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1.
Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus in the order Nidovirales, family Arteriviridae, genus Arterivirus. The virus induces a prolonged viremia, replicates in macrophages, and produces persistent infection. The purpose of this study was to determine if PRRSV could persist for 90 d or more in a large population of breeding-age gilts housed under environmental conditions typical of commercial swine production and to determine if experimentally infected gilts could shed virus to naïve sentinel gilts beyond 90 d postinfection. Using the intranasal route, we inoculated 120 PRRSV-naïve gilts, 4 mo of age, with 5 mL of cell culture fluid containing a total dose of 102.4 TCID50 of a field isolate (MN-30100) of PRRSV. The index gilts were organized into 3 groups (A, B, and C), 40 gilts per group. To assess the dynamics of the experimental infection, a monitor group of 30 index gilts was blood-tested on days 0, 3, 7, 14, 30, 60, 90, 120, 150, and 180 postinfection. PRRSV viremia was detected with the polymerase chain reaction (PCR) on days 3, 7, and 14 and by virus isolation (VI) on days 7 and 14. PRRSV antibodies were detected from day 14 by enzyme-linked immunosorbent assay (ELISA). To assess shedding, 30 PRRSV-naïve sentinel gilts were commingled with the index gilts on day 90 postinfection and tested by PCR, VI, and ELISA every 15 d until 180 d postinfection; all samples were negative. To assess persistence, 40 index and 10 sentinel gilts were slaughtered at 120 (group A), 150 (group B), or 180 (group C) d postinfection. Evidence of PRRSV was not detected by PCR or VI in any tissue samples from the 120 index gilts. These results indicate that persistence and shedding of PRRSV are of short duration in breeding-age gilts.  相似文献   

2.
Embryonated chicken eggs (ECE) and the Madin-Darby canine kidney (MDCK) cell line were compared for isolation of swine influenza virus (SIV) from nasal swabs and tissue samples. Samples originated from 30 pigs experimentally inoculated with 2 × 106 to 2 × 107 embryo infectious dose 50% (EID50)/mL of swine influenza strain A/Swine/Indiana/1726/88 (H1N1). The results were analyzed with McNemar's chi-squared test for symmetry. The results indicated that more samples were SIV-positive with ECE than with tissue culture (P ≤ 0.001), suggesting that ECE remains the system of choice for isolation of SIV. It is recommend that routine use of both SIV isolation systems will increase the sensitivity of detection of virus shedding by considering the differences in growth and tropism of diverse SIV strains.  相似文献   

3.
4.
Lymphocyte subsets isolated from germ-free piglets experimentally infected with swine influenza virus (SIV), porcine reproductive and respiratory syndrome virus (PRRSV) or porcine circovirus type 2 (PCV2) were studied and the profile of these subsets among these three infections was monitored. Germ-free piglets were used since their response could be directly correlated to the viral infection. Because SIV infections are resolved even by colostrum-deprived neonates whereas PRRSV and PCV2 infections are not, SIV was used as a benchmark for an effectively resolved viral infection. PRRSV caused a large increase in the proportion of lymphocytes at the site of infection and rapid differentiation of B cells leading to a high level of Ig-producing cells but a severe reduction in CD2CD21+ primed B cells. Unlike SIV and PCV2, PRRSV also caused an increase in terminally differentiated subset of CD2+CD8α+ γδ cells and polyclonal expansion of major Vβ families suggesting that non-specific helper T cells drive swift B cell activation. Distinct from infections with SIV and PRRSV, PCV2 infection led to the: (a) prevalence of MHC-II+ T cytotoxic cells, (b) restriction of the T helper compartment in the respiratory tract, (c) generation of a high proportion of FoxP3+ T cells in the blood and (d) selective expansion of IgA and IgE suggesting this virus elicits a mucosal immune response. Our findings suggest that PRRSV and PCV2 may negatively modulate the host immune system by different mechanisms which may explain their persistence.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0091-x) contains supplementary material, which is available to authorized users.  相似文献   

