A study of peripheral blood in hedgehogs in Turkey.

The aim of this study was to determine diameters of blood cells, differential counts of peripheral blood leukocytes, alpha-naphthyl acetate esterase (ANAE), acid phosphatase (ACP-ase) activity of some leukocyte types, and enzymatic positivity percentages of peripheral blood lymphocytes in two hedgehogs species, Hemiechinus auritus, the long-eared hedgehog, and Erinaceus concolor, the southern white-breasted hedgehog. Air-dried peripheral blood smears were stained with May-Grünwald-Giemsa stain. ANAE and ACP-ase were stained in glutaraldehyde-acetone-fixed smears. ANAE-positive lymphocytes displayed a dot-like positivity pattern characterized with 1-5 reddish brown cytoplasmic granules, whereas ACP-ase positive lymphocytes displayed a dot-like positivity pattern characterized with 1-3 pinkish cytoplasmic granules. Monocytes gave a diffuse and strong reaction while neutrophils displayed a weak positive reaction for ANAE and ACP-ase. No difference was observed in mean diameters of peripheral blood cells of these species. It was found that lymphocytes made up the majority (64.3% and 65.5%) of leukocytes, followed by neutrophils (23.9% and 23.3%), eosinophils (9.0% and 7.6%), monocytes (1.8% and 2.3%), and basophils (1.0% and 1.3%) in H. auritus and E. concolor, respectively. Mean ANAE positivity oflymphocytes was 36.6% and 51.3% and ACP-ase positivity was 32.1% and 37.5% for H. auritus and E. concolor, respectively. The ANAE positivity of lymphocytes in E. concolor was significantly (P < 0.05) higher than that of H. auritus.

Blood of African Hedgehog Atelerix albiventris Contains 115-kDa Trypanolytic Protein that Kills Trypanosoma congolense.

INTRODUCTION: Protozoan parasites of the Order Trypanosomatida infect a wide range of multicellular plants and animals, causing devastating and potentially fatal diseases. Trypanosomes are the most relevant members of the order in sub-Saharan Africa because of mortalities and morbidities caused to humans and livestock. PURPOSE: There are growing concerns that trypanosomes are expanding their reservoirs among wild animals, which habours the parasites, withstand the infection, and from which tsetse flies transmit the parasites back to humans and livestock. This study was designed to investigate the potentials of the African hedgehog serving as reservoir for African animal trypanosomes. METHODS: Five adult hedgehogs alongside five laboratory mice were intraperitoneally inoculated with 106 and 104 of Trypanosoma congolense cells, respectively, and monitored for parasitemia and survival. Serum from twenty hedgehogs was subjected to trypanocidal activity-guided fractionation by successive ion-exchange and gel-filtration chromatographies, followed by characterization with Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). RESULTS: Hedgehogs were resistant to the infection as no parasite was detected and none died even after 60 days, while all the mice died within 12 days. Both the serum and plasma prepared from hedgehogs demonstrated trypanocidal activity- rapidly killed trypanosomes even when diluted 1000 times. The trypanolytic factor was identified to be proteinaceous with an estimated molecular weight of 115-kDa. CONCLUSION: For the first time, it is here demonstrated that hedgehog blood has significant trypanolytic activity against T. congolense. The potential application of the hedgehog protein for the breeding of trypanosomosis-resistant livestock in tsetse fly belt is discussed.

Germ cell desquamation-based testis regression in a seasonal breeder, the Egyptian long-eared hedgehog, Hemiechinus auritus.

Testes of seasonally breeding species experience a severe functional regression before the non-breeding period, which implies a substantial mass reduction due to massive germ-cell depletion. Two alternative mechanisms of seasonal germ-cell depletion have been described in mammals, apoptosis and desquamation (sloughing), but their prevalence has not been determined yet due to reduced number of species studied. We performed a morphological, hormonal, and molecular study of the mechanism of seasonal testicular regression in males of the Egyptian long eared-hedgehog (Hemiechinus auritus). Our results show that live, non-apoptotic, germ cells are massively depleted by desquamation during the testis regression process. This is concomitant with both decreased levels of serum testosterone and irregular distribution of the cell-adhesion molecules in the seminiferous epithelium. The inactive testes maintain some meiotic activity as meiosis onset is not halted and spermatocytes die by apoptosis at the pachytene stage. Our data support the notion that apoptosis is not the major testis regression effector in mammals. Instead, desquamation appears to be a common mechanism in this class.

