functions, except for IL6, which was down-regulated. Furthermore, neutrophil apoptosis can be modulated by altering apoptosis-associated genes in response to harmful stimuli. These findings provide valuable information on how neutrophils react to major and minor mastitis-causing bacteria. However, future research should explore the interplay between minor pathogens and the host’s responses.

The objective of this study was to investigate the therapeutic effects of inactivated Propionibacterium acnes and lipopolysaccharide derived from Escherichia coli cells in cats affected by feline panleukopenia virus (FPV). A retrospective study of 80 FPV-positive cats was divided into two groups: a treatment group receiving inactivated Propionibacterium acnes and lipopolysaccharide derived from Escherichia coli cells along with supportive treatment and a no-treatment group receiving only supportive treatment. There was no significant difference in the total white blood cell counts between the two groups. However, the total white blood cell counts of both groups were low on day 0 and increased significantly on days 3 and 6 of treatment. Additionally, the white blood cell counts in the treatment group significantly increased during days 3 to 6 compared with those of the no-treatment group (p < 0.01). The mortality rate was not significantly different between the two groups. In a prospective study, the serum and fecal immunoglobulin A (IgA) levels were measured in both groups. There were no significant differences in IgA levels between the two groups in either the serum or feces.

Traditionally, non-aureus staphylococci and mammaliicocci (NASM) were not considered significant players in bovine mastitis. This study investigated the involvement of NASM (Staphylococcus hominis and Staphylococcus chromogenes) and lactic acid bacteria (LAB) strains (Weissella paramesenteroides) through bovine neutrophil responses. Bovine neutrophils displayed minimal apoptosis upon NASM and LAB challenge. Neutrophils expressed high TLR2 after challenge, but TLR6 expression varied and remained low in NASM pathogen recognition. Bovine neutrophils effectively engulfed and killed LAB, but their activity was significantly impaired against NASM. This was evident in S. chromogenes, where reduced TLR6 recognition and a weakened phagocytic response likely contributed to a lower bactericidal effect. Regardless of the bacteria encountered, intracellular ROS production remained high. S. chromogenes-challenged neutrophils displayed upregulation in genes for pathogen recognition (TLRs), ROS production, and both pro- and anti-apoptotic pathways. This response mirrored that of Weissella. except for CASP9 and BCL2, suggesting these bacteria have divergent roles in triggering cell death. Our findings suggest that S. chromogenes manipulates bovine neutrophil defenses through coordinated changes in functional responses and gene expression, while LAB strains have a weaker influence on apoptosis.

Non-typhoidal Salmonella (NTS) is still one of the most infectious foodborne pathogens causing problematic health issues worldwide in both human and veterinary medicine. Poultry meat was one of the important sources of NTS spreading and tended to be highly resistant to antibiotics. The aim of this study was to determine the prevalence, serotypes, and antimicrobial-resistant patterns of Salmonella circulating in broiler farms and poultry slaughterhouses in an intensive farming area in the upper northern part of Thailand from August to October 2019. Fifty samples of boot swabs were collected from 50 broiler farms, 50 cecal samples, and 250 neck skin samples from slaughterhouses. Salmonella was identified by culture method and serum-agglutination and antimicrobial susceptibility testing was tested using the automated VITEK-2 compact system. This study's total prevalence of Salmonella was 53.71% (n=188/350). We found that 53% (159/300) of cecal and neck skins samples were collected from slaughterhouses and 58% (29/50) of boot swabs collected from broiler farms were positive for NTS. Twenty-four serotypes of NTS were identified, the most encountered was S. Kentucky. The antimicrobial-resistant patterns showed that all the strains were non-susceptible to amikacin, cefalexin, cephalothin, and gentamicin and were susceptible to imipenem, neomycin, and nitrofurantoin. The NTS prevalence in samples from broiler farms was slightly higher than in poultry slaughterhouses indicating that there was contamination in the farming and slaughtering process. Therefore, both hygienic measurements in poultry production and antimicrobial usage in the poultry industry should be considered.