5.
Animal manure can be a source of antibiotic-resistant genes (ARGs) and pharmaceutical residues; however, few studies have evaluated the presence of ARG in pasture-raised animal production systems. The objective of this study was to examine changes in microbiome diversity and the presence of antibiotic residues (ABRs) on three farms that contained a diverse range of animal species: pasture-raised poultry (broiler and layer), swine, and beef cattle. Total bacterial communities were determined using 16S rRNA microbiome analysis, while specific ARGs (sulfonamide [Sul; Sul1] and tetracycline [Tet; TetA]) were enumerated by qPCR (real-time PCR). Results indicated that the ARG abundances (Sul1 [P < 0.05] and TetA [P < 0.001]) were higher in layer hen manures (16.5 × 10−4 and 1.4 × 10−4 µg kg−1, respectively) followed by broiler chickens (2.9 × 10−4 and 1.7 × 10−4 µg kg−1, respectively), swine (0.22 × 10−4 and 0.20 × 10−4 µg kg−1, respectively) and beef cattle (0.19 × 10−4 and 0.02 × 10−4 µg kg−1, respectively). Average fecal TetA ABR tended to be greater (P = 0.09) for broiler chickens (11.4 µg kg−1) than for other animal species (1.8 to 0.06 µg kg−1), while chlortetracycline, lincomycin, and oxytetracycline ABRs were similar among animal species. Furthermore, fecal microbial richness and abundances differed significantly (P < 0.01) both among farms and specific species of animal. This study indicated that the microbial diversity, ABR, ARG concentrations, and types in feces varied from farm-to-farm and from animal species-to-animal species. Future studies are necessary to perform detailed investigations of the horizontal transfer mechanism of antibiotic-resistant microorganisms (ARMs) and ARG.  相似文献   

6.
1. The significance of airborne transmission in epidemics of infectious diseases in the livestock production industry remains unclear. The study therefore investigated the shedding route (faeces vs. exhaled air) of a vaccine strain of infectious bursal disease virus (IBDV) by broilers and the emission of airborne virus.

2. The experimental room contained 526 broilers which were orally inoculated at the age of 20?d. The airborne virus was sampled by three different bioaerosol samplers: Andersen six-stage impactor, all-glass impinger (AGI-30) and OMNI-3000.

3. Infected broilers started to shed virus in faeces on d 5 post inoculation (PI), and stopped shedding on d 12 PI. The faecal virus remained detectable for at least two d after drying under broiler room conditions. No virus was detected in the air exhaled by broilers.

4. Airborne virus was collected on d 5, 8 and 12 PI at 20?cm above the floor, and on d 8 and 12 PI in exhausted air. The emission rates of IBDV were 4·0 log10 50% tissue culture infectious dose (TCID50)/bird/d on d 8 PI, and 4·5 log10 TCID50/bird/d on d 12 PI.

5. We concluded that broilers shed IBDV mainly through their faeces. The presence of indoor airborne virus is associated with the viral presence in faeces. The successful recovery of airborne virus in exhausted air indicates there is a potential risk of virus spreading to the ambient environment via air.  相似文献   

7.
8.
Among the reported probiotic Bacillus strains, B. subtilis C-3102 has the unique potential to improve feed uptake under stress conditions in the broilers, piglets, and cows. In this study, we sought to evaluate the protective effect of feed additive probiotic Bacillus subtilis C-3102 against Salmonella enterica infection of specific pathogen-free (SPF) chicks in floor pens in two experiments. In the experiment-1, the chicks in the control group (n=32) were fed a basal diet and those in the C-3102 group (n=32) were fed a basal diet supplemented with 1×106 CFU/g of feed for 28 days. On day 7 post-challenge with S. enterica, there was no significant change in the body weight between both the groups throughout the test period, whereas detection rates of S. enterica in the C-3102 group were significantly lower in the cecum and liver on days 21 and 14 post-challenge, respectively. In the experiment-2, minimum dosage of C-3102 cells required to protect Salmonella infection was evaluated using 3 dosages. Chicks were divided into four groups, fed with different dosages of C-3102 (1×106, 5×105, 3×105, and 0 CFU/g of feed), and challenged with S. enterica (2.8×108 CFU/chicken). S. enterica infection was completed within 7 days post- challenge and was almost excluded from the liver and spleen on day 21 post- challenge in the control group. Average values showed a trend for higher infection rates in the control group >3×105>5×105>1×106 CFU/g on days 14 and 21 post-challenge. These results suggest that B. subtilis C-3102 supplementation has the potential to reduce S. enterica infection rates and/or to accelerate the exclusion of S. enterica from the chicks.  相似文献   

