Morphological and Cytochemical Studies of Peripheral Blood Cells of Schizothorax prenanti

The morphological and cytochemical studies of peripheral blood cells of Schizothorax prenanti were studied by light and electron microscopy. Erythrocytes, thrombocytes and three types of leucocytes, lymphocytes, neutrophils and monocytes, were distinguished and characterized. In addition to mature erythrocytes, immature and dividing erythrocytes were observed. A few organelles such as mitochondria were distributed in the cytoplasm of erythrocytes. Lymphocytes with heavily clumped heterochromatic nucleus and minimal cytoplasm were classified into small and large lymphocytes. Three different populations of granules, with distinctive ultrastructural aspect, were observed in neutrophils. Monocytes were the fewest leucocytes possessing rich organelles, phagocytized materials and vacuoles. Thrombocytes with various types were the most abundant blood cells among leucocytes and contained a prominent nucleus with dense bands of heterochromatin and many cytoplasmic vacuoles. Periodic acid‐Schiff staining was positive in neutrophils, monocytes, lymphocytes and thrombocytes, but not in erythrocytes. Peroxidase‐positive staining was observed in neutrophils and monocytes, but not in erythrocytes, lymphocytes and thrombocytes. Only neutrophils were positive for oil red O. Except for erythrocytes, the other blood cells stained positively for acid phosphatase. Only neutrophils and monocytes were positive for α‐naphthyl acetate esterase. None of the cells studied were positive for alkaline phosphatase. The morphologic and cytochemical features of blood cells of S. prenanti are similar to those of other fish. This investigation may be helpful as a tool to monitor the health status of cultured S. prenanti and will grant early detection of clinical pathology.     

Ultrastructure of the Spleen and Head Kidney of Striped Bass

Abstract

The spleen and head kidney of striped bass Morone saxatilis were examined by electron microscopy, including examination of ultrastructural localization of peroxidase. The spleen is composed of a loosely organized reticulum that supports the red and white pulp. The principal hematopoietic and mature blood cells observed were erythrocytes, erythroblasts, lymphocytes, macrophages, thrombocytes, and aggregates of pigmented macrophages. In addition, limited numbers of neutrophils, neutrophilic myelocytes (immature neutrophils), plasma cells, eosinophils and monocytes were identified. Reticular cells and fibers appeared to provide a structural framework for both cells and blood sinuses in the spleen. Like the spleen, the head kidney contained erythrocytes, macrophages, monocytes, lymphocytes, and other less frequently observed cell types. A distinct difference in the cytologic composition of the two organs, however, was in the greater number of neutrophilic myelocytes found in the head kidney, where these cells were often arranged in groups. Positive peroxidase reactions were observed for neutrophils, neutrophilic myelocytes, macrophage phagolysosomes, and pigmented macrophages in both the spleen and head kidney.     

THE TRANSMISSION OF BABESIA BOVIS IN HERDS OF EUROPEAN AND ZEBU X EUROPEAN CATTLE INFESTED WITH THE TICK, BOOPHILUS MICROPLUS

SUMMARY Transmission of Babesia bovis by the tick Boophilus microplus was studied in 4 breeding herds of European and Zebu x European cattle under different levels of tick infestation. The observations consisted of weekly counts of female ticks on the cattle, monthly serological tests for antibodies to B. bovis, examination of tick larvae from the pasture to determine B. bovis infection rates and comparison of the suitability of paddocks for tick reproduction. The rate of transmission (inoculation rate) was estimated in terms of the daily probability of infection and consisted of the product of the mean daily tick infestation head^-1 and the babesial infection rate in tick larvae. The theoretical value of the minimum inoculation rate required to produce stability of babesiosis is 0.01. This value was exceeded only in a herd of European cattle with minimal tick control and grazing on a pasture favourable for tick reproduction. Instability of babesiosis occurred in the other herds of European cattle after tick numbers had been reduced by pasture spelling and strategic dipping and after reduction in the babesial infection rate in ticks apparently caused by unfavourable environmental conditions. Over a period of years, the tick infection rates also declined as a consequence of the reduction in numbers of ticks. The Zebu x European cattle failed to generate inoculation rates greater than the minimum level, even though no tick control measures were applied. This was attributed to lower babesial infection rates in the ticks than those observed in a comparable herd of European cattle and to the high tick resistance of the Zebu crosses which maintained the tick populations at low levels. Both factors combined to produce a low inoculation rate.     

