Experimental dual infection of pigs with an H1N1 swine influenza virus (A/Sw/Hok/2/81) and Mycoplasma hyopneumoniae

Dual infection of pigs with swine influenza virus (SIV) and Mycoplasma hyopneumoniae was carried out to compare the clinical and pathological effects of dual infection in caesarian derived and colostrums deprived (CDCD) pigs, with that of a single infection with M. hyopneumoniae. In Experiment 1, 40-day-old CDCD pigs were inoculated only with SIV (A/Sw/Hok/2/81, H1N1). The virus was isolated from nasal swabs for 5-6 days. None of these pigs showed clinical signs of infection throughout the experimental period. These results suggested that this strain can infect pigs but is only slightly pathogenic when it is inoculated singly to a CDCD pig. In Experiment 2, 60-day-old CDCD pigs were inoculated with M. hyopneumoniae and then were inoculated with SIV (A/Sw/Hok/2/81) at 1 week (MHYO-7d-SIV-7d group) or 3 weeks (MHYO-21d-SIV-7d group) after M. hyopneumoniae inoculation. Macroscopically, dark red-to-purple lung lesions were observed in all of pigs at 14 or 28 days post-inoculation. Percentages of dark red-to-purple lung lesions in dual infection groups (MHYO-7d-SIV-7d group: 18.7 +/- 4.2%, MHYO-21d-SIV-7d group: 23.0 +/- 8.0%) were significantly (P < 0.05) increased compared to those of each control group in which pigs were inoculated only with M. hyopneumoniae (MHYO-14d group: 4.7 +/- 2.9%, MHYO-28 group: 3.3 +/- 2.4%). Microscopically, bronchial epithelial lesions (epithelial disruption, degeneration, hyperplasia and formation of microabscess) were frequently observed in dark red-to-purple lung lesions of only the dual infection groups. These results demonstrate that the lung lesion of pigs inoculated with M. hyopneumoniae and SIV is more severe than that of pigs inoculated only with M. hyopneumoniae.     

Detection of Cryptosporidium muris type oocysts from beef cattle in a farm and from domestic and wild animals in and around the farm

Cryptosporidium muris type oocysts were detected from 21 of 516 beef cattle in a farm. Then we surveyed Cryptosporidium oocysts in 348 beef and dairy cattle, 500 pigs, 101 dogs, 38 wild animals and 11 zoo-kept animals in and around the farm. Oocysts were detected from only 2 of 25 Japanese field mice, Apodemus speciosus in the same farm. Gene analysis suggested that the oocysts were different from the C. muris type bovine isolate.     

Effects of option mitigating ammonia volatilization on CH4 and N2O emissions from a paddy field fertilized with anaerobically digested cattle slurry

A lysimeter experiment was carried out to evaluate the effects of the NH₃ volatilization mitigation by adding anaerobically digested cattle slurry (ADCS) alone, with wood vinegar (WV) or with a higher level of floodwater (HFW), on emissions of CH₄ and N₂O from a paddy soil planted with fodder rice. We have carried out the following treatments: (1) chemical fertilizer, (2) ADCS, (3) ADCS + WV, and (4) ADCS + HFW; the height of floodwater was 10 cm in the latter treatment, and it was 3 to 4 cm in the other treatments just before fertilizer applications. Nitrogen fertilizer rate added to soil in each treatment was 30 g NH₄⁺–N m⁻² (split in one basal and two top-dressing additions). Ammonia volatilization in the ADCS treatment was 2.7 g NH₃–N m⁻² throughout the growing season, and it was significantly reduced by 79% and 55% in the ADCS + WV and ADCS + HFW treatments, respectively. The total amount of CH₄ emitted in the ADCS treatment in the growing season was not significantly enhanced by the mitigation of NH₃ volatilization either by adding wood vinegar or by increasing the height of the floodwater. Negligible N₂O emissions were observed in all treatments during the growing period.     

