This study aimed to investigate the effects of selenium on the ion profiles in the heart, liver, spleen, and kidney through the oral administration of hexavalent chromium. Approximately 22.14 mg/kg b.w. K2Cr2O7 was added to water to establish a chronic poisoning model. Different selenium levels (0.00, 0.31, 0.63, 1.25, 2.50, and 5.00 mg Na2SeO3/kg b.w.) around the safe dose were administered to the experimental group model. Ca, Mg, Mn, Fe, Cu, and Zn were detected in the organs through flame atomic absorption spectrometry after these organs were exposed to K2Cr2O7 and Na2SeO3 for 14, 28, and 42 days. Results showed that these elements exhibited various changes. Ca contents declined in the heart, liver, and spleen. Ca contents also decreased on the 28th day and increased on the 42nd day in the kidney. Mn contents declined in the heart and spleen but increased in the kidney. Mn contents also decreased on the 28th day and increased on the 42nd day in the liver. Cu contents declined in the heart and spleen. Cu contents increased on the 28th day and decreased on the 42nd day in the liver and kidney. Zn contents declined in the heart and spleen. Zn contents increased on the 28th day and decreased on the 42nd day in the liver and kidney. Fe contents decreased in the heart and liver. Fe contents increased on the 28th day and decreased on the 42nd day in the spleen and kidney. Mg contents did not significantly change in these organs. Appropriate selenium contents enhanced Mn and Zn contents, which were declined by chromium. Conversely, appropriate selenium contents reduced Ca, Fe, and Cu contents, which were increased by chromium. In conclusion, the exposure of chickens to K2Cr2O7 induced changes in different trace elements, and Na2SeO3 supplementation could alleviate this condition.

•Cr(VI) administration lead to kidney oxidative damage in male chickens.•Moderate doses of Selenium could alleviate nephrotoxicity induced by Cr(VI).•Excessive additive Se even exacerbated kidney damage.Our study aimed to explore whether Na2SeO3 (Se) can alleviate the nephrotoxicity induced by K2Cr2O7 [Cr(VI)]. One hundred and five male chickens were randomly divided into seven groups with 15 chickens each group: The 6 experimental groups received K2Cr2O7 alone or in combination with 0.31, 0.63, 1.25, 2.50, and 5.00 mg/kg for 42 days, respectively, while control group was treated with equivalent water. Exposure to Cr(VI) significantly increased MDA contents and organ coefficient, whereas decreased T-SOD activities, Ca2+-ATPase activities, mitochondrial membrane potential and GSH contents, and histological studies demonstrated renal damage. Above indicators were restored by Se supplement (0.31, 0.63, and 1.25 mg/kg), in which supplement with 0.63 mg/kg Se developed more effectively than the other two groups; on the contrary, in the groups of Se supplement with 2.50 and 5.00 mg/kg, the above indicators were not ameliorated and even exacerbated. This study demonstrated that Cr(VI) can result in kidney oxidative damage in male chickens, and Se of certain dose has the protective effects against Cr(VI)-induced nephrptoxicity.

As a traditional Chinese multiherbal formula, Yu-Ping-Feng (YPF) is frequently used to treat cold, flu and inflammation-associated diseases. We aimed to evaluate the immunostimulatory effects of polysaccharide isolated from YPF (YPF-PS) in vitro and in vivo. In in vitro experiment, macrophage cell proliferation, phagocytosis rate, cytokine and costimulatory molecule release, T lymphocyte proliferation, cell cycle distribution, CD4+ and CD8+ T cell percentages were determined. To investigate the in vivo effects of YPF-PS treatment, different doses YPF-PS were administered to chicken vaccinated against Newcastle disease. The immune organ index, lymphocyte proliferation, antibody titer, cell cycle distribution, and the cell percentage of CD4+ and CD8+ were assessed. In vitro results indicated that YPF-PS at 15.62 μg mL−1 could increase the LPS-induced macrophage cell proliferation and phagocytosis rate significantly. The levels of cytokine (nitric oxide, tumor necrosis factor-alpha, interleukin-1 beta, interleukin-6, and interferon beta) and costimulatory molecules (CD80 and CD86) were also considerably enhanced. Moreover, YPF-PS could significantly enhance T lymphocyte proliferation individually or synergistically with phytohemagglutinin. It promoted lymphocyte entry into S and G2/M phases and increased the percentages of CD4+ and CD8+ T cells effectively. In addition, in vivo experiments showed that YPF-PS could enhance serum HI antibody titer. The results about T lymphocyte proliferation, cell cycle distribution, CD4+ and CD8+ cell percentages in chickens were also confirmed. YPF-PS has efficacious immunomodulatory properties and could be used as a new potential immune stimulator for food and medical purposes.