Monkeypox zoonotic associations: insights from laboratory evaluation of animals associated with the multi-state US outbreak.

At the onset of the 2003 US monkeypox outbreak, virologic data were unavailable regarding which animal species were involved with virus importation and/or subsequent transmission to humans and whether there was a risk for establishment of zoonotic monkeypox in North America. Similarly, it was unclear which specimens would be best for virus testing. Monkeypox DNA was detected in at least 33 animals, and virus was cultured from 22. Virus-positive animals included three African species associated with the importation event (giant pouched rats, Cricetomys spp.; rope squirrels, Funisciuris sp.; and dormice, Graphiuris sp.). Virologic evidence from North American prairie dogs (Cynomys sp.) was concordant with their suspected roles as vectors for human monkeypox. Multiple tissues were found suitable for DNA detection and/or virus isolation. These data extend the potential host range for monkeypox virus infection and supports concern regarding the potential for establishment in novel reservoir species and ecosystems.

Seroprevalence of Severe Fever with Thrombocytopenia Syndrome Virus in Hedgehog from China.

Severe fever with thrombocytopenia syndrome, an emerging hemorrhagic fever, is caused by severe fever with thrombocytopenia syndrome virus (SFTSV), a tick-borne bunyavirus. Information regarding SFTSV animal hosts is very limited. In this study, we showed that 64% (9/14) of hedgehogs in Shandong Province, China were seropositive to SFTSV antibody, suggesting that hedgehog could be a vertebrate parasitifer for SFTSV.

Seasonal variations of plasma lipids and lipoproteins in the hedgehog, an animal model for lipoprotein (a) metabolism: relation to plasma thyroxine and testosterone levels.

We describe a study of the seasonal variations of hedgehog plasma lipids and lipoproteins and their correlation with changes in the activities of the thyroid and testis. In ten male hedgehogs, plasma concentrations of lipids, thyroxine and testosterone were assayed each month for 1 year beginning in September, while plasma lipoproteins from five of these animals were analyzed at the same dates using density gradient ultracentrifugation. All classes of plasma lipids (cholesterol, total glycerol and phospholipids) exhibited statistically significant seasonal variations in their respective concentrations, with simultaneous maxima (cholesterol: 207 +/- 39 mg/100 ml; total glycerol: 50 +/- 9 mg/100 ml; phospholipids: 266 +/- 25 mg/100 ml) during late fall-early winter, i.e., during the period of the year when plasma levels of both thyroxine and testosterone were minimal. Plasma lipids subsequently decreased to minimal levels either in early summer (cholesterol: 129 +/- 18 mg/100 ml; phospholipids: 178 +/- 20 mg/100 ml) or in late winter (total glycerol: 22 +/- 9 mg/100 ml). Very low density lipoproteins (d less than 1.015 g/ml) were found at low levels (less than 15 mg/100 ml) during the cold months, and then became detectable as trace components only. The total concentration of the mixed lipoprotein population (i.e., low density lipoproteins, Lp(a), and high density lipoprotein (HDL)-like particles) in the d 1.015-1.065 g/ml interval decreased by almost 50% from January to February (from 164.3 to 89.2 mg/100 ml), i.e., following a 10-fold increase in the level of plasma testosterone, and immediately before the rapid doubling in plasma thyroxine concentration. The staining intensity of the electrophoretic band with migration characteristics corresponding to those of Lp(a) decreased considerably during winter. At the same period of the year, lower density (1.032-1.055 g/ml) HDL-like particles disappeared. The concentration of lipoproteins with d 1.065-1.162 g/ml, which included Lp(a) particles in addition to typical HDL, equally underwent seasonal variations. These variations consisted of two successive maxima in late fall (426.4 mg/100 ml) and late winter (458.3 mg/100 ml) with two subsequent decreases leading to minima in February (327.8 mg/100 ml) and August (257.1 mg/100 ml). Finally, very high density lipoproteins (d 1.162-1.259 g/ml) were heterogeneous, containing both cholesterol-rich (d 1.162-1.227 g/ml) and phospholipid-rich (d 1.194-1.259 g/ml) subpopulations.(ABSTRACT TRUNCATED AT 400 WORDS)

Distribution and partial characterization of the plasma lipoproteins in the hedgehog (Erinaceus europaeus L.), a hibernator with potential use in the study of the hormonal regulation of lipoprotein metabolism.

