Eperythrozoon ovis — A blood parasite of sheep

Eleven goats given dog-derived Sarcocystis sporocysts, showed illness from day 18. Doses of 5 × 10⁶ sporocysts caused a progressive temperature rise, with peaks at 6,11 and 18 days. Necropsy at 18 and 19 days revealed multiple petechiation and schizogony in endothelial cells. In goats given smaller doses, Hb, PCV and TP levels fell from day 17. IFAT titres rose from day 28.     

The porcine acute phase protein response to acute clinical and subclinical experimental infection with Streptococcus suis

The pig acute phase protein (APP) response to experimental Streptococcus suis (S. suis) infection was mapped by the measurement of the positive APPs C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp) and major acute phase protein (pig-MAP) and the negative APPs albumin and apolipoprotein (Apo) A-I. The aim was to elucidate the differences in the acute phase behaviour of the individual APPs during a typical bacterial septicaemic infection. Pigs were inoculated subcutaneously with live S. suis serotype 2 and blood was sampled before and on various days post inoculation (p.i.), until the pigs were killed and autopsied on day 14 p.i. Clinical signs (fever and lameness) were observed in four of the five inoculated pigs from day 2 p.i., and these pigs also had arthritic lesions at autopsy. CRP and SAA showed fast increases in serum concentrations, CRP being elevated from days 1 to 12 p.i. and peaking at 10 times the day 0-levels on day 1 p.i. SAA rose quickly to peak levels of 30-40 times the day 0-level on days 1-2 and returned to pre-inoculation level on day 5 p.i. Hp and pig-MAP showed slightly slower responses, both peaking around 5 days p.i. Hp was increased throughout the experiment with maximum levels around 10 times the day 0-levels, and pig-MAP was elevated on days 1-12 p.i. with peak levels of around seven times the day 0-levels. Apo A-I was decreased from days 1 to 8 and showed minimum levels of about 40% of day 0-levels around 1-2 days p.i. No clear pattern of changes in albumin levels could be identified. One pig, showing clinical signs on day 2 only, also showed an APP response, although of a relatively short duration, whereas three pigs presenting clinical signs for several days had a more protracted acute phase response. Remarkably, the one pig showing no clinical signs and no arthritic lesions showed an APP response comparable to that of the other, clinically affected pigs. Thus, both acute clinical and subclinical S. suis infection could be revealed by the measurement of one or more of the APPs CRP, SAA, Hp, pig-MAP and Apo A-I. The combined measurement of two or three APPs, including proteins with slow and fast kinetics, should be used to achieve the highest sensitivity for the detection of ongoing S. suis infection during a prolonged time period. A diagnostic tool based on such APP-measurements could considerably improve strategic control procedures for this important infection.     

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.     

Observations on histamine, serotonin, cortisol and cortisol binding globulins in bovine anaplasmosis

Changes in histamine, serotonin, cortisol and cortisol binding globulin (CBG) levels were observed in Anaplasma marginale-infected bovine calves. In addition, packed cell volume (PCV), platelet number and percentage parasitaemia were also recorded. The whole blood histamine and serotonin values rose significantly (P less than 0.01 and P less than 0.025, respectively) during the acute stage of Anaplasma infection. Higher serum cortisol and CBG levels (P less than 0.05) were observed in acute and carrier infections, respectively. A sharp drop in thrombocyte count (59%) and PCV (33%) was also noticed in clinical anaplasmosis. The results suggest that the higher levels of biogenic amines which are known to produce increased vasodilation, capillary permeability and tissue anoxia and hypercortisolaemia to protect animals from stress and cell damage may play a similar role in the pathogenesis of acute anaplasmosis.     

