Prevalence of anti-Yersinia antibodies in European brown hares in North-Rhine Westphalia, Germany

Yersinia (Y.) pseudotuberculosis infections may lead to significant lethality in European brown hare (Lepus europaeus, Pallas) populations especially during the cold and wet seasons. In recent decades, also Y. enterocolitica was isolated from hares found dead. Consequently, a Western-blot technique proved to be valuable for the detection of antibodies against all pathogenic Yersinia isolates was applied to monitor the prevalence of antibodies in hare populations in North-Rhine Westphalia, Germany. A total of 89.6% of the 230 animals tested was seropositive. Further investigations should be performed to elucidate the role of subclinical yersiniosis in the decline of European brown hare populations in Germany.     

Tuberculosis in a free-living brown hare (Lepus europaeus occidentalis)

A free-living brown hare (Lepus europaeus occidentalis) trapped during a survey of tuberculous possums in the Ahaura Valley, Westland, was found to have both microbiological and histopathological evidence of infection with Mycobacterium bovis. The mesenteric lymph nodes were enlarged and showed extensive caseation, tuberculous nodules were present in the liver and kidney, and there was a proliferative pleurisy. Histologically, characteristic tuberculous granulomatous foci were seen in the mesenteric lymph nodes, liver, kidney, pleura and lungs, often in association with small numbers of acid-fast organisms. This is the first report of naturally occurring tuberculosis due to M. bovis in the brown hare.     

Hepatitis E virus in wild rabbits and European brown hares in Germany

Recently, a change of hepatitis E from being a typical travel‐associated disease to an autochthonous zoonosis in Germany was observed. An increasing number of autochthonous infections with the hepatitis E Virus (HEV) have been recognized in developed countries. Venison from wild boar is already known to be a potential source of infection, if not prepared properly by the consumer. In Germany, certain wild animals are known to be a reservoir for HEV. However, current information is missing about European brown hares (Lepus europaeus) and wild rabbits (Oryctolagus cuniculus). Thus, a total of 833 hunting‐harvested animals (European brown hares n = 669; wild rabbits n = 164) were tested for the occurrence of HEV RNA and HEV antibodies. For this, liver and blood specimens were taken after hunts in six German federal states. HEV antibodies were found by ELISA in 2.2% (624/14) of European brown hares, but no HEV RNA was detectable by nested real‐time RT‐PCR. In contrast, a seroprevalence of 37.3% (126/47) was observed for wild rabbits, and 17.1% (164/28) of the samples were HEV RNA positive. Genomic analysis revealed that these partial sequences clustered within the rabbit clade of HEV‐3 genotype. In addition, one rabbit sequence segregated into subtype 3g of HEV‐3. Highest seroprevalences for hares and rabbits were detected in the federal states of Bavaria and of Schleswig‐Holstein, respectively. Comparing urban, rural and insular areas, the highest seroprevalence was shown for wild rabbits in rural areas and for European brown hares on the northern island Fehmarn. This study provides evidence that European brown hares and wild rabbits from Germany can be infected with HEV. The different prevalences indicate that wild rabbits are a potential reservoir for HEV in Germany, whereas European brown hares seem to be only of minor importance for the epidemiology of HEV.     

Tuberculosis in a free-living brown hare (Lepus europaeus occidentalis)

A free-living brown hare (Lepus europaeus occidentalis) trapped during a survey of tuberculous possums in the Ahaura Valley, Westland, was found to have both microbiological and histopathological evidence of infection with Mycobacterium bovis. The mesenteric lymph nodes were enlarged and showed extensive caseation, tuberculous nodules were present in the liver and kidney, and there was a proliferative pleurisy. Histologically, characteristic tuberculous granulomatous foci were seen in the mesenteric lymph nodes, liver, kidney, pleura and lungs, often in association with small numbers of acid-fast organisms. This is the first report of naturally occurring tuberculosis due to M. bovis in the brown hare.     

Blood coagulation times in the European brown hare (Lepus europaeus)

Background:  Many causes of mortality in the European brown hare, such as bacterial and viral infections, anticoagulant poisoning, and trauma, may result in hemorrhage. There are, however, no reference values concerning blood clotting in this species.  Objectives:  The aim of this study was to determine reference values for blood coagulation times and related parameters in healthy European brown hares.  Methods:  Blood samples from 30 clinically healthy adult hares (15 males and 15 females) were obtained. Hares were physically restrained for blood collection from the cephalic vein into tubes containing citrate and EDTA.  Results:  Mean ± SD were obtained for thrombin time (TT) (13.97 ± 1.37 seconds), prothrombin time (PT) (13.32 ± 2.15 seconds), activated partial thromboplastin time (APTT) (16.73 ± 1.86 seconds), fibrinogen concentration (2.98 ± 1.06 g/L), and platelet count (355.28 ± 128.73 × 10⁹/L). Conclusions: Reference values for blood coagulation times and other parameters associated with blood clotting will be useful in the laboratory evaluation of hemorrhage in the European brown hare.     

