Cellular Immunity to <Emphasis Type="Italic">Toxoplasma gondii</Emphasis> in Congenitally Infected Newborns and Immunocompetent Infected Hosts

The aim of this study was to determine the frequency of anergy to Toxoplasma gondii in congenitally infected newborns and immunocompetent infected individuals. Specific anergy to Toxoplasma has been reported previously, especially in cases of congenital toxoplasmosis. Whole blood from 592 immunocompetent patients with suspected toxoplasmosis was cultured in the presence of soluble Toxoplasma antigen for 7 days. Activated T lymphocytes were detected by flow cytometry. In patients over 1 year of age, the percentage of soluble Toxoplasma antigen-stimulated T cells expressing the interleukin-2 receptor CD25 was higher in infected patients than in uninfected subjects (40.0±18.3% vs. 1.8±2.0%, P<0.0001). No differences were detected between seroconverters, i.e. those with recent rises in IgM and IgG antibodies, and those with acquired or congenital toxoplasmosis. Similar results were observed when congenitally infected (n=38) and uninfected (n=89) children under 1 year of age were compared (17.6±9.2% vs. 3.0±4.9%, P<0.0001). The sensitivity and specificity values of CD25 detection for diagnosis of congenital toxoplasmosis in infants were 95% and 89%, respectively, at a threshold value of 7% above control culture. The results show that specific cellular immunity is detectable in virtually all Toxoplasma-infected patients, including newborns. Detection of CD25 constitutes a simple, sensitive and specific test for diagnosis of congenital toxoplasmosis. Electronic Publication     

Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of <Emphasis Type="Italic">Toxoplasma gondii</Emphasis> expressed in the yeast <Emphasis Type="Italic">Pichia pastoris</Emphasis>

In this study, we successfully expressed a chimerical surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii in Pichia pastoris. Eighty human serum samples, including 60 from confirmed cases of toxoplasmosis, were tested against the purified recombinant SAG1/2 in Western blots. Results of Western blots targeted at Toxoplasma IgG and IgM showed that the recombinant SAG1/2 reacted with all sera from the toxoplasmosis cases but none with the Toxoplasma-negative serum samples. These results showed that the P. pastoris-derived recombinant SAG1/2 was sensitive and specific and suitable for use as antigen for detecting anti-Toxoplasma antibodies. To further investigate the immunological characteristic of the recombinant protein, the recombinant SAG1/2 was injected subcutaneously into BALB/c mice, and their serum was tested against total protein lysate of T. gondii. Mice immunized with the recombinant SAG1/2 reacted specifically with the native SAG1 and SAG2 of T. gondii. Significant proliferation of splenocytes stimulated with tachyzoite total protein lysate was observed in vaccinated BALB/c mice but not in those from negative control mice. Specific production of IFN-γ, the Th1-type cytokines, was also found in stimulated splenocytes from vaccinated mice. These results show that the chimeric protein recombinant SAG1/2 can elicit a Th1-associated protection against T. gondii infections in mice. Finally, vaccinated mice were significantly protected against lethal challenge with live T. gondii RH strain tachyzoites (P < 0.005), and their survival time increased significantly compared to the negative control.     

<Emphasis Type="Italic">Toxoplasma gondii</Emphasis> antibody profile in HIV-infected pregnant women and the risk of congenital toxoplasmosis

The purpose of this study was to investigate the antibodies to Toxoplasma gondii in human immunodeficiency virus (HIV)-infected pregnant women and to determine the association between serological profile and the risk of congenital toxoplasmosis. The study, conducted in a public maternity ward from May 2002 to April 2005, included all HIV-infected women who delivered live infants during the 36 months, and, as a control group, all HIV-negative women that delivered live infants in the first 12 months of the study. Antibodies to T. gondii were detected in 1,624 of 2,421 HIV-negative women (67%; 95% confidence interval [CI] 65–69%) and in 121 of 168 HIV-infected patients (72%; 95% CI 65–79%). A total of 547 HIV-negative and 103 HIV-infected patients were tested at delivery and had positive T. gondii-specific IgG. In HIV-negative women, the median of the specific IgG concentration was 79 (interquartile range 38–160), and in HIV-infected patients, it was 283 (interquartile range 94–704) (P < 0.001). In the group of co-infected women, the only infant with congenital toxoplasmosis was born to a mother with acute toxoplasmosis infection acquired during pregnancy who did not have a high specific IgG concentration or a positive result for specific IgM. We concluded that high T. gondii-specific IgG values were much more frequent among HIV-infected pregnant women, but it did not translate into an increased risk of maternal–fetal transmission of toxoplasmosis.     