Bovine mastitis caused by Staphylococcus aureus may exacerbate by resulting in significant economic losses and impacting milk quality. To date, the use of gallic acid, a phenolic compound naturally occurring in various plants, holds promise due to its potent anti-oxidant and anti-inflammatory effects in many pieces of literature, thus, making it a subject of interest in bovine innate immunity research. Here we used gallic acid to assess its potential immunomodulation on milk phagocytes in vitro challenges with mastitis-causing bacteria. Our findings indicated that cells exposed to gallic acid showed no harm to cell viability but might maintain the longevity of cells during the bacterial infection. Gallic acid-treated cells displayed reduced cell migration, phagocytosis, and bacterial killing ability, while showing an increase in ROS production, all of which are undoubtedly linked to the intracellular killing abilities of the cells. Nonetheless, the extracellular structure called neutrophil extracellular traps (NETs) was significantly released after receiving gallic acid, representing extracellular killing. We also reported that gallic acid neutralizes inflammation by regulating specific pro-inflammatory genes (IL1B, IL6, TNF) and ROS-generating genes (CYBA, LAMP1, RAC1), subsequently preventing tissue damage. Regarding apoptosis-related genes and proteins, the increased production of caspase-3 and Bcl-2 family proteins could potentially promote the longevity of cells, implicated in the mechanism of combating bacterial invasion during udder inflammation and infection. The novel role of gallic acid on milk phagocytes highlights its potential immunomodulatory properties and contributes to our understanding of its effects on bacterial-host interactions, and provides valuable molecular insights.

Rapid influenza diagnostic tests (RIDTs) could considerably assist in clinical management resolving outbreaks of diseases in humans and animals. The RIDT for animals is rarely available and considerably expensive, especially in Thailand. In this study, therefore, the objective of this study was to determine the capacity of a commercial human RIDT “QuickNavi™Flu2”, an RIDT for influenza viruses A and B, to detect influenza viruses A antigen including 2 inactivated swine influenza viruses: H1N1 and H3N2, canine parainfluenza virus, and feline calicivirus. In addition, the efficacy of QuickNavi™Flu2 was compared with the real-time polymerase chain reaction assay (real-time rt-PCR). Both of SIV were used as stock solutions, and were serially diluted 6 concentrations before testing. The stock and diluted solutions of both SIV, specimens of canine parainfluenza virus, feline calicivirus and 96 unknown samples were tested by QuickNavi™Flu2 according to the guideline. Then all of samples were repeated testing with real-time rt-PCR assay. Results showed that Quick Navi-Flu could not give positive results for dilutions lower than 1:10 and 1:100, respectively. The sensitivity and specificity values of QuickNavi™Flu2 test in comparison to real-time rt-PCR assay for influenza A virus were 36.1% and 100%, respectively. In conclusion, a human RIDT can detect swine influenza H1N1 and H3N2 in high viral concentration, and appeared to be a useful tool for screening in animals when the viral load is sufficient.

Myxomatous mitral valve disease (MMVD) is the most common heart disease in small-breed dogs, often leading to heart failure. Oxidative stress in MMVD can harm mitochondria, decreasing their DNA content. This study assesses dogs’ oxidative stress and mitochondrial DNA at different MMVD stages. Fifty-five small-breed dogs were categorized into four groups, including: A—healthy (n = 15); B—subclinical (n = 15); C—heart failure (n = 15); and D—end-stage MMVD (n = 10). Serum malondialdehyde (MDA) and mitochondrial DNA in peripheral blood were analyzed. Quantitative real-time PCR measured mitochondrial DNA, and PCR data were analyzed via the fold-change Ct method. Serum MDA levels were assessed using competitive high-performance liquid chromatography (HPLC). Mitochondrial DNA was significantly lower in group B (−0.89 ± 2.82) than in group A (1.50 ± 2.01), but significantly higher in groups C (2.02 ± 1.44) and D (2.77 ± 1.76) than B. MDA levels were notably elevated in groups B (19.07 ± 11.87 µg/mL), C (23.41 ± 12.87 μg/mL), and D (19.72 ± 16.81 μg/mL) in comparison to group A (9.37 ± 4.67 μg/mL). Nevertheless, this observed difference did not reach statistical significance. It is noteworthy that mitochondrial DNA content experiences a decline during the subclinical stage but undergoes an increase in cases of heart failure. Concurrently, oxidative stress exhibits an upward trend in dogs with MMVD. These findings collectively suggest a potential association between mitochondrial DNA, oxidative stress, and the progression of MMVD in small-breed dogs.