The new French 2010 Rabbit Hemorrhagic Disease Virus causes an RHD-like disease in the Sardinian Cape hare (Lepus capensis mediterraneus)

Lagovirus is an emerging genus of Caliciviridae, which includes the Rabbit Hemorrhagic Disease Virus (RHDV) of rabbits and the European brown hare syndrome virus (EBHSV) of hares that cause lethal hepatitis. In 2010, a new RHDV related virus (RHDV2) with a unique genetic and antigenic profile and lower virulence was identified in France in rabbits. Here we report the identification of RHDV2 as the cause in Sardinia of several outbreaks of acute hepatitis in rabbits and Cape hare (Lepus capensis mediterraneus). This is the first account of a lagovirus that causes fatal hepatitis in both rabbits and hares.     

The Occurrence of European Brown Hare Syndrome in Finland

European brown hare syndrome (EBHS) is a viral hepatitis that affects European brown hares (Lepus europaeus) and varying hares (Lepus timidus). The disease occurs today in free-living and farmed hares in most European countries (Gustafsson et al. 1989, Gavier-Widén & Mörner 1991). In Sweden and Denmark the EBHS was first observed at the beginning of the eighties, however, in Finland and Norway the disease has not been reported yet.     

Haemorrhagic disease of lagomorphs: evidence for a calicivirus.

Studies on the aetiological agents of rabbit haemorrhagic disease (RHD) and European brown hare syndrome show that the viruses responsible for these infections can be placed in the family Caliciviridae. Established members of this group are vesicular exanthema virus (prototype), San Miguel sea lion virus and feline calcivirus. The human hepatitis E virus and the Norwalk agent may soon be included. The RHD virus genome consists of a positive stranded RNA molecule composed of 7437 nucleotides. A major subgenomic RNA of 2.2 kb, colinear with the 3' end of the genomic RNA, can also be recovered from infected liver tissue, and both RNAs are enclosed within viral capsids formed by a single major protein of approximately 60 kDa. Electron microscopic examination of organ suspensions from diseased animals shows two types of particle; 35-40 nm complete virions have the regularly arranged cup-shaped depressions typical of calcivirus morphology, and 23-25 nm smooth particles resulting from degradation of the outer surface structures of the complete virions.     

Detection of Francisella tularensis subsp. holarctica in a European brown hare (Lepus europaeus) in Thuringia, Germany

The isolation of Francisella tularensis subsp. holarctica biovar II (strain 06T0001) from a European brown hare (Lepus europaeus) from Thuringia, Germany, is described for the first time. Identification of the microorganism was carried out by phenotypic characterisation, partial sequencing of the 16S rRNA gene and specific PCR using the primers TUL4-435/TUL4-863 and FtC1/FtC4. The epidemiology of tularemia in Germany is discussed and a risk assessment for humans is made.     

Biochemical responses and oxidative stress in Francisella tularensis infection: a European brown hare model

Background

The aim of the present study was to investigate biochemical and oxidative stress responses to experimental F. tularensis infection in European brown hares, an important source of human tularemia infections.

Methods

For these purposes we compared the development of an array of biochemical parameters measured in blood plasma using standard procedures of dry chemistry as well as electrochemical devices following a subcutaneous infection with a wild Francisella tularensis subsp. holarctica strain (a single dose of 2.6 × 10⁹ CFU pro toto).

Results

Subcutaneous inoculation of a single dose with 2.6 × 10⁹ colony forming units of a wild F. tularensis strain pro toto resulted in the death of two out of five hares. Plasma chemistry profiles were examined on days 2 to 35 post-infection. When compared to controls, the total protein, urea, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were increased, while albumin, glucose and amylase were decreased. Both uric and ascorbic acids and glutathione dropped on day 2 and then increased significantly on days 6 to 12 and 6 to 14 post-inoculation, respectively. There was a two-fold increase in lipid peroxidation on days 4 to 8 post-inoculation.

Conclusions

Contrary to all expectations, the present study demonstrates that the European brown hare shows relatively low susceptibility to tularemia. Therefore, the circumstances of tularemia in hares under natural conditions should be further studied.     

Arbovirus antibodies in wild game caught in Moravia]

The HIT method was used to examine blood serums of the game in Moravia (roebuck, red deer, fallow deer, mouflon, wild boar, brown hare) for the presence of antibodies to arboviruses of these groups: alphavirus (Sindbis-SIN), flavivirus (West Nile-WN), tick-borne encephalitis (TBE) and Bunyamwera (Tahyna-TAH, Calovo-CVO). Antibodies to all viruses were detected, and namely in these frequencies: SIN 0.9%, WN 16.9%, TAH 41.5%, CVO 23.1% and TBE 8.5%.     