Ultrastructural and Cytochemical Properties of Peripheral Blood Cells of Piebald Naked Carp (Gymnocypris eckloni)

The ultrastructural and cytochemical properties of peripheral blood cells of Gymnocypris eckloni were investigated by transmission electron microscopy and a range of cytochemical techniques to provide clear insight into the structure and function of blood cells from this fish. Ultrastructurally, erythrocytes, leucocytes (neutrophils, eosinophils, lymphocytes, monocytes), thrombocytes and plasma cells were identified in the peripheral blood of G. eckloni. The most special ultrastructural characteristics of blood cells in this fish were that neutrophils exhibited only one type of cytoplasmic granules containing an eccentric, spherical or oval electron‐dense core, and eosinophils presented two types of granules with non‐uniform electronic density and without crystalloids in their cytoplasm. Neutrophils, eosinophils, lymphocytes, monocytes and thrombocytes were positive for periodic acid–Schiff and α‐naphthyl acetate esterase staining. Intense peroxidase positive staining was observed in neutrophils and monocytes, but not in eosinophils, lymphocytes and thrombocytes. Neutrophils, eosinophils and monocytes were stained positively for acid phosphatase, whereas lymphocytes and thrombocytes did not stain. Leucocytes and thrombocytes were negative for alkaline phosphatase and Sudan black B staining. Erythrocytes were negative for all cytochemical staining. The cytochemical and ultrastructural features of peripheral blood cells of G. eckloni were similar to those of other fish species. However, some important differences were identified in G. eckloni.     

Effects of direct exposure to cold weather under grazing in winter on the physiological,immunological, and behavioral conditions of Japanese Black beef cattle in central Japan

This study aimed to reveal the effects of direct exposure to cold weather under grazing in winter (GW) on the health of Japanese Black beef cattle, as assessed using physiological, immunological, and behavioral parameters. Ten Japanese Black beef cattle (328 ± 45 kg, 7.6 ± 3.4 years of age) were used in this experiment. In winter, five of the 10 cattle grazed for 2 months in a 1.8 ha pasture (GW), and the remaining cattle were fed under confined conditions with tethering (control [CT]). The two groups were fed similar feed during the experiment, except for the grazing forage. Blood samples were collected approximately every 2 weeks. The numbers of neutrophils and monocytes and antioxidant enzyme activity were higher in the GW group than in the CT group (p < 0.05). The proportions of CD4‐single‐positive cells were lower in GW cattle than in CT cattle (p = 0.06). This study showed that direct exposure of beef cattle to cold weather under GW enhanced the levels of circulating neutrophils and monocytes and contributed to the kinetic homeostasis of lymphocytes but also activated antioxidant enzymes due to an increase in oxidative stress.     

Identification of feline monocytes and neutrophils as effector cells in antibody-dependent cellular cytotoxicity: sequential analysis, using light microscopy, histochemistry, and scanning electron microscopy

Feline monocytes and neutrophils functioned as effector cells in antibody-dependent cellular cytotoxicity (ADCC) against antibody-coated chicken erythrocytes. Using light microscopy, effector cell populations were identified in effector-target cell interactions, with further characterization of these identical individual effector cells by histochemical evaluations and scanning electron microscopy. Monocytes and neutrophils, but not lymphocytes, were observed attacking target cells. Carbonyl iron depletion of monocytes and neutrophils from peripheral blood leukocytes caused a marked reduction from a mean of 62% to 3.6% lysis in ADCC as measured by a 4-hour 51Cr release assay. Effector cells functioning in the ADCC reaction were visualized, using sequential analysis and light microscopy, histochemistry, and scanning electron microscopy.     

