Histopathological characteristics of Kupffer cells and ito cells in the porcine and bovine liver

We previously reported that no Kupffer cells reacted with the antibody against lysozyme, and Ito cells contained a large cytoplasmic vacuole in the feline liver. In this report, we further examined the characteristics of porcine and bovine hepatic non-parenchymal cells. In the liver of both animals, Kupffer cells were positive for lysozyme, and cytoplasmic vacuoles in Ito cells were small. The histopathological characteristics of porcine and bovine hepatic non-parenchymal cells were different from those of the feline liver.     

Comparison of the response of bovine and human neutrophils to various stimuli.

Elastase release, oxidant production and cytoplasmic Ca2+ fluxes by bovine and human neutrophils were compared using sensitive kinetic assays on a photon-counting spectrofluorometer. The stimulants used were phorbol myristate acetate (PMA), cytochalasin B, zymosan opsonized with bovine complement (bOZ) or human complement (hOZ), calcium ionophore, formyl-methionyl-leucyl-phenylalanine (FMLP) and concanavalin A (Con A). The respiratory burst of bovine and human neutrophils was stimulated by PMA and OZ but not by cytochalasin B, or calcium ionophore. Con A weakly stimulated this response in human neutrophils but not bovine. FMLP stimulated the respiratory burst of human but not bovine neutrophils. For evaluation of elastase release, human neutrophils were pretreated with cytochalasin B for 5 min and then stimulated. Cytochalasin B alone did not stimulate elastase release from human neutrophils. Phorbol myristate acetate, calcium ionophore, hOZ, FMLP and Con A did stimulate human neutrophils pretreated with cytochalasin B to release elastase. Human serum OZ was also able to stimulate elastase release from human neutrophils not pretreated with cytochalasin B. Some bovine neutrophils released elastase in response to cytochalasin B alone. Those bovine neutrophils that did not release elastase in response to cytochalasin B alone released elastase when stimulated with Con A or calcium ionophore after cytochalasin B pretreatment. Bovine neutrophils did not release elastase in response to FMLP or PMA with or without cytochalasin B pretreatment, but did release elastase in response to bOZ alone. Total elastase activity of bovine neutrophils was determined to be about 50 times less than that of human neutrophils. Intracellular calcium fluxes were stimulated in human neutrophils by calcium ionophore, FMLP, hOZ and Con A but not by PMA or cytochalasin B. Bovine neutrophil calcium fluxes were stimulated by calcium ionophore, Con A and bOZ; cytochalasin B also stimulated bovine neutrophils to increase cytoplasmic calcium concentration. Cytoplasmic calcium fluxes were not stimulated in bovine neutrophils by PMA or FMLP. In summary, human and bovine neutrophils respond similarly to calcium ionophore and OZ, but differently to PMA, cytochalasin B, Con A and FMLP.     

Leukocyte response of bovine mammary gland to injection of killed cells and cell walls of Staphylococcus aureus.

Injection of mammary glands of cows with heat-killed staphylococcal cells, staphylococcal cell walls, or distilled water induced leukocytosis. The magnitude of leukocytic response to staphylococcal antigens in cows of each treatment group depended on the extent of previous experience with staphylococcal cell materials. The 2nd intramammary injection produced greater reactions than did the 1st, and the 1st injection in cows previously parenterally vaccinated with killed cells in oil-water adjuvant produced responses comparable with those elicited by the 2nd intramammary injection in nonvaccinated cows. The implications that these changes have toward understanding the pathogenesis of, and immunity to, staphylococcal mastitis were discussed.     

Effect of Ostertagia ostertagi secretions and various putative secretagogues and inhibitors on aminopyrine accumulation in dispersed bovine abomasal gland cells

Aminopyrine accumulation in suspensions of isolated gastric glands was used to determine the effect of Ostertagia ostertagi secretions and putative secretagogues and inhibitors on abomasal parietal cells. Parasite secretions did not affect acid production nor did histamine. Dibutyryl cyclic adenosine monophosphate (cAMP) and pentagastrin significantly increased aminopyrine accumulation by gastric glands and cimetidine, omeprazole and thiocyanate significantly decreased aminopyrine accumulation confirming their roles as stimulators and inhibitors of gastric acid production, respectively.     

