Haematology, cytochemical and ultrastructural characteristics of blood cells in leopard (Panthera pardus)

The leopard (Panthera pandus) is an endangered Asian big cat found in Thailand and also listed in the CITES, Appendix 1. Blood samples from 17 leopards (six males and 11 females) were collected from the cephalic vein for haematology, cytochemical and ultrastructural studies. Red blood cells (RBCs) were slightly anisocytosis, ranged 5–6.5 μm with 5.6 μm mean diameter. They were easy to form rouleaux, blunt end crenation and some RBCs contained Heinz bodies. The male leopards had a significantly higher packed cell volume (39.3 ± 7.5%) values and absolute eosinophil number (1.658 ± 1.483 × 10⁹/l) than the females (30.7 ± 3.2%; 0.965 ± 0.611 × 10⁹/l). The cytochemical results were: sudanophilia and myeloperoxidase in neutrophils and some monocytes; nonspecific esterase (alpha-naphthyl acetate esterase, ANAE) in the granules of eosinophils, some lymphocytes, some monocytes and platelets; beta-glucuronidase (βG) in granules of basophils, monocytes and some lymphocytes. The ANAE and βG reaction were detected inter-granular of eosinophils. More detailed morphological aspects of all blood cells were observed by means of scanning electron microscope and transmission electron microscope (TEM). The many round granules and homogeneous content under TEM were characteristics of leopard eosinophils. The engulfed RBCs by neutrophils were detected under TEM in one male leopard. This study provides a guide for the haematology, identification of the morphology, cytochemistry and ultrastructure of blood cells in leopards that is useful for zoological veterinarians in leopard conservation.     

Haematological and biochemical parameters and the serum concentrations of phosphorus, lead, cadmium and chromium in flamingo (Phoenicopterus ruber) and black-headed gull (Larus ridibundus) in Iran

Blood samples of nine flamingos and 12 black-headed gulls from Fars province of Iran were used to determine the haematological and biochemical factors and the concentrations of phosphorus, lead, chromium and cadmium in serum. Haematological parameters in flamingo—packed cell volume (PCV), haemoglobin (Hb) concentration, red blood cell (RBC) number, white blood cell (WBC) count, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), MCH concentration (MCHC), heterophiles, lymphocytes and thrombocytes—were found to be 35.21±1.6 (%), 117.8±59 (g/l), 2.27±0.29 (×10¹²/l), 5.93±1.25 (×10⁹/l), 201.84±86 (fl), 62.54±5.73 (pg), 329±1.6 (g/l), 64.71±4.47 (%), 35.14±2.1 (%) and 76.4±9.2 (10⁹/l), respectively. Haematological parameters in black-headed gull—PCV, Hb, RBC, WBC, MCV, MCH, MCHC, heterophiles, lymphocytes and thrombocytes—were found to be 39±2.52 (%), 123±13.3 (g/l), 2.89±0.45 (×10¹²/l), 2.25±0.42 (×10⁹/l), 184±17.32 (fl), 60.33±6.74 (pg), 327.6±3.8 (g/l), 57.33±12.2 (%), 42.66±4.7 (%) and 61.44±8.25 (10⁹/l), respectively. The results of blood serum biochemistry in flamingo indicated that the concentrations of glucose, cholesterol, total protein, albumin, uric acid, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatine phosphokinase (CPK), phosphorus, cadmium, lead and chromium were 8.45±1.65 (mmol/l), 10.4±0.01 (mmol/l), 55±4.7 (g/l), 17.1±2.7 (g/l), 528.99±172.4 (μmol/l), 70.83±19.77 (IU/l), 4.2±0.2 (IU/l), 19.78±5.38 (IU/l), 197.16±57.45 (IU/l), 2.01±0.4 (mmol/l), 2.55±0.98 (μmol/l), 11.14±3.95 (μmol/l) and 4.08±1.41 (μmol/l), respectively. The results of blood biochemistry in black-headed gull indicated that the serum concentrations of glucose, cholesterol, total protein, albumin, uric acid, AST, ALT, ALP, CPK, phosphorus, cadmium, lead and chromium were 10.78±1.39 (mmol/l), 7.37±0.63 (mmol/l), 51±8.1 (g/l), 18.3±2 (g/l), 707.8±210.55 (μmol/l), 92.66±17.14 (IU/l), 9.21±1.2 (IU/l), 27.73±5.37 (IU/l), 164.33±48.81 (IU/l), 2.09±0.59 (mmol/l), 3.26±1.1 (μmol/l), 10.32±2.49 (μmol/l) and 5.91±1.25 (μmol/l), respectively. The results showed high concentrations of heavy metals in both species, which could be an indication of environmental pollution.     

