Effects of dietary corticosterone on peripheral blood lymphocyte and granulocyte populations in immature domestic fowl

Four-week-old chickens were fed on diets containing 0, 5, 10, 20 and 40 mg corticosterone per kg for eight days. Packed cell volume (PCV), the numbers of granulocytes (G) and lymphocytes (L), the G:L ratio and plasma corticosterone concentration were measured. Plasma corticosterone concentration was increased by dietary corticosterone from day 2 onwards. The number of lymphocytes was increased by serial bleeding but dietary corticosterone depressed the response and caused a dose-related lymphocytopenia from day 1. There was a dose related increase in granulocytes from day 4 and a dose-related increase in G:L ratio from day 1. A dose-related increase in PCV was evident only on days 2 and 4. Correlation coefficients showed little evidence of relationships between plasma corticosterone concentrations and the recorded blood variables.     

Staphylococcal infections in rabbit does on two industrial farms

The main reasons for culling adult rabbit does on two Spanish rabbit farms were investigated for a year. The most important conditions were mastitis (33.3 per cent), followed by subcutaneous abscesses (9.9 per cent) and pyometra (8.7 per cent). Staphylococcus aureus infections were the most severe problem, the organism being isolated from 69.2 per cent of infected animals. Pasteurella species were more prevalent in cases of pyometra and pneumonia. Two strains of S aureus were identified by using polymorphism of the coagulase gene as the criterion. One of these strains was responsible for the majority of the staphylococcal infections and was isolated from several pathological processes.     

Canine lymphocyte cultures in vitro: Evaluation of peripheral blood lymphocyte response to mitogens

Lymphocytes from dog peripheral blood have been stimulated in vitro with 3 different mitogens (Con A, PHA and PWM). Culture medium was RPMI 1640 enriched with either autologous plasma, fetal calf serum or a newly described defined serum substitute. In such cultures the number of surviving and activated cells was measured by cytofluorometry and the proliferation was assessed by thymidine incorporation. In unstimulated cultures, up to 70% of all cells had disappeared (died) during the first 42 hours of incubation, whereas the number of viable cells was reduced to 50–60% in mitogen stimulated cultures. Of the surviving lymphocytes, between 25–40% of the cells appeared to have an elevated RNA-content (activated or G₁ cells). By comparison between thymidine incorporation and number of mitogen induced G₁ cells, a very high correlation was found (r=0.92). However, the slope of the regression line was much lower than expected. The low thymidine incorporation per activated cell was primarily related to the high cell death and a resulting dilution of tritiated thymidine. Indeed, preliminary results suggested that the same thymidine incorporation per G_1b cells could be obtained if peripheral blood lymphocytes were washed immediately before pulsing as could be obtained with lymphnode cells without washing.     

An improved method for the isolation of enriched canine peripheral blood mononuclear cell and peripheral blood lymphocyte preparations

A two-step purification method was developed for obtaining (1) peripheral blood mononuclear cell preparations of greater than 97% purity and (2) peripheral blood lymphocyte preparations of greater than 95% purity from canine whole blood with yields similar to or greater than those obtained by conventional techniques.     

Cytochemical characterization of peripheral blood cell populations of two Cyprinidae,Carassius auratus and Ctenopharyngodon idellus

Fish are the most diverse species of all vertebrate groups, and their blood cells have shown variable characteristics in terms of morphology. Cytochemical staining for enzyme activity in blood leukocytes will help assess the immune function of fish. We characterize blood cells from crucian carp (Carassius auratus) and grass carp (Ctenopharyngodon idellus) by using a Diff‐Quick stain as well as different cytochemical methods. Blood specimens obtained from crucian carp and grass carp were evaluated after cytochemical staining for acid phosphatase (ACP), alkaline phosphatase (ALP), naphthol AS chloroacetate esterase (AS‐DNCE), naphthyl acetate esterase (NAE), α‐naphthyl butyrate esterase (NBE), peroxidase (MPO) and periodic acid–Schiff's reaction (PAS) using commercial kits. Blood cell types were evaluated based on their morphological characteristics and the presence or absence of specific chromogen. The expression pattern of enzymes was similar between the two Cyprinidae and was also broadly consistent with other fish species. However, there were some interesting differences detected between crucian carp and grass carp, including naphthol AS chloroacetate esterase activity in monocytes, peroxidase activity and location in thrombocytes. The ACP, ALP and MPO expressions of different leukocytes of the two Cyprinidae were evaluated by Image Pro Plus and were analysed for statistical significant differences. This investigation provides basic haematology and enzyme activity analyses for crucian carp and grass carp and serves as an approach to evaluating the immune response of fish.     