This study aimed to investigate the effects of chromic chloride (CrCl3) on Ca, Mg, Mn, Fe, Cu, and Zn contents in the brain and serum of chicken. Seventy-two chickens were randomly divided into four groups and treated with different doses of CrCl3 via drinking water: 0, 1/8, 1/4, and 1/2 LD50 for 42 days. The contents of the elements were evaluated through inductively coupled plasma mass spectrometry. Results showed that Cr contents in the brain and serum were higher than those in the control groups, although no significant dose-dependent changes (P > 0.05) in brain of the Cr-treated groups were observed at 42 days. As exposure time was prolonged and CrCl3 dosage was increased, Ca contents increased (P < 0.05). Mg and Cu contents in serum decreased; by contrast, Mg and Cu contents initially increased and then decreased in the brain. Fe and Zn contents in the serum increased; conversely, Fe and Zn contents in the brain decreased. CrCl3 exposure did not significantly affect Mn contents at 14 or 28 days, but significantly decreased (P < 0.05) at 42 days. Therefore, excess Cr3+ intake can disrupt absorption and deposition of other trace elements in the brain and serum; the blood–brain barrier may prevent the accumulation of these elements in the brain exposed to CrCl3.

•RAPS could increase cell proliferation and phagocytosis rate significantly.•RAPS could elevate the levels of cytokine: NO, TNF-α, and IFN-β obviously.•RSPS had stronger expression level on CD80 and CD86 than PAPS and RAMPS.•Compared to PAPS and RAMPS, PSPS had different structure characterization.•RAPS would be anticipated as a component of new-type immunostimulant.In this study, we evaluated structure characterization and immunomodulatory activity of polysaccharides from Astragalus aboriginum Richardson (RAPS), Atractylodes macrocephala Koidz (RAMPS) and Rumia seseloides Hoffm (RSPS) in vitro on chicken macrophage. We found that molecular weight of RAPS and RAMPS was 122.4 and 109.4 kDa higher than 64.71 kDa of RSPS. Glucose occupied 83.95% and 66.39% in RAPS and RAMPS, respectively. RSPS mainly contained glucose and galacturonic acid, which accounted for 32.35% and 29.25%, respectively. The NMR results displayed that RAPS and RAMPS contained β- glucose, β-galactose, and β-galacturonic acid. The backbone was 1 → 6 linked glucose. RSPS showed at least six monosaccharide response signals. In vitro experiment, the results showed that RAPS at dosage of 15.62 μg mL−1 exhibited significant immunological on chicken macrophage compared to RAMPS and RSPS. Interestingly, costimulatory molecules levels in RSPS group were higher than that of RAPS, which may associated with the special structure of RSPS.