Haematological indicators of inflammation exhibited by Australian Falconiformes

Numerous studies have reported some haematological characteristics of diurnal raptors; however, few of these have characterised the haematological response to disease. We investigated the haematological characteristics, exhibited in response to a range of injuries and naturally occurring inflammatory disease, of seven birds from six species of Falconiformes. A spectrum of leukocyte responses was observed. Some form of morphological atypia was evident in the heterophils of all birds and was the most reliable indicator of inflammation. The concentration of leukocytes varied between each case, with the least concentration of heterophils observed to be 0.18 × 10⁹/l and the greatest concentration of heterophils observed to be 6.00 × 10⁹/l. This study illustrates the heterogeneous haematological response exhibited by Falconiformes to different inflammatory challenges.     

Determination of the effect of probiotic (Saccharomyces cerevisiae) on growth performance and hematological parameters of rabbits

Insufficient supply of animal protein is a major problem in developing countries including Nigeria. Rabbits are adjudged to be a convenient source of palatable and nutritious meat, high in protein, and contain low fat and cholesterol. A doe can produce more than 15 times her own weight in offspring in a year. However, its productivity may be limited by inadequate nutrition. The objective of this study was to determine the effect of probiotic (Saccharomyces cerevisiae) supplementation on growth performance and some hematological parameters of rabbit. The appropriate level of the probiotic inclusion for excellent health status and optimum productivity was also determined. A total of 40 male rabbits were randomly divided into four groups (A–D) of ten rabbits each. Each group was subdivided into two replicates of five rabbits each. They were fed pelleted grower mash ad libitum. The feed for groups A to C were supplemented with bioactive yeast (probiotic) at inclusion levels of 0.08, 0.12, and 0.16 g yeast/kg diet, respectively. Group D had no yeast (control). Daily feed intake was determined. The rabbits were weighed weekly. The packed cell volume (PCV), hemoglobin concentration, white blood cell total, and differential counts were determined at the 8th week, 16th week, and 22nd week following standard procedures. The three results which did not have any significant difference were pooled together. Group A which had 0.08 g yeast/kg of diet had a significantly lower (P ≤ 0.05) PCV than groups B (which had 0.12 g yeast/kg of diet) and C (which had 0.16 g yeast/kg of diet) as well as D (the control). Total WBC count for groups B and C (14.35 ± 0.100 × 10³/μl and 14.65 ± 0.786 × 10³/μl, respectively) were significantly higher (P ≤ 0.05) than groups A and D (6.33 ± 0.335 × 10³/μl and 10.40 ± 0.296 × 10³/μl, respectively). Also the absolute neutrophils and lymphocytes counts were significantly higher (P ≤ 0.05) in groups B and C than in groups A and D. Group B had significantly higher (P ≤ 0.05) weight gain (1.025 ± 0.006 kg/rabbit) followed by group A (0.950 ± 0.092 kg/rabbit). The control (group D) had the least weight gain of 0.623 ± 0.0.099 kg/rabbit. These results showed that like most probiotics, bioactive yeast at an appropriate level of inclusion had a significant beneficial effect on health status and growth rate of rabbit. Probiotic supplementation level of 0.12 g yeast/kg of diet was recommended for optimum rabbit production.     

Transient Extremity Ischemia Augments CD34+ Progenitor Cell Availability

Peripheral blood is an easily accessed source for stem cell production; however, the number of cells produced is relatively low. We hypothesized that ischemic preconditioning may serve as a safe method to increase the number of CD34+ cells that can be harvested and cultured in a short period. This study was conducted to test this hypothesis by examining the safety and efficacy of brief, transient ischemia of the lower limbs to augment the number of cells that can be produced from blood of healthy volunteers. Following induction of ischemia, blood samples were withdrawn at baseline, 30 min, 12 h and 24 h. The number of progenitor cells was determined by flow cytometry after the harvested cells were cultured for 5 days. We also analyzed the blood samples to determine IL-8 and VEGF concentrations. No serious adverse events were observed. The total number of cells increased from 0.46 ± 0.1 × 10⁶ cells/ml in the pretreatment blood samples to 0.7 ± 0.1 × 10⁶ cells/ml in blood taken 12 h after the conclusion of transient ischemia, p = 0.0029. The number of CD34+ cells increased from 4.23 ± 0.8 × 10⁴ cells/ml in the pretreatment samples to 7.17 ± 1.34 × 10⁴ cells/ml in blood taken 12 h after ischemia, p = 0.0001. The harvested stem cells maintained their ability to construct tubular structures. The augmentation in the number of CD34+ cells was positively correlated with the increase of IL-8, but not with VEGF concentrations. Ischemic preconditioning is a safe and effective technique to increase the availability of stem cells for therapeutic purposes.     

