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.     

Thrombocyte morphology and morphometric observations in two vulture species

Background: The important role of thrombocytes in hemostasis is well documented, but little information is available on the thrombocyte morphology of avian species, including vultures. Objective: The objective of this study was to describe and compare the morphology and morphometric parameters of thrombocytes in 2 vulture species. Methods: Blood samples were collected into tubes containing acid‐citrate‐dextrose from 5 Cape vultures (Gyps coprotheres) and 6 white‐backed vultures (Gyps africanus) at the De Wildt Breeding Center, Northwest Province, South Africa. Wright's‐stained blood smears were examined by light microscopy. Samples were processed and examined by scanning and transmission electron microscopy (TEM) using standard techniques. Morphometric parameters (perimeter, area, minimum and maximum diameter, and aspect ratio) were measured on 140 thrombocytes using imaging software. Results: Thrombocytes were predominantly oval to elliptical, with few pseudopodia. The nucleus was the most prominent feature of the cells. Large vacuoles were visible in the cytoplasm by both light and TEM. Ultrastructurally, microtubules and dense bodies were seen in most cells. Other cytoplasmic organelles seen by electron microscopy included mitochondria, endoplasmic reticulum, a surface connecting canalicular system, Golgi complex, lipid droplets, and glycogen. Thrombocytes of Cape vultures had a significantly (P=.005) higher aspect ratio compared with white‐backed vultures. Thrombocyte estimates in blood smears were similar in both species, with a combined mean of 31.6 × 10⁹/L. Conclusion: The morphologic features of thrombocytes in southern African vultures are similar in most ways to those of other avian species. Although thrombocytes in white‐backed vultures were slightly more spherical than those of Cape vultures, no other significant differences were found between the 2 species.     

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.     

Comparative pharmacokinetics of norfloxacin nicotinate in common carp (Cyprinus carpio) and crucian carp (Carassius auratus) after oral administration

Comparative pharmacokinetics of norfloxacin nicotinate (NFXNT) was investigated in common carp (Cyprinus carpio) and crucian carp (Carassius auratus) after a single oral dose of 10 mg/kg body weight (b.w.). Analyses of plasma samples were performed using ultra‐performance liquid chromatography (UPLC) with fluorescence detection. After oral dose, plasma concentration–time curves of common carp and crucian carp were best described by a two‐compartment open model with first‐order absorption. The pharmacokinetic parameters of common carp were similar to those of crucian carp. The distribution half‐life (t_1/2α), elimination half‐life (t_1/2β), peak concentration (C_max), time‐to‐peak concentration (T_max), and area under the concentration–time curve (AUC) of common carp were 1.58 h, 26.33 h, 6069.79 μg/L, 1.08 h, and 103072.36 h·μg/L, respectively, and those corresponding to crucian carp were 1.36 h, 26.55 h, 9586.06 μg/L, 0.84 h, and 126604.4 h·μg/L, respectively. These studies demonstrated that 10 mg NFXNT/kg body weight in common carp and crucian carp following oral dose presented good pharmacokinetic characteristics.     

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.     

Comparative pharmacokinetics and tissue distribution of quinocetone in crucian carp (Carassius auratus), common carp (Cyprinus carpio L.), and grass carp (Ctenopharyngodon idella) following the same experimental conditions

The pharmacokinetics and tissue distribution of quinocetone (QCT) in crucian carp (Carassius auratus), common carp (Cyprinus carpio L.), and grass carp (Ctenopharyngodon idella) were compared after oral administration of QCT (50 mg/kg body weight) at water temperature of 24 ± 1 °C. Similar QCT plasma concentration–time profiles were found in the three species of cyprinid fish at the same dosage regimen and water temperature, which were all fitted two‐compartment open pharmacokinetic model. However, different pharmacokinetic parameters were observed in crucian carp, common carp, and grass carp. The absorption rate constants (K_a) of QCT were 1.65, 1.40 and 1.74/h, respectively and absorption half‐lives (t_1/2kα) were 0.42, 0.49, and 0.40/h, respectively. The distribution half‐life (t_1/2α) was 2.83, 0.67, and 0.88 h, respectively, and elimination half‐lives (t_1/2β) of QCT were 133.97, 63.55, and 40.76 h, respectively. The maximum concentrations (C_max) of QCT in plasma were 0.315, 0.182, and 0.139 μg/mL and the time to peak concentrations (T_p) were 1.45, 0.96, and 1.08 h, respectively. The area under the plasma concentration‐time curves (AUC) were 12.35, 5.99, and 4.52 μg·h/mL, respectively. The distribution volumes (V_d/F) of QCT were calculated as 117.81, 128.71, and 220.10 L/kg, respectively. The tissue analysis showed that a similar regularity was obtained in the three species of cyprinids with a single dose of 50 mg/kg body weight after oral administration at the same water temperature. The tissue concentration of QCT in each fish was in order of liver>kidney>muscle, while the residues of QCT in the three species of cyprinid fish were in order of crucian carp>common carp>grass carp.     