9.
Feline leukemia virus (FeLV) causes a range of neoplastic and degenerative diseases in cats. To obtain a more sensitive and convenient diagnosis of the disease, we prepared monoclonal antibodies specific for the FeLV p27 to develop a rapid diagnostic test with enhanced sensitivity and specificity. Among these antibodies, we identified two clones (hybridomas 8F8B5 and 8G7D1) that specifically bound to FeLV and were very suitable for a diagnostic kit. The affinity constants for 8F8B5 and 8G7D1 were 0.35 × 109 and 0.86 × 109, respectively. To investigate the diagnostic abilities of the rapid kit using these antibodies, we performed several clinical studies. Assessment of analytical sensitivity revealed that the detection threshold of the rapid diagnostic test was 2 ng/mL for recombinant p27 and 12.5 × 104 IU/mL for FeLV. When evaluating 252 cat sera samples, the kit was found to have a kappa value of 0.88 compared to polymerase chain reaction (PCR), indicating a significant correlation between data from the rapid diagnostic test and PCR. Sensitivity and specificity of the kit were 95.2% (20/21) and 98.5% (257/261), respectively. Our results demonstrated that the rapid diagnostic test would be a suitable diagnostic tool for the rapid detection of FeLV infection in cats.  相似文献   

10.
Understanding the dynamics of porcine reproductive and respiratory syndrome virus (PRRSV) vertical transmission is important to enhance the accuracy of monitoring protocols for endemically infected breeding herds. The objectives of this study were to determine the prevalence of PRRSV within infected litters, to quantify viremia, and to identify specific attributes of infected individuals. Eight gilts were intramuscularly inoculated with 101 TCID50 of a mildly virulent PRRSV strain (MN-30100) at 90 d of gestation. All inoculated gilts transmitted the virus in utero. The proportion of PRRSV PCR-positive piglets and the level of viremia in the piglets were higher at 4 d of age than at birth or at weaning. No specific attributes were associated with PRRSV infection in the piglets. This is the first report, that we are aware of, documenting the efficient in utero transmission of an extremely low dose of a mildly virulent strain of PRRSV. The results support the sampling of piglets late during lactation as a tool to monitor PRRSV shedding from sow-herds.  相似文献   

11.
Twenty 6‐week‐old specific pathogen‐free pigs were divided into four groups. On day 0 of the experiment, PRRSV–PRV (n = 6) and PRRSV (n = 4) groups were intranasally inoculated with porcine reproductive and respiratory syndrome virus (PRRSV) (105.6 TCID50). On day 7, the PRRSV–PRV and PRV (n = 6) groups were intranasally inoculated with pseudorabies virus (PRV) (103.6 TCID50). Control pigs (n = 4) were kept as uninoculated negative controls. Half of the pigs in each group were euthanized and necropsied on day 14 or 21. Clinical signs such as depression and anorexia were observed in the PRRSV–PRV and PRV groups after inoculation with PRV. Although febrile response was observed after virus inoculations, the duration of that response was prolonged in the PRRSV–PRV group compared with the other groups. The lungs in the PRRSV–PRV group failed to collapse and were mottled or diffusely tan and red, whereas the lungs of the pigs in the other groups were grossly normal. Histopathologically, interstitial pneumonia was present in all PRRSV‐inoculated pigs, but the pneumonic lesions were more severe in the PRRSV–PRV group. Mean PRRSV titres of tonsil and lung in the PRRSV–PRV group were significantly (P < 0.05) higher than that in the PRRSV group on day 21. These results indicate that dual infection with PRRSV and PRV increased clinical signs and pneumonic lesions in pigs infected with both viruses, as compared to pigs infected with PRRSV or PRV only, at least in the present experimental conditions.  相似文献   