Time course differential gene expression in response to porcine circovirus type 2 subclinical infection

This study was aimed at characterizing the potential differences in gene expression in piglets inoculated with Porcine circovirus type 2 (PCV2), the essential causative agent of postweaning multisystemic wasting syndrome. Seven-day-old caesarean-derived, colostrum-deprived piglets were distributed into two groups: control (n = 8) and pigs inoculated with 10^5.2 TCID₅₀ of the Burgos PCV2 isolate (n = 16). One control and three inoculated pigs were necropsied on days 1, 2, 5, and 8 post-infection (p.i.). The remaining pigs (four of each group) were sequentially bled on days 0, 7, 14, 21, and 29 p.i. (necropsy). Total RNA from the mediastinal lymph node (MLN) and lysed whole blood (LWB) samples were hybridized to Affymetrix Porcine GeneChip^®. Forty-three probes were differentially expressed (DE) in MLN samples (FDR < 0.1, fold change > 2) and were distributed into three clusters: globally down-regulated genes, and up-regulated genes at early (first week p.i.) and late (day 29 p.i.) stages of infection. In LWB samples, maximal differences were observed at day 7 p.i., with 54 probes DE between control and inoculated pigs. Main Gene Ontology biological processes assigned to up-regulated genes were related to the immune response. Six common genes were found in both types of samples, all of which belonged to the interferon signaling antiviral effector pathway. Down-regulated genes were mainly related to cell adhesion and migration in MLN, and cellular organization and biogenesis in LWB. Microarray results were validated by quantitative real-time PCR. This study provides, for the first time, the characterization of the early and late molecular events taking place in response to a subclinical PCV2 infection.     

Studies on the pathogenesis of bovine ephemeral fever in sentinel cattle. II. haematological and biochemical data

Twenty-two non-lactating dairy cattle from a sentinel herd previously described (St. George, 1985) were monitored daily during an outbreak of ephemeral fever. Nine developed clinical ephemeral fever between 25 December 1981 and 30 January 1982. There were no subclinical infections with bovine ephemeral fever virus in the group. There were, however, subclinical infections with CSIRO Village, Akabane, Aino, Tinaroo and Kimberley viruses as described by St. George et al. (1984). Six of the nine affected cattle showed a neutrophilia with a concurrent lymphopaenia on the day of pyrexia; however, the differential white cell profile had begun to change up to 24 h prior to leucocytosis. Serum carboxypeptidase values fell by 24 h following the febrile response. Plasma fibrinogen rose rapidly in all six cows. The peak concentration (15.6 ± 2.70 g l⁻¹) occurred 3 days after pyrexia with the highest individual increase being from 6.05 to 19.6 g l⁻¹. Plasma fibrinogen levels remained elevated for at least 7 days.

Serum calcium fell significantly during Day 1 of the disease, the mean decline being 0.22 ± 0.08 mmol l⁻¹. The greatest individual fall was from 2.33 to 1.92 mmol l⁻¹. None of the affected cattle showed any compensatory change in serum magnesium. There was no change in the normal values of creatinine, urea, γ-GT, AST and alkaline phosphatase. Bovine ephemeral fever virus was isolated from only four of the six cases, whereas specific antibody was detected in all cattle 3–4 days after recovery.     

Pathogenesis of bovine viral diarrhoea-mucosal disease (BVD-MD) virus infection in tracheal organ cultures

Bovine foetal tracheal organ cultures were infected with two strains of BVD-MD virus and observed for 35 days. The effect of the virus was assessed by observing ciliary activity in gross tissue explants and histological changes in haematoxylin and eosin stained sections. Decrease in ciliary activity and mild epithelial degeneration were first observed at 4 days post infection (p.i.), the epithelial degeneration progressing to complete destruction by day 35 p.i. Viral titres in extracellular fluids rose sharply from day 4 p.i., reached peak levels (10⁵ TCID₅₀/ml) between 15 and 18 days p.i., and eventually declined but persisted at lower titres up to day 35 p.i., when observations were terminated.     

Relative resistance of Dorper crossbred ewes to gastrointestinal nematode infection compared with St. Croix and Katahdin ewes in the southeastern United States