The development of assisted reproductive technologies (ART) in mammalian species such as in vitro embryo production (IVEP) has the potential to provide great benefits for significant population increase, improve genetic performance and advancement, and reduce transmission of venereal diseases. Correspondingly, in vitro capacitation of sperm is also paramount, related to the ability of sperm to fertilize oocytes, and was created to imitate in vivo conditions in the female reproductive tract. Amid in vitro capacitation developments, studies on how far in vitro capacitation has progressed in mimicking in vivo scenes have not been thoroughly reviewed as a comparative form. Therefore, the present study outlined the series of alterations in mammalian sperm capacitation during their journey in the female reproductive tract by exploring and juxtaposing processes under in vivo and in vitro conditions. Several essential aspects that become gaps between in vivo and in vitro were also identified and elaborated comprehensively in this systematic literature review. We noted that although in vitro capacitation procedures in certain mammalian species have made promising progress and improvements, it is still poorly successful in other species like horses. Our findings further postulated that the occurrence of cryocapacitation, the high ratio of capacitated sperm/oocyte required for successful fertilization, and the incidence of polyspermy cause capacitation under in vitro settings is less efficient and not yet fully comparable to in vivo. This work is therefore proposed several aspects that need to be bettered from in vitro milieu to make it analogous to in vivo environments in modulating sperm capacitation.

Chronic kidney disease (CKD) is a frequent condition in elderly cats. Bcl-2 is linked to kidney disease through the processes of apoptosis and fibrosis. The purpose of this study is to examine Bcl-2 levels in CKD and clinically healthy age-matched cats in order to evaluate the relationship between Bcl-2 levels, signalment, and blood parameters in cats with CKD. The circulating levels of Bcl-2 were determined using an immunoassay in twenty-four CKD cats and eleven clinically healthy age-matched cats by the utilization of the general linear model (GLM), Pearson correlation, principal component analysis (PCA), ROC curves, the Cox hazard model, and Kaplan–Meier survival analysis. These were all conducted in order to explore Bcl-2 levels and their connection with other variables. The Bcl-2 immunohistochemical intensity was graded in each glomerulus and tubulointerstitium. McNemar's test was performed in order to compare the expression of Bcl-2 in the two renal tissue sites. The circulating Bcl-2 of CKD cats was significantly lower than those of clinically healthy age-matched cats (P = 0.034). The presence of circulating Bcl-2 (P < 0.01) and the severity of CKD (P = 0.02) were both linked with the survival time of cats with CKD. The area under the curve (AUC) of Bcl-2 for detection of CKD was 0.723. In cats, decreased circulating Bcl-2 was associated with increased blood BUN, creatinine levels, and CKD severity. Bcl-2 protein expression was reduced in the renal tissues of CKD cats as the disease progressed, resulting in a decrease in their survival time. This study demonstrated that Bcl-2 may be effective in diagnosing feline CKD.

Background Elephant endotheliotropic herpesvirus causes a hemorrhagic disease (EEHV-HD) that is a major cause of death in juvenile Asian elephants with EEHV1 and EEHV4 being the most prevalent. Aim To perform a retrospective clinical data analysis. Methods Records of a total of 103 cases in Thailand confirmed by polymerase chain reaction (PCR) on blood and/or tissue samples. Results The severity of clinical signs varied among EEHV subtypes. EEHV1A was the most prevalent with 58%, followed by EEHV4 with 34%, EEHV1B with 5.8% and EEHV1&4 co-infection with 1.9%. Overall case fatality rate was 66%. When compared among subtypes, 100% case fatality rate was associated with EEHV1&4 co-infection, 83% with EEHV1B, 75% with EEHV1A, and the lowest at 40% for EEHV4. Calves 2- to 4-year old were in the highest age risk group and exhibited more severe clinical signs with the highest mortality. Majority of cases were found in weaned or trained claves and higher number of cases were observed in rainy season. A gender predilection could not be demonstrated. Severely affected elephants presented with thrombocytopenia, depletion of monocytes, lymphocytes and heterophils, a monocyte:heterophil (M:H) ratio lower than 2.37, hypoproteinemia (both albumin and globulin), severe grade of heterophil toxicity, and low red blood cell counts and pack cell volumes. Survival was not affected by antiviral drug treatment in the severely compromised animals. Conclusion Early detection by laboratory testing and aggressive application of therapies comprising of supportive and anti-viral treatment can improve survival outcomes of this disease.