Evaluation of a recombinant measles virus expressing hepatitis C virus envelope proteins by infection of human PBL-NOD/Scid/Jak3null mouse

In this study, we infected NOD/Scid/Jak3null mice engrafted human peripheral blood leukocytes (hu-PBL-NOJ) with measles virus Edmonston B strain (MV-Edm) expressing hepatitis C virus (HCV) envelope proteins (rMV-E1E2) to evaluate the immunogenicity as a vaccine candidate. Although human leukocytes could be isolated from the spleen of mock-infected mice during the 2-weeks experiment, the proportion of engrafted human leukocytes in mice infected with MV (10³–10⁵ pfu) or rMV-E1E2 (10⁴ pfu) was decreased. Viral infection of the splenocytes was confirmed by the development of cytopathic effects (CPEs) in co-cultures of splenocytes and B95a cells and verified using RT-PCR. Finally, human antibodies against MV were more frequently observed than E2-specific antibodies in serum from mice infected with a low dose of virus (MV, 10⁰–10¹ pfu, and rMV-E1E2, 10¹–10² pfu). These results showed the possibility of hu-PBL-NOJ mice for the evaluation of the immunogenicity of viral proteins.     

Comparison of the immunosuppressive effects of dexamethasone, flunixin meglumine and meloxicam on the in vitro response of calf peripheral blood mononuclear cells

This study compared the immunosuppressive effects of dexamethasone (DEX), flunixin meglumine (FLU) and meloxicam (MEL) on the peripheral blood mononuclear cells (PBMCs) of seven healthy Holstein calves in vitro. DEX significantly inhibited lymphocyte proliferation and expression of interferon (IFN)-γ, interleukin (IL)-2 and IL-4 messenger RNA (mRNA) in comparison with FLU and MEL. FLU and MEL dose-dependently inhibited lymphocyte proliferation, but did not significantly reduce mRNA expression. Our in vitro study indicates that steroidal anti-inflammatory drugs (SAIDs) as well as nonsteroidal anti-inflammatory drugs (NSAIDs) have immunosuppressive effects on calf PBMCs. These findings are important for assessing the indications and complications of NSAIDs in calves.     

Studies on the organo-mineral colloidal complexes of paddy soil (Part 2) Humus contained in G1 of the degraded paddy soil

Regarding paddy soil colloids as the so-called “organo-mineral colloidal complexes” of A. F. Tyulin, the authors separated the colloidal fractions after his method, and reported the characrzstics of each fraction, and content of humus and some characteristics thereof in the previous paper¹⁾.     

An outbreak of rabbit hemorrhagic disease (RHD) caused by Lagovirus europaeus GI.2/rabbit hemorrhagic disease virus 2 (RHDV2) in Ehime, Japan

A total of ten 1–2-year-old rabbits died within 2 weeks at a facility in Ehime prefecture in May 2019. Necropsy revealed liver discoloration and fragility, hemorrhage of some organs and blood coagulation failure. On histopathologic examination, necrotizing hepatitis was a common finding, together with fibrin thrombi in the small vessels and hemorrhage in some organs. Rabbit hemorrhagic disease (RHD) virus gene was detected in liver samples, and viral particles of approximately 32 nm in diameter were found in the cytoplasm of degenerated hepatocytes by electron microscopy. Phylogenetic analysis based on the partial VP60 gene sequence classified it as Lagovirus europaeus GI.2/RHDV2. This is the first confirmed outbreak of RHD caused by globally emerging GI.2/RHDV2 in Japan.     

Seasonal variation of domoic acid in a bivalve Spondylus versicolor in association with that in plankton samples in Nha Phu Bay,Khanh Hoa,Vietnam

Previously we reported that a significant level of domoic acid, a causative toxin for amnesic shellfish poisoning, was detected in bivalves belonging to a genus Spondylus from various tropical Asian countries. These findings indicate that causative plankton species for domoic acid widely distribute in these areas. In the present study, we monitored seasonal change of domoic acid level of Spondylus versicolor in association with that of plankton samples in Nha Phu Bay, Khanh Hoa, Vietnam from December 2004 to October 2005. The toxicity of S. versicolor showed distinct seasonal variation. During the period when domoic acid level of S. versicolor was increasing, a significant level of domoic acid was detected in the plankton samples, showing the correlation between these two parameters. These findings show that plankton causative for domoic acid occurred in the bay, and S. versicolor accumulate domoic acid during a bloom of the toxic plankton by food web transfer.