Hematology and clinical chemistry of adult yellow-headed temple turtles (Hieremys annandalii) in Thailand

Background: Yellow‐headed temple turtles (YHT), Hieremys annandalii, native to Thailand, are protected from exploitation under the Wild Animal Reservation and Protection Act, also listed under Appendix II of the Convention on International Trade of Endangered Species and the International Union for the Conservation of Nature red list. Objectives: The objectives of this study were to describe quantitative, morphologic, and cytochemical features of blood cells and plasma biochemical analytes of clinically healthy YHT. Methods: Blood samples were collected from 40 adult YHT from October 2007 to February 2008. Hematologic and biochemical analyses, cytochemical staining, and ultrastructural evaluation were performed using standard methods. Results: Hematologic results (mean ± SD) included: RBC count, 0.275 ± .094 × 10⁶ cells/μL; WBC count, 11.7 ± 6.6 × 10³ cells/μL; heterophils, 29.4 ± 6.9%; eosinophils, 23.7 ± 5.3%; basophils, 21.2 ± 1.9%; lymphocytes, 14.8 ± 5.9%; and azurophils, 10.7 ± 5.3%. Erythrocytes stained dark red with peroxidase‐staining. Periodic acid‐Schiff stain could not differentiate between thrombocytes and lymphocytes. Thrombocytes contained cytoplasmic vacuoles, similar to mammalian platelets and those of birds and snakes. Heterophils and eosinophils were similar in structure and cytochemical staining characteristics to those of other turtles and reptiles. Structure of basophils was similar to avian basophils. Lymphocytes and azurophils had similar cytochemical staining compared with mammalian lymphocytes and monocytes. Mean MCHC, WBC counts, absolute azurophil counts, and plasma alanine aminotransferase activity were higher in male turtles than in females. Conclusion: Blood characteristics of YHT are species‐specific, and this study can be served as a reference for future clinical studies and medical care of YHT.     

Morphology of blood cells in carp (Cyprinus carpio L.)

The morphology of blood cells in the carp was investigated by light and electron microscopy. Erythrocytes, thrombocytes, lymphocytes, granulocytes and monocytes were identified as the peripheral blood cells. Thrombocytes were round to long oval, each containing vesicular and microtubular structures and an oval nucleus with abundant heterochromatins. Lymphocytes were divided into three types in size, small, medium and large. Some of the small and medium lymphocytes were alpha-naphthyl-acetate esterase (ANAE) positive, while large lymphocytes were pyroninophilic. Granulocytes were distinguished into three types (type I, type II and type III) according to the morphology of the nucleus and granules. Type I granulocytes possessed lobulated nuclei and a large number of cytoplasmic granules, each of which was oval and contained electron-dense materials and a crystalloid. Type II granulocytes had small eccentric nuclei and were subdivided into IIa and IIb granulocytes by electron microscopic analysis. Granules of type IIa granulocytes were furnished with an electron-dense rim. Granules of type IIb granulocytes were larger than those of type IIa, containing randomly distributed electron-dense and electron-lucent materials. Type III granulocytes possessed round nuclei and a few large granules. The granules were filled with regularly arranged fibriform materials and some needle-like structures. Monocytes were morphologically similar to those of mammals.     

Comparative pharmacokinetics of enrofloxacin in healthy and Aeromonas hydrophila‐infected crucian carp (Carassius auratus gibelio)

The comparative pharmacokinetics of enrofloxacin (ENR) and its metabolite ciprofloxacin (CIP) were investigated in healthy and Aeromonas hydrophila‐infected crucian carp after a single oral (p.o.) administration at a dose of 10 mg/kg at 25 °C. The plasma concentrations of ENR and of CIP were determined by HPLC. Pharmacokinetic parameters were calculated based on mean ENR concentrations by noncompartmental modeling. In healthy fish, the elimination half‐life (T_1/2λz), maximum plasma concentration (C_max), time to peak (T_max), and area under the concentration–time curve (AUC) values were 64.66 h, 3.55 μg/mL, 0.5 h, and 163.04 μg·h/mL, respectively. In infected carp, by contrast, the corresponding values were 73.70 h, 2.66 μg/mL, 0.75 h, and 137.43 μg·h/mL, and the absorption and elimination of ENR were slower following oral administration. Very low levels of CIP were detected, which indicates a low extent of deethylation of ENR in crucian carp.     