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.     

Haematology, morphology and blood cells characteristics of male and female Siamese fighting fish (Betta splendens)

The aim of the present study was to obtain baseline data on blood cell size, morphology and haematological parameters in Siamese fighting fish (Betta splendens) since there is limited information in the published literature. Blood samples from the caudal vein of apparently healthy Siamese fighting fish (male: n = 40 and female: n = 36) were collected. Haematological values of the blood samples were determined using standard techniques. The morphological features of blood cells were described according to observations made by light microscopy. The various types of blood cells measurement were carried out with the help of a stage and an ocular micrometre at a magnification of ×1,000. Erythrocytes, thrombocytes and four types of leucocytes: lymphocytes, monocytes, heterophils and eosinophils, were distinguished and characterised. The average size of the erythrocyte cell and nucleus was 97.33 and 16.28 μm², respectively. Results showed a positive correlation between erythrocyte size and nucleus size for Siamese fighting fish (r = 0.470, p < 0.01). We also found sex-dependent differences for total white blood cell count, lymphocytes and heterophils in Siamese fighting fish (p < 0.05). Statistical analysis revealed that differences in other haematological parameters and blood cell morphology, between male and female fish were not statistically significant (p > 0.05).     

Multichannel oscillatory-flow multiplex PCR microfluidics for high-throughput and fast detection of foodborne bacterial pathogens

In the field of continuous-flow PCR, the amplification throughput in a single reaction solution is low and the single-plex PCR is often used. In this work, we reported a flow-based multiplex PCR microfluidic system capable of performing high-throughput and fast DNA amplification for detection of foodborne bacterial pathogens. As a demonstration, the mixture of DNA targets associated with three different foodborne pathogens was included in a single PCR solution. Then, the solution flowed through microchannels incorporated onto three temperature zones in an oscillatory manner. The effect factors of this oscillatory-flow multiplex PCR thermocycling have been demonstrated, including effects of polymerase concentration, cycling times, number of cycles, and DNA template concentration. The experimental results have shown that the oscillatory-flow multiplex PCR, with a volume of only 5 μl, could be completed in about 13 min after 35 cycles (25 cycles) at 100 μl/min (70 μl/min), which is about one-sixth of the time required on the conventional machine (70 min). By using the presently designed DNA sample model, the minimum target concentration that could be detected at 30 μl/min was 9.8 × 10⁻² ng/μl (278-bp, S. enterica), 11.2 × 10⁻² ng/μl (168-bp, E. coli O157: H7), and 2.88 × 10⁻² ng/μl (106-bp, L. monocytogenes), which corresponds to approximately 3.72 × 10⁴ copies/μl, 3.58 × 10⁴ copies/μl, and 1.79 × 10⁴ copies/μl, respectively. This level of speed and sensitivity is comparable to that achievable in most other continuous-flow PCR systems. In addition, the four individual channels were used to achieve multi-target PCR analysis of three different DNA samples from different food sources in parallel, thereby achieving another level of multiplexing.     

High pentraxin-3 plasma levels associate with thrombocytopenia in acute Puumala hantavirus-induced nephropathia epidemica

Our aim was to investigate whether plasma levels of the long pentraxin-3 (PTX3) associate with the severity of Puumala hantavirus-induced nephropathia epidemica (NE). Sixty-one prospectively identified consecutively hospitalized NE patients were examined. Plasma PTX3, interleukin (IL)-6, terminal complement complex SC5b-9, complement component C3, C-reactive protein (CRP), creatinine, sodium, kynurenine, and tryptophan levels, as well as the blood cell count, were determined for up to five consecutive days after hospitalization. Receiver operating characteristic (ROC) analysis revealed that the maximum PTX3 level >101.6 ng/ml (high PTX3) showed a sensitivity of 71% and a specificity of 89% for detecting platelet level <50 × 10⁹/l, with an area under the curve (AUC) value of 0.78 (95% confidence interval [CI] 0.63–0.94). High PTX3 level was also associated with several other variables reflecting the severity of the disease: patients with high PTX3 level had higher maximum blood leukocyte (16.1 vs. 9.7 × 10⁹/l, p < 0.001), plasma IL-6 (16.9 vs. 9.0 pg/ml, p = 0.007), and creatinine (282 vs. 124 μmol/l, p = 0.007) levels than patients with low maximum PTX3 level. They also had longer hospital stays (8 vs. 5 days, p = 0.015) compared to patients with low PTX3 level. High plasma PTX3 levels are associated with thrombocytopenia and the overall severity of NE.     