Morphologic, cytochemical staining, and ultrastructural characteristics of blood cells from eastern diamondback rattlesnakes (Crotalus adamanteus)

OBJECTIVE: To evaluate light microscopic, cytochemical, and ultrastructural characteristics of blood cells from eastern diamondback rattlesnakes. ANIMALS: 10 healthy snakes. PROCEDURE: Various stains, including Wright-Giemsa, benzidine peroxidase, Sudan black B, chloroacetate esterase, alpha-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff with diastase, and toluidine blue, were used to stain leukocytes differentially on multiple blood smears. Electron microscopy also was performed. RESULTS: Lymphocytes were the most commonly observed leukocyte and could be distinguished from thrombocytes, using periodic acid-Schiff stain with diastase. Azurophils also were commonly observed; their granules stained with peroxidase. Eosinophils were not identified; however, 2 morphologic variations of heterophils were seen in the blood of all snakes and were considered the same cell type at different stages of cytoplasmic granule development. Heterophil granules were better preserved, using a one-step Wright-Giemsa method that did not require alcohol fixation prior to staining. Degranulated heterophils were observed in all preparations. CONCLUSIONS: Most leukocytes of eastern diamondback rattlesnakes can be identified easily on Wright-Giemsa-stained preparations. However, hematologic stains that do not require alcohol fixing prior to staining may be preferred for leukocyte evaluation in certain reptiles. A limited degree of heterophil maturation may continue in the blood of healthy snakes. This, along with degranulation of heterophils, may result in a variable staining pattern in this cell type, regardless of the stain used. CLINICAL RELEVANCE: Results provide baseline data for use in hematologic testing in diagnosis of disease and monitoring of treatment of sick or injured snakes.     

Effects of active immunization against GnRH on LH, FSH and prolactin storage, secretion and response to their secretagogues in pony geldings

Six pony geldings were actively immunized against GnRH conjugated to bovine serum albumin (BSA) to study 1) the relative dependency of LH and FSH storage, secretion and response to GnRH analog on GnRH bioavailability and 2) the effects of reduced GnRH bioavailability on GnRH storage in the hypothalamus. Five geldings were immunized against BSA. Geldings were immunized in December and 4, 8, 14, 20, 26 and 32 wk later. Ponies immunized against GnRH had increased (P less than .01) GnRH binding in plasma within 6 wk. By June, plasma concentrations of LH and FSH in ponies immunized against GnRH had decreased (P less than .02) by 86 and 59%, respectively, relative to ponies immunized against BSA. The LH response to an injection of GnRH analog, which did not bind to anti-GnRH antibodies, was reduced (P less than .005) by 90% in ponies immunized against GnRH relative to ponies immunized against BSA. In contrast, the FSH response to GnRH analog was similar (P greater than .1) for both groups. Immunization against GnRH reduced (P less than .05) weight of the anterior pituitary (AP) by 31%, LH content in AP by 91% and FSH content in AP by 55% relative to ponies immunized against BSA. There was no effect of GnRH immunization on prolactin characteristics or on GnRH concentrations in the median eminence, preoptic area or body of the hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cellular interactions regulating the in vitro response of bovine lymphocytes to ovalbumin.