12.
BackgroundPseudorabies virus (PRV) infection leads to high mortality in swine. Despite extensive efforts, effective treatments against PRV infection are limited. Furthermore, the inflammatory response induced by PRV strain GXLB-2013 is unclear.ObjectivesOur study aimed to investigate the inflammatory response induced by PRV strain GXLB-2013, establish an inflammation model to elucidate the pathogenesis of PRV infection further, and develop effective drugs against PRV infection.MethodsKunming mice were infected intramuscularly with medium, LPS, and different doses of PRV-GXLB-2013. Viral spread and histopathological damage to brain, spleen, and lung were determined at 7 days post-infection (dpi). Immune organ indices, levels of reactive oxygen species (ROS), nitric oxide (NO), and inflammatory cytokines, as well as levels of activity of COX-2 and iNOS were determined at 4, 7, and 14 dpi.ResultsAt 105–106 TCID50 PRV produced obviously neurological symptoms and 100% mortality in mice. Viral antigens were detectable in kidney, heart, lung, liver, spleen, and brain. In addition, inflammatory injuries were apparent in brain, spleen, and lung of PRV-infected mice. Moreover, PRV induced increases in immune organ indices, ROS and NO levels, activity of COX-2 and iNOS, and the content of key pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6, tumor necrosis factor-α, interferon-γ and MCP-1. Among the tested doses, 102 TCID50 of PRV produced a significant inflammatory mediator increase.ConclusionsAn inflammatory model induced by PRV infection was established in mice, and 102 TCID50 PRV was considered as the best concentration for the establishment of the model.  相似文献   

13.
The minimum intravenous infectious dose for ovine progressive pneumonia virus (OPPV) WLC1 was determined using twenty-four 6 month-old lambs. Twelve groups of two 6 month-old lambs were inoculated intravenously (i.v.) with tissue culture fluid containing ovine progressive pneumonia virus (OPPV) WLC1 titers ranging from 107.6 TCID50/lamb down to 10−3.4 TCID50/lamb and were monitored for seroconversion using the OPPV agar gel immunodiffusion assay (AGID). Fifteen of the 16 lambs given equal or greater than 100.6 TCID50 seroconverted, and virus could be isolated from peripheral blood leukocytes in 13 out of the 15 of these lambs. None of the eight lambs receiving less than 100.6 TCID50 seroconverted during the 12 months. The results of this study indicated that 100.6 or 4 TCID50/lamb given i.v. was capable of establishing infection.  相似文献   

14.
The urinary corticoid to creatinine ratio (UCCR) is one of the most commonly used screening tests for canine hypercortisolism (HC). In this study, a reference interval was established for UCCR using IMMULITE 2000 XPi, the latest chemiluminescence enzyme immunoassay. The diagnostic performance of this method for UCCR in canine HC was also evaluated. The median UCCR was 1.06 × 10−5 (range: 0.28–2.49) for 58 healthy dogs, and an upper reference limit of 1.98 × 10−5 (90% confidence interval: 1.76–2.15) was determined. The median UCCR in the 12 dogs with HC (7.38 × 10−5, range 1.86–29.98) was significantly higher than that in the 16 dogs with mimic-HC (1.59 × 10−5, range 0.47–3.42, P<0.001). The area under the curve for UCCR to differentiate HC dogs from mimic-HC dogs was 0.971, with a sensitivity of 91.7% and specificity of 100% when the cut-off value was set at 3.77 × 10−5. The UCCR of 16 paired urine samples collected at home and in hospital showed that the UCCR of samples collected in the hospital was significantly higher than that of samples collected at home (mean difference 3.30 × 10−5, 95% confidence interval: 0.70–5.90, P=0.001). In summary, we established the upper reference limit for UCCR using IMMULITE 2000 XPi in dogs and confirmed that UCCR is a useful diagnostic test for HC in dogs if urine samples are collected at home.  相似文献   