青海豆科植物根瘤及固氮特性的研究

对青海野生及栽培豆科植物结瘤以及固氮状况进行了调查研究,栽培豆科植物均能自然结瘤,野生豆科植物结瘤率近为45%。青海地区豆科植物根瘤大多圆形,棒状或指状,形状较为规则。栽培豆科植物根瘤多数为粉红色,而野生豆科植物多数为白色,棕色或黄色。125种根瘤样品用ARA测定结果表明,30%根瘤为无效根瘤,不同豆科植物不同地区根瘤固氮活性相差较大,一般固氮活力都较低,乙炔还原活性小于1umolC2H2/gFw.h.的检测样品有40%,最高达30umolC2H2/gFw.h,根瘤固氮活性与生育期,生活习性及生态条件有关。     

Hepatitis E Virus Antibody Prevalence in Wildlife in Poland

Hepatitis E is an important public health problem mostly in developing but occasionally also in industrialized countries. Domestic and wildlife animals are considered reservoirs of the hepatitis E virus (HEV). Since no information on the prevalence of autochthonous HEV infections in human and animal in Poland is available, the aim of the study was to investigate the HEV seroprevalence of different wildlife species as potential virus reservoirs in the country. No HEV antibodies were found in any of the sera collected from the red deer (Cervus elaphus), European bison (Bison bonasus), roe deer (Capreolus capreolus), elk (Alces alces), fallow deer (Dama dama), sika deer (Cervus nippon), Tatra chamois (Rupicapra rupicapra tatrica) or brown bear (Ursus arctos). HEV‐specific antibodies were detected in 44.4% (95% CI 38.3–50.7) serum samples originated only from wild boars. The percentage of seropositive wild boars differed significantly between the provinces and was positively correlated with the wild boar density and rurality of the area. This study showed that HEV circulates among wild boar population in Poland, and this species should be considered as an important reservoir of the virus.     

Contents of selected heavy metals in the liver, kidneys and abdominal muscle of the brown hare (Lepus europaeus Pallas, 1778) in Central Pomerania, Poland

The content of iron, zinc, copper, manganese, lead, and cadmium was determined in the kidney, liver, and abdominal muscle of the brown hare (Lepus europaeus), a species known for its bioindicative potential in environmental quality assessment. The animals assayed were divided into immature (in their first year of life; n=25) and adult (n=39). The hares were acquired in an area situated far away from major cities and direct impacts of industrial and transport pollution. The zinc content found in the assayed hare's muscles (25.9 microg/g w.w.) and that of manganese in the kidneys, liver, and muscles (2, 2.51 i 0.85 microg/g w.w., respectively) tended to be higher than those reported from the brown hare elsewhere in Europe. The content of cadmium (particularly in the kidneys) and lead proved substantial and close to those typical of the hare inhabiting industry-affected areas. Analysis of the data pooled for all the individuals showed Zn and Cd contents to increase with age in the kidneys, the liver Cd content increasing with age as well; on the other hand, the age-Cu content was negative. In addition, a number of significant correlations between the metals themselves were revealed, particularly with respect to Zn-Cd correlation in kidneys (r(s)=0.47), Fe-Cd, Zn-Cu, Mn-Zn, Mn-Cu, and Mn-Pb in the liver (r(s) of 0.42, 0.86, 0.72, 0.79, and 0.42, respectively), and Zn-Cu in the muscle (r(s)=0.56). The kidney cadmium content was higher by 81% in the adult than in the immature hare, the adult hare muscle copper content being lower by 15.5%.     

The first appearance of infectious hemorrhagic disease of rabbits in Austria

In three Austrian rabbit units (two in the west, one in the east) heavy losses occurred in April/May 1989. The clinical course (deaths in adult animals, haemorrhagic diathesis) of the disease was indicative for an infection with the RHD-virus. Necropsy confirmed the suggestive clinical diagnosis as bleeding of the nostrils, hyperaemia in the respiratory tract, spleen tumor, partly decolorized livers, hyperaemia of kidneys with rare petechias could be observed. Histologic examination revealed centrolobular liver necrosis, lung bleeding and edema, tumor of spleen and atrophy of spleen follicles. Rabbits infected with organ suspensions died within 48 hours. The organ suspensions and the suspensions of the already necropsied animals showed a haemagglutination titer for above 1:100. This reaction could be inhibited with a specific RHD-antiserum. The intramuscular application of a RHD-reference strain in one hare, two wild rabbits and a rabbit did not induce clinical disease or death of the leporids during a six week observation period. The rabbit died within 48 hours post infection. However the hare and wild rabbits showed high antibody titers by ELISA at the end of the observation period. Before infection the two wild rabbits were serologically negative.     