The use of metal oxide nanoparticles as an alternative antimicrobial agent has gained attention due to the increasing problem of antimicrobial resistance. Understanding its properties and potential benefits can contribute to the development of more effective and sustainable treatments in veterinary medicine. The aim of this study was to characterize TiO2-NP formulations and evaluate their antibacterial and wound healing abilities. The diameters and zeta potentials were determined using the Zetasizer in conjunction with dynamic light scattering. The agar-well diffusion method, time-kill kinetic assay and crystal violet assay were used to evaluate their antimicrobial activities. Wound healing assays were conducted both in vitro and in vivo. The study demonstrated that TiO2-NP formulations exhibit significant antimicrobial properties against various bacterial strains such as S. aureus and E. coli. No measurable E. coli growth was observed within a 15-min period following exposure to TiO2-NP formulations. The TiO2-NP formation can improve wound healing by enhancing cell migration and collagen formation in both in vitro and in vivo conditions. In summary, our study suggests that TiO2-NP has the potential for use as an antimicrobial agent for animal wound treatment due to its ability to suppress bacterial growth and biofilm formation, as well as to enhance wound healing.

The recent advances in sperm cryopreservation transcend cryobanking and other assisted reproductive technologies. Since its discovery, cryopreservation has contributed positive impacts on animal breeding as well as in genetic exchange, improvement, and conservation efforts. However, cryoinjury and variabilities in cryopreservation outcomes remain as key challenges to sperm cryobiology. The present work explored the molecular bases for such freezability differences and freezing-thawing injuries in the ruminant sperm. Relevant biomarkers identified in the seminal plasma and the spermatozoa were highlighted, including lipids, proteins, metabolites, transcripts, and genes. Specific molecular mechanisms concerning sperm structures and functions were also examined relative to their association to cryotolerance, and spermiogram or seminogram modifications following cryopreservation procedures. Current conflicts and gaps in the knowledge base on ruminant spermatozoa were also emphasized. Further investigation of these areas using the available breakthrough molecular tools such as omics technologies is therefore proposed to improve, optimize, or even predict the overall quality of frozen-thawed ruminant semen towards reproductive efficiency.

Background and Aim: Inappropriate overuse of antimicrobials might be associated with the spreading of antimicrobial-resistant bacteria in animal-based food products. Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli have been recognized as an emerging global problem in a One Health approach. This study aimed to assess the occurrence and antimicrobial-susceptible profiles of ESBL-producing E. coli among post-weaned calves and lactating cows in a parallel animal husbandry area. Materials and Methods: Seventy-two pool fecal samples were collected from 36 smallholder dairy farms registered in Ban Hong Dairy Cooperatives, Lamphun Province, Thailand. Pre-enriched fecal samples were cultured in MacConkey agar supplemented with cefotaxime. The potential E. coli isolates were identified by not only biochemical tests but also polymerase chain reaction assay of the 16S rRNA gene. ESBL production was confirmed by the combination disk test. Antimicrobial susceptibility testing was performed by the Kirby–Bauer disk diffusion method. Results: The occurrence of ESBL-producing E. coli at the farm level was 80.56%. The different phenotypic antibiogram of ESBL-producing E. coli was observed among post-weaned calf and lactating cow specimens. The most frequent resistance patterns of ESBL-producing isolates from both groups were amoxicillin-ceftiofur-cephalexin-cephalothin-cloxacillin-streptomycin-oxytetracycline-sulfamethoxazole/trimethoprim. For the median zone diameter, enrofloxacin-resistant isolates with narrow zone diameter values from lactating cow specimens were particularly more than post-weaned calf specimens (p