15.
16.
Following the recent use of a live vaccine against porcine reproductive and respiratory syndrome virus (PRRSV) in Denmark, both American (vaccine) and European-type PRRSV now coexist in Danish herds. This situation highlighted a requirement for supplementary tests for precise virus-typing. As a result, we developed a RT-PCR assay able to detect as well as type PRRSV. To provide maximal sequence information, complete viral open reading frames (ORFs 5 and 7) were targeted for amplification. The RT-PCR test was able to amplify complete PRRSV ORFs from complex materials such as boar semen containing as little as 1 TCID50 ml−1 of PRRSV. Typing of viruses was accomplished by any one of three strategies: (i) use of type-specific PCR primers, (ii) size determination of ORF 7 amplicons, (iii) DNA sequencing. All three typing strategies showed complete concordance with the currently used method of typing with monoclonal antibodies (MAbs) when used on a panel of PRRSV field isolates covering the period 1992–1997. The ORF 7-based test had particularly desirable characteristics, namely, highly sensitive detection of PRRSV without apparent type bias, typing of the detected virus, discrimination between pure and mixed virus populations, and semi-quantitative assessment of type ratios in mixed populations, all in a single PCR reaction. In addition, the obtained sequence data were used to predict two simple and rapid strategies (single-enzyme restriction length polymorphy analysis and oligonucleotide hybridization) for confirmation of the specificity of ORF 7 RT-PCR reactions. As such, the RT-PCR assay provides a new, powerful diagnostic tool to study the population dynamics between present and emerging PRRSV-types.  相似文献   

17.

Background

This study was conducted to investigate effect of exogenous melatonin on the development of mouse mature oocytes after cryopreservation.

Results

First, mouse metaphase II (MII) oocytes were vitrified in the open-pulled straws (OPS). After warming, they were cultured for 1 h in M2 medium containing melatonin at different concentrations (0, 10−9, 10−7, 10−5, 10−3 mol/L). Then the oocytes were used to detect reactive oxygen species (ROS) and glutathione (GSH) levels (fluorescence microscopy), and the developmental potential after parthenogenetic activation. The experimental results showed that the ROS level and cleavage rate in 10−3 mol/L melatonin group was significantly lower than that in melatonin-free group (control). The GSH levels and blastocyst rates in all melatonin-treated groups were similar to that in control. Based on the above results, we detected the expression of gene Hsp90aa1, Hsf1, Hspa1b, Nrf2 and Bcl-x1 with qRT-PCR in oocytes treated with 10−7, or 10−3 mol/L melatonin and untreated control. After warming and culture for 1 h, the oocytes showed higher Hsp90aa1 expression in 10−7 mol/L melatonin-treated group than in the control (P < 0.05); the Hsf1, Hsp90aa1 and Bcl-x1 expression were significantly decreased in 10−3 mol/L melatonin-treated group when compared to the control. Based on the above results and previous research, we detected the development of vitrified-warmed oocytes treated with either 10−7 or 0 mol/L melatonin by in vitro fertilization. No difference was observed between them.