Genistein Alters the Release of Oxytocin,Prostaglandins, Cortisol and LH during Insemination in Gilts

Soya products containing phytooestrogens are widely used as feed for pigs. However, limited data are available on the effects of phytooestrogen on the endocrine status of pigs. The aim of this work was to study the impact of the phytooestrogen genistein added to a soya‐free diet on the hormonal pattern in gilts during oestrus and artificial insemination (AI). Ten gilts were fed a soya‐free diet and fitted with jugular vein catheter through vena auricularis. The gilts were randomly divided into two groups (G‐ and C‐group) where the G‐group was given pure genistein, 1 mg/kg body weight (BW) twice daily, per os. Blood samples were collected before, during and after AI. Oxytocin, prostaglandin E_2, prostaglandin F_2α, 13,14‐dihydro‐15‐keto‐prostaglandin F_2α (PGFM), cortisol and LH concentrations in blood plasma were analysed. Oxytocin concentrations were almost twice as high in the G‐group as in C‐group after the AI. Prostaglandin E₂ concentrations were higher in G‐group than in C‐group during the entire sampling period. After AI, the concentrations of prostaglandin E₂ increased in G‐group but not in C‐group. Prostaglandin F_2α concentration had a pulsatile pattern, with increasing pulses after AI in G‐group. Plasma PGFM concentrations increased after AI with a small variation between the groups. Plasma cortisol concentration increased after AI in C‐group. LH decreased after AI in G‐group. Genistein stimulated elevations of plasma oxytocin and prostaglandin E₂ concentrations and a pulsative pattern in prostaglandin F_2α concentration. The possible involvement of genistein in plasma cortisol and basal LH concentrations in gilts given genistein may also be suggested.     

Characterisation of duck thromhocytes

Duck thrombocytes were initially identified in peripheral blood mononuclear cells (PBMCS) purified from whole blood on Ficoll-Paque density gradients by examining stained smears of these cells. These thrombocytes could be readily purified from lymphocytes on the FACStar cell sorter by their increased side-scatter. They were similar to chicken thrombocytes in both appearance and function; they had a diameter of 4.5–6 μm and contained large vacuoles and were able to phagocytose carbon and Staphylococcus aureus. A monoclonal antibody (mAb) BA3 was generated which binds specifically to duck thrombocytes and has facilitated the characterisation of these cells which comprise up to 50–60 per cent of cells in Ficoll-Paque purified duck PBMCS.     

Pharmacokinetics of cefquinome in crucian carp (Carassius auratus gibelio) after oral,intramuscular, intraperitoneal,and bath administration

The pharmacokinetics (PK) of cefquinome (CEQ) was studied in crucian carp (Carassius auratus gibelio) after single oral, intramuscular (i.m.), and intraperitoneal (i.p.) administration at a dose of 10 mg/kg body weight and following incubation in a 5 mg/L bath for 5 hr at 25°C. The plasma concentration of CEQ was determined using high‐performance liquid chromatography (HPLC). PK parameters were calculated based on mean CEQ concentration using WinNonlin 6.1 software. The disposition of CEQ following oral, i.m., or i.p. administration was best described by a two‐compartment open model with first‐order absorption. After oral, i.m., and i.p. administration, the maximum plasma concentration (C_max) values were 1.52, 40.53, and 67.87 μg/ml obtained at 0.25, 0.23, and 0.35 hr, respectively, while the elimination half‐life (T_1/2β) values were 4.68, 7.39, and 6.88 hr, respectively; the area under the concentration–time curve (AUC) values were 8.61, 339.11, and 495.06 μg hr/ml, respectively. No CEQ was detected in the plasma after bath incubation. Therapeutic blood concentrations of CEQ can be achieved in the crucian carp following i.m. and i.p. administration at a dosage of 10 mg/kg once every 2 days.     

