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Marek disease (MD) is a viral disease characterized by the development of lymphoma in poultry. Although morphologic confirmation of lymphoma is used to diagnose MD, immunohistochemical detection of MD virus- EcoRI-Q (Meq), which is a viral protein that is expressed exclusively in MD tumor cells, would further improve the accuracy of diagnosis. We developed monoclonal antibodies (mAbs) that specifically detect Meq by immunohistochemistry (IHC) using formalin-fixed, paraffin-embedded (FFPE) sections. We evaluated the sensitivity and specificity of 14 mAbs that we produced, using FFPE samples of MDCC-MSB1 cells, MD tumor tissues, and tissues of uninfected chickens. Four different antigen retrieval conditions were investigated. Thirteen mAbs reacted with Meq in FFPE sections, but immunohistochemical reactivity and specificity varied depending on the mAb and antigen retrieval condition; heat-induced antigen retrieval (HIAR) was more effective at detecting Meq than the other tested conditions. HIAR pH 9 tended to increase immunoreactivity and decrease specificity. Of the 5 mAbs that immunoreacted strongly with Meq without nonspecific reactions under the optimal antigen retrieval conditions, 3 mAbs (1C1-121, 3A3-112, 5F7-82) did not produce background staining of tumor or non-tumor tissues; 2 mAbs (2C5-11, 4A5-54) produced background staining. The mAb 6B5-128 reacted moderately with Meq without nonspecific reactions and background staining. The remaining mAbs showed weak immunoreactivity or problematic nonspecific reactions. Our results suggest that some of our developed mAbs can be used in IHC to detect Meq in FFPE sections with high specificity, and that the use of IHC may greatly improve the diagnosis of MD.

In this study, a total of nine chicken samples obtained from two broiler flocks in Oita and Tottori prefectures in 2020 were examined for Chicken anemia virus (CAV) infection. The samples were collected from clinically suspected flocks and diseased chickens. The CAV genome was detected in all nine samples tested by real-time PCR. Phylogenetic analyses and sequence comparisons of the full-length VP1 gene sequences indicated that all the Japanese CAV strains obtained in this study formed a similar cluster of genotype III and shared high nucleotide (99.62-100%) identity. The current Japanese CAV strains were closely related to Chinese CAV strains but not related to vaccine strains. One positive selection site of VP1 was detected among the Japanese CAV strains.

Marek disease (MD) is a viral disease characterized by the development of lymphoma in poultry. Although morphologic confirmation of lymphoma is used to diagnose MD, immunohistochemical detection of MD virus-EcoRI-Q (Meq), which is a viral protein that is expressed exclusively in MD tumor cells, would further improve the accuracy of diagnosis. We developed monoclonal antibodies (mAbs) that specifically detect Meq by immunohistochemistry (IHC) using formalin-fixed, paraffin-embedded (FFPE) sections. We evaluated the sensitivity and specificity of 14 mAbs that we produced, using FFPE samples of MDCC-MSB1 cells, MD tumor tissues, and tissues of uninfected chickens. Four different antigen retrieval conditions were investigated. Thirteen mAbs reacted with Meq in FFPE sections, but immunohistochemical reactivity and specificity varied depending on the mAb and antigen retrieval condition; heat-induced antigen retrieval (HIAR) was more effective at detecting Meq than the other tested conditions. HIAR pH 9 tended to increase immunoreactivity and decrease specificity. Of the 5 mAbs that immunoreacted strongly with Meq without nonspecific reactions under the optimal antigen retrieval conditions, 3 mAbs (1C1-121, 3A3-112, 5F7-82) did not produce background staining of tumor or non-tumor tissues; 2 mAbs (2C5-11, 4A5-54) produced background staining. The mAb 6B5-128 reacted moderately with Meq without nonspecific reactions and background staining. The remaining mAbs showed weak immunoreactivity or problematic nonspecific reactions. Our results suggest that some of our developed mAbs can be used in IHC to detect Meq in FFPE sections with high specificity, and that the use of IHC may greatly improve the diagnosis of MD.

【要旨】A 大学栄養学科 2016 年度入学生を対象に記名自記式質問紙調査を実施し，一人暮らしの者 20 名（女性 17 名，男性 3 名）の回答より 1～2 年次から 4 年次までの食事作りへの意識・実践の変化を分析した。食事作り意識の変化として，何も見ずに作ることができると考える料理が 35 項目中 5 項目有意に増加し（p＜0.05），食事の「栄養バランス」「料理・味付けの組み合わせ」等に気を付けている者が増加した。また各年次で共通して，食品選択で重視していることに「価格」，食事作りで気を付けていることに「作り方が簡単」「短時間で 食事を準備」，料理の参考に「インターネット」「高校までの家庭料理」と回答する者が多く，大学での習得知 識や教科書等を参考とする者は少なかった。食事作り実践の変化として，泡立て器，スケール，おろし器，ゴムベラの調理器具所有者は有意に増加した（p＜0.05）一方，月 1 回以上作る料理の種類は減少が見られた。食事作りを「ほぼ毎日」する者は 5 割，主食・主菜・副菜の揃った食事を「毎食食べる」者は 3 食いずれも 15% 以下と年次による大きな変動はなかった。4 年次までに料理のレパートリーの広がりや食事バランスへの意識 向上が見られた反面，実際の食事作り実践や食事内容へ反映する者は少ないことが課題として示されたことから，学生の生活状況等を考慮し，日常において継続的な食事作りと食事内容の充実に繋がる教育上の工夫が必要と考えられる。, A self-administered questionnaire survey was conducted on students enrolled in the Department of Nutrition at University A in 2016. On the basis of the answers provided by 20 students living alone (17 women, 3 men), in first and second year and then to the fourth year, we analyzed changes in awareness of and practice around meal preparation. As a change in awareness of meal preparation, the number of students who believe they can cook meals without using a recipe increased significantly for 5 out of 35 dishes (p