Occurrence of antibodies anti-Neospora caninum, anti-Toxoplasma gondii, and anti-Leishmania chagasi in serum of dogs from Pará State, Amazon, Brazil

A cross-sectional study was conducted to determine the occurrence of anti-Toxoplasma gondii, anti-Neospora caninum, and anti- Leishmania chagasi antibodies in dogs of the state of Pará, Brazil. For this purpose, 129 blood samples were collected from dogs of different ages and gender. Samples of 72 dogs were collected from 39 rural properties from 19 municipalities, and 57 samples were from stray dogs, collected after captivity by the Center of Zoonosis Control from the municipality of Santarém. The sera were analyzed for anti-T. gondii and anti-N. caninum antibodies by indirect fluorescent antibody tests with cutoff values of 1:16 and 1:50, respectively. For the presence of L. chagasi antibodies, enzyme-linked immunosorbent assay was used and positive results were confirmed by immunochromatographic method using the recombinant antigen K39. Of the total of 129 dogs, 90 (69.8%) were positive for T. gondii, 16 (12.4%) for N. caninum, and 30 (23.3%) for L. chagasi. Antibodies for all three parasites were found simultaneously in seven dogs (5.4%), mostly in urban dogs (six of seven). No association was observed related to gender and location (urban or rural) of dogs and occurrence of N. caninum and T. gondii antibodies although, regarding L. chagasi, higher prevalence was found in females (P < 0.02) and in dogs from urban location (P < 0.001). From the 39 farms, in 30 (76.9%) at least one dog was positive for T. gondii or N. caninum or both. Higher occurrence of Leishmania antibodies was observed in N. caninum-negative dogs (P < 0.05).     

Prevalence and haemato-biochemical parameters of trypanosome-infected pigs at Nsukka, Nigeria

The haematological and biochemical parameters of pigs slaughtered at the Nsukka Abattoir between March and May, 2009 with trypanosome infection were evaluated. A total of 300 pigs were screened for trypanosome infection using parasitological diagnostic methods. Twelve (4%) of the pigs were found to be infected. All infections were due to Trypanosoma brucei. The infected blood samples, and an equivalent non-infected (control) blood were subjected to haematological and biochemical analysis. The packed cell volume (PCV) of the infected and non-infected pigs ranged between 14–50% (mean 27.8 ± 2.8) and 23–42 (36.25 ± 0.97) and differed significantly (P < 0.05). Infection had no significant (P > 0.05) effect on the other blood parameters. Total protein (TP), albumin (ALB) and creatine showed no significant difference (P > 0.05) whereas aspertate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), urea and total bilirubin differed significantly (P < 0.05) between the infected and non-infected pigs. All these parameters increased in the infected animals with the exception of ALP which decreased. This study confirms the preponderance of T. brucei in the pig population of Nsukka area, Nigeria. It further reveals that significant haematocrit and serum biochemical changes occurred in the infected pigs. These findings are thought to have potential public health and some diagnostic significance.     

In vivo evaluation of the improved MCMS-0102 pacemaker with a rapid pacing mode for induction of experimental heart failure in animals