The hedgehog is a hibernator in which yearly cycles of several endocrine activities and seasonal variations of plasma lipids have already been demonstrated. We have consequently undertaken a study of plasma lipids and lipoproteins in this animal, bled during late April and May. Plasma cholesterol levels (178 +/- 30 mg/100 ml) were comparable with those in normal humans, while triacylglycerol was lower (46 +/- 17 mg/100 ml) and phospholipids higher (252 +/- 35 mg/100 ml). The main characteristics of the plasma lipoprotein spectrum, as determined by sequential and density gradient preparative ultracentrifugation, analytical ultracentrifugation and gel filtration chromatography, were (1) a low concentration of very-low-density components (d less than 1.006 g/ml, about 20 mg/100 ml); (2) a continuity between the low (1.006-1.063 g/ml) and high (1.063-1.21 g/ml) density components, the former (about 150 mg/100 ml) exhibiting a considerable heterogeneity upon polyacrylamide gel electrophoresis while the latter were largely predominating (570 mg/100 ml); (3) the presence, at a density of 1.087 g/ml, of a band migrating electrophoretically like human low-density lipoproteins, a finding consistent with the results of apolipoprotein electrophoresis, which showed the presence of a high-molecular-weight counterpart to apolipoprotein B, in both the low- and high-density ranges defined above; (4) the presence, throughout the entire density spectrum, of an apolipoprotein with molecular weight and mobility in polyacrylamide/urea gels similar to human apolipoprotein A-I, and (5) the presence of very-high-density lipoproteins (d 1.178-1.259 g/ml) responsible for the transport of approximately 15% of plasma cholesterol and 20% of phospholipids.

Amino acid sequences of the alpha and beta chains of adult hemoglobin of the European hedgehog, Erinaceus europaeus.

Globin prepared from hemoglobin of the European hedgehog (Erinaceus europaeus) was separated into alpha and beta polypeptide chains by chromatography on a CM 52 column. The S-aminoethylated alpha and beta chains were each digested with trypsin and the resulting peptides were isolated. The sequences of all the tryptic peptides were established. The ordering of these peptides in the alpha and beta chains was deduced from their homology with the primary structures of alpha and beta chains of human adult hemoglobin. Comparing the primary structures of the alpha and beta chains of adult hemoglobin of the European hedgehog thus obtained with those of adult hemoglobin of the tupai (Tupaia glis), 35 amino acids substitutions in the alpha chains and 30 in the beta chains were recognized.

Venom resistance in the hedgehog, Erinaceus europaeus: purification and identification of macroglobulin inhibitors as plasma antihemorrhagic factors.

Hedgehog (Erinaceus europaeus) plasma contains factor(s) which neutralize the hemorrhagic activity of European viper (Vipera berus) venom. These antihemorrhagic factor(s) were purified by gel filtration on Sephacryl S-200 and chromatography on Cibacron Blue Sepharose. The macroglobulin fraction obtained was characterized for proteinase inhibiting activity, molecular weight and purity by polyacrylamide gel electrophoresis and immunological cross-reactivity and was found to contain the three macroglobulin proteinase inhibitors--alpha 2-macroglobulin, alpha 2-beta-macroglobulin and beta-macroglobulin. The purified macroglobulins were totally able to neutralize Vipera berus venom hemorrhagic activity, but no distinction between them in this respect was possible.

Purification and characterization of alpha 2-, alpha 2-beta- and beta-macroglobulin inhibitors in the hedgehog, Erinaceus europaeus: beta-macroglobulin identified as the plasma antihemorrhagic factor.

