Effects of Simple and Disposable Chicken Cages for Experimental Eimeria Infections

During experimental Eimeria infections in chickens, facilities are often contaminated by fecal oocysts known to be highly resistant to both chemical and enzymatic treatments. Thus, studies using experimental Eimeria infections have been limited due to the difficulty of complete elimination of residual oocysts from both cages and facilities. To overcome this limitation, simple, inexpensive, and disposable cages were constructed from cardboard boxes and tested during experimental Eimeria maxima infections. The cages were used in animal rooms with only a 1.7% evidence of coccidia contamination between adjacent cages. No significant differences in fecal oocyst output and body weight gain were noted between animals housed in disposable cages and animals housed in wire control cages. This cage design is a useful means for preventing oocyst contamination during experimental conditions, suggesting that this disposable cage design could be used for other avian infectious disease studies

Identification and Comparative Expression Analysis of Interleukin 2/15 Receptor β Chain in Chickens Infected with E. tenella

BACKGROUND: Interleukin (IL) 2 and IL15 receptor β chain (IL2/15Rβ, CD122) play critical roles in signal transduction for the biological activities of IL2 and IL15. Increased knowledge of non-mammalian IL2/15Rβ will enhance the understanding of IL2 and IL15 functions. METHODOLOGY/PRINCIPAL FINDINGS: [corrected] Chicken IL2/15Rβ (chIL2/15Rβ) cDNA was cloned using 5'/3'-RACE. The predicted protein sequence contained 576 amino acids and typical features of the type-I cytokine receptor family. COS-7 cells transfected with chIL2/15Rβ produced proteins of approximately 75 and 62.5 kDa under normal and tunicamycin-treated conditions, respectively. The genomic structure of chIL2/15Rβ was similar to its mammalian counterparts. chIL2/15Rβ transcripts were detected in the lymphoblast cell line CU205 and in normal lymphoid organs and at moderate levels in bursa samples. Expression profiles of chIL2/15Rβ and its related cytokines and receptors were examined in ConA-stimulated splenic lymphocytes and in ceca-tonsils of Eimeria tenella-infected chickens using quantitative real-time PCR. Expression levels of chIL2/15Rβ, chIL2Rα, and chIL15Rα were generally elevated in ceca-tonsils and ConA-activated splenic lymphocytes. However, chIL2 and chIL15 expression levels were differentially regulated between the samples. chIL2 expression was upregulated in ConA-activated splenic lymphocytes, but not in ceca-tonsils. In constrast, chIL15 expression was upregulated in ceca-tonsils, but not in ConA-activated splenic lymphocytes. CONCLUSIONS/SIGNIFICANCE: We identified an avian form of IL2/15Rβ and compared its gene expression pattern with those of chIL2, chIL15, chIL2Rα, and chIL15Rα. Our observations suggest that chIL15 and its receptors, including chIL2/15Rβ, play important roles in mucosal immunity to intestinal intracellular parasites such as Eimeria

The impact of COVID-19 on cancer care in a tertiary hospital in Korea: possible collateral damage to emergency care

OBJECTIVES We investigated the impact of the COVID-19 pandemic on cancer care in a tertiary hospital in Korea without specific lockdown measures. METHODS A retrospective cohort of cancer patients from one of the largest tertiary hospitals in Korea was used to compare healthcare utilization in different settings (outpatient cancer clinic, the emergency department [ED], and admissions to the hematology/oncology ward) between January 1 and December 31, 2020 and the same time period in 2019. The percent changes in healthcare utilization between the 2 periods were calculated. RESULTS A total of 448,833 cases from the outpatient cohort, 26,781 cases from the ED cohort, and 14,513 cases from the admission cohort were reviewed for 2019 and 2020. The total number of ED visit cases significantly decreased from 2019 to 2020 by 18.04%, whereas the proportion of cancer patients remained stable. The reduction in ED visits was more prominent in patients with symptoms suspicious for COVID-19, high-acuity cases, and those who lived in non-capital city areas. There were no significant changes in the number of total visits, new cases in the outpatient clinic, or the total number of hospitalizations between the 2 periods. CONCLUSIONS During the pandemic, the number of ED visits significantly decreased, while the use of the outpatient clinic and hospitalizations were not affected. Cancer patients’ ED visits decreased after the COVID-19 outbreak, suggesting the potential for collateral damage outside the hospital if patients cannot reach the ED in a timely manner