The MCMS-0102 cardiac pacemaker for rapid ventricular pacing to induce heart failure in animals has been improved in terms of miniaturization and performance. To determine the performance of the new MCMS-0102, six devices were implanted in beagle dogs, and two of these devices were reimplanted for continued pacing in a total of eight beagle dogs. The hearts were paced at 260 beats per minute for 4 weeks (P group: n = 8). The hemodynamic status of the P group was examined and compared with nonpaced dogs (NP group: n = 8). The neurohumoral status of the P group was evaluated before and after rapid pacing. Stable operation of the six devices during rapid pacing was confirmed using the telemetry system. Postmortem examinations revealed features similar to clinical heart failure characterized by massive ascites, pleural effusion, cardiomegaly, and liver congestion in all the paced dogs. Cardiac output was 1.1 ± 0.2 l/min in the NP group and 0.5 ± 0.1 l/min in the P group (P < 0.0001). The left atrial pressure and the central venous pressure of the P group and the NP group were 23 ± 6 versus 6 ± 2 mmHg (P < 0.0001) and 10 ± 3 versus 4 ± 3 mmHg (P < 0.001), respectively. In the paced dogs, plasma renin activity increased from 0.5 ± 0.4 to 8.5 ± 7.4 ng/ml/h (P < 0.05) and atrial natriuretic peptide levels increased from 69 ± 41 to 229 ± 72 pg/ml (P < 0.001). The improved MCMS-0102 was successfully implanted in beagle dogs and it succeeded in inducing the congestive heart failure model.     

Detection of <Emphasis Type="Italic">Toxoplasma gondii</Emphasis> in water by an immunomagnetic separation method targeting the sporocysts

An immunomagnetic separation (IMS) method was developed to detect Toxoplasma gondii in fresh waters by using the monoclonal antibody 4B6 targeting the sporocyst wall of T. gondii, Hammondia hammondi, Hammondia heydorni, and Neospora caninum. Water concentrates obtained by filtering 10- to 20-l samples samples were spiked with Toxoplasma oocysts, sonicated to release the sporocysts, and analyzed by IMS-4B6. Mean sporocyst recoveries were 74.5 ± 5.3% in drinking water, 30.6 ± 2.4 and 37.1 ± 3.2% in surface waters, and 81.6 ± 2.1% in IMS buffer. Then, this IMS method was integrated in a multistep procedure (i.e., filtration, IMS, immunofluorescence and autofluorescence) to detect Toxoplasma in unspiked and spiked water samples (10–30 l) of various qualities. Sporocyst recoveries ranged from 14.4 to 44.7% in drinking water samples spiked with 1–10 oocysts/l, and from 17.8 to 32.5% in surface water samples spiked with 10 oocysts/l. Sporocysts were not detected in 25 unspiked water samples. A sporocyst-like structure was seen in one of these unspiked samples, but its coccidian nature could not be proved by three polymerase chain reaction (PCR) methods targeting sequences of coccidian small and large subunit rRNA genes and Toxoplasma repetitive elements. In conclusion, IMS-4B6 is relevant for the detection of Toxoplasma in water generating small concentrates (<1 ml). Due to 4B6 cross-reactions, a PCR would be useful to further characterize coccidian sporocysts found microscopically.     

RNAi-mediated silencing of a novel Ascaris suum gene expression in infective larvae

In the present study, the potential of RNA interference (RNAi) as a gene silencing tool and the resultant effects on Ascaris suum larval development was examined by targeting a gene (represented by the EST 06G09) specifically expressed in the infective larvae of A. suum. BALB/c mice were infected with RNAi-treated larvae. The results showed that the target gene was silenced after soaking for 72 h, and the survival rate of the RNAi-treated larvae was reduced by 17.25% (P < 0.01). A significant difference (P < 0.05) was detected in the numbers of larvae collected from the livers and lungs of infected mice 4 days after infection with untreated larvae (164.29 ± 21.51) and RNAi-treated larvae (71.43 ± 14.35). Significant differences (P < 0.01) were also found in the body length and width between untreated larvae (480 ± 105.77 μm for length and 23.93 ± 3.72 μm for width) and RNAi-treated larvae (400.57 ± 71.31 μm for length and 20.20 ± 2.43 μm for width). These results show that the gene represented by EST 06G09 may play a role in the development of A. suum larvae.     

Seroprevalence of and risk factors for Toxoplasma gondii antibodies among asymptomatic blood donors in Egypt

A cross-sectional study was conducted to evaluate the seroprevalence of and risk factors for Toxoplasma gondii antibodies in 260 blood donors seen at blood banks in Mansoura University Hospital, Egypt. Blood donors were interviewed about sociodemographic characteristics and risk factors for T. gondii infection. A blood sample was taken to document their T. gondii antibody status using enzyme-linked immunosorbent assay. Overall, 155 (59.6%) of 260 blood donors were positive for anti-T. gondii IgG antibodies. Multivariate logistic regression analysis showed a significant association between T. gondii seropositivity and eating meat by-products (luncheon/shawerma) (adjusted odds ratio [OR] 80.82 [95% CI 18.62–350.81], P < 0.0001) or being non-educated (adjusted OR 32.25 [95% CI 7.46–139.44], P < 0.0001). These findings highlight that T. gondii is prevalent among blood donors in Egypt.     