Nutrient digestibility of rabbit does in different stages of reproductive cycle under tropical conditions

Effect of stages of reproductive cycle on body weight, feed intake and apparent nutrient digestibility of rabbits was investigated. Pregnant/lactating (PL) does increased their feed intake above pregnant lactating does. Dry matter (DM) digestibility was higher in pregnant does than in lactating, PL or open does. Digestibility of crude fibre, crude protein (CP) and nitrogen-free extractives (NFE) were significantly higher for open and pregnant does than lactating and PL does. Ether extract (EE) digestibility was similar for pregnant, lactating and PL does. Digestible CP requirement of does increased with physiological state from open to PL states. Open and pregnant does were meeting their protein requirement while lactating and PL does were not meeting their requirement. Feed intake was higher for first week pregnant, fourth week pregnant, first week lactating, first week pregnant/fourth week lactating and second week pregnant/fourth week lactating does than open does. DM digestibility was higher for first and fourth week pregnant does than the other physiological stages. EE digestibility was similar for all physiological stages. CP digestibility was higher in fourth week pregnant does than the other physiological stages. Fourth week pregnant does had higher NFE digestibility than first and fourth week lactating and the various stages of concurrent pregnant and lactating does. Open does, first and fourth week pregnant does were meeting their protein requirement while does in the first and fourth week of lactating and stages of concurrent PL were not meeting their requirement.     

左旋咪唑对鸡离体外周血淋巴细胞的作用

本文报道了左旋咪唑对鸡离体外周血淋巴细胞穴PBL雪增殖及伴刀豆球蛋白A穴ConA雪诱导细胞穴CIC雪活性的作用:左旋咪唑以剂量依赖性方式促进鸡PBL的增殖;与ConA对促进鸡PBL增殖呈现协同作用;对脂多糖穴LPS雪激活鸡PBL具有明显促增殖作用;可逆转磷酸组胺和肾上腺素对ConA和LPS激活鸡PBL增殖的抑制作用;低浓度左旋咪唑不能逆转氢化可的松对ConA激活鸡PBL增殖的抑制作用,但高浓度穴100～400μg/ml雪可明显逆转氢化可的松的抑制作用;左旋咪唑可明显逆转氢化可的松的对LPS激活鸡PBL增殖的抑制作用;可促进CD+4细胞增殖,抑制CD+8细胞增殖,使CD+4/CD+8细胞比值升高,CIC活性增强。     

Changes in peripheral blood leukocyte populations in pigs with naturally occurring exudative epidermitis

The objective of the study was to analyze changes in peripheral blood leukocyte subsets in cases of naturally occurring exudative epidermitis (EE) in pigs. Five of ten piglets developed the chronic clinical form of EE 2-5 days after weaning (PW). Blood samples were obtained at 7, 14 and 21 days from both normal and clinically affected piglets for routine haematology and for the determination of CD45, CD21, CD4, CD8 and gammadeltaTCR cell markers by flow cytometry. When compared with clinically normal piglets EE affected pigs showed significantly decreased values of monocytes at 14 and 21 days PW, and increased numbers of neutrophils and leukocytes at 21 days PW. The EE affected pigs also had an early significant CD4(+) and CD8(high+) T lymphocyte proliferative response at 7 days PW. However affected pigs had a significantly reduced number of B (CD21(+)) and gammadeltaTCR(+) T lymphocytes in blood at 21 days PW. Although all values remained within the normal range, the significant differences in some peripheral blood leukocyte subsets between the two groups of piglets suggest that the generalised cutaneous infection with Staphylococcus hyicus is severe enough to induce a systemic inflammatory and immune responses.     

Early-life longitudinal survey of peripheral blood lymphocyte subsets in Beagle dogs

A longitudinal study of peripheral blood lymphocyte subsets from 7 to 15 months of age was performed in Beagle dogs employing a multiparametric flow cytometry. The data were compared with data obtained from adult Beagle dogs that were housed in the same animal facilities and that were subjected to the same controls during the 34 weeks of the study. Absolute counts of total lymphocytes and CD3+ T, CD3+CD4+ Th and CD21+ B lymphocytes decreased during the entire 34 weeks period of the study in the young dogs group. The same was observed with regard to the percentage of CD3+CD4+ Th lymphocytes and the CD4/CD8 ratio, while the percentage of CD3+CD8+ Tc lymphocytes increased from 7 to 15 months of age. These age-related changes found in lymphocyte subsets distribution of young dogs led to level the absolute and relative values of adult dog lymphocytes. The observations of this longitudinal study illustrate the changes related to maturation of lymphocyte subsets that occur during early life in Beagle dogs.     