Pharmacokinetics of sulphadimidine in carp (Cyprinus carpio L.) and rainbow trout (Salmo gairdneri Richardson) acclimated at two different temperature levels

Summary

The influence of temperature (10° C and 20° C) on pharmacokinetics and metabolism of sulphadimidine (SDM) in carp and trout was studied.

At 20° C a significantly lower level of distribution (Vd_area ) and a significantly shorter elimination half‐life (T _(½>) β) was achieved in both species compared to the 10° C level. In carp the body clearance parameter (Cl_B (SDM) was significantly higher at 20° C compared to the value at 10° C, whereas for trout this parameter was in the same order of magnitude for both temperatures.

N₄‐acetylsulphadimidine (N₄‐SDM) was the main metabolite of SDM in both species at the two temperature levels. The relative N₄‐SDM plasma percentage in carp was significantly higher at 20° C than at 10° C, whereas there was in trout no significant difference.

In neither species was the peak plasma concentration of N_4‐SDM (C_maxN4‐SDM)) significantly different at two temperatures.

The corresponding peak time of this metabolite (T_{max (N4‐SDM)}) was significantly shorter at 20° C compared to 10° C in both carp and trout.

In carp at both temperatures, acetylation occurs to a greater extent than hydroxylation. Only the 6‐hydroxymethyl‐metabolite (SCH₂OH) was detected in carp, at a significant different level at the two temperatures. Concentrations of hydroxy metabolites in trout were at the detection level of the HPLC‐method (0.02‐μg/ml). The glucuronide metabolite (SOH‐gluc.) was not detected in either species at the two temperatures.     

Pharmacokinetics of orbifloxacin in crucian carp (Carassius auratus) after intravenous and intramuscular administration

The pharmacokinetics of orbifloxacin was studied after a single dose (7.5 mg/kg) of intravenous or intramuscular administration to crucian carp (Carassius auratus ) reared in freshwater at 25°C. Plasma samples were collected from six fish per sampling point. Orbifloxacin concentrations were determined by high‐performance liquid chromatography with a 0.02 μg/ml limit of detection, then were subjected to noncompartmental analysis. After intravenous injection, initial concentration of 5.83 μg/ml, apparent elimination rate constant (λ_z) of 0.039 hr^?1, apparent elimination half‐life (T_1/2λz) of 17.90 hr, systemic total body clearance (Cl) of 75.47 ml hr^?1 kg^?1, volume of distribution (Vz) of 1,948.76 ml/kg, and volume of distribution at steady‐state (Vss) of 1,863.97 ml/kg were determined, respectively. While after intramuscular administration, the λ_z, T _1/2λz, mean absorption time (MAT ), absorption half‐life (T _1/2ka), and bioavailability were determined as 0.027 hr^?1, 25.69, 10.26, 7.11 hr, and 96.46%, respectively, while the peak concentration was observed as 3.11 ± 0.06 μg/ml at 2.0 hr. It was shown that orbifloxacin was completely but relatively slowly absorbed, extensively distributed, and slowly eliminated in crucian carp, and an orbifloxacin dosage of 10 mg/kg administered intravenously or intramuscularly would be expected to successfully treat crucian carp infected by strains with MIC values ≤0.5 μg/ml.     

Characterization of the Blood Cells of Australian Crocodiles (Crocodylus porosus [SCHNEIDER] AND C. johnstoni [KREFFT])

Circulating blood of eight crocodiles (four Crocodylus porosus [SCHNEIDER], four Crocodylus johnstoni [KREFFT]) was examined by light microscopy, cytochemistry and transmission electron microscopy. The results were correlated and on that basis erythrocytes, thrombocytes, lymphocytes, monocytes and three main types of granulocytes were characterized. Erythrocytes, thrombocytes, lymphocytes and monocytes were similar in appearance to those described for other reptiles. In addition, lymphocytes and monocytes had ultrastructural details similar to those described for the cells in man. The three main granulocytes were designated types I, II and III. Type III had characteristics which were similar to those described for the basophil. Types I and II were eosinophilic on blood films stained with Giemsa. On ultrastructural detail, and by comparison with ultrastructural reports for the granulocytes of the fowl and lizard (Agamia stellio), type I appeared to be similar to the heterophil but type II could not be characterized definitely. However, the crystalline array present in granules of type II has been recorded for eosinophil granules in man.     