Detection of toxoplasma antibodies in sera of Salmonidae by ELISA

Toxoplasma gondii is a protozoan parasite with a worldwide distribution. It is capable of infecting all warm-blooded animals. Toxoplasmosis was not considered a waterborne zoonoses, but recently, it has been reported in many marine mammals. Coastal pollution by sewage from humans and pets has been suggested as a source for toxoplasma infection in these animals. Recent reports of toxoplasmosis in marine mammals raise concern that cold-blooded marine animals are potential sources of T. gondii infection. Conversely, the increasing proclivity for eating fish, crabs, shrimp, and mollusks—raw, undercooked, smoked, or dried—facilitates zoonoses infections caused by protozoan microorganisms; and one of them is toxoplasma. Detection of antibodies against T. gondii can be achieved by different serological tests such as enzyme-linked immunosorbent assay (ELISA). To determine whether toxoplasmosis has a role in Salmonidae infection, which is the most common seafood in Shahrekord district, this research was carried out on 50 Salmonidae aged 4 months (weight 700 ± 200 g). ELISA was performed on serum samples for detecting T. gondii-specific immunoglobulin M (IgM) and immunoglobulin G (IgG). As a result, toxoplasmic IgM antibody was detected in five of 50 samples (cut-off value of ≥0.183). Based on these findings, we believe that Salmonidae may be susceptible to primary T. gondii infection. While there is still no evidence of T. gondii transmission from cold-blooded sea animals to human via consuming their meat or other products, further research can be done to prove the possibility of this hypothesis.     

Transplacental transmission of Toxoplasma gondii in reinfected pregnant female canines

Twelve pregnant female canines, naturally infected with Toxoplasma gondii, were reinfected with T. gondii: three (GI) received tachyzoites subcutaneously (1.0 × 107), three (GII) were orally inoculated with oocysts (1.5 × 104), and six (GIII) were kept as a nonreinfected control group. All the reinfected female canines (GI and GII) miscarried or presented fetal death, while only one GIII female presented a stillborn in a litter of four pups (P < 0.01). Fever, lymphoadenopathy, miscarriage, and fetal death were the main clinical alterations observed. The highest serological titers detected through the indirect fluorescence antibody test (IFAT) were 1,024 (GI) and 4,096 (GII). In group III, the titers ranged between 64 and 256. By bioassays in mice, T. gondii was isolated in 17 organs of the reinfected adult canines, in 11 of the control group, and in 20 of the neonates. Positive immunostaining of cysts and/or tachyzoites were observed in 26 canine tissues (14 from GI and GII and ten from GIII). The agent was detected by immunohistochemistry in the encephalon of a neonate and in the spinal cord of a stillborn, thus, confirming that T. gondii infected canine fetuses, provoking miscarriages, even in bitches that presented primoinfection.     

Effects of artemether, artesunate and dihydroartemisinin administered orally at multiple doses or combination in treatment of mice infected with Schistosoma japonicum

