• Journal of Microbiology
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• 제44권5호
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• pp.548-555
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• 2006

FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

• Journal of Animal Science and Technology
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• 제65권5호
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• pp.951-970
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• 2023

This study utilized Italian ryegrass silage (IRGS) - based total mixed ration (TMR) as feedstuff and evaluated its effects on rumen fermentation, growth performance, blood parameters, and bacterial community in growing Hanwoo heifers. Twenty-seven Hanwoo heifers (body weight [BW], 225.11 ± 10.57 kg) were randomly allocated to three experimental diets. Heifers were fed 1 of 3 treatments as follows: TMR with oat, timothy, and alfalfa hay (CON), TMR with 19% of IRGS (L-IRGS), and TMR with 36% of IRGS (H-IRGS). Feeding high levels of IRGS (H-IRGS) and CON TMR to heifers resulted in a greater molar proportion of propionate in the rumen. The impact of different TMR diets on the BW, average daily gain, dry matter intake, and feed conversion ratio of Hanwoo heifers during the growing period did not differ (p > 0.05). Furthermore, the blood metabolites, total protein, albumin, aspartate aminotransferase, glucose, and total cholesterol of the heifers were not affected by the different TMR diets (p > 0.05). In terms of rumen bacterial community composition, 264 operational taxonomic units (OTUs) were observed across the three TMR diets with 240, 239, and 220 OTUs in CON, L-IRGS, and H-IRGS, respectively. IRGS-based diets increased the relative abundances of genera belonging to phylum Bacteroidetes but decreased the abundances of genus belonging to phylum Firmicutes compared with the control. Data showed that Bacteroidetes was the most dominant phylum, while Prevotella ruminicola was the dominant species across the three TMR groups. The relative abundance of Ruminococcus bromii in the rumen increased in heifers fed with high inclusion of IRGS in the TMR (H-IRGS TMR). The relative abundance of R. bromii in the rumen significantly increased when heifers were fed H-IRGS TMR while P. ruminicola increased in both L-IRGS and H-IRGS TMR groups. Results from the current study demonstrate that the inclusion of IRGS in the TMR is comparable with the TMR containing high-quality forage (CON). Thus, a high level of IRGS can be used as a replacement forage ingredient in TMR feeding and had a beneficial effect of possibly modulating the rumen bacterial community toward mainly propionate-producing microorganisms.

• Asian-Australasian Journal of Animal Sciences
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• 제31권12호
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• pp.1881-1889
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• 2018

Objective: In dryland areas of China, alfalfa hay (AH) is a possible substitute for concentrate feed for beef cattle. To evaluate the potential benefits of this substitution, we studied the effect of the ratio of AH intake to total dry matter (DM) intake on average daily body-weight gain (ADG), dietary energy utilization status, and economic benefit in Gansu province. Methods: In each of two feeding trials in 2016 (trial 1 [T1], July 3 to 17; trial 2 [T2], August 15 to September 23), crossbred male Simmental calves were allocated to low AH (LA), medium AH (MA), and high AH (HA) feeding groups (n = 4 per group). The target ADG was set as 1 kg for both trials. In a one-way-layout design based on conventional feeding practices in the province, calves received diets containing the different AH amounts, with a constant ratio of corn stover:total DM and decreasing rations of concentrate feed proportional to the increase in AH. Calves in T1 received AH at 15% (T1-LA), 23% (T1-MA), or 31% (T1-HA) of their dietary DM allowances; those in T2 received 9% (T2-LA), 24% (T2-MA), or 34% (T2-HA) AH. Results: Among the T1 groups, both ADG and economic benefit were highest in T1-LA; whereas in T2, they were higher in the T2-LA and T2-MA groups than in T2-HA. Energy digestibility did not significantly differ among the groups in either trial. The dietary AH inclusion ratios of 14% in the warm season and 8% to 21% in the cool season appeared to yield optimal ADG, metabolizable energy intake, and economic benefit. Conclusion: Low-level inclusion of AH, ranging from 8% to 21%, is a practical approach for beef cattle feeding. This modified feeding regimen likely will promote increased growth performance during the fattening stage of beef steers in dryland areas of Gansu province, China.