Molecular evidence of Anaplasma infection in naturally affected domestic cats of Pakistan

Feline anaplasmosis is considered as an emerging tick-borne disease of zoonotic potential. The aim of current study was to investigate the molecular prevalence of anaplasmosis, associated risk factors, and alterations in hematological parameters of domestic cats from Lahore, Pakistan. Blood samples of 100 domestic cats from district Lahore were examined microscopically and the extracted genomic DNA from each sample was processed for the amplification of 16 S rRNA gene of Anaplasma. PCR confirmed isolates were purified for sequencing. The data regarding the risk factors was collected in a predesigned questionnaire and statistically analyzed by logistic regression analysis. The study found a molecular prevalence of 13% (13/100) among analyzed blood samples. The nucleotide analysis of Anaplasmataceae species sequences amplified by PCR showed high resemblance (99%) with isolates from Korea, Japan, Malaysia, Philippines, and India. The potential risk factors found to be significantly associated (p < 0.05) with disease dynamics based on the Chi-Square test were tick infestation on studied animals, previous tick history, tick control status, house hygiene, and housing type. A significant (p < 0.05) decrease in the number of platelets, erythrocytes, hemoglobin level, and pack cell volume was observed in cats suffering from anaplasmosis compared to the healthy ones. The current study is the first report of Anaplasma infection in domestic cats of Pakistan. This study will be effectual in designing the control strategies for this disease.     

Enhanced Lipid Peroxide Levels in the Erythrocytes of Calves with Haemoglobinuria

Eight 6–9 month old calves, showing clinical signs of intermittent haemoglobinuria, even after treatment with an antipiroplasmal drug (4,4-diamidinodiazoaminobenzene diaceturate), were examined for oxidative damage to their erythrocytes and the presence of hemoprotozoa in blood smears. Four calves without signs of haemoglobinuria served as controls. The blood smears from three of the eight calves contained piroplasms for Theileria annulata. Irrespective of the presence of piroplasms in their blood smears, the calves with haemoglobinuria had significantly (p<0.01) lower haemoglobin concentrations (Hb) and packed cell volumes (PCV). The lipid peroxide level in the erythrocytes, but not in the plasma, of calves with red urine was significantly (p<0.05) higher than that for the controls. It is concluded that haemoglobinuria, irrespective of the presence of piroplasms in blood smears, is associated with oxidative stress to erythrocytes and peroxidation of polyunsaturated fatty acids of cell membrane.     

Studies on correlations among parasitaemia and some hemolytic indices in two tropical diseases (theileriosis and anaplasmosis) in Fars province of Iran

This study was carried out in two observational clinical studies. Study 1 comprised 50 adult crossbred cattle naturally infected by Theileria annulata. Infected animals were divided into 4 subgroups with different parasitaemia (<1%, 1–3%, 3–5% and >5%). Study 2 comprised 20 adult crossbred cattle naturally infected by Anaplasma marginale. Infected animals were divided into 3 subgroups with different parasitaemia (<10%, 10–20% and 20–30%). In study 1, a significant negative correlation (P < 0.001) was observed between parasitaemia and superoxide dismutase (SOD). Positive correlations (P < 0.001) were observed between parasitaemia and lactate dehydrogenase (LDH) and mean corpuscular fragility (MCF). In study 2 positive correlations (P < 0.05) were observed among parasitaemia and MCF and LDH activity. SOD activity had a negative correlation with parasitaemia in cattle with parasitaemia lower than 10% but no significant correlation (P > 0.05) was observed between SOD activity and parasitaemia in cattle with 10–20 and 20–30% parasitaemia. In comparison of both studies we came to the conclusion that in theileriosis as the severity of disease increased the anaemia, MCF and LDH activity increased and SOD activity decreased at any parasitaemia, but in anaplasmosis the anaemia, MCF and LDH activity increased at any parasitaemia but SOD activity decreased only in early but not in advanced stages of disease.     