Effective use of the TSPY gene‐specific copy number in determining fetal DNA in the maternal blood of cynomolgus monkeys

Since the available concentration of single‐copy fetal genes in maternal blood DNA is sometimes lower than detection limits by PCR methods, the development of specific and quantitative PCR detection methods for fetal DNA in maternal blood is anticipated, which may broaden the methods that can be used to monitor pregnancy. We used the TaqMan qPCR amplification for DYS14 multi‐copy sequence and the SRY gene in maternal blood plasma (cell‐free DNA) and fractional precipitated blood cells (cellular DNA) from individual cynomolgus monkeys at 22 weeks of pregnancy. The availability of cell‐free fetal DNA was higher in maternal blood plasma than that of cellular DNA from fractional precipitated blood cells. There was a significantly higher (P < 0.001) mean copy number of fetal male DYS14 from maternal plasma (4.4 × 10⁴ copies/mL) than that of detected fetal cellular DNA from fractional blood cell pellets. The sensitivity of the DYS14 PCR assay was found to be higher than that of the SRY assay for the detection of fetal DNA when its presence was at a minimum. The DYS14 assay is an improved method for quantifying male fetal DNA in circulating maternal blood in the primate model.     

Pharmacokinetics and tissue residues of moroxydine hydrochloride in gibel carp,Carassius gibelio after oral administration

The pharmacokinetics and tissue residues of moroxydine hydrochloride were studied in gibel carp at water temperature of 15 and 25 °C. Samples (blood, skin, muscle, liver, and kidney) were collected over 10 days after the treatment and analyzed by high‐performance liquid chromatography with an ultraviolet detector. The results indicated that the influence of water temperature on the metabolism of the drug was significant. The plasma concentration–time data of moroxydine hydrochloride conformed to single‐compartment open model at the two water temperatures. There were higher absorption rate (t_1/2ka) and longer elimination half‐lives (t_1/2ke) at 15 °C (4.29 and 15.87 h, respectively) compared with those at 25 °C (3.02 and 4.22 h, respectively). The maximum plasma concentration (C_max) and the time‐point of maximum plasma concentration (T_p) were 2.98 μg/mL and 10.35 h at 15 °C and 3.12 μg/mL and 4.03 h at 25 °C, respectively. The distribution volume (V_d/F) of moroxydine hydrochloride was estimated to be 4.55 L/kg at 15 °C and 2.89 L/kg at 25 °C. The total body clearance (CL_b) of moroxydine hydrochloride was determined to be 0.25 and 0.49 L/(h·kg) at 15 °C and 25 °C, respectively; the areas under the concentration–time curve were 75.89 μg·h/mL at 15 °C and 42.33 μg·h/mL at 25 °C. The depletion of moroxydine hydrochloride in gibel carp was slower with a longer half‐life period, especially at lower water temperature that was tested.     

Assessment of the Hemocue-b for measuring hemoglobin in horse blood

Our purpose was to assess the accuracy and precision of a point of care hemoglobinometer (HemoCue‐B hemoglobin photometer) for measuring hemoglobin concentration in horse blood. Samples of jugular venous blood from 12 healthy adult horses were collected in EDTA. In order to test the device over a wide range of values, each sample was divided into nine aliquots, and autologous plasma was added or removed from the aliquots to produce blood with PCV values that approximated 5, 10, 20, 30, 40, 50, 60, 70, and 80%, respectively. The aliquots were rocked to ensure mixing of plasma and cells. Then hemoglobin by HemoCue‐B (Hb_HQ) and hemoglobin by the cyanmethemoglobin method (Hb_CY) were measured on each aliquot. The PCV of each aliquot was also measured and this value was used for subsequent analyses. To test repeatability, hemoglobin was measured twice by the HemoCue‐B on approximately 40% samples. Samples with Hb_HQ >25.4 g dL^?1 required dilution prior to analysis. Hb_CY ranged from 1.6 to 33.4 g dL^?1. After regression, Hb_CY = ?0.16 + 1.04 Hb_HQ (n = 101; r² = 99.6%). By inspection of a modified Bland‐Altman plot, Hb_HQ values <16 g dL^?1 closely approximated Hb_CY; however, at greater values, Hb_HQ underestimated Hb_CY by as much as 3.2 g dL^?1. The difference between repeated measurements with the HemoCue‐B was 0.02 ± 0.16 g dL^?1 (mean ± SD; n = 10) and nonsignificant. After regression, PCV = ?0.76 + 2.78 Hb_HQ (n = 101; r² = 99.4%). We conclude that HemoCue‐B can be used to measure hemoglobin concentration in horse blood, and that it is accurate when hemoglobin is <16 g dL^?1. PCV can be estimated by multiplying Hb_HQ by 2.8 and then subtracting 0.8.     