A study was conducted to evaluate relative resistance of Dorper crossbred (DO), Katahdin (KA), St. Croix (SC), and Hampshire (HA) ewes to natural and experimental gastro-intestinal (GI) nematode infection over a 20-month period. The objective of Experiment 1 was to evaluate breeds for resistance to infection acquired naturally from mixed grass pastures. In Year 1 (May-December 2000) de-worming of ewes occurred during wet, hot conditions in July and during late pregnancy in December. In Year 2 (January-December 2001), ewes were de-wormed after fecal egg count (FEC) for a breed group rose above 1000 eggs per gram (epg) or after blood packed cell volume (PCV) of an individual ewe fell below 20. FEC was determined every 28 days and PCV every 14-28 days. In both the years, ewes were pastured together, except during the 28-days breeding periods, on tall fescue, bermudagrass, or ryegrass, and rotated among pastures dependent on forage availability. Ewes were in good or excellent condition (body condition score of 3-4 out of 5) throughout the study. The objective of Experiment 2 was to evaluate the breeds for relative resistance to an experimental infection with Haemonchus contortus infective larvae. Both PCV and FEC were determined every 7 days from 14 to 42 days after inoculation with 30000 infective larvae per ewe. In Experiment 1, Year 1, FEC was slightly greater and PCV was lower from July to September in DO ewes (breed x time, P<0.001). In Year 2, de-worming occurred 14 days later in DO ewes compared with other breed types. Otherwise PCV and FEC were similar among the hair breeds and higher and lower, respectively, compared with HA ewes (breed x time, P<0.001). In Experiment 2, FEC and PCV were similar among hair breeds; FEC was lower and PCV higher in hair breeds compared with that of HA ewes (P<0.01). Relative resistance of mature Dorper crossbred ewes was comparable to that of Katahdin and St. Croix ewes and superior to that of Hampshire ewes.     

Effects of induced fescue toxicosis on plasma and tissue catecholamine concentrations in sheep

Sixteen crossbred wethers were distributed among four treatments and fed a control ration based on annual rye-orchardgrass (R-O) for 8 days. Indwelling jugular cannulae were installed and experimental regimes begun the following day (experimental day 1). One-half of the wethers were fed a ration based on endophyte-infected Kentucky-31 fescue while the remainder continued to receive the R-O control diet for 10 days. Spiperone, a dopamine antogonist, was administered to one-half of the wethers receiving each ration on days 8 and 9. Plasma prolactin (PRL), dopamine (DA), norepinephrine (NE) and epinephrine (E) were measured in jugular venous blood on days 1, 3, 5 and 7-10 of the trial. On day 10, the animals were decapitated; and DA, NE, E and 3,4-dihydroxyphenylacetic acid (DOPAC) and monoamine oxidase (MAO) were determined in hypothalamic and pituitary tissue. Plasma DA was elevated (P less than .05) following day 8 in wethers fed infected fescue over those fed (R-O), while plasma PRL was reduced (P = .08). Wethers receiving Spiperone had lowered (P less than .05) plasma DA and elevated (P less than .01) plasma PRL. Plasma DA was negatively correlated (P less than .01) with plasma PRL (r = -0.50) following day 8. Plasma NE and E levels and NE, E, DA and DOPAC tissue concentrations were not affected by Spiperone administration or diet. MAO levels in pituitaries were higher (P less than .01) for Spiperone-treated wethers. Wethers receiving the toxic fescue ration exhibited elevated plasma DA concentrations which was associated with depressed prolactin secretion in wethers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Peritoneal Fluid Analysis in Ponies after Abdominal Surgery

Ten student surgery ponies were subjected to exploratory laparotomy. Abdominal paracentesis was performed preoperatively and daily postoperatively for 6 days, then the ponies were euthanatized and necropsied. Initial baseline peritoneal fluid parameters were within established reference limits. Postoperatively, the total leukocyte count and total protein in the peritoneal fluid rose and remained elevated for the 6 days of the study. Complete blood counts (CBCs) were performed preoperatively and on days 1 and 4 postoperatively. On day 1, a stress leukogram with a mild inflammatory component developed, but by day 4, the CBCs were within normal limits. The mean plasma fibrinogen levels, which were determined daily, peaked on day 4.     

Plasma histamine concentration and histamine detection in peripheral blood eosinophils in cats

Plasma histamine levels were measured in 11 clinically healthy cats and 15 cats with allergic dermatitis. Histamine levels were markedly elevated in 5/15 allergic cats. A calcium ionophore, A23187, stimulates histamine release from feline peripheral blood cells. Immunostaining of blood smears from clinically healthy cats revealed that approximately 10% of eosinophils possessed histamine-containing granules. These results indicate that some peripheral eosinophils in cats contain histamine and can release histamine by appropriate stimulation.     