Haematological and serum biochemical profile of the upside-down catfish, Synodontis membranacea Geoffroy Saint Hilaire from Jebba Lake, Nigeria

Haematological and serum biochemical studies of natural population of Synodontis membranacea from Jebba Lake, North Central Nigeria were investigated in order to establish their mean and reference values. Bi-monthly collection of 1,408 live fish samples was carried out between April 2002 and March 2004, using gill nets of various mesh sizes ranging from 5.08 to 10.16 cm. The mean baseline value established for species-specific haematological and serum biochemical parameters were red blood cell (RBC) 3.83 ± 1.49 × 10¹² l⁻¹, haemoglobin (HB) 8.38 ± 1.96 g dl⁻¹, and packed cell volume (PCV) 25.65 ± 5.89%; mean cell volume 78.25 ± 37.90 fl; mean cell haemoglobin (MCH) 33.04 ± 12.50 pg; mean cell haemoglobin concentration 26.53 ± 15.18 g dl⁻¹; white blood cell (WBC) 315.65 ± 95.37 × 10⁻⁹; agranulocytes (Agr) 82.07 ± 11.38%; monocytes (Mon) 6.37 ± 3.01%; lymphocytes (Lym) 76.49 ± 10.81%; granulocytes (Gran) 40.28 ± 17.48%; neutrophils (Neut) 24.42 ± 10.68%; eosinophils (Eos) 16.14 ± 8.25%; basophils 0.09 ± 0.04%; protein 40.19 ± 7.45 g l⁻¹; albumin 19.78 ± 5.67 g l⁻¹; creatinine 49.71 ± 16.15 μmol l⁻¹; urea 3.05 ± 0.67 nmol l⁻¹; uric acid 0.76 ± 0.33 nmol l⁻¹; glucose 4.24 ± 1.74 mmol l⁻¹; cholesterol 8.46 ± 2.27 mmol l⁻¹; calcium 2.35 ± 0.94 mmol l⁻¹; potassium 13.36 ± 4.45 mmol l⁻¹; sodium 139.39 ± 23.19 mmol l⁻¹; alanine aminotransferase (ALT) 11.79 ± 2.67 U l⁻¹; aspartate aminotransferase 16.80 ± 4.73 U l⁻¹; and alkaline phosphatase 63.01 ± 20.44 U l⁻¹. Only three of these parameters (i.e. neutrophil, glucose and potassium) differed significantly (P > 0.05) on gender basis. Pearson’s correlation coefficients indicated significant relationship of standard length and total weight with RBC, PCV, HB, WBC, Agr, Mon, Lym, Gran, Neut, Eos, sodium, and ALT only. The study has provided baseline haematological and biochemical data for use in health monitoring and productivity of S. membranacea, which would be of great value for future comparative surveys in this era of increased fish culture in Nigeria.     

Serum biochemical parameters of Huso huso

Serum biochemical parameters are important aspects in the management of endangered species such as Huso huso. The values of these parameters can be used for confirming the sex and any changes in the quality of waters and related soils. Serum samples of 15 H. huso fish were analysed and their serum parameter values were determined as mean±SD in both sexes. Age, weight, total length and fork length were the same between groups. We compared the levels of calcium (Ca; 2.13 ± 0.69–2.37 ± 0.38 mmol/l), total protein (TOP; 4.51 ± 1–5.50 ± 0.94 mg/dl), blood urea nitrogen (BUN; 1.32 ± 0.23–1.35 ± 0.31 mmol/l), albumin (Alb; 0.88 ± 0.18–1.26 ± 0.29 mg/dl), globulin (Glb; 3.63 ± 0.84–4.5 ± 0.69 mg/dl), uric acid (UA; 1.66 ± 0.18–1.79 ± 0.27 mmol/l), creatinine (CREA; 30.48 ± 4.31–30.48 ± 6.10 mmol/l), phosphorous (P; 2.18 ± 0.38–2.91 ± 0.67 mmol/l), magnesium (Mg; 1.15 ± 0.26–1.51 ± 0.35 mmol/l), glucose (Glc; 3.42 ± 0.84–6.69 ± 1.54 mmol/l), alkaline phosphatase (ALP; 4.48 ± 1.82–5 ± 1.04 IU/l) and amylase (156.19 ± 34.07–166.25 ± 64.27-IU/l). We have shown that there were no differences in the Ca, TOP, BUN, UA, CREA, Mg, ALP and amylase between sexes. However, male fish have higher Glc, P, Alb and Glb than females.     

