Bovine colostral CD8-positive cells are potent IFN-gamma-producing cells

IFN-gamma plays an important role in cellular immunity contributing to microorganism elimination. We have previously reported that bovine colostrum contains high levels of IFN-gamma as well as immunoglobulin. Lymphocytes are potent producers of IFN-gamma, so establishing the lymphocyte population in colostrum will help to identify the source of colostral cytokines. In this study, we used flow cytometric analysis to quantify the population of three types of lymphocytes found in colostrum; namely, CD4 (Th) cells, CD8 (cytotoxic T) cells, and gammadelta-T cells. We also quantified the concentration of colostral IFN-gamma using ELISA. IFN-gamma concentrations were measured in colostrum obtained from 96 healthy Holstein cows; the levels tended to decrease on the first day post-parturition. Flow cytometric analysis showed that many gammadelta-T- and CD8-positive cells were present in the colostrum, and that the CD4/CD8 ratio was low. The ratios of CD8- and gammadelta-T-positive cells to cells of other types decreased during the 5 days after parturition, but that of CD4-positive cells showed no change during the observation period. To identify IFN-gamma-expressing colostral lymphocytes, we used magnetic separation technology to separate the lymphocytes (CD4, CD8 and gammadelta-T) from colostral cells, then examined them for IFN-gamma mRNA expression with real-time PCR (RT-PCR). RT-PCR analysis revealed potent expression of the IFN-gamma gene in CD8-positive cells, reaching higher levels than in CD4- or gammadelta-T-positive cells. These results suggest that the CD8-positive T cells in colostrum play a role as producers of IFN-gamma.     

Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.     

External factors that regulate movement in the marine rotifer Brachionus plicatilis

Fisheries Science - There are two types of movement pattern in the Brachionidae (rotifers), swimming and attachment, although the factors that induce a shift between them have not been adequately...     

Fish‐processing wastes as an alternative diet for culturing the minute rotifer Proales similis de Beauchamp

We evaluated the suitability of fish waste diet (FWD) for culturing the minute rotifer Proales similis through the observation of their population growth and particle size selective feeding. A total of five treatments either with Nannochloropsis oculata or FWD (0.75 and 0.50 g/L) or the combination of N. oculata and FWD were set up in triplicates. P. similis were cultured in diluted natural seawater (8 g/L salinity) and 26 ± 1°C with the diet treatments being applied randomly. We daily monitored the rotifer density, the number of bacteria and the water quality in all the cultures. The population density of P. similis increased exponentially in all treatments, while the mean growth in FWD 0.75 g/L was significantly higher (p < .05) than that in the control. Addition of N. oculata to FWD resulted to lowered P. similis population growth. Bacterial colony count was high in FWD and introduction of P. similis to the diet decreased their density. The estimated bacteria ingestion rates were generally in the range of 6.03 × 10²–1.24 × 10⁴ bacteria/rotifer/hr and there was a positive linear relationship between bacterivory and rotifer population growth. We also observed a shift in the particle size distribution with a reduction in the frequency and concentration of small‐sized particles (<2.5 μm) at day 6. These results accentuate the potential of fish‐processing waste as diet for culturing P. similis which feeds on bacteria and small particles (≤2.5 μm) that are by‐products of degradation of this diet.     

Concordance between Results of Medium-term Liver Carcinogenesis Bioassays and Long-term Findings for Carcinogenic 2-Nitropropane and Non-carcinogenic1-Nitropropane in F344 Rats

This study was conducted to determine the concordance of results for a pair of structural isomers, 2-nitropropane (2-NP) and 1-nitropropane (1-NP), using the rat medium-term liver carcinogenesis bioassay (Ito test) and previously published long-term carcinogenicity tests. Male F344 rats were given a single intraperitoneal injection of DEN (200 mg/kg b.w.) to initiate hepatocarcinogenesis. After 2 weeks, they received per os 0, 0.8, 4 or 20 mg/kg/day of 2-NP or 1-NP six times a week and were subjected to two-thirds partial hepatectomy at week 3. Non-initiated groups receiving 0 or 20 mg/kg/day were also included. The animals were sacrificed for quantitative analysis of GST-P-positive foci at week 8. With the highest dose of 2-NP, significantly increased numbers and areas of GST-P-positive foci were demonstrated as compared with the respective control but were not noted with 1-NP. In the non-DEN-initiated groups, many small GST-P-positive foci of less than 0.2 mm in diameter were also induced in the rats treated with 2-NP at 20 mg/kg/day but were lacking with 1-NP. These results strongly support that 2-NP is a complete hepatocarcinogen with a potent initiation activity, whereas 1-NP is not.     

Accuracy of selected neurological clinical tests in diagnosing MRI-detectable forebrain lesion in dogs

This retrospective case study aims to evaluate the accuracy of menace response, response to nasal stimulation and proprioceptive placing in diagnosing forebrain lesion in dogs. A total of 145 client-owned dogs investigated by magnetic resonance imaging study of the brain between December 2017 and June 2019 were evaluated. Seventy-one dogs with no magnetic resonance imaging-detectable intracranial and significant cerebrospinal fluid abnormality or recent history of seizure (<48 h) served as controls. Binary regression analysis was performed to determine the sensitivity, specificity and likelihood ratios of each selected test. Older age at presentation was a significant risk factor for the presence of a forebrain lesion. Menace (62.5%) and proprioceptive deficits (40.5%) were common findings in all dogs. They were also significantly associated with the presence of forebrain abnormality. Moreover, they were more sensitive (77.3% and 82.2%, respectively) and specific (50.0% and 62.5%, respectively) when applied to dogs aged 6 years or older. Nonetheless, all of these tests' likelihood ratios, and thus reliability are poor. These neurological tests are commonly employed for diagnosing forebrain disease in dogs, yet are not highly accurate in diagnosing forebrain abnormality. Clinicians should interpret these clinical test results along with the patient history when designing a diagnostic plan.     

Pneumocystis carinii pneumonia in a Cavalier King Charles Spaniel

Pneumocystis carinii pneumonia was diagnosed by postmortem examination of a one-year-old Cavalier King Charles Spaniel with four-week history of dyspnea. Cytologic and histologic examination of lung tissues revealed numerous P. carinii trophozoites and cysts, and P. carinii specific DNA was detected by polymerase chain reaction. The dog showed hypogammagloblinemia and extremely low levels of serum IgG. It was considered that P. carinii pneumonia in this case was associated with an immunodeficient condition which has already been reported in Miniature Dachshunds.