Improving the reproductive,immunity and health status of rabbit does using honey bee venom

The aim of the present study was to investigate the long-term effect of using bee venom (BV) on the reproductive performance, immune, and health status of rabbit does and its effect on their litters. Sixty mature does, from Spanish V-line rabbit stock, were randomly assigned to four homogeneous groups with 15 does each. The 1^st, 2^nd and 3^rd groups were injected twice weekly under the neck skin with 0.1 ml solution contains 0.1, 0.2 and 0.3 mg BV/rabbit respectively. The 4^th group served as a control group. From the results, litter size at birth, litter weight and survival rate at weaning age as well as milk yield were significantly (p ≤ 0.05) increased in BV groups than in the control group. Serum estradiol 17-β (E2) was significantly (p ≤ 0.05) higher (15%) in the rabbit does treated with BV compared to the control group. The treated does with BV at any study doses showed a gradual and significant (p ≤ 0.05) decrease (12%) in serum progesterone levels (P4) compared to the control. They also showed a significant (p≤0.05) increase in conception (17%) and fertility rates (10%) compared to the control does. Treatment of rabbit does with BV caused a gradual and significant (p ≤ 0.05) reduction in both aspartate aminotransferase (AST) (16%) and alanine aminotransferase (ALT) (37%) liver enzyme activities. Additionally, results have shown that BV resulted in a gradual and significant (p ≤ 0.05) increase in total antioxidant capacity (TAC), antioxidative enzymes such as glutathione S-transferase (GST) and glutathione peroxidase (GPx), serum IgG, IgM and IgA levels with significant (p ≤ 0.05) decrease in malondialdehyde (MDA) and thiobarbituric acid reactive substances (TBARS) in BV groups compared to the control group. Results suggest that BV can be used in rabbit farming as an effective and safe alternative to artificial chemical drugs (sexual-stimulants) to improve certain reproductive traits, immune response and health.     

替米考星和甲硝唑对猪外周血淋巴细胞增殖的影响

采用细胞形态学观察、淋巴细胞标记和噻唑兰(MTT)法,探究替米考星和甲硝唑单独用药和联合用药对猪外周血淋巴细胞转化率、α-醋酸萘酯酶法阳性率和淋巴细胞增殖的影响。试验结果显示甲硝唑具有提高细胞免疫的功能。在体外试验时,终浓度为0.25~100μg/mL的甲硝唑可以促进淋巴细胞的增殖和转化;在体内试验时,浓度为2.5~10 mg/kg的甲硝唑可以增加成熟T淋巴细胞的数量。但是替米考星在体内和体外试验的结果却与阴性对照差异不显著,表明它不具有促进或抑制淋巴细胞增殖的能力,对机体的细胞免疫无影响。     

Alterations in lymphocyte subpopulations in peripheral blood of sheep persistently infected with border disease virus.

Fourteen sheep persistently infected with border disease virus were investigated to examine the effects of persistent viraemia on lymphocyte subpopulations in peripheral blood and their responses to the mitogen phytohaemagglutinin (PHA) in vitro. Persistently infected sheep had significantly more CD8+ (cytotoxic-suppressor) T-lymphocytes than uninfected sheep of the same age (P less than 0.001). The total number of CD4+ (helper) T-lymphocytes were not significantly different but there were more T-lymphocytes (CD5+) which were CD4- and CD8- in normal sheep than persistently infected sheep (P less than 0.001). Peripheral lymphocytes obtained from persistently infected sheep showed significantly reduced blastogenesis induced by PHA than those obtained from normal sheep (P less than 0.001).     