Rapid communication Monoclonal antibodies reactive with chicken interleukin-17

Abstract Chicken interleukin-17 (chIL-17) gene was previously characterized through cloning from a chicken intestinal expressed sequence tag (EST) cDNA library. To further investigate the biological properties of chIL-17, six monoclonal antibodies (mAbs) against a bacterially expressed chIL-17 recombinant protein were produced and their binding specificities characterized. Antibodies which were initially selected on the basis of their specific binding reactivity with recombinant chIL-17 in ELISA were further characterized by Western blot analysis. Monoclonal antibodies specific for chIL-17 identified 20 and 21 kDa protein bands in the culture supernatant and cell lysate of CU205 cells. These mAbs also recognized specific bands for chIL-17 in the cell lysate from conconavalin A (Con A)-activated, but not from normal splenic lymphocytes. Furthermore, these mAbs detected a 16 kDa protein in the lysate of CU205 cells treated with tunicamycin and stained an intracellular protein in CU205 cells in flow cytometric analysis. Together, these results indicate that these new mAbs are specific for chIL-17 and will be a useful tool for structural and immunological studies of IL-17 in poultry.

List of primers used in quantitative real-time PCR.

<p>List of primers used in quantitative real-time PCR.</p

Distribution and molecular weight of chicken IL2/15Rβ.

<p>(A) Expression of chIL2/15Rβ transcripts in various chicken tissues and cell lines. Total RNA was isolated from various tissues of 10-day-old chickens and analyzed with quantitative real-time PCR. Tissue samples were pooled from five chickens. Expression levels were normalized to those of β-actin from the same samples. Data represent means of triplicate samples. Data are representative of two independent experiments with similar pattern results. CU205, REV-transformed lymphoblast cell line; HD11, macrophage cell line; ND, not detected. (B) Detection of chicken IL2/15Rβ protein with Western blot analysis. Whole-cell lysates of COS-7 cells were collected 48 h (lanes 1 and 2) after transient transfection with a chIL2/15Rβ-HA construct (lane 2) or empty pcDNA 3.1 (lane 1). To determine the size of the chIL2/15Rβ backbone, transfected cells were incubated for 24 h and then treated with 5 µg/ml tunicamycin as an inhibitor of N-linked glycosylation followed by incubation for an additional 6 h (lane 3) and 24 h (lane 4). Cell lysates from COS-7 cells were separated by SDS-PAGE under reducing conditions. Arrows indicate specific bands. Data are representative of three independent experiments with similar pattern results.</p

Determination of gut lesion scores, serum carotenoid levels, and IFN-γ transcript levels.

<p>Ten-day-old chickens were orally infected with 1×10⁴ sporulated <i>E. tenella</i> oocysts. Nine chickens were randomly chosen for serum samples and gut lesion scoring 7 days after <i>Eimeria</i> infection. (A) Lesion scores (0–4) were based on scoring techniques previously described (Johnson and Reid, 1970). (B) Serum samples were extracted with ten volumes of acetone to precipitate proteins. Absorbencies of the supernatants were determined spectrophotometrically at 456 nm using a β-carotene standard. Bars represent the means ± standard error from nine chickens. (C) Expression of IFN-γ mRNA in cecal-tonsils of chickens infected with <i>E. tenella</i>. Tissue samples were pooled from five chickens and subjected to quantitative real-time PCR. Expression levels were normalized to those of β-actin from the same samples. The <i>y</i> axis represents the fold change in expression of IFN-γ gene from <i>E. tenella</i>-infected chickens as compared to uninfected chickens. Data represent means ± standard error of triplicate samples. * <i>P</i><0.05, ** <i>P</i><0.01 or *** <i>P</i><0.001 was considered significant compared to uninfected chickens. Data are representative of two independent experiments with similar pattern results.</p