Artemether and artesunate, derivatives of the antimalarial artemisinin, as well as their main metabolite, dihydroartemisinin, all exhibit antischistosomal activities. The purpose of the current study was to compare the effects of artemether, artesunate and dihydroartemisinin administered orally at multiple doses or combination in treatment of mice infected with Schistosoma japonicum. We carried out experiments with mice, infected with 40 cercariae of S. japonicum, and treated with artemether, aretesunate and dihydroartemisinin (all at a single dose of 300 mg/kg, and the dose of the mixed three drugs is also 300 mg/kg) at multiple doses or combination therapy on days 6–8 or 34–36 post-infection. Administration with artemether, artesunate or dihydroartemisinin for 3 successive days reduced total worm burdens by 79.5−86% (30.86 ± 4.98 of mean total worm burden in control), female worm burdens by 79.4−86.7% (11.29 ± 2.63 of mean female worm burden in control) (all P values <0.01 vs. control), depending on different treatment protocols given on days 6–8 post-infection. However, no differences were seen between each treatment group (all P > 0.05). While the same treatment was given on days 34–36 post-infection, total worm burden reductions of 73.8−75.8% were achieved (29.44 ± 3.36 of mean total worm burden in control), which were significant when compared with the untreated control group (all P values <0.01). In all different treatment groups, female worm reductions (ranging from 88.7% to 93.1%, while the mean female worm burden in control is 10.33 ± 1.80) were consistently higher than the total worm reductions, resulting always in significantly lower female worm burdens when compared to the corresponding control (all P values < 0.01). However, there were no significant differences found between each treatment group (all P values >0.05). It is concluded that artemether, artesunate and dihydroartemisinin can be used to control schistosomiasis japonica, as a strategy to prevent S. japonicum infection. Administration with artemether, artesunate and dihydroartemisinin at multiple doses or in combined treatment damages both juvenile and adult S. japonicum, without statistically significant differences among the three drugs at the same dose.     

Serodiagnosis of experimental sparganum infections of mice and human sparganosis by ELISA using ES antigens of Spirometra mansoni spargana

We conducted a study of serodiagnosis of experimental sparganum infections of mice and human sparganosis by enzyme-linked immunosorbent assay (ELISA) using excretory–secretory (ES) antigens of Spirometra mansoni spargana and compared the sensitivity and specificity of crude and ES antigens for detecting the specific anti-sparganum IgG antibodies. By crude antigen ELISA and ES antigen ELISA, anti-sparganum IgG was detected in all of 30 serum samples of the infected mice; no cross-reactions were observed in serum samples of the mice infected with Trichinella spiralis, Schistosoma japanicum, Toxoplasma gondii, and normal mice. Anti-sparganum IgG was detected by ES antigen ELISA in sera of mice infected with one, two, four, six, and eight spargana at 3 weeks post-infection (wpi), with a detection rate of 100%, and lasted to 18 wpi when the experiment was ended. The difference in anti-sparganum antibody levels among five groups of the infected mice was statistically significant (F = 245.296, p < 0.05); the antibody levels were correlated with infecting doses of spargana (r = 0.323, p < 0.05). The sensitivity of both ELISA in detecting the serum samples of patients with sparganosis was 100% (20/20), but 96.72% (59/61) of specificity of ES antigen ELISA in detecting serum samples of patients with cysticercosis, echinococcosis, paragonimiosis, clonorchiosis, and schistosomiasis, and healthy persons was significantly greater than 72.13% (44/61) of crude antigen ELISA (χ ² = 14.027, p < 0.05). Our finding indicates that ELISA using ES antigens of S. mansoni spargana may be applied to the specific early serodiagnosis of sparganosis.     

Neuromuscular adaptation during prolonged strength training, detraining and re-strength-training in middle-aged and elderly people