Relapse infection after chemotherapy in dogs experimentally infected with Trypanosoma brucei brucei

Studies on relapsing Trypanosoma brucei brucei infections in dogs after Berenil treatment revealed that the first relapse occurred 13 to 64 days after chemotherapy and 36 to 79 days after inoculation. A second relapse infection was observed in two dogs 43 and 60 days after a second Berenil treatment. During the aparasitaemic period following chemotherapy in four dogs, successful transmission (as evidenced by subsequent parasitaemia) following the intraperitoneal inoculation of homogenate of brain from two of the dogs into recipient rats was obtained. Transmission with blood collected just before the animals were sacrificed was, however, negative. Hornogenates of other organs (liver, spleen, eyes, testes, kidneys, heart and lymph node) were also non-infective. One dog inoculated with relapsed trypanosomes and treated with Berenil soon after showing parasitaemia was completely cured of the infection. It was considered that the brain is the source of relapse in T b brucei infection after Berenil therapy and that the relapse was not due to drug resistance.     

A morphometric and functional study of the toxicity of atmospheric ammonia in the extrathoracic airways in pigs

The effects of atmospheric ammonia (NH₃) on the nasal and tracheal mucosa of pigs were investigated by morphometric and functional methods. Pigs were exposed to four concentrations of NH₃ [5 (control), 25, 50 and 100 ppm] for 6 days in a specially designed air-pollutant exposure chamber. Samples were taken from the turbinates and the trachea, and the respiratory mucosa was examined by light and scanning electron microscopy. Dose-response curves to carbachol and isoproterenol were constructed using isolated strips of tracheal smooth muscle, with or without epithelium. In pigs exposed to ammonia, considerable mucosal injuries were observed in the turbinates but not in the trachea. The number of neutrophils in the epithelial layer and in the lamina propria, and epithelial hyperplasia were closely and significantly correlated with the concentrations of ammonia (r=0.894, p<0.001; r=0.727, p<0.001; and r=0.818, p<0.001, respectively). Except for the lamina propria, all these changes were significant (p<0.05) at ammonia concentrations as low as 25 ppm. The percentage of the surface of the turbinate mucosa that was ciliated tended to decrease with increasing ammonia concentration (r=0.439, p<0.082). Ammonia induced smooth-muscle hyperresponsiveness to carbachol with a close linear correlation between individual values of the carbachol-induced maximal effect and the NH₃ concentrations (r=0.526, p<0.003). While mechanical destruction of the epithelium induced an increase in E _max in the control group, no difference was observed between the intact and denuded strips from animals exposed to ammonia. The response to isoproterenol was not influenced by ammonia. It was concluded that quantitative histological analysis of the inflammatory infiltration and epithelial hyperplasia in the turbinates is a useful tool for quantifying the effects of atmospheric pollutants in pigs; a 6-day exposure to ammonia induces nasal irritation and functional disturbances of the tracheal smooth-muscle contractions at concentrations as low as 25 ppm.Abbreviations E _max maximal effect - IM intramuscularly - IV intravenously - NAL nasal lavage - pD ₂ negative logarithm of the molar concentration of agonist which induces an increase in tension equivalent to 50% of the maximal effect E _max expressed as grams force per mg of dry weight - r correlation coefficient     

Humoral immune response of sheep to infection with Eperythrozoon ovis

Circulating antibody was detected by an indirect fluorescent antibody test (IFAT) in the serum of sheep infected experimentally with Eperythrozoon ovis. Antibodies were first detected 15 to 32 days after infection with E ovis and titres peaked at 41 days. This antibody may be associated, at least in part, with protection against infection with E ovis since the initial increase in antibody titre coincided with a fall in the primary parasitaemia. A role for antibody is suggested further by the fact that the prepatent period of infection was prolonged by one day and the parasitaemia initially remained at low levels in infected sheep protected by passively transferred hyperimmune serum. Moreover, following primary infection, acquired immunity was manifest by a lack of parasitaemia following challenge infections while increased IFA titres were observed. No evidence of opsonic activity was observed in an in vitro erythrophagocytosis test in that neither mouse macrophages nor sheep monocytes phagocytosed E ovis infected or uninfected erythrocytes sensitised with hyperimmune serum.     