The effects of drug detection training on behavioral reactivity and blood neurotransmitter levels in drug detection dogs: A preliminary study

The aim of the present study was to analyze the effects of drug detection training on behavior and blood neurotransmitter levels in drug detection dogs so as to investigate some variables influencing dog reactivity and responsiveness to training. In all, 20 dogs were sampled out of the Guardia di Finanza canine population. All the subjects were born, reared, housed, and trained in the same facility and followed the same training sessions. Dogs’ behavioral reactivity was scored according to a standardized working dogs test to evaluate natural dog attitudes. Plasma samples were analyzed by the high-performance liquid chromatography method to evaluate adrenaline, noradrenaline, L-3,4-dihydroxyphenylalanine, homovanillic acid (HVA), 3-methoxy-4-hydroxyphenylglycol acid (MHPG), 5-hydroxyindole acetic acid (5-HIAA), and 5-hydroxytryptamine (5-HT) levels. 5-HT and 5-HIAA were also analyzed from platelets. The analysis was carried out considering training, breed, and sex as independent variables. From a behavioral point of view, significant differences were recorded before and after training in “sociability,” “playfulness,” “predatory instinct,” and “aggressiveness” scores. Lower levels of platelet 5-HT and 5-HIAA were found after training. Plasma L-3,4-dihydroxyphenylalanine levels differed between sexes, with males showing higher concentrations. These results underline the importance of complete and objective evaluations protocols of the dogs before, during, and after drugs search training to determine effective and successful selection strategies and training procedures.     

Prostaglandin F2 alpha and prolactin in experimental hypogalactia in sows

Peripheral plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F2alpha(PGFM), progesterone, prolactin and oestrone were determined in 20 sows for two days before and three weeks after parturition. Groups of four sows each received one of the following five treatments post partum: 30 ml sterile 0.9 per cent saline solution intrauterinely; ovariectomy and 30 ml saline solution intrauterinely; 10 ml Lugol's iodine plus 20 ml saline solution intrauterinely; ovariectomy and 10 ml Lugol's iodine plus 20 ml saline solution intrauterinely, or progesterone (0.5 mg [kg bodyweight]-1 intramuscularly). Saline solution and iodine were administered every 48 hours, starting immediately after parturition, for one week. Ovariectomy was performed within eight hours of delivery. Progesterone was given every third day for 12 days. Piglet weight gains were used as a reflection of milk yield. In all sows, oestrone values were elevated before parturition, but fell by the end of delivery and were very low during lactation. PGFM concentrations rose during the last two days of pregnancy to reach maximal values at the time of delivery. Plasma progesterone levels declined concomitantly with the rise in PGFM values before parturition. Basal values of progesterone were achieved within 24 hours after delivery in control sows receiving saline treatment. Progesterone values fell immediately after ovariectomy in sows receiving saline or iodine treatment but were slightly elevated for one week in sows that received only intrauterine iodine treatment, suggesting that complete regression of corpora lutea is prevented by suppression of post parturient uterine prostaglandin production. Sows injected with progesterone maintained plasma values of about 5 to 15 nmol litre-1.(ABSTRACT TRUNCATED AT 250 WORDS)     

Studies on the pathogenesis of bovine ephemeral fever in experimental cattle III. Virological and biochemical data

In an attempt to define the nature of the response of cattle to ephemeral fever infection, a number of indicators of inflammation were monitored during clinical disease. The total Ca, Zn, Fe, Cu, glucose and phosphate in plasma, together with blood ammonia, were assayed relative to changes in the rectal temperature. Ca_T levels fluctuated markedly and hypocalcaemia occurred in 4 of 8 cattle. Plasma Zn and Fe values fell while plasma Cu levels rose markedly in all cattle. Mean levels of serum NH₃ of 20–30 μmol 1⁻¹ rose to a peak value of 56 μmoll⁻¹. Plasma glucose levels rose to a peak of 4.6 ± 0.5 mMl⁻¹ and the plasma phosphate levels fell from 2.4 ± 0.1 mMl⁻¹ to 1.17 ± 0.2 mMl⁻¹ during fever. Values of pCO₂ fell from a mean of 46.9 ± 3.6 mmHg to 36.4 ± 3.1 mmHg and coincided with a rise in pH. Virus was isolated 73 h (± 23) after inoculation and persisted until 130 h (± 21). The common role of these parameters in generalised inflammation and ephemeral fever is discussed.     