We examined the contribution of MHC class II-restricted T cells (CD4+), MHC class I-restricted T cells (CD8+), gamma/delta T cell receptor (TCR)+ T cells, B cells and macrophages to the development and control of in vitro proliferative responses of bovine lymphocytes to ovalbumin (OA). Cell populations for in vitro assay were obtained from peripheral blood (peripheral blood leukocytes, PBL) of OA-primed cattle. Specific cell populations were depleted or purified from PBL by staining with monoclonal antibodies (MAbs) against the appropriate differentiation antigens and sorting on a Fluorescence Activated Cell Sorter (FACS). OA-specific in vitro responses of in vivo primed PBL were dependent on the presence of CD4+ T cells. Their presence could not be replaced by the inclusion of T cell growth factor (TCGF) in the culture system, indicating that CD4+ T cells probably actively proliferate in response to antigenic stimulation. Bovine CD8+ T cells and gamma/delta TCR+ T cells appeared to exert a suppressive effect on proliferative responses. No proliferation was observed in PBL after the depletion of MHC class II+ cells. In this case, the response could be restored by the addition of macrophages or LPS-activated B cells to the MHC class II- population.     

Stimulation techniques and response characteristics of the M and F waves and H reflex in dogs

The voltage and duration of electrical rectangular pulsed stimuli needed to produce an F wave and a monosynaptic reflex (H wave) and the characteristics of these responses were recorded in clinically normal dogs. Optimal stimulus to produce H waves was 0.1 to 0.2 ms and less than 80 volts. F waves were variable in appearance and were most evident following 0.5 ms and 125 to 150 volt stimulation. F waves had shorter latency than comparable H waves.     

An evaluation of the mononuclear cells derived from bovine mammary gland dry secretions using leukocyte antigen specific monoclonal antibodies, light scattering properties and non-specific esterase staining.

The distribution of mononuclear cells isolated from the bovine mammary gland during the nonlactating (dry) period was examined using monoclonal antibodies against leukocyte cell surface antigens, cellular light scattering properties, and the presence of nonspecific esterase. Most of the mononuclear cells isolated during the dry period were lymphocytes. T cells predominated until about 1 week prior to parturition. During the week prior to calving, the percentage of B cells increased until it approximated T cells. The ratio of CD4:CD8 cells was 2-3:1 for mammary gland T cells. This was similar to the ratio found in peripheral blood. At dry-off, about 12% of mammary mononuclear cells were macrophages. The macrophage percentage increased (to about 30%) at mid-dry and remained at this levels until parturition. PMN's were isolated with the mononuclear cells during the first 2 weeks dry and the week prior to calving. Three methods were used to identify mammary macrophages. Esterase staining (as an enzymatic method), forward angle/90 degrees light scatter (based on size and internal complexity), and MHC class II/forward angle light scatter (based on size and surface markers) were compared. Each method yielded similar specificity for macrophage identification. Non-adherent cell fractions, obtained by passage of the cells over Sephadex G-10 columns, were enriched in CD4 positive T cells, somewhat depleted of B cells, and depleted of macrophages and PMN's. Cells eluted from G-10 columns, with lidocaine, were mostly lymphocytes, but reflected the cells loaded onto the column.     

Humoral immune response of the bovine fetus to in utero vaccination with attenuated bovine coronavirus

A fetal response to in utero vaccination with attenuated bovine coronavirus (9 to 49 days before parturition) was determined in 8 calves, 5 vaccinated and 3 controls. Calves were derived by hysterotomy before parturition and were maintained in a closed gnotobiotic environment. The IgA, IgM, and IgG values and coronavirus-neutralizing antibody titers were higher in the sera and intestinal loop fluid from vaccinated calves than in those from control calves. Sections of ileum and ileal lymph nodes from 1-day-old vaccinated calves, when stained with monospecific anti-bovine IgG, IgM, and IgA had numerous positively stained plasma cells. Positive fluorescence was not detected in comparable tissues from controls. When the 8 calves were given virulent coronavirus orally at 6 days of age, vaccinated calves did not become ill, whereas control calves had diarrhea in 19 to 22 hours. All calves were killed at 10 days of age. Control calves had lesions characteristic of coronavirus infection, and intestinal epithelial cells were positive by fluorescent antibody tests. In vaccinated calves, lesions of coronavirus infection were absent, and results of fluorescent antibody tests were negative. Although in utero vaccination with a coronavirus vaccine stimulated immunity in the newborn calf, the frequency of abortions (2 of 14 cows inoculated intra-amniotically) and premature births (4 of 14) precluded practical application.     