A Comparison of the Concentrations of C-reactive Protein and α1-Acid Glycoprotein in the Serum of Young and Adult Dogs with Acute Inflammation

The concentrations of C-reactive protein (CRP) and ₁-acid glycoprotein (AAG) were evaluated in 1-, 3- and 18-month-old dogs (four of each age) that had been inoculated with turpentine oil. The CRP and AAG in 3-month-old and younger dogs subjected to surgery or inoculated with either Staphylococcus aureus or a viral vaccine were also evaluated. The average CRP concentration in the sera peaked 2 days after inoculation of turpentine oil. The peak CRP concentrations in 3- and 18-month-old dogs were significantly (p<0.05) greater than those in 1-month-old dogs. The average AAG concentration in the sera peaked 4 days after inoculation of turpentine oil. No significant difference was found in AAG concentrations between any of the age groups. When experimentally inoculated with S. aureus or subjected to oophorohysterectomy, the CRP and AAG concentrations increased in 3-month-old dogs, but they increased little in 1-month-old dogs. The CRP and AAG in dogs inoculated with the viral vaccine did not increase. In dogs with fractures or subjected to percutaneous gastrostomy, the CRP and AAG concentrations correlated with the condition of dogs.     

Complement Receptor Type 3 (CR3)- and Fc Receptor (FcR)-Mediated Matrix Metalloproteinase 9 (MMP-9) Secretion and Their Intracellular Signalling of Bovine Neutrophils

Complement receptor type 3 (CR3)- and Fc receptor (FcR)-mediated metalloproteinase-9 (MMP-9) secretion and their intracellular signalling of bovine neutrophils were evaluated. Relative density of MMP-9 secreted by neutrophils stimulated with opsonized zymosan (OPZ, stimulant for CR3) was significantly (p < 0.05) increased when the OPZ concentration was increased from 0 to 0.4 mg/ml. Similar results were obtained for neutrophils stimulated with heat-aggregated IgG (Agg-IgG, stimulant for Fc receptor) at concentrations from 0 to 0.40 mg/ml. Preincubation of neutrophils with 1–30 nmol/L wortmannin (phosphoinositide 3-kinase inhibitor) resulted in inhibition of MMP-9 secretion induced by stimulation with OPZ and Agg-IgG in a concentration-dependent manner, 30 nmol/L wortmannin causing complete inhibition. Similarly, preincubation of neutrophils with 0–100 μmol/L genistein (tyrosine kinase inhibitor) also resulted in inhibition of OPZ- and Agg-IgG-induced MMP-9 secretion in a concentration-dependent manner, with 100 μmol/L genistein causing complete inhibition. Significant (p < 0.05) positive correlations were found between MMP-9 and luminal-dependent chemiluminescent response (LDCL) in the case of stimulation with OPZ (r = 0.754) and in the case of stimulation with Agg-IgG (r = 0.728). Our findings suggested that CR3 and FcR play a critical role in production of MMP-9 and may be regulated by intracellular signal transduction, including that by phosphoinositide 3-kinase (PI3K) and tyrosine kinase (TK).     

Fasting hyperglycaemia with oral glucose tolerance in acute Trypanosoma congolense infection of rats

Intraperitoneal inoculation of rats with Trypanosoma congolense (Federe strain) produced a sustained parasitaemia from days 7 to 23 post-infection (pi). The fasting tail-blood glucose (FBG) concentrations in the infected animals increased (p<0.05) from 3.8+/-0.2 mmol/l on day 0 pi to 4.6+/-0.2, 4.9+/-0.2 and 5.8+/-0.3 mmol/l on days 7, 10 and 17 pi and decreased (p<0.05) to 3.1+/-0.8 and 2.9+/-0.7 mmol/l on days 20 and 23 pi, but the values in the uninfected controls varied between 3.8+/-0.3 mmol/l on day 0 pi and 3.9+/-0.2 mmol/l on day 23 pi. After oral glucose intake (1.0 g/kg) and determination of tail-blood glucose (BG) concentrations after 2 h, the percentage increase in BG from FBG was either comparable in infected and uninfected groups (days 7, 20, 23 pi) or lower (p<0.05) in the infected group (days 10, 17), suggesting the same rate of tissue glucose delivery in both groups or a faster rate in the infected group. Therefore, oral glucose tolerance in the infected rat was not impaired, but there was initial fasting hyperglycaemia followed by fasting hypoglycaemia in the later stage.     