Effect of praziquantel prolonged administration on granuloma formation around Schistosoma japonicum eggs in lung of sensitized mice

Schistosomiasis remains a major public health problem and it is an immune disease. The schistosome egg is the primary parasite factor responsible for the overt disease. The eggs release the soluble antigen, which induces intensive tissue reaction, a granulomatous reaction to the eggs. If granuloma formation could be suppressed, overt disease might not develop. Praziquantel is an effective antischistosomal drug especially for adult worms. However, whether praziquantel has a suppressing effect on granuloma formation around schistosome eggs directly remains unclear. The purpose of the present study was to investigate the effect of praziquantel, especially administered persistently, on granuloma formation around Schistosoma japonicum eggs in the lung of sensitized mice. Thirty-six mice were divided into three groups averagely. Group A was a control group. First, the mice were injected with schistosomal eggs hypodermically in abdomen, and 10 days later injected with schistosomal eggs intravenously via a tail vein. Group B was a praziquantel short administration group. In addition to the injections of schistosomal eggs as the same of Group A, the mice were administered with praziquantel in a daily dose of 300 mg/kg for 3 days, from 1 day before the intravenous injection of the eggs. Group C was a praziquantel prolonged administration group. In addition to the injections of schistosomal eggs as the same of Group A, the mice were administered with praziquantel in a daily dose of 150 mg/kg for 5 days weekly until the mice were sacrificed. Three mice of each group were sacrificed on days 7, 14, 28, and 56, respectively after the intravenous injection of the eggs, and the lung tissues were fixed with formalin and the slices were HE stained. The granulomas containing eggs in their centers were selected, and 25–30 granulomas from the animals of each group were measured at each time period. The mean areas of egg granulomas of each group were calculated, and the neutrophilic granulocytes, eosinocytes, lymphocytes, fibroblasts, and macrophages within the egg granulomas were counted. The mean numbers of them of each group were calculated. All the data of each group were analyzed and compared statistically. On day 56 after the intravenous injection of the eggs, the mean area of schistosomal egg granulomas in group B was (227.4 ± 728.0) × 10³ μm², less than that of [(297.9 ± 153.3) × 10³ μm²] in group A, and the suppression rate was 23.7% (P < 0.05). On days 7, 14, 28, and 56, the mean areas of schistosomal egg granulomas in group C were (575.8 ± 155.6) × 10³ μm², (310.5 ± 854.0) × 10³ μm², (267.7 ± 513.3) × 10³ μm², and (214.9 ± 446.4) × 10³ μm², respectively, significantly less than those of [(692.7 ± 232.6) × 10³ μm², (439.4 ± 165.0) × 10³ μm², (385.7 ± 129.3) × 10³ μm², and (297.9 ± 153.3) × 10³ μm²] in group A. The suppression rates were 16.9%, 29.3%, 30.6%, and 27.9%, respectively (P values <0.05). On day 56, the mean numbers of neutrophilic granulocytes were 11.4 ± 5.0 in group A and 5.2 ± 3.1 in group C, respectively, with the suppression rate of 54.4% in group C (P < 0.05). On day 56, the mean numbers of eosinocytes within the egg granulomas were 2.3 ± 2.0, 0.1 ± 0.3, and 0.3 ± 0.6 in groups A, B, and C, respectively, with the suppression rate of 95.7% in group B and 87.0% in group C (P values <0.05). On day 56, the mean numbers of macrophages within egg granulomas were 14.3 ± 6.9 in group C, compared with 18.6 ± 8.2 in group A, the suppression rate was 23.1% (P < 0.05). On day 56, the mean numbers of fibroblasts within the egg granulomas were 6.6 ± 4.4 and 5.8 ± 2.6 in groups B and C, respectively, and compared with 14.3 ± 7.8 in group A, the increasing extents decreased by 53.8% and 59.4%, respectively (P values <0.05). Therefore, the administration of praziquantel, especially the prolonged administration, can suppress the formation of schistosomal egg granulomas, including reduction in the areas of granulomas and suppression of the inflammatory cells and the hyperplasia of fibroblasts within granulomas.     