•RAMPStp and RAMPS60c could significantly enhance T lymphocyte proliferation and increase antibody titers.•RAMPStp and RAMPS60c could improve the percentages of CD4+ and CD8+ T cells.•RAMPStp and RAMPS60c could promote lymphocytes enter into S and G2/M phases.•The glucose side chains of the polymer could be more responsible for the immune-enhancing activity.•RAMPStp would be anticipated as a component of new-type immunopotentiator.Build on our previous research, polysaccharides from the rhizome of Atractylodis macrocephalae Koidz (RAMPS), RAMPStp and RAMPS60c were prepared and the structural characterization and immune response of ND vaccine in chicken were investigated. Immune organ index, Lymphocyte proliferation, antibody titers, cell cycle distribution, and percentages of CD4+ and CD8+ cells were determined. GPC analysis showed that the Mn of RAMPS with two peaks were 1.29 × 105 and 1.74 × 103, respectively. GC–MS analysis revealed that RAMPS was composed of glucose, mannose, arabinose, galactose, xylose, d-Ribose and rhamnose, with mass percentages of 66.39%, 21.24%, 5.64%, 2.65%, 2.30%, 1.15% and 0.64%, respectively. NMR spectroscopic analysis demonstrated that a preliminary structure of RAMPS was proposed as 1,3-linked β-d-Galp and 1,6-linked β-d-Galpresidues. In vivo test showed that RAMPStp and RAMPS60c could promote peripheral lymphocytes proliferation and entering into S and G2/M phases, enhance serum HI antibody titer and effectively improve the percentages of CD4+ and CD8+ T cells in chickens vaccinated with ND vaccine at most time points. The actions of RAMPStp and RAMPS60c were stronger than that of Lev, and RAMPStp presented the best efficacy. These results indicated that RAMPStp and RAMPS60c characterize of the immune-enhancing activity and RAMPStp possessed the strongest activity. It would be anticipated as a component of new-type immunopotentiator.

•RAMPStp and RAMPS60c enhanced T lymphocyte proliferation.•RAMPS60c and RAMPStp could promote lymphocytes enter into S and G2/M phases and improve the percentages of CD4+ and CD8+ T cells.•RAMPStp presented the best efficacy.•RAMPStp would be expected as a component of new-type immunopotentiator.This study evaluated the immune-enhancing activity of polysaccharides from the rhizoma of Atractylodis macrocephalae Koidz (RAMPS) in vitro. Lymphocyte proliferation, cell cycle distribution, and percentages of CD4+ and CD8+ cells were determined. Different concentrations of RAMPS were added to peripheral blood T lymphocytes. Results showed that RAMPStp and RAMPS60c could significantly enhance T lymphocyte proliferation individually or synergistically with phytohemagglutinin at most concentrations. The active sites of RAMPStp and RAMPS60c were then selected. Lymphocyte cell cycle distribution and percentages of CD4+ and CD8+ cells were determined by flow cytometry. At most time points, RAMPS60c and RAMPStp could promote lymphocytes enter into S and G2/M phases. RAMPStp and RAMPS60c effectively improved the percentages of CD4+ and CD8+ T cells. RAMPStp produced optimal effects. Therefore, RAMPStp could be used as a component of novel immunopotentiators.

•This study is about developing the colloidal gold-based immunochromatographic assay for rapid detection of Mycoplasma suis in porcine plasma.•In order to develop the colloidal gold-based immunochromatographic strips, polyclonal antibody (pAb) against Mycoplasma suis was used in this study.•The one-step colloidal gold immunochromatographic strip is easy, rapid, and convenient to use and requires little equipment.A one-step immunochromatographic assay using gold nanoparticles coated with polyclonal antibody (pAb) against Mycoplasma suis (M. suis) was developed in this study for the detection of M. suis in porcine plasma. The colloidal gold was prepared by the reduction of gold salt with sodium citrate coupled with pAb against M. suis. The pAb was produced by immunizing the BALB/c mice with recombinant MSG1 (rMSG1) protein from M. suis expressed in Escherichia coli. The optimal concentrations of the capture antibody and the coating antibody were 12 μg/ml and 1.5 mg/ml, respectively, and that of the blocking buffer was 1% bovine serum albumin. The lower detection limit of the immunochromatographic assay test was 100 ng/ml with visual detection under optimal conditions of analysis. Classical swine fever virus, porcine reproductive and respiratory syndrome virus, swine pneumonia mycoplasma, swine toxoplasma, and porcine parvovirus were used to evaluate the specificity of the immunochromatographic strips. No cross-reaction of the antibodies with other related swine pathogens was observed. This qualitative test based on the visual evaluation of the results did not require any equipment. The assay time for M. suis detection was less than 10 min, suitable for rapid detection at the grassroots level. The one-step colloidal gold immunochromatographic strips that we developed had high specificity and sensitivity. Therefore, this method would be feasible, convenient, rapid, and effective for detecting M. suis in porcine plasma.