Infectivity rate and transmission potential of Hyalomma anatolicum anatolicum ticks for Babesia equi infection

The infectivity rate of Babesia equi in the salivary glands of Hyalomma anatolicum anatolicum was assessed. The hungry nymphs were fed on a donkey experimentally infected with B. equi. The engorged dropped-off nymphs were collected at different levels of parasitaemia and kept in BOD incubator. After ecdysis, the hungry adults were prefed on rabbits for different time intervals, thereafter the salivary glands were dissected out and acini were examined after methyl green pyronin (MGP) staining. A total of 134 male and 139 female ticks were dissected out. Average infected acini per tick were found to be significantly higher (p<0.05) in male as compared to the female ticks. Further, maximum infected acini in both male and female ticks were found at 24h of prefeeding on rabbits and overall infected acini per tick increased with rise in parasitaemia. The release of infected ticks on susceptible donkeys resulted in development of clinical babesiosis.     

The effects of ACTH,prednisolone and Escherichia coli endotoxin on some clinical haematological and blood biochemical parameters in dwarf goats

Summary

ACTH (μg kg ^‐1 i.v.) and prednisolone (1 μg^‐1 i.v.) caused a moderate but statistically significant inhibition of rumen contractions, whereas no effects on heart rate and body temperature were observed. Both hormones induced hyperglycaemia and leucocytosis, characterised by moderate lymphopenia and a profound increase in the number of circulating neutrophils. A significant decrease in plasma iron and increase in plasma zinc concentrations were observed.

After 3 daily i.m. injections of ACTH (10 μg ^‐1 day ^‐1) decreases were seen in both serum Alkaline phosphatase (ALP) activity and plasma trace metal concentrations; heart rate was significantly higher. Intraveneous injection of E. coli endotoxin (0.1 μg kg ^‐1) caused shivering, fever, inhibition of rumen contractions, changes in heart rate, lymphopenia, neutropenia followed by neutrophilic leucocytosis, hypoferraemia, hypozincaemia, hypoglycaemia and a decline in serum ALP activity. ACTH, given for 3 days, reduced the febrile responses to E. coli endotoxin, modified the changes in heart rate, intensified the inhibition of rumen contractions, and induced a more marked decrease in the number of circulating neutrophils. ACTH pretreatment did not affect the endotoxin‐induced decrease in blood glucose concentrations nor the drop in plasma zinc and iron values.

These results suggest that (1) glucocorticosteroids are not primarily involved in the fall in plasma iron and zinc concentrations during E. coli endotoxin‐induced fever, (2) the effects of endotoxin released glucocorticosteroids on white blood cells and blood glucose are masked by some other effect(s) of endotoxin, and (3) in dwarf goats, ACTH has antipyretic properties without influencing normal body temperature. This effect is probably not dependent on adrenal cortical activity.     

Sows Intramammarily Inoculated with Escherichia coli: Influence of Time of Infection,Hormone Concentrations and Leucocyte Numbers on Development of Disease