Effect of age on immunocytochemical staining characteristics of adenohypophyseal cells in Mongolian pony mares and stallions

OBJECTIVE: To determine the effect of age on immunocytochemical staining characteristics of adenohypophyseal cells of Mongolian pony mares and stallions. ANIMALS: 35 Mongolian ponies. PROCEDURE: Pituitary glands from 1- to 22-year-old horses of both sexes were collected at a commercial slaughterhouse in China and allocated into 7 groups according to age and sex: prepubertal stallions (n = 5; 1 to 2 years old), young stallions (6; 3 to 7 years old), middle-aged stallions (4; 10 to 12 years old), old stallions (5; 15 to 22 years old), young mares (3; 3 to 7 years old), middle-aged mares (5; 10 to 12 years old), and old mares (7; 15 to 22 years old). Pituitary glands were processed for microscopy, and sections were immunocytochemically stained for various pituitary hormones. The percentages and cell areas of 6 types of adenohypophyseal cells were determined by use of morphometry. RESULTS: Age-related alterations in adenohypophyseal cells were detected in horses > 15 years old. Percentage of somatotrophs decreased with age regardless of sex, and percentage of lactotrophs increased with age in stallions. In old mares, percentage of follicle-stimulating hormone cells increased, whereas cell area of follicle-stimulating hormone and luteinizing hormone cells decreased. Differences between sexes for percentage of somatotrophs and lactotrophs, and cell area of follicle-stimulating hormone and luteinizing hormone cells were not evident in old horses. CONCLUSIONS AND CLINICAL RELEVANCE: Differences among percentages and cell areas of pituitary cell types in old horses may be associated with degeneration of the sex organs.     

Expression of cytokines and respiratory burst activity of milk cells in response to Azadirachta indica during bovine mastitis

The immuno therapeutic potential of hydro-methanolic extract of Azadirachta indica (A. indica) was studied during bovine clinical mastitis (CM). The somatic cell count (SCC), total bacterial count (TBC), milk differential leukocyte count (DLC), hydrogen peroxide (H₂O₂), superoxide anion (O₂ ⁻) production and interleukin- 2 (IL-2) and gamma interferon (IFN-γ) cytokines expression were studied before and after intramammary infusion of A. indica extract in diseased cows. The results revealed that A. indica treatment significantly (P < 0.05) decreased the SCC, TBC, milk neutrophil percent and significantly (P < 0.05) enhanced milk lymphocyte percent, H₂O₂ and O₂ ⁻ production by milk cells. The IL-2 and IFN-γ were expressed in normal healthy cows and diseased cows after A. indica treatment, whereas both the cytokines could not be expressed in cows treated with antibiotic and in untreated diseased cows. The results of the present study indicated anti inflammatory, antibacterial and immunomodulatory potential of the herb, these activities could be due to the presence of bioactive principle in the extract. This is a preliminary trial indicated beneficial effect of the herb against bovine mastitis it can be developed as an alternative therapy where the use of antibiotics is normally restricted.     

Relationship between the milk yield response to short-term bovine somatotropin treatment and the lipolytic response to adrenaline in dairy cows

The aim of this experiment was to determine if the milk yield response of dairy cows to short-term treatment with bovine somatotropin (bST) was correlated with the non-esterified fatty-acid (NEFA) response to an adrenaline challenge. Twenty-six multiparous Holstein cows (58+/-5.4 days postpartum) received daily sub-cutaneous injections of saline for 7 days followed by sub-cutaneous injections of 20mg/day of bST for 14 days. On day 7 of the saline treatment and day 14 of the bST treatment the cows were given an intravenous injection of adrenaline (1.4 microg/kg body weight). Blood samples were taken before and after the adrenaline challenge. The difference in milk yield between the saline and the second week of bST treatment (MYR) varied considerably between animals (from -0.4 to +8.0 kg/day). MYR was positively correlated with the change in the basal concentration of NEFA between the saline and second week of bST treatment, as well as with the change in the area under the profile of NEFA above basal values following the adrenaline challenge. It remains to be established whether the greater lipolytic responses to adrenaline of the cows with the greater MYR reflects the deeper negative energy that these animals also experienced or a fundamental difference in the physiology of their adipose tissue.     