This study aimed to detect Theileria annulata infection using polymerase chain reaction (PCR) amplification in Hyalomma asiaticum ticks. Sequence analysis showed that the 28 analyzed sequences obtained from 81 H. asiaticum ticks had an identical length and sequence which were closely related to that of T. annulata. This study is the first to report on the presence of T. annulata in H. asiaticum ticks in China.

The aim of the present study was to investigate the effects of Fu Zi on changes in the body heat of dogs. Twelve clinically healthy dogs were divided into two groups: the control group (six dogs) and the experimental group (six dogs). The control group was made to ingest normal saline mixed with canned meat, while the experimental group was made to ingest the Fu Zi solution mixed with canned meat. The infrared thermographic system was used to determine the level of body heat generated by these dogs. These areas include the dorsocranial (DCr), dorsocaudal (DCd), ventrocranial (VCr), and ventrocaudal (VCd) regions at pretreatment and were determined at 10, 20, 30, 50, 90, 120, 240, and 360 minutes after treatment for each of these areas. The results showed a tendency toward increased body heat until 30 minutes after ingestion of the Fu Zi powder mixed with canned meat. The significant differences in the changes of body heat were detected at 360 minutes in the DCd regions, 20 minutes in the VCr regions, and 30 minutes in the VCd regions between the experimental and control groups (p < 0.05). Based from our results, we find that Fu Zi can increase and maintain the dogs' body heat for at least 6 hours.

Sixteen shorthorn cows from Xiazhuang farm were admitted to the Veterinary Teaching Hospital at the College of Animal Science and Veterinary Medicine, Shandong Agricultural University for evaluation of poor appetite, listlessness, fever, tachycardia, tachypnea, lethargy, positive jugular venous pulse and anemia. Blood smear examination and polymerase chain reaction analysis in these cows revealed an infection with Mycoplasma wenyonii. The subjects were divided into two groups: control group (three cows) treated with intramuscular injection with imidocarb dipropionate (3 mg/kg/day for 2 days) and the experimental group (13 cows), treated with injection-acupuncture (Imidocarb Dipropionate, 1 mg/kg, once every 3 days for 6 days) at BL17, BL18, BL20, BL25, ST36, SP06 and CV04. At day 15, negative results were found using blood smear examination in all control and experimental groups.

•NDV two major genotypes had high sequence homology.•Polymorphism of Class II NDV genes evolve by positive selection.•The dN/dS for F gene were higher than those for HN gene.In our research, the molecular evolutions of NDV F and HN genes were analyzed. The phylogenetic analyses of NDV sequences indicated that NDV could be divided into two genotypes: Class I (lentogenic strains) and Class II (velogenic or mesogenic strains). Each genotype possesses high gene homology. Furthermore, the selected pressure analysis showed that the dN/dS of velogenic, mesogenic NDV strains F gene was significantly high and the ω(dN/dS) is 1.1725 > 1. These results imply that mutations in velogenic, mesogenic NDV F gene are favored by positive natural selection and it has acted to diversify NDV virulence at the nucleotide and amino acid level. We estimated that the subsequent rapid adaptation of the Newcastle disease virus to chickens were likely dependent on a high rate of mutation and the positive selection of mutations in the major F gene.