• Asian-Australasian Journal of Animal Sciences
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• 제32권5호
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• pp.614-628
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• 2019

Objective: The inhibitory leukocyte immunoglobulin-like receptors (LILRBs) play an important role in innate immunity. The present study represents the first description of the cloning and structural and functional analysis of LILRB1 and LILRB3 isolated from two genetically disparate chicken lines. Methods: Chicken LILRB1-3 genes were identified by bioinformatics approach. Expression studies were performed by transfection, quantitative polymerase chain reaction. Signal transduction was analyzed by western blots, immunoprecipitation and flow cytometric. Cytokine levels were determined by enzyme-linked immunosorbent assay. Results: Amino acid homology and phylogenetic analyses showed that the homologies of LILRB1 and LILRB3 in the chicken line 6.3 to those proteins in the chicken line 7.2 ranged between 97%-99%, while homologies between chicken and mammal proteins ranged between 13%-19%, and 13%-69%, respectively. Our findings indicate that LILRB1 and LILRB3 subdivided into two groups based on the immunoreceptor tyrosine-based inhibitory motifs (ITIM) present in the transmembrane domain. Chicken line 6.3 has two ITIM motifs of the sequence LxYxxL and SxYxxV while line 7.2 has two ITIM motifs of the sequences LxYxxL and LxYxxV. These motifs bind to SHP-2 (protein tyrosine phosphatase, non-receptor type 11) that plays a regulatory role in immune functions. Moreover, our data indicate that LILRB1 and LILRB3 associated with and activated major histocompatibility complex (MHC) class I and ${\beta}2-microglobulin$ and induced the expression of transporters associated with antigen processing, which are essential for MHC class I antigen presentation. This suggests that LILRB1 and LILRB3 are transcriptional regulators, modulating the expression of components in the MHC class I pathway and thereby regulating immune responses. Furthermore, LILRB1 and LILRB3 activated Janus kinase2/tyrosine kinase 2 (JAK2/TYK2); signal transducer and activator of transcription1/3 (STAT1/3), and suppressor of cytokine signaling 1 genes expressed in Macrophage (HD11) cells, which induced Th1, Th2, and Th17 cytokines. Conclusion: These data indicate that LILRB1 and LILRB3 are innate immune receptors associated with SHP-2, MHC class I, ${\beta}2-microglobulin$, and they activate the Janus kinase/signal transducer and activator of transcription signaling pathway. Thus, our study provides novel insights into the regulation of immunity and immunopathology.

• Animal Bioscience
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• 제35권7호
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• pp.964-974
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• 2022

Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry and economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for studies on HPAIV resistance. Therefore, in this study, we investigated gene expression related to the mitogen-activated protein kinase (MAPK) signaling pathway by comparing non-infected, HPAI-infected resistant, and susceptible Ri chicken lines. Methods: Resistant (Mx/A; BF2/B21) and susceptible Ri chickens (Mx/G; BF2/B13) were selected by genotyping the Mx and BF2 genes. Then, the tracheal tissues of non-infected and HPAIV H5N1 infected chickens were collected for RNA sequencing. Results: A gene set overlapping test between the analyzed differentially expressed genes (DEGs) and functionally categorized genes was performed, including biological processes of the gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. A total of 1,794 DEGs were observed between control and H5N1-infected resistant Ri chickens, 432 DEGs between control and infected susceptible Ri chickens, and 1,202 DEGs between infected susceptible and infected resistant Ri chickens. The expression levels of MAPK signaling pathway-related genes (including MyD88, NF-κB, AP-1, c-fos, Jun, JunD, MAX, c-Myc), cytokines (IL-1β, IL-6, IL-8), type I interferons (IFN-α, IFN-β), and IFN-stimulated genes (Mx1, CCL19, OASL, and PRK) were higher in H5N1-infected than in non-infected resistant Ri chickens. MyD88, Jun, JunD, MAX, cytokines, chemokines, IFNs, and IFN-stimulated expressed genes were higher in resistant-infected than in susceptible-infected Ri chickens. Conclusion: Resistant Ri chickens showed higher antiviral activity compared to susceptible Ri chickens, and H5N1-infected resistant Ri chickens had immune responses and antiviral activity (cytokines, chemokines, interferons, and IFN-stimulated genes), which may have been induced through the MAPK signaling pathway in response to H5N1 infection.

• Animal Bioscience
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• 제36권6호
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• pp.851-860
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• 2023

Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.

• Journal of Microbiology and Biotechnology
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• 제26권8호
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• pp.1446-1451
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• 2016

Clostridium difficile toxin A causes acute gut inflammation in animals and humans. It is known to downregulate the tight junctions between colonic epithelial cells, allowing luminal contents to access body tissues and trigger acute immune responses. However, it is not yet known whether this loss of the barrier function is a critical factor in the progression of toxin A-induced pseudomembranous colitis. We previously showed that NADH:quinone oxidoreductase 1 (NQO1) KO (knockout) mice spontaneously display weak gut inflammation and a marked loss of colonic epithelial tight junctions. Moreover, NQO1 KO mice exhibited highly increased inflammatory responses compared with NQO1 WT (wild-type) control mice when subjected to DSS-induced experimental colitis. Here, we tested whether toxin A could also trigger more severe inflammatory responses in NQO1 KO mice compared with NQO1 WT mice. Indeed, our results show that C. difficile toxin A-mediated enteritis is significantly enhanced in NQO1 KO mice compared with NQO1 WT mice. The levels of fluid secretion, villus disruption, and epithelial cell apoptosis were also higher in toxin A-treated NQO1 KO mice compared with WT mice. The previous and present results collectively show that NQO1 is involved in the formation of tight junctions in the small intestine, and that defects in NQO1 enhance C. difficile toxin A-induced acute inflammatory responses, presumably via the loss of epithelial cell tight junctions.