Effects of a 24-week strength training performed twice weekly (24 ST) (combined with explosive exercises) followed by either a 3-week detraining (3 DT) and a 21-week re-strength-training (21 RST) (experiment A) or by a 24-week detraining (24 DT) (experiment B) on neural activation of the agonist and antagonist leg extensors, muscle cross-sectional area (CSA) of the quadriceps femoris, maximal isometric and one repetition maximum (1-RM) strength and jumping (J) and walking (W) performances were examined. A group of middle-aged (M, 37–44 years, n=12) and elderly (E, 62–77, n=10) and another group of M (35–45, n=7) and E (63–78, n=7) served as subjects. In experiment A, the 1-RM increased substantially during 24 ST in M (27%, P < 0.001) and E (29%, P < 0.001) and in experiment B in M (29%, P < 0.001) and E (23%, P < 0.01). During 21 RST the 1-RM was increased by 5% at week 48 (P < 0.01) in M and 3% at week 41 in E (n.s., but P < 0.05 at week 34). In experiment A the integrated electromyogram (IEMG) of the vastus muscles in the 1-RM increased during 24 ST in both M (P < 0.05) and E (P < 0.001) and during 21 RST in M for the right (P < 0.05) and in E for both legs (P < 0.05). The biceps femoris co-activation during the 1-RM leg extension decreased during the first 8-week training in M (from 29 ± 5% to 25 ± 3%, n.s.) and especially in E (from 41 ± 11% to 32 ± 9%, P < 0.05). The CSA increased by 7% in M (P < 0.05) and by 7% in E (P < 0.001), and by 7% (n.s.) in M and by 3% in E (n.s.) during 24 ST periods. Increases of 18% (P < 0.001) and 12% (P < 0.05) in M and 22% (P < 0.001) and 26% (P < 0.05) in E occurred in J. W speed increased (P < 0.05) in both age groups. The only decrease during 3 DT was in maximal isometric force in M by 6% (P < 0.05) and by 4% (n.s.) in E. During 24 DT the CSA decreased in both age groups (P < 0.01), the 1-RM decreased by 6% (P < 0.05) in M and by 4% (P < 0.05) in E and isometric force by 12% (P < 0.001) in M and by 9% (P < 0.05) in E, respectively, while J and W remained unaltered. The strength gains were accompanied by increased maximal voluntary neural activation of the agonists in both age groups with reduced antagonist co-activation in the elderly during the initial training phases. Neural adaptation seemed to play a greater role than muscle hypertrophy. Short-term detraining led to only minor changes, while prolonged detraining resulted in muscle atrophy and decreased voluntary strength, but explosive jumping and walking actions in both age groups appeared to remain elevated for quite a long time by compensatory types of physical activities when performed on a regular basis. Accepted: 2 May 2000     

Eosinophil Cationic Protein Stimulates TGF-β<Subscript>1</Subscript> Release by Human Lung Fibroblasts In Vitro

Eosinophilic inflammation and airway remodeling are features of asthma. Eosinophil cationic protein (ECP) is released by activated eosinophils and transforming growth factor (TGF)-β₁ has major functions in the fibrotic process. We therefore hypothesized that ECP stimulates TGF-β₁ release by human lung fibroblasts. Fibroblasts in monolayer displayed a constitutive release of TGF-β₁, which increased in presence of ECP (436 ± 60 vs. 365 ± 48 pg/ml at 48 h; P < 0.01). mRNA expression of TGF-β₁ was almost twofold in ECP-stimulated fibroblasts. ECP in three-dimensional cultures stimulated both TGF-β₁ release (180 ± 61 vs. 137 ± 54 pg/ml; P < 0.01) and fibroblast-mediated collagen gel contraction (28 vs. 39% of initial gel area at 48 h; P < 0.001). ECP stimulates TGF-β₁-release by human lung fibroblasts, suggesting a potential mechanism for eosinophils in the fibrotic response. This may be an important mechanism by which ECP promotes remodeling of extra cellular matrix leading to airway fibrosis in asthmatics.     

Restitution analysis of alternans and its relationship to arrhythmogenicity in hypokalaemic Langendorff-perfused murine hearts

Alternans and arrhythmogenicity were studied in hypokalaemic (3.0 mM K⁺) Langendorff-perfused murine hearts paced at high rates. Epicardial and endocardial monophasic action potentials were recorded and durations quantified at 90% repolarization. Alternans and arrhythmia occurred in hypokalaemic, but not normokalaemic (5.2 mM K⁺) hearts (P < 0.01): this was prevented by treatment with lidocaine (10 μM, P < 0.01). Fourier analysis then confirmed transition from monomorphic to polymorphic waveforms for the first time in the murine heart. Alternans and arrhythmia were associated with increases in the slopes of restitution curves, obtained for the first time in the murine heart, while the anti-arrhythmic effect of lidocaine was associated with decreased slopes. Thus, hypokalaemia significantly increased (P < 0.05) maximal gradients (from 0.55 ± 0.14 to 2.35 ± 0.67 in the epicardium and from 0.67 ± 0.13 to 1.87 ± 0.28 in the endocardium) and critical diastolic intervals (DIs) at which gradients equalled unity (from −2.14 ± 0.52 ms to 50.93 ± 14.45 ms in the epicardium and from 8.14 ± 1.49 ms to 44.64 ± 5 ms in the endocardium). While treatment of normokalaemic hearts with lidocaine had no significant effect (P > 0.05) on either maximal gradients (0.78 ± 0.27 in the epicardium and 0.83 ± 0.45 in the endocardium) or critical DIs (6.06 ± 2.10 ms and 7.04 ± 3.82 ms in the endocardium), treatment of hypokalaemic hearts with lidocaine reduced (P < 0.05) both these parameters (1.05 ± 0.30 in the epicardium and 0.89 ± 0.36 in the endocardium and 30.38 ± 8.88 ms in the epicardium and 31.65 ± 4.78 ms in the endocardium, respectively). We thus demonstrate that alternans contributes a dynamic component to arrhythmic substrate during hypokalaemia, that restitution may furnish an underlying mechanism and that these phenomena are abolished by lidocaine, both recapitulating and clarifying clinical findings.     