Both influenza C (ICV) and influenza D (IDV) viruses were recently included as bovine respiratory disease (BRD) causes, but their role in BRD has not been evaluated. Therefore, the mortality and reproductive performances of BRD calves with different isolated viruses were determined in this study. Data on 152 BRD calves with bovine viral diarrhoea virus (BVDV), bovine respiratory syncytial virus (BRSV), bovine coronavirus (BCoV), bovine parainfluenza virus 3 (BPIV-3), ICV, or IDV from nasal swab samples using real-time rt-PCR were used. The general data and respiratory signs were recorded immediately, and thereafter, the data on dead or culling calves due to BRD and reproductive performance were collected. The percentages of the BRD calves were 71.7%, 52.6%, 40.8%, 10.5%, 68.4%, and 65.8% for BVDV, BRSV, BCoV, BPIV-3, ICV, and IDV, respectively. Mucous secretion (OR = 4.27) and age ≤ 6 months (OR =14.97) had higher risks of mortality than those with serous secretion and older age. The calves with IDV had lower risks of culling than those without IDV (OR = 0.19). This study shows that most viral infections in BRD calves are a combination of viruses with BVDV, ICV, and IDV. In addition, IDV might have a role in reducing the severity of BRD calves.

Background MicroRNAs (miRNAs) play an essential role in gene regulators in many biological and molecular phenomena. Unraveling the involvement of miRNA as a key cellular factor during in vitro canine parvovirus (CPV) infection may facilitate the discovery of potential intervention candidates. However, the examination of miRNA expression profiles in CPV in tissue culture systems has not been fully elucidated. Method In the present study, we utilized high-throughput small RNA-seq (sRNA-seq) technology to investigate the altered miRNA profiling in miRNA libraries from uninfected (Control) and CPV-2c infected Crandell Reese Feline Kidney cells. Results We identified five of known miRNAs (miR-222-5p, miR-365-2-5p, miR-1247-3p, miR-322-5p and miR-361-3p) and three novel miRNAs (Novel 137, Novel 141 and Novel 102) by sRNA-seq with differentially expressed genes in the miRNA repertoire of CPV-infected cells over control. We further predicted the potential target genes of the aforementioned miRNAs using sequence homology algorithms. Notably, the targets of miR-1247-3p exhibited a potential function associated with cellular defense and humoral response to CPV. To extend the probing scheme for gene targets of miR-1247-3p, we explored and performed Gene Ontology (GO) enrichment analysis of its target genes. We discovered 229 putative targets from a total of 38 enriched GO terms. The top over-represented GO enrichment in biological process were lymphocyte activation and differentiation, marginal zone B cell differentiation, negative regulation of cytokine production, negative regulation of programed cell death, and negative regulation of signaling. We next constructed a GO biological process network composed of 28 target genes of miR-1247-3p, of which, some genes, namely BCL6, DLL1, GATA3, IL6, LEF1, LFNG and WNT1 were among the genes with obviously intersected in multiple GO terms. Conclusion The miRNA-1247-3p and its cognate target genes suggested their great potential as novel therapeutic targets or diagnostic biomarkers of CPV or other related viruses.

Chicken heterophils serve as the first-line defence in chicken innate immunity. The present study aimed to explore the effects of quercetin, a herbal phenolic substance known for its inflammatory modulation, on mRNA expression patterns in chicken heterophils. Isolated heterophils from two genetically distinct chicken lines, hybrid commercial and Thai indigenous chickens, Pradu Hang Dam, were in vitro co-cultured with live intestinal pathogenic Salmonella enteritidis under Phosphate-buffered saline (PBS) or quercetin supplementation. Quantitative real-time RT-PCR was performed to investigate the effects of quercetin on mRNA transcripts of 12 genes. The results showed that heterophils from hybrid chickens treated with quercetin had higher relative expression levels of IL6, IL8L1, IL8L2, IL18, CCL4, LITAF, and MIF. Unlike hybrid chickens, Pradu Hang Dam chickens had down-regulation of cytokine genes, IL1B, IL6, IL8L1, IL8L2, IL12B, and IL18, and chemokines, CCL4 and MIF. In addition, NOS2, IL17A, and TGFB1 were up-regulated in this lineage. In conclusion, our findings emphasize the modulatory effects of quercetin to down-regulate most pro-inflammatory genes, whereas it up-regulates anti-inflammatory gene (TGFB1) in Pradu Hang Dam chickens.