Haematology of Persian Fallow Deer ( Dama mesopotamica )

The haematology of Persian fallow deer was studied and the means of various parameters were determined for sex and age groups. For total samples, the mean ± standard deviation of haematological parameters were: red blood cells (RBC), 7.42 ± 1.27 × 10¹²/l; haematocrit (PCV), 38.83 ± 7.38%; haemoglobin (Hb), 148.0 ± 17.3 g/l; mean cell volume (MCV), 50.84 ± 7.26 fl; mean corpuscular haemoglobin (MCH), 20.17 ± 1.98 pg; mean corpuscular haemoglobin concentration (MCHC), 38.71 ± 3.88%; white blood cells (WBC) 3.200 ± 1.697 × 10⁹/l; neutrophils, 1.410 ± 0.446 × 10⁹/l, lymphocytes, 1.382 ± 1.116 × 10⁹/l, monocytes, 0.048 ± 0.100 × 10⁹/l, eosinophils, 0.341 ± 0.339 × 10⁹/l, basophils, 0.009 ± 0.005 × 10⁹/l, neutrophil:lymphocyte ratio (N:L), 2.11 ± 2; platelets, 354.8 ± 116.62 × 10⁹/l and fibrinogen, 2.855 ± 1.126 g/l. Three types of haemoglobin were separated by electrophoresis: HbC with higher migration, HbB and HbA with lowest movement. Significant differences were seen for RBC, MCV, MCH, HbC, and HbB concentrations between sexes.     

Insecticidal, acaricidal and repellent effects of DEET- and IR3535-impregnated bed nets using a novel long-lasting polymer-coating technique

A novel long-lasting repellent-treated net (LLRTN) has been designed by binding the skin repellents N,N-diethyl-m-toluamide (DEET), or IR3535, onto the fibres of bed net fabric using a new polymer-coating technique. The repellent toxicological effectiveness and residual activity of a factory-based repellent-impregnated fabric has been evaluated by laboratory testing against adult Aedes aegypti mosquitoes and nymphal Ixodes ricinus ticks. By using this repellent-embedding impregnation technique, concentrations exceeding 10 g/m² could be achieved with one single polymer layer. Both DEET- and IR3535-impregnated fabrics revealed a dose-dependent insecticidal as well as acaricidal activity. One hundred percent knockdown times of DEET-treated bed nets ranged from 187.5 ± 31.8 to 27.5 ± 3.5 min against A. aegypti, and between 214 ± 47 and 22.6 ± 5 min against nymphal I. ricinus, linked to a DEET concentration of 1.08 and 10.58 g/m², respectively. With IR3535, A. aegypti produced dose-dependent 100% knockdown times varying from 87.5 ± 10.6 to 57.5 ± 3.5 min and between 131.4 ± 6.5 and 33.8 ± 5 min against nymphal I. ricinus, respectively, linked to concentrations between 1.59 and 10.02 g/m². One hundred percent repellency measured by complete landing and biting protection of impregnated fabric by using the arm-in-cage test could be achieved at DEET concentrations exceeding 3.7 to 3.9 g/m², and for IR3535 concentrations over 10 g/m². One hundred percent landing and biting protection could be preserved with DEET-treated fabrics for 29 weeks at an initial concentration of 4.66 g/m², 54 weeks at 8.8 g/m², 58 weeks at 9.96 g/m² and 61 weeks at 10.48 g/m² for DEET, and 23 weeks for IR3535-treated fabric at a concentration of 10.02 g/m². Unlike repellent-treated fabric, a brand of a commercially available long-lasting insecticide-treated net tested containing 500 mg permethrin/m² did not protect from mosquito bites. First results on bioactivity and long-lasting efficacy show that the new LLRTN technique is highly promising as a potential candidate for future malaria control strategies, especially in areas where pyrethroid resistance occurs.     

Determination of some normal serum parameters in starry sturgeon (Acipenser stellatus Pallas, 1771) during spring season

Hematology can be a useful tool for monitoring health status, detecting illnesses, and following the progress of diseases and responses to therapy. Despite advances in fish medicine in recent years, interpretation of fish hematology is often hampered by lack of meaningful reference values and the bewildering diversity of fish species. Serum samples of 40 Acipenser stellatus fish were analyzed (20 male and 20 female) and their serum parameter values were measured in both sexes. Serum biochemical values were determined (mean ± SEM) for sodium (Na; 149.2 ± 1.917 mmol/l), potassium (K; 2.75 ± 0.097 mmol/l), calcium (Ca; 8.293 ± 0.282 mg/dl), phosphorus (P; 12.39 ± 0.267 mg/dl), glucose (Glc; 166.40 ± 8.264 mg/dl), triglyceride (trig; 699.6 ± 22.94 mg/dl), bilirubin (bilirubin; 0.616 ± 0.0234 mg/dl), total protein (TOP; 2.988 ± 0.0842 g/dl), albumin (Alb; 1.218 ± 0.0415 g/dl), cholesterol (CHO; 238.2 ± 11.24 mg/dl), creatinine (CREA; 0.1085 ± 0.0048 mg/dl), and blood urea nitrogen (BUN; 15.32 ± 0.5104 mg/dl). The serum values for bilirubin, Na, P, and CREA were significantly higher in females, whereas BUN and Alb were significantly higher in males. The correlations of coefficients between measured parameters were also determined.     