•NDV two major genotypes had high sequence homology.•Polymorphism of Class II NDV genes evolve by positive selection.•The dN/dS for F gene were higher than those for HN gene.In our research, the molecular evolutions of NDV F and HN genes were analyzed. The phylogenetic analyses of NDV sequences indicated that NDV could be divided into two genotypes: Class I (lentogenic strains) and Class II (velogenic or mesogenic strains). Each genotype possesses high gene homology. Furthermore, the selected pressure analysis showed that the dN/dS of velogenic, mesogenic NDV strains F gene was significantly high and the ω(dN/dS) is 1.1725 > 1. These results imply that mutations in velogenic, mesogenic NDV F gene are favored by positive natural selection and it has acted to diversify NDV virulence at the nucleotide and amino acid level. We estimated that the subsequent rapid adaptation of the Newcastle disease virus to chickens were likely dependent on a high rate of mutation and the positive selection of mutations in the major F gene.

This study focused on the effect of silica nanoparticles as adjuvant for vaccine applications comprised of gp85, a dominating structural protein of J Subgroup Avian Leukosis Virus (ALV-J), and which was evaluated by comparing with the responsiveness induced by that emulsified in Freund adjuvant. Thirty-six chickens were inoculated twice with gp85 adjuvanted with the silica nanoparticles or Freund’s adjuvant at the 2nd and 3rd week old. Two weeks later, the inoculated chickens were challenged with a 102.2 50% tissue culture infective dose (TCID50) of ALV-J. The blood samples were collected weekly to detect the serum antibodies and viremia. Results showed that positive serum antibodies (S/P value > 0.6) against gp85 emerged at the third week in the inoculated chickens, while the antibodies level persisted longer in silica nanoparticles adjuvanted-group to Freund’s adjuvanted-group. Furthermore, viremia in silica nanoparticles adjuvanted-group was recovered more quickly compared with Freund’s adjuvanted-group. Hence our study revealed that silica nanoparticles can effectively improve the protection of gp85 vaccine against ALV-J and present a better performance than Freund’s adjuvant.

To obtain an effective vaccine candidate against duck Tembusu viral (DTMUV) disease which causes egg-drop and great economical loss in the Chinese duck industry, liposome vaccines containing recombinant E protein were prepared and assessed in this study. The recombinant plasmid (PET28a-E) was constructed and transformed into BL21 (DE3) cells to produce E proteins. The recombinant E proteins were purified and entrapped by liposomes through reverse-phase evaporation. Eighty-four cherry valley ducks were randomly divided into seven groups and inoculated intramuscularly at one- or seven-day-old with liposomes-E protein or Freund's adjuvant-E protein vaccine. Blood samples were collected from the first week to the tenth week for serum antibody, plasma for viremia, as well as oropharyngeal and cloacal swabs for virus shedding analyses after being challenged with a 102.4 50% tissue culture infective dose (TCID50) of duck Tembusu virus. Results showed that serum antibody level of the liposomes vaccine was higher than the Freund's adjuvant vaccine, and inoculating twice was superior to once; furthermore, the viremia and virus shedding tests also proved that the liposomes vaccine can provide complete protection against DTMUV challenge. These results demonstrated that the liposomes-E protein vaccine could be used as a potential candidate vaccine to prevent DTMUV infection in ducks.

The objective of this study is to evaluate the effects of chromic chloride (CrCl3) on chick embryo fibroblast (CEF) viability. The cells were incubated with CrCl3 (0.02, 0.1, 0.5, 2.5, 12.5, and 62.5 μM), and the viability was determined using MTT assay, morphological detection and flow cytometry. The results show that lower concentrations of CrCl3 (0.02, 0.1, and 0.5 μM) did not damage CEF viability. At 0.1 μM, CrCl3 can increase CEF viability (P < 0.05). However, at higher concentrations of CrCl3 (2.5, 12.5, and 62.5 μM), the number of apoptotic and necrotic cells (P < 0.01) and intracellular reactive oxygen species (P < 0.01) increased. In addition, decreased mitochondrial membrane potential (P < 0.01) and enhanced intracellular calcium levels (P < 0.01) were observed after the exposure. Moreover, apoptotic morphological changes induced by these processes in CEF were confirmed using Hoechst 33258 staining. Cell death induced by higher concentrations of CrCl3 was caused by an apoptotic and a necrotic mechanism, whereas the main mechanism of oxidative stress and induced mitochondrial dysfunction was apoptotic death. The induced apoptotic death in CEF is concentration- and time-dependent.