Herbal phytochemicals featuring active ingredients including quercetin and curcumin have shown potential in treating human and animal diseases. The current study investigated their potential function in vitro for host immunomodulation associated with Streptococcus agalactiae subclinical bovine mastitis via milk-isolated neutrophils. Our results showed a positive influence on cellular migration, reactive oxygen species (ROS) generation, phagocytosis, and bacterial killing as well as neutrophil extracellular traps (NETs) release. This study also highlighted several important molecular aspects of quercetin and curcumin in milk-isolated neutrophils. Gene expression analyses by RT-PCR revealed significant changes in the expression of proinflammatory cytokines (IL1B, IL6, and TNF), ROS (CYBA), phagocytosis (LAMP1), and migration (RAC). The expression levels of apoptotic genes or proteins in either pro-apoptosis (CASP3 and FAS) or anti-apoptosis (BCL2, BCL2L1, and CFLAR) were significantly manipulated by the effects of either quercetin or curcumin. A principal component analysis (PCA) identified the superior benefit of quercetin supplementation for increasing both cellular and molecular functions in combating bacterial mastitis. Altogether, this study showed the existing and potential benefits of these test compounds; however, they should be explored further via in vivo studies.

Canine parvovirus (CPV), the etiology of hemorrhagic enteritis in dogs, was first isolated as CPV type 2 (CPV-2) almost 40 years ago, and was soon replaced by the emergence of new variant types. The major viral capsid proteins encoded by the VP2 gene are the sites where amino acids are often substituted, accounting for the unusual nature of this type of DNA virus. The alteration of specific residues has contributed to different antigenic variants which have affected the evolution of virus binding and host immunity to this virus. Sequence analysis of the VP2 gene and subsequent characterization have revealed three circulating CPV-2 strains, CPV-2a, CPV-2b, and CPV-2c, identified by mutations at amino acid residue 426. The latter strain displays increased pathogenicity in dogs and an extended host range. The present review article aimed at updating contemporary information on epidemiological studies and surveys from CPV field work. Moreover, we pointed out some sensitive and rapid diagnostic tools for detecting CPV in clinical samples, techniques which will be useful for health monitoring and management of CPV with currently available vaccines.

Aim: To investigate gene expression of microRNA (miRNA) milieus (MIRLET7E, MIR17, MIR24-2, MIR146A, and MIR181C), inflammatory cytokine genes (interleukin 1β [IL1B], IL6, CXCL8, and tumor necrosis factor [TNF]), and the pathogen receptor toll-like receptor (TLR4) in bovine neutrophils under quercetin supplementation. Materials and Methods: Isolated bovine neutrophils were incubated with bacterial lipopolysaccharide under quercetin treatment or left untreated. Real-time polymerase chain reaction was performed to determine the expression of the miRNAs and messenger RNA (mRNA) transcripts in neutrophils. Results: Quercetin-treated neutrophils exhibited a remarkable suppression in MIR24-2, MIR146A, and MIR181C expression. Similarly, mRNA expression of IL1B, IL6, CXCL8, TLR4, and TNF genes noticeably declined in the quercetin group. Many proinflammatory genes (IL1B, IL6, and CXCL8) and the pathogen receptor TLR4 had a negative correlation with MIR146A and MIR181C as revealed by Pearson correlation. Conclusion: Interaction between cognate mRNAs and miRNAs under quercetin supplementation can be summarized as a positive or negative correlation. This finding may help understand the effects of quercetin either on miRNA or gene expression during inflammation, especially as a potentially applicable indicator in bovine mastitis.