新疆部分地区兔球虫种类调查研究

对新疆部分地区家兔球虫的种类进行调查,发现兔的艾美耳球虫检出率为100%,并检出10种兔艾美耳球虫,即斯氏艾美耳球虫(Eimera stiedai)、大型艾美耳球虫(E.magna)、无残艾美耳球虫(E.irrestidua)、梨型艾美耳球虫(E.piriformis)、小型艾美耳球虫(Eexigua)、肠艾美耳球虫(E.intestinalis)、盲肠艾美耳球虫(E.coecicola)、穿孔艾美耳球虫((E.perforans)、野兔艾美耳球虫(E.leporis)、黄艾美耳球虫(E.flsvescens),其中穿孔艾美耳球虫、小型艾美耳球虫和斯氏艾美耳球虫为优势虫种.     

Preliminary reference values for electrocardiography,echocardiography and myocardial morphometry in the European brown hare (Lepus europaeus)

The study aimed at defining reference values for electrocardiographic (ECG) and echocardiographic parameters as well as macroscopic dimensions of the heart and microscopic dimensions of cardiomyocytes in the European brown hare. The studies were conducted on 30 adult, clinically healthy hares of either sex caught in Poland. ECG and echocardiography were performed supravitally on anaesthetized hares. After euthanasia, gross and microscopic myocardial and cardiomyocyte dimensions were determined. Heart rate amounted to 140 ± 37.5 beats/min, the leading rhythm involved the sinus rhythm. P wave time was 26 ± 5 ms, PQ time was 80 ms, QRS time was 29 ± 3.5 ms, and ST was 97.5 ± 7 ms. Echocardiography determined a left ventricular wall end-diastolic diameter of 8.6 ± 2.0 mm and an intraventricular septum end-diastolic diameter of 5.75 ± 1.0 mm. The thickness of the interventricular septum corresponded to that of the free wall of the left ventricle, a finding consistent with physiological hypertrophy. Preliminary reference values were established for echocardiography. The findings were similar to those obtained at necropsy. The ECG and echocardiographic studies represent the first supravital examination of cardiac function in the hare. The obtained results illustrate adaptation of hare''s myocardium to its mode of life. The cardiac findings resemble the athlete''s heart syndrome described in humans. The findings may prove useful in further studies on the physiology of the cardio-vascular system in the hare.     

Occurrence of Baylisascaris transfuga in wild populations of European brown bears (Ursus arctos) as identified by a new PCR method

The European brown bear (Ursus arctos) is a species present in limited areas of Europe and several small populations are considered endangered. This species can be affected by a range of parasites. In particular, the genus Baylisascaris is an emerging parasite of wild animals which can cause severe larva migrans syndrome in aberrant hosts, which include 100 species of birds, mammals and also humans. Baylisascaris transfuga is the species reported from bears, and with the exception of a few laboratory trials, little is known about the capacity of this species to infect other animals. Furthermore, the identification of this species has traditionally been based on light microscopy, using either morphology of the adults at necropsy or detection of the eggs in faeces, which are methods limited by the experience and the efforts of the observer. The current work was aimed at developing a specific PCR to detect the parasite directly from faecal samples of naturally infected brown bears in the field, without the need for previous flotation. Using eggs and adults of B. transfuga collected in Croatia, we first developed a PCR to detect a portion of the second internal transcribed spacer region (ITS-2) of ribosomal DNA and then applied it to bear faecal samples spiked with a known number of B. transfuga eggs. We show here for the first time that this method allows the detection of a minimum of two Baylisascaris eggs in 25mg of faecal material, thus it demonstrates a high diagnostic capacity that could be applied to evaluate the prevalence of the parasite in faecal samples from wild populations of brown bears.     

Genetic profile and serosurvey for virus infections of Japanese wild boars in Shikoku Island

To estimate the genetic background of Japanese wild boars inhibiting the 4 prefectures of Shikoku Island (Kagawa, Tokushima, Kouchi and Ehime), we examined haplotypes of mitochondrial DNA (mtDNA) and genotypes of the nuclear glucosephosphate isomerase-processed pseudogene (GPIP) in 189 wild boars. Of the 8 different mtDNA haplotypes (J5, J10, J12, J13, J15, J19, J20 and E33) detected in Shikoku Island, the first to be identified were J19 and J20 (of Japanese wild boar lineage) and E33 (of European domestic pig lineage). The presence of haplotype E33 indicates local crossbreeding between wild boar and escaped domestic pigs and/or crossbred Inobuta. Four kinds of the GPIP genotypes were detected from 189 wild baors in Shikoku Island, but no European GPIP alleles were found. In 113 wild boars, no antibodies against Classical Swine Fever or Aujeszky's disease were detected, suggesting that they had not been exposed to those pathogens.