The aim of this study was to identify factors that influence the development of disease in sows inoculated with Escherichia coli in the mammary gland. Ten cross‐bred primiparous sows were intramammarily inoculated with living E. coli bacteria at different time points before parturition: seven sows within 48 h before parturition and three sows approximately 96 h before parturition. Before and after inoculation, blood samples and mammary gland biopsy specimens were collected and clinical observations were made. All seven sows inoculated close to parturition developed a rectal temperature of >39.5°C during the first 48 h post‐partum and two of them also showed other signs of clinical disease. In the sows inoculated 4 days before parturition, the rectal temperature never exceeded 39.5°C during the first 48 h post‐partum and none of them showed any other sign of clinical disease. There was a tendency (P < 0.1) that histological signs of mastitis were more frequent in the sows inoculated close to parturition. There were no overall differences between the two groups of sows in plasma concentrations of cortisol, oestradiol‐17β and 15‐ketodihydro‐PGF_2α before inoculation. Before inoculation, the number of neutrophils in the blood was overall higher (P < 0.05) in the group of sows that were inoculated close to parturition. In comparison, the number of lymphocytes before inoculation had a tendency (P < 0.1) to be lower in that group. The data suggest that the time of infection of the mammary gland relative to parturition and the number of circulating neutrophils at the time of infection influence the development of clinical coliform mastitis in the sow.     

Aspergillus awamori‐fermented mung bean seed coats enhance the antioxidant and immune responses of weaned pigs

The potential benefits of Aspergillus‐fermented mung bean seed coats (FMSC) for weaned pigs remain unexplored. Both in vitro and in vivo experiments were employed to evaluate the potential of FMSC supplement on the growth, antioxidant and immune responses of weaned pigs. The total polyphenols and DPPH scavenging capability of ethanol extract of FMSC exhibited a greater (p < 0.01) increase than those of pre‐fermentation. With the addition of the polyphenol of FMSC extract, an increase in phagocytosis by neutrophils and proliferation of peripheral blood mononuclear cells (PBMC) were found. However, these observations were significantly inhibited (p < 0.05) in those activated cells. Next, 96 weaned pigs were allotted with a randomized complete block design into four dietary treatments, including 0 (control), 600, 1200 or 1800 mg/kg FMSC in a corn–soya bean meal basal diet for a 35‐day trial. The pigs were injected with swine enzootic pneumonia (SEP) vaccines at day 3 and day 21 respectively. The results showed that dietary treatment failed to affect growth performance or serum SEP titre. The diet supplemented with 600–1800 mg/kg FMSC decreased faecal lactoferrin on day 21 and increased plasma trolox equivalent antioxidant capacity (TEAC) and erythrocytes catalase activity, as well as decreased (p < 0.01) plasma malondialdehyde (MDA) concentration on day 35. Diet supplementation of 1800 mg/kg FMSC increased phagocytosis by neutrophils and PBMC proliferation induced by pokeweed mitogen (PWM). However, the polymorphonuclear leucocytes (PMN)‐positive respiratory burst cells were decreased in the supplementation of 1200 or 1800 mg/kg FMSC respectively. In addition, the serum haptoglobin concentration was decreased in the supplementation with 1200 mg/kg FMSC. Taken together, FMSC enriches polyphenols with antioxidative and immune modulated properties. After feeding FMSC, an improvement in antioxidative capability and immunocompetence was found, implying that FMSC could provide as a feed additive at optimal level 1200 mg/kg for weaned pigs.     

Expression in Escherichia coli and purification of the functional feline granulocyte colony-stimulating factor

Feline granulocyte colony-stimulating factor (G-CSF) with an N-terminal histidine hexamer tag was expressed as inclusion bodies in E. coli. The G-CSF solubilized in 6 M guanidine solution was absorbed onto a Ni-NTA column and, after washing with decreasing concentrations of guanidine, eluted with imidazole in a soluble and apparently pure form. The activity of the recombinant feline G-CSF was 3×10⁶ U/mg protein, as assayed by its stimulatory effect on NFS-60 cell proliferation. When a low level of purified feline G-CSF was administered once a day for two successive days to cats, the number of neutrophil increased 4-fold while the levels of other blood cell types remained virtually unchanged. Daily administration of G-CSF for a total of 11 days led to a more than 10-fold increase in neutrophils, an 8-fold increase in the number of monocytes and 2-fold increase in lymphocytes. No severe side effects or antibody production was observed in cats after administration of G-CSF.