Elephant endotheliotropic herpesvirus-hemorrhagic disease (EEHV-HD) is the primary cause of acute, highly fatal, hemorrhagic diseases in young Asian elephants. Although monocytopenia is frequently observed in EEHV-HD cases, the role monocytes play in EEHV-disease pathogenesis is unknown. This study seeks to explain the responses of monocytes/macrophages in the pathogenesis of EEHV-HD. Samples of blood, frozen tissues, and formalin-fixed, paraffin-embedded (FFPE) tissues from EEHV1A-HD, EEHV4-HD, co-infected EEHV1A and 4-HD, and EEHV-negative calves were analyzed. Peripheral blood mononuclear cells (PBMCs) from the persistent EEHV4-infected and EEHV-negative calves were also studied. The results showed increased infiltration of Iba-1-positive macrophages in the inflamed tissues of the internal organs of elephant calves with EEHV-HD. In addition, cellular apoptosis also increased in the tissues of elephants with EEHV-HD, especially in the PBMCs, compared to the EEHV-negative control. In the PBMCs of persistent EEHV4-infected elephants, cytokine mRNA expression was high, particularly up-regulation of TNF-α and IFN-γ. Moreover, viral particles were observed in the cytoplasm of the persistent EEHV4-infected elephant monocytes. Our study demonstrated for the first time that apoptosis of the PBMCs increased in cases of EEHV-HD. Furthermore, this study showed that monocytes may serve as a vehicle for viral dissemination during EEHV infection in Asian elephants.

Elephant endotheliotropic herpesvirus (EEHV) infection is known to cause acute fatal hemorrhagic disease, which has killed many young Asian elephants (Elephas maximus). Until recently, in vitro isolation and propagation of the virus have not been successful. This study aimed to isolate and propagate EEHV using continuous cell lines derived from human and/or animal origins. Human cell lines, including EA. hy926, A549, U937, RKO, SW620, HCT-116 and HT-29, and animal cell lines, including CT26.CL25 and sp2/0-Ag14, were investigated in this study. Mixed frozen tissue samples of the heart, lung, liver, spleen and kidney obtained from fatal EEHV1A- or EEHV4-infected cases were homogenized and used for cell inoculation. At 6, 24, 48 and 72 h post infection (hpi), EEHV-inoculated cells were observed for cytopathic effects (CPEs) or were assessed for EEHV infection by immunoperoxidase monolayer assay (IPMA) or quantitative PCR. The results were then compared to those of the mock-infected controls. Replication of EEHV in the tested cells was further determined by immunohistochemistry of cell pellets using anti-EEHV DNA polymerase antibodies or re-inoculated cells with supernatants obtained from passages 2 or 3 of the culture medium. The results reveal that no CPEs were observed in the tested cells, while immunolabeling for EEHV gB was observed in only U937 human myeloid leukemia cells. However, quantitation values of the EEHV terminase gene, as well as those of the EEHV gB or EEHV DNA polymerase proteins in U937 cells, gradually declined from passage 1 to passage 3. The findings of this study indicate that despite poor adaptation in U937 cells, this cell line displays promise and potential to be used for the isolation of EEHV1 and EEHV4 in vitro.

The in vitro virustatic and virucidal tests of the crude extract of Cynodon dactylon against infection with porcine reproductive and respiratory syndrome virus (PRRSV), a cause of major devastating pig disease, were described. Crude extract of C. dactylon was prepared for cytotoxicity on tissue-culture cells that were used to measure virustatic and virucidal activities against PRRSV. Crude extract of C. dactylon at 0.78 mg/mL showed no cytotoxicity on the cell line, and at that concentration significantly inhibited replication of PRRSV as early as 24 hours post infection (hpi). C. dactylon also inactivated PRRSV as determined by immunoperoxidase monolayer assay (IPMA) compared to the control experiments. In summary, the present study may be among the earliest studies to describe virustatic and virucidal activities of C. dactylon crude extract against PRRSV in vitro. Extracts of C. dactylon may be useful for PRRSV control and prevention on pig farms.