Three macroglobulin inhibitors were purified from hedgehog (Erinaceus europaeus) plasma by sequential chromatography on Cibacron Blue Sepharose, Sephacryl S-200 and preparative agarose gel electrophoresis. Each macroglobulin was characterized for proteinase inhibiting activity, molecular weight by polyacrylamide gel electrophoresis (PAGE), subunit size by sodium dodecyl sulfate (SDS)-PAGE, immunological cross-reactivity to other macroglobulins and antihemorrhagic activity against European viper (Vipera berus) venom. Hedgehog alpha 2-macroglobulin is a tetramer (Mr 800,000) composed of identical monomers (Mr 200,000) that inhibits all proteinases tested and is the homologue of human alpha 2-macroglobulin, rat alpha 2-acute phase globulin, dog alpha 1-macroglobulin and swine alpha 2-macroglobulin fast. Hedgehog alpha 2-beta-macroglobulin is a dimer (Mr 450-550,000) composed of identical monomers (Mr 200,000) that inhibits all proteinases tested and appears to be structurally similar to other animal 'half-molecule' macroglobulins. Hedgehog beta-macroglobulin (Mr 700,000) gave subunits of 34,000 and 39,000 after SDS-PAGE and showed cross-reactivity with swine alpha 2-macroglobulin slow. It inhibits all proteinases tested and is the only macroglobulin with antihemorrhagic activity against V. berus venom. This antihemorrhagic activity may be due to beta-macroglobulin's different structure as compared to other macroglobulins, which may make it less susceptible to inactivation by venom proteinases.

Normal haematological values of European hedgehogs (Erinaceus europaeus) from an English rehabilitation centre.

Blood samples were taken from 29 male and 21 female clinically normal European hedgehogs (Erinaceus europaeus) that had been overwintered in an English rehabilitation centre, and the mean (sd) and ranges of their haematological values were determined. The mean cellular volume and lymphocyte counts of the female hedgehogs were slightly but significantly higher than those of the male hedgehogs.

Clinical pathology and sample collection of exotic small mammals.

The clinical pathology of some of the less common and newly emerging small mammal species is detailed in this article. The species covered here include the chinchilla, prairie dog, African hedgehog, and sugar glider. Venipuncture sites and sampling techniques are discussed in general and for each species. Detailed information on the hematology and serum biochemistry values of these animals is presented in numerous tables. Specific information is also provided for urinalysis, fecal analysis, dermatologic sampling, and cytology.

Hematologic and biochemical variables of hedgehogs (Erinaceus europaeus) after overwintering in rehabilitation centers.

BACKGROUND: Information about laboratory reference intervals (RIs) of European Hedgehog (Erinaceus europaeus) hospitalized at rehabilitation centers is scarce. OBJECTIVE: The purpose of this study was to establish hematologic and biochemical RIs for rehabilitated hedgehogs before the release into the wild, and to assess whether sex and management of the center influence laboratory results. METHODS: Blood was collected from 50 hedgehogs at 3 centers. Thirty-eight animals were included in the study based on normal body weight, absence of clinical signs of disease, Bunnell index > 0.80, and absence of hibernation during overwintering. CBCs were performed using an automated laser cell counter followed by morphologic analysis of blood smears. Clinical biochemistry was performed using an automated spectrophotometer. RIs were determined as recommended by the ASVCP guidelines. RESULTS: Hematology profiles revealed a prevalence of lymphocytes, a constant presence of nucleated RBCs, Howell-Jolly bodies and basophils, and bilobed nuclei in neutrophils and eosinophils. Biochemistry profiles were characterized by higher creatinine and urea concentrations, and higher ALP and GGT activities compared with other domestic species. The sex did not influence the results. Conversely, numbers of eosinophils, activated and large granular lymphocytes, and concentrations of total protein, glucose and cholesterol were different among the centers, likely due to different management practices (eg, antiparasitic treatments, environmental exposure to microorganisms, diet). CONCLUSION: The RIs established in this study can be used to monitor the health status of hedgehogs in rehabilitation centers. As management practices appeared to influence some variables, it is recommended to standardize the management protocols to minimize their influence on laboratory data.

Accumulation of background levels of persistent organochlorine and organobromine pollutants through the soil-earthworm-hedgehog food chain.