Immunolocalization and pathological alterations following Strongyloides venezuelensis infection in the lungs and the intestine of MHC class I or II deficient mice

The present study, investigated the mechanisms involved in the immune responses of Major Histocompatibility Complex class I or class II knockout mice, following Strongyloides venezuelensis infection. Wild-type C57BL/6 (WT), MHC II(-/-) and MHC I(-/-) mice were individually inoculated with 3000 larvae (L3) of S. venezuelensis and sacrificed on days 1, 3, 5, 8, 13 and 21 post-infection (p.i.). Samples of blood, lungs and small intestines were collected. The tissue samples were stained with hematoxylin-eosin for the pathological analysis. The presence of the parasite was demonstrated by immunoperoxidase analysis. MHC II(-/-) mice presented a significantly higher number of adult worms recovered from the small intestine on day 5p.i. and presented elevated numbers of eggs in the feces. The infection by S. venezuelensis was completely eliminated 13 days after infection in WT as well as in MHC I(-/-) mice. In MHC II(-/-) mice, eggs and adult worms were still found on day 21 p.i., however, there was a significant reduction in their numbers. In the lung, the parasite was observed in MHC I(-/-) on day 1 p.i. and in MHC II(-/-) mice on days 1 and 5 p.i. In the small intestine of WT mice, a larger number of parasites were observed on day 8 p.i. and their absence was observed after day 13 p.i. Through immunohistochemistry analysis, the parasite was detected in the duodenum of WT on days 5 and 8 p.i., and in knockout mice on days 5, 8 and 13 p.i.; as well as in posterior portions of the small intestine in MHC I(-/-) and MHC II(-/-) on day 13 p.i., a finding which was not observed in WT mice. We concluded that immunohistochemistry analysis contributed to a more adequate understanding of the parasite localization in immunodeficient hosts and that the findings aid in the interpretation of immunopathogenesis in Strongyloides infection.     

Detection of Porcine circovirus type 2 viremia and seroconversion in naturally infected pigs in a farrow-to-finish barn

Porcine serum was assayed by 2 polymerase chain reaction (PCR) protocols (nested PCR [nPCR] and non-nested PCR) and a competitive enzyme-linked immunosorbent assay to determine when Porcine circovirus type 2 (PCV2) viremia and a rise in the serum level of PCV2-specific antibody occurred in pigs raised in a large Canadian farrow-to-finish barn. Eight serial blood samples were collected from each of 40 pigs from 5 to 156 (+/- 1.5) d of age; 6 pigs were removed from the study for various reasons at various times. Viremia was not detected in the samples collected before 72 d of age but was detected in those collected on or after 72 d: of 33 pigs, 7 (21%) had only 1 serum sample positive for PCV2 DNA by nPCR after day 72; 11 (33%) were intermittently positive by nPCR, non-nested PCR, or both between 72 and 156 d; and the remaining 15 (45%) were repeatedly positive (in 2 to 4 samples). The level of serum antibody against PCV2 declined after weaning and increased between 72 and 107 d of age, only after PCV2 was detected in serum. Our results show that PCV2 viremia persists in the presence of elevated levels of PCV2-specific antibody.     

CHANGES OF BLOOD METABOLITES IN HORSES AFTER RACING, WITH PARTICULAR REFERENCE TO URIC ACID

Changes in concentration of a number of blood metabolites in 30 thoroughbred horses were recorded after an 1110 metre race. No significant changes occurred in blood urea or aspartate aminotransferase during the three hours after racing. Plasma sodium, potassium and calcium levels were increased immediately after racing but had returned to normal one hour after racing. Plasma phosphate showed a significant fall in concentration one hour after racing. Creatinine and lactic acid concentrations were elevated ten minutes after racing and although they subsequently decreased, the level of lactic acid was still significant one hour later. Uric acid levels were well above resting levels at ten minutes after racing but rose even more in the subsequent hour. Urinary uric acid levels were also elevated during this time. Three hours after racing some horses still had elevated plasma uric acid levels and all of them showed a significant rise in creatine phosphokinase. The possible physiological basis of these findings is discussed.     