•In this study, we prepared the subunit vaccines of liposome containing gp85 protein of ALV-J.•Liposomal vaccines were able to stimulate chickens to produce high level of serum antibody and reduce the presence of viremia effectively.•Liposome can enhance immune response of gp85 protein more than Freund's adjuvant.To study the potential of liposome vaccines in the clinical prevention of ALV-J, the effect of recombinant gp85 protein of subgroup J avian leukosis virus (ALV-J) entrapped by liposomes in chickens against ALV-J infection was investigated in this paper. A recombinant plasmid (PET28a-gp85) containing the PET28a vector and gp85 gene was constructed and then expressed in Rosetta (DE3) cells with 0.5 mM IPTG to produce recombinant gp85 proteins that could be entrapped by liposomes through reverse-phase evaporation. The chickens were inoculated intramuscularly either once or twice with the liposomes or with Freund's adjuvant emulsion containing recombinant gp85 protein. Sixty chickens were raised to one week old for the first inoculation and to three weeks old for the second inoculation. Chickens raised to five weeks old were challenged with a 102.4 50% tissue culture infective dose (TCID50) of ALV-J. Blood samples were collected from each chicken at weekly intervals for serum antibody and viremia analyses. Changes in serum antibodies showed that positive serum antibodies (S/P value >0.6) could be induced in all groups regardless of the frequency of inoculation but improved significantly in the twice-inoculated groups. As well, high levels of antibodies emerged earlier in the Freund's adjuvant groups but persisted longer in the liposome groups. Detection of viremia indicated that the liposomes provide better protection against ALV-J than Freund's adjuvant emulsion and that this protection is directly influenced by serum antibody levels. Overall, this study reveals the potential of liposome vaccines containing recombinant gp85 protein in the clinical prevention of ALV-J.

•In this study, we prepared the subunit vaccines of liposome containing gp85 protein of ALV-J.•Liposomal vaccines were able to stimulate chickens to produce high level of serum antibody and reduce the presence of viremia effectively.•Liposome can enhance immune response of gp85 protein more than Freund's adjuvant.To study the potential of liposome vaccines in the clinical prevention of ALV-J, the effect of recombinant gp85 protein of subgroup J avian leukosis virus (ALV-J) entrapped by liposomes in chickens against ALV-J infection was investigated in this paper. A recombinant plasmid (PET28a-gp85) containing the PET28a vector and gp85 gene was constructed and then expressed in Rosetta (DE3) cells with 0.5 mM IPTG to produce recombinant gp85 proteins that could be entrapped by liposomes through reverse-phase evaporation. The chickens were inoculated intramuscularly either once or twice with the liposomes or with Freund's adjuvant emulsion containing recombinant gp85 protein. Sixty chickens were raised to one week old for the first inoculation and to three weeks old for the second inoculation. Chickens raised to five weeks old were challenged with a 102.4 50% tissue culture infective dose (TCID50) of ALV-J. Blood samples were collected from each chicken at weekly intervals for serum antibody and viremia analyses. Changes in serum antibodies showed that positive serum antibodies (S/P value >0.6) could be induced in all groups regardless of the frequency of inoculation but improved significantly in the twice-inoculated groups. As well, high levels of antibodies emerged earlier in the Freund's adjuvant groups but persisted longer in the liposome groups. Detection of viremia indicated that the liposomes provide better protection against ALV-J than Freund's adjuvant emulsion and that this protection is directly influenced by serum antibody levels. Overall, this study reveals the potential of liposome vaccines containing recombinant gp85 protein in the clinical prevention of ALV-J.