The bioaccumulation of persistent organic pollutants (POPs), such as polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and DDT and metabolites, was investigated in the soil-earthworm-hedgehog food chain. Concentrations of selected POPs were measured in soil and earthworms collected in grassland and open woodland and in hair and blood of hedgehogs foraging in two parks containing these habitats. Despite background concentrations in soil (ranging from 1.3 to 9.3 ng/g for DDTs, 2.3 to 6.5 ng/g for PCBs and 0.08 to 0.20 ng/g for PBDEs), biota-soil accumulation factors (BSAFs) indicated that earthworms accumulated POPs (0.48-1.70 for DDTs, 1.09-2.76 for PCBs and 1.99-5.67 for PBDEs) and that animals feeding on earthworms are potentially exposed to higher concentrations of pollutants. BSAFs decreased with increasing soil concentrations for the three groups of compounds, suggesting that steady-state equilibrium was not reached in soil or earthworms. Positive, but low, log-linear relationships were found for DDT (r(2)=0.23, p<0.05 for Brasschaat and r(2)=0.63, p<0.01 for Hoboken) and PCB (r(2)=0.13, p<0.05 for both parks) concentrations between soil and earthworms. In order to relate earthworm to hedgehog POP concentrations, the foraging behavior of each individual was taken into account. The use of hair as a potential biomonitoring tissue in exposure and risk assessment of POPs was evaluated by examining the relationship between PCB and p,p'-DDE levels in hedgehogs' hair and blood. Contaminant profiles were used to gain insight into biotransformation of the studied compounds in each step of the investigated food chain and in the blood of hedgehogs, as well as the consequences thereof for their incorporation in hair. The absence of a discernable relationship between POP concentrations in earthworms and hair is possible due to variation in individual foraging behavior and POP uptake. Our results suggest that POPs in tissues should be measured from an adequate number of individuals per population instead of relying on indirect estimates from levels in soil or prey items.

Demonstration of a phospholipase A2 inhibitor in human plasma and in plasma from the European hedgehog (Erinaceus europaeus).

1. Endogenous phospholipase A2 (PLA2) inhibitors in human plasma and in plasma from the hedgehog (Erinaceus europaeus) were demonstrated. 2. The PLA2 activity increased 45-fold in human plasma when a PLA2 binding factor was removed. Furthermore, two peaks of PLA2 inhibitory activity were found after DEAE-chromatography. 3. High levels of PLA2 inhibitory activity was found in plasma from the European hedgehog, E. europaeus. 4. The molecular weight was estimated to 140,000. 5. On DEAE-chromatography two peaks were found which were chromatographically similar to the PLA2 inhibitors in human plasma.

Plasma glucose and insulin in the hibernating hedgehog.

Plasma glucose and insulin have been studied during lethargy and spontaneous arousal of hibernating hedgehogs. During lethargy, plasma glucose and insulin were low and showed no variation. Glucose and insulin injections given during lethargy showed no effects on plasma insulin and glucose respectively but confirmed a very low plasma clearance of glucose and insulin. During spontaneous arousal, the increase in plasma insulin occured before the increase in blood glucose and at about the time that utilization of blood glucose was restored.

Daily and seasonal variations in plasma LH and testosterone concentrations in the adult male hedgehog (Erinaceus europaeus).

A double-antibody heterologous radioimmunoassay (RIA) was developed to measure plasma LH values in hedgehogs. This RIA system used anti-rat LH serum and rabbit LH (AFP-559B) for radioiodination and as standard. The accuracy of the method was evaluated and indicated the ability to detect various relative concentrations of LH in plasma. The minimum detectable dose was 0.2 ng/ml. The intra- and inter-assay coefficients of variation were 4.2 and 7.9% respectively. Biological tests, e.g. effect of castration, effect of castration + testosterone implant and GnRH administration, confirmed that this method was suitable to determine subsequent changes in pituitary gonadotrophic activity in the hedgehog. LH concentrations were determined in blood samples obtained during 1 year: (a) each month, at 4-h intervals during 24 h, from different groups of unanaesthetized animals fitted with a catheter and (b) twice a month, under a light anaesthesia, from the same group of 6 animals. During the year: (1) the range of LH change was narrow (minimum values congruent to 0.25 ng/ml and maximum values congruent to 2.00 ng/ml); (2) the 24-h LH patterns did not exhibit any daily rhythm; (3) a clear annual rhythm was observed with the highest values from February to April and the lowest values in October and November. LH decreased rapidly at the end of summer and increased progressively from December to February, during hibernation. In these experiments, it was not possible to determine the characteristics of LH release patterns in the hedgehog but individual profiles indicated clearly the episodic secretion of LH, particularly during the highest pituitary activity period. During the year, a close relationship between the seasonal cycles of plasma LH and testosterone was observed.