Efficacy of a single oral dose of isorsorbide 5-mononitrate in normal dogs and in dogs with congestive heart failure

Isosorbide 5-mononitrate (5-ISMN) was evaluated in normal dogs and dogs with congestive heart failure (CHF) in a randomized, blinded, and placebo-controlled study. Equilibrium blood pool imaging was used to detect changes in regional blood volume distribution. Six normal dogs were administered placebo, 2, 3, and 4 mg/kg 5-ISMN PO on separate days with a 1-week washout period between randomized dosings. Six dogs with CHF were administered placebo or 4 mg/kg 5-ISMN on separate days with a 1-week washout period between randomized dosings. Data were collected at baseline and at 15, 30, 45, 60, 90, 120, 150, 180, 210, and 240 minutes after dosing. Measured variables included indirect arterial blood pressure (BP), heart rate (HR), packed cell volume (PCV), scintigraphic count rates for normal dogs, and scintigraphic count rates for CHF dogs. Blood for plasma 5-ISMN concentration determination was collected at 60 minutes. Scintigraphic counts were corrected for decay and expressed as a percentage of the whole. No differences were detected in BP, HR, PCV, thoracic blood volume percentage (TBVP), or abdominal blood volume percentage (ABVP) between placebo and 5-ISMN in normal dogs at any dose. No differences were detected in TBVP or ABVP between placebo and 5-ISMN in dogs with CHF Plasma 5-ISMN concentration exceeded the minimum therapeutic concentration in all dogs and at all doses 60 minutes after drug administration. Equilibrium blood pool imaging failed to detect a shift in blood volume with oral 5-ISMN administration at any dose tested in normal dogs and dogs with CHF, despite adequate drug absorption. On the basis of the results of this study, 5-ISMN may not be beneficial in the treatment of dogs with CHF.     

Changes in serum oestrone sulphate and progesterone levels of red deer hinds during pregnancy

Serum oestrone sulphate (E1S) and progesterone (P) levels were recorded from six red deer hinds throughout the first two-thirds of gestation. E1S was generally undetectable until 30 days post conception when levels rose, peaking between 60 and 80 days, and fell to lower values between 80 and 170 days post conception. P levels rose after conception and remained elevated for the remainder of the study. This study indicates that in general the presence of measurable E1S levels in serum of hinds is associated with pregnancy.     

Clinical, haematological and plasma biochemical changes in specified-pathogen-free (sporozoa) lambs experimentally infected with low numbers of Sarcocystis tenella sporocysts

Two groups of lambs raised free of sporozoan infection were inoculated with Sarcocystis tenella sporocysts and compared with controls. Lambs from Group 1 were inoculated with 5000 sporocysts and those in Group 2 were given 20,000. Transient increases in rectal temperatures occurred between 23 and 39 days post-inoculation (dpi), although the lambs appeared normal and retained their appetites. Packed cell volumes (PCV) of lambs given 20,000 sporocysts decreased dramatically from 28 to 38 dpi after which they slowly returned to near pre-inoculation levels by 99 dpi. The anaemia was normocytic/normochromic. White cell counts (WCC) rose in infected lambs from 49 dpi, reflecting principally an increase in lymphocyte numbers. Plasma albumin of Group 2 decreased at 28 dpi and remained depressed until the experiment was terminated at 99 dpi. Plasma globulin of infected groups increased from 31 (Group 2) and 35 dpi (Group 1). Plasma alkaline phosphatase (ALP) of Group 2 decreased from 28 dpi and remained depressed to 99 dpi. Lactate dehydrogenase (LDH) of Group 2 was elevated at 24 and 28 dpi and from 42 to 78 dpi, while aspartate aminotransferase (AST) of the same group was elevated from 45 to 66 dpi. Creatine kinase (CK) of Group 2 was elevated from 52 to 71 dpi.