• Asian-Australasian Journal of Animal Sciences
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• 제26권2호
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• pp.189-194
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• 2013

The goal of this study was to evaluate the effects of lactation stage and individual performance on milk cis-9, trans-11 conjugated linoleic acid (CLA) content in dairy cows. In experiment 1, the milk cis-9, trans-11 CLA content from dairy cows in early ($0.33{\pm}0.014%$), middle ($0.37{\pm}0.010%$), and late stages ($0.44{\pm}0.020%$) showed significant differences (p<0.05); and the individual contents of the major fatty acids, especially cis-9, trans-11 CLA in cows of the same lactation were also variable. In the second experiment design as a validation test, our results once again showed that the individual contents of cis-9, trans-11 CLA were various, and a difference of about 2-fold (0.55% vs 0.95%) was observed, although the animals were offered same diet. These data demonstrated that lactation stage and individual performance have considerable effects on milk cis-9, trans-11 CLA contents.

• Asian-Australasian Journal of Animal Sciences
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• 제16권2호
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• pp.189-197
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• 2003

Soluble glass bolus with selenium (Se), copper (Cu) and cobalt (Co) was administered intraruminally to Philippine grazing does fed under backyard farming conditions to determine its effect on blood mineral status. Forty-five does were dosed with SGB intraruminally every 6 months, whereas 15 were without SGB during this experimental period of 10 months. Blood of does in both treatment groups were collected every other month and blood Se, plasma Cu, Ca, P and Mg were determined in this study. All does did not show clinical Se, Cu, Ca, P or Mg deficiency during this experimental period. Selenium concentration of treated does increased (p<0.01) after beginning of this experiment, whereas the level of control does decreased slowly (p<0.01). Two months after SGB administration, all treated does had higher blood Se than the lower limit of $20{\mu}g/l$ suggested by NRC (1981), whereas some control does had lower blood Se concentration than the lower limit of $20{\mu}g/l$. On the other hand, plasma Cu concentration of treated does started to increase more remarkably than control does 4 months after this experiment had started although the difference was insignificant. There were no significant differences between plasma Ca, P or Mg concentration of does in both treatment groups. Soluble glass bolus had no harmful effects on plasma macro mineral concentrations of grazing does. This study showed that SGB was available as mineral supplement to improve imbalances of selected mineral of grazing goats in the traditional backyard conditions in Luzon Island, Philippines.

• Asian-Australasian Journal of Animal Sciences
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• 제11권6호
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• pp.648-654
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• 1998

Twenty-four Javanese thin-tail ewes (11, 9, and 4 ewes giving birth to 1, 2, and 3 lambs, respectively) with similar body weight and age at breeding were used to study serum progesterone concentrations during pregnancy, milk production during lactation, and mammary gland indices at the end of lactation (3 months postpartum). The results of the experiment showed that averages serum progesterone concentrations during pregnancy in the ewes giving birth to twin and triplet lambs were higher (p < 0.01) than those giving birth to a single lamb. Ewes giving birth to 3 lambs had higher (p < 0.01) mammary dry fat-free tissue (DFFT) (by 31 and 34%), DNA concentration (by 25 and 16%) and RNA concentration (by 29 and 16%) at the end of lactation than those giving birth to 1 and 2 lambs. There was no difference in mammary collagen, protein and glycogen concentrations at the end of lactation among litter sizes. Ewes giving birth to 3 lambs had higher (p < 0.01) total mammary DNA content (by 64 and 61%) and RNA content (by 69 and 53%) at the end of lactation than those giving birth to 1 and 2 lambs. There was no difference in total mammary collagen, protein and glycogen contents at the end of lactation among litter sizes. Even though ewes with higher litter size had numerically higher milk production, there was no significant difference in milk production per 4 h among litter sizes. The results of the experiment indicated that ewes having higher litter size had greater mammary cell number and synthetic activities at the end of lactation. The results suggested that ewes with higher progesterone concentrations and better developed mammary glands during pregnancy could maintain higher cell number and activities throughout lactation.