围产科尔沁牛外周淋巴细胞HSP70的表达

选择经同期发情处理的妊娠科尔沁牛5头(2³胎),于分娩-14、-6、-2、0、2、6、14d颈静脉采血,分离淋巴细胞,采用RT-PCR、Western blotting法分别检测淋巴细胞的HSP70mRNA与蛋白表达。结果显示,各受试牛的HSP70mRNA与蛋白表达变化趋势基本一致,即从-14d开始逐渐下调,0d最低,继而再逐渐上调。结果表明,HSP70作为淋巴细胞内重要伴侣分子,随着分娩的启动和发生其表达抑制显著增强,但在分娩后被迅速解除,其表达逐渐上调。     

围产科尔沁牛外周淋巴细胞HSP70的表达

选择经同期发情处理的妊娠科尔沁牛5头（2～3胎）,于分娩-14、-6、-2、0、2、6、14d颈静脉采血,分离淋巴细胞,采用RT～PCR、Westernblotting法分别检测淋巴细胞的HSP70mRNA与蛋白表达。结果显示,各受试牛的HSP70mRNA与蛋白表达变化趋势基本-致,即从-14d开始逐渐下调,0d最低,继而再逐渐上调。结果表明,HSP70作为淋巴细胞内重要伴侣分子,随着分娩的启动和发生其表达抑制显著增强,但在分娩后被迅速解除,其表达逐渐上调。     

富硒女贞子对山羊外周血淋巴细胞增殖活性的影响

通过提取富硒女贞子多糖、蛋白质和齐墩果酸,采用MTT法测定三大类物质体外对山羊外周血淋巴细胞增殖活性的影响。方差分析的多重比较结果显示富硒女贞子多糖、蛋白质和齐墩果酸体外均具有促进ConA刺激山羊外周血淋巴细胞分裂增殖的作用,且富硒女贞子蛋白质组分、齐墩果酸组分的作用效果明显优于普通女贞子组,各相应浓度比较差异显著比例分别达到63.64%、40.00%。说明富硒女贞子能进一步提高山羊外周血淋巴细胞的增殖活性。     

Study of peripheral blood cell populations involved in the immune response of goats naturally infected with Mycobacterium caprae

Tuberculosis in goats caused by Mycobacterium bovis and Mycobacterium caprae has noteworthy sanitary and economic implications. Current diagnostic assays are based on cellular immunity and although they have demonstrated a high sensitivity, some animals remain undetected. In the present study, flow cytometry has been used to determine changes in CD4+, CD8+ and CD25+ T cell populations in peripheral blood from naturally infected goats. Proportion of lymphocytes producing PPD-specific interferon-gamma (IFN-γ) was calculated and an ELISA for detection of PPD-specific IFN-γ was performed to measure the cytokine in plasma. The infected goats showed percentages of CD4+ T cells between 27.31% and 47.23% and there were not significant differences (p=0.113) with the non-infected control goats although the mean percentage was lower in this group. Regarding CD8+ T cells, a higher percentage was observed in healthy goats compared to controls (p=0.081). The mean percentage of lymphocytes expressing CD25 without antigen stimulation (30.65±3.91) was higher in lesion and/or culture-positive animals than in the controls (21.84±1.21; p=0.053). The percentage of CD4+/IFN+ T cell population stimulated with bovine PPD was a reliable marker of infection, since the mean percentage in the infected goats was significantly higher than in the controls (p<0.05). Tuberculosis in goats caused by M. caprae induced changes in cellular populations similar to those described for M. bovis in cattle.     

Changes in blood and spleen lymphocyte populations following antigen challenge in immature male chickens

1. The effects of antigen (Ag) injection on the distribution of lymphocyte populations of Cornell K‐strain male chickens were studied.

2. Two experiments were conducted. In the first, chickens were injected with Brucella abortus (BA), a purported T‐independent antigen. In the second, chickens were injected with sheep red blood cells (SRBC), a T‐dependent antigen. Peripheral blood lymphocytes (PBL) and spleen lymphocytes isolated at 0, 3, 6, 9, 12 and 24 h following Ag injection were stained with monoclonal antibodies (mAb) detecting B‐lymphocytes, CD4⁺ and CD8⁺ cells.

3. B‐lymphocytes in the blood or spleen showed no significant changes following either BA or SRBC injection. In contrast, CD4⁺ cells were decreased in the blood and increased in the spleen following BA and SRBC injections. CD8⁺ cells were decreased in both blood and spleen following BA injection but were unchanged in either blood or the spleen following SR8C injection.

4. These results indicate that there is a change in both spleen and circulating lymphocyte populations, especially T‐helper cells, following Ag injection. T‐helper cells are apparently the primary population involved in the initiation of humoral immunity.