A concurrent infection of chicken anemia virus (CAV) and infectious bronchitis virus (IBV) was detected in Japanese native chicks in 2017, in which a high mortality rate (97.7%) was recorded in a small flock of 130 chicks exhibiting poor growth. Histological examination revealed that the affected chicks exhibited two different pathological entities: one was severe hematopoietic and lymphocytic depletion with abnormally large cells containing intranuclear inclusion bodies of CAV, whereas the other was renal tubular necrosis due to IBV infection. Immunohistochemistry detected CAV antigens in the bone marrow, liver, and spleen as well as IBV antigens in the kidneys, trachea, and air sacs. CAV was isolated from the liver sample of the chicks, and the isolated strain was designated as CAV/Japan/HS1/17. A phylogenetic analysis of the CAV VP1 gene revealed that CAV/Japan/HS1/17 is genetically similar to Chinese strains collected from 2014 to 2016. An experimental infection was performed using CAV/Japan/HS1/17 and specific-pathogen-free chicks to determine the pathogenicity of CAV/Japan/HS1/17. The isolate caused 100% anemia and 70% mortality to chicks inoculated at one day old, 80% of chicks inoculated at seven days old also developed anemia, and 10% died from CAV infection. These results suggest that the unusually high mortality in Japanese native chicks can be attributed to dual infection with both CAV and IBV. The results of the experimental infection suggest that CAV/Japan/HS1/17 has a pathogenic potential to specific-pathogen-free chicks and a relatively higher pathogenicity than previous Japanese CAV strains.

Avian erythroblastosis (AE; erythroid leukosis) was detected in a 78-day-old Japanese native chicken. At necropsy, the liver was enlarged and diffusely dark red in color. Moderate splenomegaly was observed. Histologically, round to polygonal tumor cells were observed only in the blood vessels of the liver and other organs. Immunohistochemistry (IHC) was performed on formalin-fixed, paraffin-embedded (FFPE) sections to characterize tumor cells. Tumor cells, as well as normal erythrocytes as positive controls, were consistently positive for IHC by using the commercially available anti-human hemoglobin antibody. Exogenous avian leukosis virus (ALV) subgroup B and endogenous ALV-E genes were detected by PCR, although ultrastructural observation revealed no viral particles associated with tumor cells. Results suggest that the commercial anti-human hemoglobin antibody used in the study cross-react to chicken erythrocytes on FFPE sections by IHC and can also be used for definitive diagnosis of the spontaneous case of AE in chickens.

管理栄養士養成施設であるA大学2年生を対象として調理技能調査および質問紙調査を行い,切砕技術に及ぼす要因を検討した。調理技能調査は,きゅうりの輸切り,だいこんのせん切りおよびキャベツのせん切りを課題とし,所要時間の計測と切断物の評価(調査者評価および自己評価)を行った。切断物の調査者評価はいずれも約9割の学生が「やや不揃い」「不揃い」であった。一方,自己評価は調査者評価より高い傾向にあり,学生と調査者が考える技術レベルに違いがあることが推察された。切砕操作における意識は,調査時では「揃えて切ること」,日常生活では「特に意識していない」「速く切ること」「速く揃えて切ること」を回答した学生が多かった。今後は食事提供に携わる職を目指す学生に求められる技術レベルを理解させ,日常生活の切砕操作においても均一さを意識するよう促す指導が必要である。, To investigate influential factors, a knife skills survey and questionnaire were conducted among second‐year students at a university running a registered dietitian training course. The skills for slicing cucumber, Japanese radish, and cabbage were examined by both investigators and students assessing the time taken and the slices produced. For all three items, about 90% of the investigators' evaluations recorded "unevenness" or “a little unevenness. "On the other hand, the self-evaluations tended to be more positive, suggesting a difference between the perceptions of students and investigators. Many students reported that they remained conscious during the survey “ to cut same-sized slices. "In response to the questionnaire item about cutting same‐sized slices on an everyday basis, many students reported that they: "were not particularly conscious," "cut quickiy," or "cut same-sized slices quickly.“ In future, therefore, lt is essential to educate students about the level of knife skills required of those wishing to work in catering and encourage an awareness of cutting food evenly in everyday life.

Immunohistochemical method to detect avian lymphocytes is an efficient and reliable tool for accurate diagnosis, and immunological analysis of avian diseases. However, there are scarce studies reporting immunohistochemistry (IHC) using commercially available antibodies in formalin-fixed paraffin-embedded (FFPE) chicken tissues. In the present study, we established an immunohistochemical method to identify chicken T and B lymphocytes in FFPE chicken tissues using commercial antibodies against chicken or human antigens. For this IHC method, the five tested anti-T lymphocyte antibodies reacted with chicken T lymphocytes on the FFPE sections. Further, 10 commercial anti-B lymphocyte antibodies were tested; of these, three successfully detected chicken B lymphocytes for IHC. In particular, anti-human CD3 (clone F7.2.38) antibody was most suitable for the detection of chicken T lymphocytes, whereas anti-chicken B cell activating factor receptor (BAFF-R) antibody (clone 2C4) was most suitable for the detection of chicken B lymphocytes under our IHC staining conditions. These two antibodies reacted with numerous lymphocytes of all representative lymphoid tissues without problematic background staining and nonspecific reactions. Our results indicate that T and B lymphocytes in FFPE chicken tissues can be immunohistochemically detected using commercial antibodies.