Adulticidal and larvicidal activity of Beauveria bassiana and Metarhizium anisopliae against housefly, Musca domestica (Diptera: Muscidae), in laboratory and simulated field bioassays

The susceptibility of the adult and larval stage of housefly, Musca domestica L. (Diptera: Muscidae), to two entomopathogenic fungi, Metarhizium anisopliae (Metsch.) Sor. and Beauveria bassiana (Bals.) Vuill., was evaluated under laboratory and simulated field bioassays. Bioassays on adult houseflies were carried out at different conidial concentrations ranging from 10³ to 10⁹ conidia/ml in petri plate and minichamber assays. Absolute mortality was observed within 4–5 days at all the concentrations tested. M. anisopliae was found to be more effective with LC₅₀ of 6.75 × 10⁷ conidia/ml compared with 1.21 × 10⁸ conidia/ml of B. bassiana in petri plate bioassay. Similar trend was observed in minichamber bioassay. Larvicidal activity evaluated through petri plate bioassay also indicated that M. anisopliae was more effective larvicide with LC₅₀ of 4.1 × 10⁸ conidia/ml as against 3.31 × 10⁹ conidia/ml of B. bassiana. Larvicidal activity was further evaluated in simulated field condition of decaying waste matrix using dry conidial formulations (10⁸ conidia/g) of both the fungi. Larval mortality obtained in this assay was 43% (B. bassiana) and 63% (M. anisopliae). Remarkably better performance of M. anisopliae as an adulticidal and larvicidal agent over B. bassiana in laboratory bioassays as well as simulated field conditions suggests that it may have good potential to become part of an integrated housefly control program.     

Effects of Metarhizium anisopliae conidia mixed with soil against the eggs of Aedes aegypti

The effectiveness of Metarhizium anisopliae IP 46 conidia mixed with soil was tested against Aedes aegypti eggs. Mycelium and new conidia developed first on eggs between 4.8 and 15 days respectively after incubation of fungus-treated soils at 3.3 × 10³ up to 3.3 × 10⁵ conidia/g soil at 25°C and relative humidities close to saturation. After 15-day incubation, 53.3% of the eggs exposed to soil with 3.3 × 10⁵ conidia/g showed external development of mycelium and conidia. Fungus-inoculated soils (but not untreated controls) showed some mycelial growth and sporulation apart from the eggs. Some eggs on treated soils hatched; those larvae died and eventually showed fungal development on their bodies. The cumulative relative eclosion of larvae after submersion of treated eggs in water decreased from 52.2% at 3.3 × 10³ conidia/g to 25.3% at 3.3 × 10⁵ conidia/g. These findings clearly showed that A. aegypti eggs can be infected by M. anisopliae when deposited on fungus-contaminated soil. The effectiveness of M. anisopliae against gravid females, larvae, and also eggs of A. aegypti underscored the possible usefulness of this fungus as a mycoinsecticide, whether naturally occurring or artificially applied, in the breeding sites of this mosquito.     

Comparative Cell Shape and Diffusional Water Permeability of Red Blood Cells from Indian Elephant ( Elephas maximus ) and Man ( Homo sapiens )