Haematology and serum biochemistry of golden eagle (Aquila chrysaetos) in Iran

Haematological and serum biochemical values were estimated in blood samples collected from 21 apparently adult golden eagles (Aquila chrysaetos) of both sexes. The mean values of red blood cells, packed cell volume, haemoglobin, white blood cells, heterophils, lymphocytes, monocytes and eosinophils were 1.63 ± 0.11 × 10¹²/l, 0.47 ± 0.009 l/l, 91.73 ± 1.52 g/l, 24.31 ± 1.97 × 10⁹/l, 4.40 ± 0.22 × 10⁹/l, 16.81 ± 0.65 × 10⁹/l, 0.99 ± 0.19 × 10⁹/l and 2.10 ± 0.30 × 10⁹/l, respectively. The leucocytes had 69.14%, 4.09%, 18.12% and 8.65% lymphocytes, monocytes, heterophils and eosinophils, respectively. The results of serum biochemistry in the golden eagle indicated that the concentrations of glucose, total protein, albumin, total globulin, cholesterol, triglyceride, uric acid, blood urea nitrogen, creatinine, calcium, phosphorous, aspartate aminotransferase, alanine aminotransferase, creatine kinase, lactate dehydrogenase and alkaline phosphatase were 16.42 ± 0.73 mmol/l, 49.76 ± 1.35 g/l, 20.46 ± 0.79 g/l, 29.30 ± 1.47 g/l, 2.14 ± 0.09 mmol/l, 2.04 ± 0.08 mmol/l, 457.67 ± 97.46 μmol/l, 2.74 ± 0.17 mmol/l, 53.27 ± 3.87 μmol/l, 2.37 ± 0.24 mmol/l, 1.73 ± 0.08 mmol/l, 293.24 ± 18.96 IU/l, 28.21 ± 2.36 IU/l, 411.29 ± 58.37 IU/l, 1,209.89 ± 21.73 IU/l and 67.31 ± 5.29 IU/l, respectively. There were no significant differences between haematological and serum biochemical parameters of male and female golden eagles (P > 0.05).     

Inducible nitric oxide synthase inhibition influenced granuloma formation with suppressed collagen expression in myositis caused by <Emphasis Type="Italic">Toxocara canis</Emphasis> in mice

The role of nitric oxide (NO) in granuloma pathology is largely unclear to date. We investigated the role of NO in fibrotic granuloma development in the musculature of mice infected with Toxocara canis from 1 day (dpi) to 8 weeks post-infection (wpi) using the NO synthase (NOS) inhibitors, L-NIL (l-N ⁶-1-iminoethyl lysine). In infected mice, elevated serum NO concentrations were seen at 1 dpi (204.1 ± 0.2 μM) and 1 wpi (145.1 ± 0.2 μM); it declined drastically from 4 wpi onwards (57.0 ± 0.1 μM). In L-NIL-treated mice, the NO concentration was drastically reduced from 15% during 1 wpi; thereafter, it was restored to almost half that in infected mice. Inducible NOS expression was enhanced in infected and L-NIL-treated mice at 4 wpi but declined at 8 wpi as assessed by immunohistochemistry. L-NIL treatment resulted in large, irregularly shaped granulomas with suppressed collagen contents at 4 wpi but not at 8 wpi. The suppressed collagen contents might have been related to decreased serum NO and Th2-type cytokine of interleukin-4 but not Th1-type cytokine of interferon-γ expression.     