Comparison of staining and concentration techniques for detection of Cryptosporidium oocysts in cat faecal specimens.

Modified Ziehl-Neelsen (MZN), auramine-phenol (A-P) and fluorescein isothiocyanate-labelled (FITC-labelled) monoclonal antibody (MAb) techniques were compared for detection of Cryptosporidium parvum oocysts in cat faecal specimens inoculated with known numbers of C. parvum oocysts. Of the three techniques, the FITC-labelled MAb technique detected more oocysts than the MZN and A-P techniques (P < 0.05), but A-P was more efficient than MZN (P < 0.05). Comparison of sucrose flotation, zinc sulphate (ZnSO4) flotation and formol-ether (F-E) sedimentation techniques revealed that F-E was the most efficient of the three (P < 0.05) for concentration of C. parvum oocysts from cat faecal specimens. On average, the F-E technique recovered 37% of oocysts from the original sample, whereas the sucrose and ZnSO4 flotation techniques recovered 33% and 11%, respectively. The findings of this study suggest that MZN and A-P staining are both useful for screening C. parvum oocysts in cat faecal materials containing 10(6) oocysts or more, but FITC-labelled MAb should be used when the number of oocysts is low. Also, the F-E sedimentation technique is recommended for concentrating oocysts in cat faecal specimens.     

RNA-Seq-based transcriptomic profiling of primary interstitial cells of Cajal in response to bovine viral diarrhea virus infection

Veterinary Research Communications - Infections with bovine viral diarrhea virus (BVDV) contribute significantly to health-related economic losses in the beef and dairy industries and are...     

The determination of the cellular volume of avian, porcine and bovine phagocytes and bovine mammary epithelial cells and its relationship to uptake of tilmicosin

In order to compare the intracellular concentration of antimicrobial agents in phagocytic and nonphagocytic cells, the knowledge of their cell volume is essential. For the first time, the determination of the avian, porcine, and bovine polymorphonuclear neutrophils (PMN), monocyte-derived macrophages, macrophages, and bovine mammary epithelial cell volume was performed using [3H]-water and [14C]-carboxyinulin. The comparison of all the cells showed that the PMN have a size range between 3.58 and 4.04 microL per mg of protein, and are smaller than the monocyte-derived macrophages and mammary epithelial cells (4.32-5.01 microL per mg of protein). The macrophages show the largest size (5.84-6.57 microL per mg of protein). The cellular uptake of tilmicosin in these cells was then determined. The examination of the intracellular/extracellular concentration ratios (Ci/Ce) after 4 h of incubation with 10 mg/mL of [14C]-labelled tilmicosin revealed that tilmicosin was well accumulated and showed a ratio of 137, 169 and 193 in avian PMN, porcine PMN, and bovine alveolar macrophages, respectively. The cellular uptake data also demonstrated that tilmicosin accumulated in nonphagocytic bovine mammary epithelial cells. The importance of the use of the appropriate species and cell type specific cell volume values for calculations was exemplified by calculating the Ci/Ce of tilmicosin using cell volume data found in the literature for human and mouse cells. The subsequent comparison of these data with the Ci/Ce calculated with the actual cell volume appropriate for the species tested revealed an under evaluation of 3-13% in monocyte-macrophages, an over evaluation of 7-18%, 16-31% and 69% in PMN, macrophages, and epithelial cells, respectively. This study highlights the importance of the proper cell volume in order to determine the Ci/Ce. Moreover, the cell volumes determined here for avian, porcine and bovine cells should facilitate further in vitro and in vivo cellular studies by veterinary researchers.