Red blood cells (RBC) from an Indian elephant (Elephas maximus) were studied by light microscopy (LM), scanning electron microscopy (SEM) and a new nuclear magnetic resonance (NMR) ‘imaging’ method based on the translational diffusion of water, NMR q-space analysis. Also, the transmembrane diffusional permeability, P _d of water in RBC was measured by using a Mn²⁺-doping NMR technique, taking human RBC as a reference. The main diameter of the elephant RBC was measured as 9.3 ± 0.7 μm by LM, 9.3 ± 0.7 μm by ‘shrinkage-corrected’ SEM, and 9.3 ± 0.4 μm by q-space anlaysis. The value is ∼1.4 μm larger than that for the human RBC. The values of P _d were, in the case of elephant RBC, 3.2 × 10⁻³ cm/s at 25 °C, 3.9 × 10⁻³ cm/s at 30 °C, 5.2 × 10⁻³ cm/s at 37 °C and 6.5 × 10⁻³ cm/s at 42 °C; all values were significantly lower than the corresponding values of P _d for human RBC, namely 4.3 × 10⁻³ cm/s at 25 °C, 5.2 × 10⁻³ cm/s at 30 °C, 6.1 × 10⁻³ cm/s at 37 °C, 7.8 × 10⁻³ cm/s at 42°C. The maximal inhibition of P _d (56%) was reached in 30 min at 37 °C with 2 mm p-chloromercuribenzene sulphonate (PCMBS) for both species of RBC. The basal permeability to water at 37 °C was estimated to be 2.3 × 10⁻³ cm/s for elephant and 2.6 × 10⁻³ cm/s for human RBC. The values of the activation energy for water permeability (E _a,d ) was significantly higher for elephant RBC (31.9 kJ/mol) than for human RBC (25.9 kJ/mol). This indicated that features other than the number of transporters per cell are likely to be important in defining the differences in water permeability in the RBC from the two species.     

Quantitative T-cell interferon-gamma responses to Mycobacterium tuberculosis-specific antigens in active and latent tuberculosis

The objective was to compare the quantitative T-cell responses measured by the commercial interferon-gamma (IFNγ) release assays (IGRAs) in active and latent tuberculosis (TB) states. T-cell responses of culture-proven TB cases were compared with those of contacts with positive IGRA results and tuberculin skin tests ≥ 15 mm. T-SPOT.TB results in 270 active TB cases and 183 community contacts showed the median spot-forming cells (SFCs) above negative control/2.5 × 10⁵ peripheral blood mononuclear cells to be 27 (−1 to 203) vs 10 (−2 to 174) in response to ESAT-6 (p < 0.001); and 37 (0 to 293) vs 13 (0 to 225) to CFP-10 (p < 0.001). The median IFNγ levels (antigen minus nil control) as measured by QuantiFERON-TB Gold In-tube in 270 cases and 142 contacts in congregate settings was 2.3 IU/ml (−0.58 to 31.44) vs 1.7 IU/ml (0.35 to 26.51, p = 0.98). Quantitative T-cell responses as measured by the T-SPOT.TB may indicate mycobacterial burden and disease activity, but cannot be used to discriminate active from latent TB.     

Comparative Cell Shape and Diffusional Water Permeability of Red Blood Cells from Indian Elephant (Elephas maximus) and Man (Homo sapiens)

Red blood cells (RBC) from an Indian elephant (Elephas maximus) were studied by light microscopy (LM), scanning electron microscopy (SEM) and a new nuclear magnetic resonance (NMR) ‘imaging’ method based on the translational diffusion of water, NMR q-space analysis. Also, the transmembrane diffusional permeability, P _d of water in RBC was measured by using a Mn²⁺-doping NMR technique, taking human RBC as a reference. The main diameter of the elephant RBC was measured as 9.3 ± 0.7 μm by LM, 9.3 ± 0.7 μm by ‘shrinkage-corrected’ SEM, and 9.3 ± 0.4 μm by q-space anlaysis. The value is ∼1.4 μm larger than that for the human RBC. The values of P _d were, in the case of elephant RBC, 3.2 × 10⁻³ cm/s at 25 °C, 3.9 × 10⁻³ cm/s at 30 °C, 5.2 × 10⁻³ cm/s at 37 °C and 6.5 × 10⁻³ cm/s at 42 °C; all values were significantly lower than the corresponding values of P _d for human RBC, namely 4.3 × 10⁻³ cm/s at 25 °C, 5.2 × 10⁻³ cm/s at 30 °C, 6.1 × 10⁻³ cm/s at 37 °C, 7.8 × 10⁻³ cm/s at 42°C. The maximal inhibition of P _d (56%) was reached in 30 min at 37 °C with 2 mm p-chloromercuribenzene sulphonate (PCMBS) for both species of RBC. The basal permeability to water at 37 °C was estimated to be 2.3 × 10⁻³ cm/s for elephant and 2.6 × 10⁻³ cm/s for human RBC. The values of the activation energy for water permeability (E _a,d ) was significantly higher for elephant RBC (31.9 kJ/mol) than for human RBC (25.9 kJ/mol). This indicated that features other than the number of transporters per cell are likely to be important in defining the differences in water permeability in the RBC from the two species.