Pharmacokinetics and selected behavioral responses to butorphanol and its metabolites in goats following intravenous and intramuscular administration

Objective To evaluate disposition of a single dose of butorphanol in goats after intravenous (IV) and intramuscular (IM) administration and to relate behavioral changes after butorphanol administration with plasma concentrations. Design Randomized experimental study. Animals Six healthy 3‐year‐old neutered goats (one male and five female) weighing 46.5 ± 10.5 kg (mean ± D). Methods Goats were given IV and IM butorphanol (0.1 mg kg^?1) using a randomized cross‐over design with a 1‐week interval between treatments. Heparinized blood samples were collected at fixed intervals for subsequent determination of plasma butorphanol concentrations using an enzyme linked immunosorbent assay (ELISA). Pharmacokinetic values (volume of distribution at steady state [Vd_SS], systemic clearance [Cl_TB], extrapolated peak plasma concentration [C₀] or estimated peak plasma concentration [C_MAX], time to estimated peak plasma concentration [T_MAX], distribution and elimination half‐lives [t_1/2], and bioavailability) were calculated. Behavior was subjectively scored. A two‐tailed paired t‐test was used to compare the elimination half‐lives after IV and IM administration. Behavioral scores are reported as median (range). A Friedman Rank Sums test adjusted for ties was used to analyze the behavioral scores. A logit model was used to determine the effect of time and concentration on behavior. A value of p < 0.05 was considered significant. Results Volume of distribution at steady state after IV administration of butorphanol was 1.27 ± 0.73 L kg^?1, and Cl_TB was 0.0096 ± 0.0024 L kg^?1 minute^?1. Extrapolated C₀ of butorphanol after IV administration was 146.5 ± 49.8 ng mL^?1. Estimated C_MAX after IM administration of butorphanol was 54.98 ± 14.60 ng mL^?1, and T_MAX was 16.2 ± 5.2 minutes; bioavailability was 82 ± 41%. Elimination half‐life of butorphanol was 1.87 ± 1.49 and 2.75 ± 1.93 hours for IV and IM administration, respectively. Goats became hyperactive after butorphanol administration within the first 5 minutes after administration. Behavioral scores for goats were significantly different from baseline at 15 minutes after IV administration and at 15 and 30 minutes after IM administration. Both time and plasma butorphanol concentration were predictors of behavior. Behavioral scores of all goats had returned to baseline by 120 minutes after IV administration and by 240 minutes after IM administration. Conclusions and Clinical Relevance The dose of butorphanol (0.1 mg kg^?1, IV or IM) being used clinically to treat postoperative pain in goats has an elimination half‐life of 1.87 and 2.75 hours, respectively. Nonpainful goats become transiently excited after IV and IM administration of butorphanol. Clinical trials to validate the efficacy of butorphanol as an analgesic in goats are needed.     

Modulatory effects of two levels of dietary Alliums on immune response and certain immunological variables,following immunization,in White Leghorn chickens

This study aimed at investigating the effects of dietary Allium sativum (garlic, G) and Allium cepa (onion, O) on immune functions in White Leghorn chicken. One‐week‐old chicks, were fed diets without (control) or with Alliums (GL and OL, 10 g or GH and OH, 30 g/kg diet). Chickens were immunized with Newcastle disease virus (NDV), sheep red blood cells (SRBC) and Brucella abortus (BA). Antibodies, lymphocyte proliferation, and ratios of CD4⁺, CD8⁺ and CD4^‐CD8^‐ lymphocytes were investigated. Histology and weights of the spleen, thymus and bursa (BF), and white blood cell (WBC) counts were studied as well. Alliums at 10 g/kg diet enhanced anti‐NDV, anti‐SRBC and anti‐BA antibody productions, whereas 30 g/kg diet had less stimulatory effects. Histology of the lymphoid organs and proliferation of peripheral blood lymphocytes (PBL) were not influenced. However, splenocyte and thymocyte proliferations were augmented with garlic. Flow cytometry analysis showed reduction in CD4⁺ and increase in CD4^‐CD8^‐ lymphocyte ratios in GH and OH groups. Garlic‐supplemented chickens had heavier spleen and thymus, and higher WBC counts, whereas BF weight increased with both Alliums at 30 g/kg diet. These results suggest that dietary Alliums have a potential to enhance the immune functions in White Leghorn chickens.