Haematological and clinical chemistry findings associated with sub-acute contamination of drinking water with varied low percentages of used engine oil

This study evaluated the haematology and clinical chemistry profile of rats given drinking water contaminated with varied low percentages of used engine oil (UEO) for a period of 21 days. Fifty female albino rats of 6–7 weeks of age were used for the study. They were divided into five groups (A–E) and given water contaminated with 5%, 1%, 0.1%, 0.01% and 0% vol/vol. of UEO respectively as the only source of drinking water for 21 days. The group E given uncontaminated water (0% contamination) served as the control. The haematological parameters and clinical chemistry profile of the rats was comprehensively evaluated after the 21 days of administration of the group-specific waters. Results showed that contamination of water with up to 5% UEO led to no significant effects (p > 0.05) on all the haematological indices and on the levels of serum alanine amino transferase, aspartate amino transferase, albumin, creatinine and calcium, blood urea nitrogen and fasting blood glucose level, feed consumption and body weight. However, the rat group given water contaminated with 5% UEO had a significantly increased serum alkaline phosphatase (AP) (p < 0.01), total bilirubin (p < 0.05) and cholesterol (p < 0.01), and a significantly decreased serum total protein and globulin (p < 0.01), and water consumption (p < 0.05). The rat group given water contaminated with 1% UEO had a significantly increased serum AP (p < 0.01), total bilirubin (p < 0.05) and cholesterol (p < 0.01), and a significantly decreased water consumption (p < 0.05), while the rat group given water contaminated with 0.1% UEO had a significantly elevated (p < 0.01) serum AP. It was concluded that sub-acute contamination of drinking water of rats with up to 5% UEO led to hepato-biliary disorders and adverse effects on hepatic secretion and excretion, including diminution of serum protein and globulin levels and elevation of serum cholesterol levels, but did not lead to any significant effects on haematology, hepatocellular integrity, kidney/renal function, pancreatic function and body weight.     

The blood picture and serum biochemistry profile of the African pied crow (Corvus albus)

The crow is commonly regarded as an indicator species for the surveillance of important diseases such as West Nile fever and avian influenza, as these diseases had been associated with significant pathology in crows and death of crows in most cases. This study evaluated the blood picture (haematology) and serum biochemistry profile of apparently healthy African pied crows trapped in Nsukka, Eastern Nigeria. A total of 25 crows were used for the study, and the evaluation of the blood picture and serum biochemistry profile followed standard procedures. Results obtained for the parameters assessed are summarised as follows (mean ± standard error): packed cell volume (%)—42.85 ± 0.90, haemoglobin concentration (g/dl)—14.09 ± 0.36, red blood cell count (10⁶/ul)—3.15 ± 0.09, mean corpuscular volume (fl)—137.53 ± 4.00, mean corpuscular haemoglobin (pg)—45.36 ± 1.68, mean corpuscular haemoglobin concentration (g/dl)—33.01 ± 0.82, total white blood cell count (10³/ul)—26.04 ± 1.35, heterophil counts (%)—67.99 ± 1.85, lymphocyte counts (%)—28.52 ± 1.85, monocyte counts (%)—1.28 ± 0.21, eosinophil counts (%)—1.59 ± 0.21, basophil counts (%)—0.36 ± 0.11, alanine amino transaminase (IU/l)—51.16 ± 5.00, aspartate amino transaminase (IU/l)—101.42 ± 3.63, serum alkaline phosphatase (IU/l)—31.34 ± 3.35, total protein (g/dl)—3.13 ± 0.13, albumin (g/dl)—1.32 ± 0.08, globulin (g/dl)—1.81 ± 0.14, cholesterol (mg/dl)—165.95 ± 6.63, blood glucose (mg/dl)—295.22 ± 11.20, urea nitrogen (mg/dl)—6.71 ± 0.63, uric acid (mg/dl)—21.44 ± 3.51 and body weight (g)—453.41 ± 9.30. There were no significant differences (p > 0.05) between the sexes in all the haematological and serum biochemistry parameters assessed, but the mean body weight of the males was significantly (p < 0.01) higher than that of the females. Data generated from this study was considered important as deviations in the normal/reference blood picture/haematology and serum biochemistry profile have a predictive value for general pathological changes in the body and in some cases specific organ damage.