Further studies of plasma protease inhibitors in the hedgehog, Erinaceus europaeus; collagenase, papain and plasmin inhibitors.

Hedgehog plasma was separated by gel filtration on Sephacryl S-200, the fractions resolved by electrophoresis and the electrophoretograms characterized for collagenase, papain and plasmin inhibiting activities with the high mol. wt substrate casein. The three inhibitors previously identified as alpha 2-, alpha 2-beta- and beta-macroglobulins were found to inhibit all three proteases. These were the only collagenase inhibitors found in plasma. Hedgehog alpha 2-chymotrypsin inhibitor and beta-protease inhibitor were both found to also inhibit papain. Three new inhibitors specific for papain (gamma-, alpha 2- and alpha 1-cysteine protease inhibitors) and one for plasmin (alpha 2-antiplasmin) were also found, bringing the number of protease inhibitors in hedgehog plasma to 14. Immunological cross-reactivity as studied by immunoelectrophoresis showed homology between hedgehog alpha 2-macroglobulin and rat murinoglobulin I and between hedgehog alpha 2-antithrombin and rat antithrombin III.

Hedgehog lipoprotein(a) is a modulator of activation of plasminogen at the fibrin surface. An in vitro study.

Lipoprotein(a) (Lp[a]), a highly atherogenic lipoprotein particle, is the prominent apolipoprotein B-containing lipoprotein in the hedgehog (Laplaud PM et al, J Lipid Res 1988;29:1157-1170). In the present work, we studied the consequences of the structural homology between the specific Lp(a) glycoprotein, apoprotein(a), and plasminogen on the generation of plasmin by fibrin-bound tissue-type plasminogen activator. The activation of plasminogen was initiated by adding either native plasma or Lp(a)-free plasma supplemented with the equivalent of 0.25 mg/ml of either purified Lp(a) or albumin to a surface of fibrin prepared on micortitration plates and to which human tissue-type plasminogen activator was specifically bound. With the Lp(a)-free plasma, an increase in the binding and activation of plasminogen as a function of time was observed. In contrast, in the presence of Lp(a) (i.e., native plasma or the reconstituted system), a significant decrease in the binding of plasmin(ogen) (approximately 60%) was obtained. These data indicate that hedgehog Lp(a) interferes with the binding and activation of plasminogen at the fibrin surface and may thereby behave as a factor regulating the extent of fibrin deposition. These results support our previous data indicating that high levels of Lp(a) may have antifibrinolytic effects in humans (Rouy D et al, Arterioscler Thromb 1991;11:629-638), are in agreement with the observation that Lp(a) is a risk factor for atherosclerotic disease, and provide further support to the view of Lp(a) as a link between atherosclerosis and thrombosis.

Protein and lipid utilization during fasting with shallow and deep hypothermia in the European hedgehog (Erinaceus europaeus).

We investigated whether the relative contributions of body protein and lipid reserves differ according to the level of energy expenditure in fasting animals. Protein and lipid utilization was therefore quantified and compared in hedgehogs which fasted with shallow and deep hypothermia, i.e. by exposure at 5 or 20 degrees C ambient temperature. Body composition was determined for every 150-g decrease in mass throughout the experiment, allowing the calculation of regression lines between body mass (independent variable, x) and body composition (dependent variable, y: water, protein, neutral lipids, phospholipids and cholesterol). There were highly significant (P < 0.001) linear decreases in all body components with decreasing body mass in both groups of hedgehogs. Neutral lipids were the main component of the total body mass loss (54%) in fasted animals with shallow and deep hypothermia, percentages of water (26-30%) and protein (10-11%) being lower, and those of phospholipid and cholesterol negligible (< 0.5%). In spite of different levels in energy expenditure (2.54 and 1.07 W.kg-1 in shallow- and deep-hypothermal fasting hedgehogs, respectively), the energy sources were identical in both groups, neutral lipid being the main fuel (91-92%) and body protein accounting for the remainder (8-9%). Prolonged fasting with shallow and deep hypothermia were marked by low alaninemia and glycemia, while plasma free fatty acids and beta-hydroxybutyrate were elevated.(ABSTRACT TRUNCATED AT 250 WORDS)