Conclusions

Our results indicate that the supplementation of melatonin (10−9 to 10−3 mol/L) in culture medium and incubation for 1 h did not improve the subsequent developmental potential of vitrified-warmed mouse MII oocytes, even if there were alteration in gene expression.  相似文献   

18.
Porcine infections with highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) cause significant morbidity and mortality and currently there are no effective vaccines for disease prevention. An attenuated strain, HuN4-F112, was obtained by passaging the HP-PRRSV HuN4 on Marc-145 cells (112th-passage). PRRSV-free pigs were inoculated intramuscularly with HuN4-F112 (102.0, 103.0, 104.0, 105.0 and 106.0 TCID50 for groups 1–5, respectively). The groups 3–5 could resist the lethal challenge and did not show any obvious changes in body temperature nor clinical signs throughout the experiment, the pathological lesions were milder and the gained weight at a greater rate (P < 0.05), compared to group 1 and control. Sequence analysis of the HuN4 passages showed a conserved epitope in GP5 protein was mutated (196QWGRL/P200 → 196RWGRL/P200), as a result the monoclonal antibody could not recognize the HuN4-F112 any more. These results suggested that the HuN4-F112 could protect piglets from lethal challenge and might be a candidate vaccine against the HP-PRRSV.  相似文献   

19.
Pigs were inoculated orally with Mycobacterium avium in doses ranging from 15.6 × 102 to 15.6 × 106 viable units daily for 15 days (Table 1). The animals were necropsied 31–32 days after the last inoculation.Pigs given doses of 15.6 × 106 and 15.6 × 105 viable units showed delayed hypersensitivity to avian tuberculin 24 days after the last inoculation (Table 2). The pig inoculated with 15.6 × 106 viable units showed macro- and/or microscopic lesions of the intestines and the liver, and of the mandibular, mesenteric and hepatic lymph nodes. Cultures showed growth of M. avium from the same tissues and from the spleen and the left tracheobronchial lymph node. The pig inoculated with 15.6 × 105 viable units showed a pronounced granulomatous infiltration in the tonsils and the mesenteric lymph nodes. Growth of M. avium was obtained from the tonsils, the intestinal mucosa (Peyer patch) and the mandibular and mesenteric lymph nodes. Viable unit counts were high in the tonsils and in the mesenteric lymph nodes (Tables 3 and 4).Lower doses gave rise to a minimal tissue reaction and/or very low viable unit counts, and are not considered to be capable of producing a progressive tuberculosis.  相似文献   

20.
The aim of these experiments was to investigate the potential antiviral effect of Saccharomyces cerevisiaeβ‐glucan on the pneumonia induced by swine influenza virus (SIV). Forty colostrum‐deprived 5‐day‐old piglets were randomly divided into four groups of 10. The 20 pigs in groups 1 and 2 were administered Saccharomyces cerevisiaeβ‐glucan orally (50 mg/day/pig; En‐Bio Technology Co., Ltd) for 3 days before SIV infection and those in groups 3 and 4 were given culture medium/diluent alone. Groups 1 and 3 were inoculated intranasally with 3 ml of tissue culture fluid containing 2 × 106 tissue culture infective doses 50% (TCID50)/ml of SIV and those in groups 2 and 4 were exposed in the same manner to uninfected cell culture supernatant. The microscopic lung lesions induced by SIV infection (group 1 pigs) were significantly more severe than those induced by infection in animals pre‐administered β‐glucan (group 3) (P < 0.05). Significantly more SIV nucleic acid was detected in the lungs of pigs experimentally infected with SIV only (group 1) at 5, 7 and 10 days post‐inoculation (dpi) compared with lungs from pigs pre‐administered β‐glucan and infected with SIV (group 3) (P < 0.05). The concentrations of interferon‐γ (IFN‐γ) and nitric oxide (NO) in bronchoalveolar lavage fluid from pigs pre‐administered β‐glucan and infected with SIV (group 3) were significantly higher than for any other group at 7 and 10 dpi for IFN‐γ, and at 5, 7 and 10 dpi for NO (P < 0.05). Saccharomyces cerevisiaeβ‐glucan reduced the pulmonary lesion score and viral replication rate in SIV‐infected pigs. These findings support the potential application of β‐glucan as prophylactic/treatment agent in influenza virus infection.  相似文献   

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