Prevalence of Coxiella burnetii in Central and Eastern European dairy herds

The aim of the present study was to assess the prevalence of Coxiella burnetii in dairy herds in Central and Eastern European countries based on ELISA and PCR tests. A total of 370 bulk tank milk samples were collected in 2019 originating from Croatia (n = 13), Czech Republic (n = 138), Hungary (n = 126), Serbia (n = 24), Slovakia (n = 53) and Slovenia (n = 16). Prevalence of C. burnetii differed according to the country of origin with Croatia showing 100.00%, the Czech Republic 98.55%, Hungary 97.61%, Serbia 70.83%, Slovakia 90.56% and Slovenia showing 62.50% average percentages of the positive herds. C. burnetii specific ELISA showed 100.00% positivity in all examined countries if herds consisted of equal or above 250 milking cows. The growing number of farms managing large number of animals, where cattle density is high correlates with the increasing prevalence of C. burnetii in the region.     

New patterns of Trichinella infection

Human and animal trichinellosis should be considered as both an emerging and reemerging disease. The reemergence of the domestic cycle has been due to an increased prevalence of Trichinella spiralis, which has been primarily related to a breakdown of government veterinary services and state farms (e.g., in countries of the former USSR, Bulgaria, Romania), economic problems and war (e.g., in countries of the former Yugoslavia), resulting in a sharp increase in the occurrence of this infection in swine herds in the 1990s, with a prevalence of up to 50% in villages in Byelorussia, Croatia, Latvia, Lithuania, Romania, Russia, Serbia, and the Ukraine, among other countries. The prevalence has also increased following an increase in the number of small farms (Argentina, China, Mexico, etc.) and due to the general belief that trichinellosis was a problem only until the 1960s. The sylvatic cycle has been studied in depth at both the epidemiological and biological level, showing the existence of different etiological agents (Trichinella nativa, Trichinella britovi, Trichinella murrelli, Trichinella nelsoni) in different regions and the existence of "new" transmission patterns. Furthermore, the role of game animals as a source of infection for humans has greatly increased both in developed and developing countries (Bulgaria, Canada, Lithuania, some EU countries, Russia, USA, etc.). The new emerging patterns are related to non-encapsulated species of Trichinella (Trichinella pseudospiralis, Trichinella papuae, Trichinella sp.), infecting a wide spectrum of hosts (humans, mammals including marsupials, birds and crocodiles) and to encapsulated species (T. spiralis, T. britovi, and T. murrelli) infecting herbivores (mainly horses). The existence of non-encapsulated species infecting mammals, birds and crocodiles had probably remained unknown because of the difficulties in detecting larvae in muscle tissues and for the lack of knowledge on the role of birds and crocodiles as a reservoir of Trichinella. On the other hand, it is not known whether horse and crocodile infections existed in the past, and their occurrence has been related to improper human behavior in breeding. The problem of horse-meat trichinellosis is restricted to France and Italy, the only two countries where horse-meat is eaten raw, whereas mutton and beef have been found to be infected with Trichinella sp. only in China.     

An overview of the epidemiology and epizootology of brucellosis in selected countries of Central and Southeast Europe

The objective of this paper is to give an overview of the epidemiologic and epizootic status of brucellosis in selected countries of Central and Southeast Europe (Balkan region). Based on dimension of the disease problem, there is a need to establish collaboration in the eradication and prevention of brucellosis between all countries in the region. Although there were no readily accessible data concerning epidemiology and epizootology of brucellosis in these countries, the limited official and published data were analyzed.

The incidence of brucellosis caused by Brucella melitensis in sheep, goats and humans is a very significant problem in Macedonia and Greece. In Greece, cattle are also affected either by B. melitensis or B. abortus. The disease is an endemic problem in some regions of Yugoslavia and includes B. suis biovar 2 in pigs and in Croatia, B. melitensis in sheep, goats and human is found occasionally. No problem appears to exist with brucellosis in Bulgaria.

Financial well-supported brucellosis control programs of the European Union that will include all countries, regardless of the magnitude of brucellosis incidence, are needed for eradication and control of brucellosis.     

Spatial distribution of Trichinella britovi, T. pseudospiralis and T. spiralis in red foxes (Vulpes vulpes) in Hungary

The red fox (Vulpes vulpes) is considered one of the main reservoir of Trichinella spp. in Europe. As limited information on Trichinella infection in wildlife of Hungary is available, 2116 red foxes, representing more than 3% of the estimated fox population of the country, were screened to detect Trichinella larvae by a digestion method. Trichinella larvae from the 35 positive foxes were identified by a multiplex PCR as Trichinella britovi (30 isolates, 85.7%), Trichinella spiralis (4 isolates, 11.4%), and Trichinella pseudospiralis (1 isolate, 2.9%). The true mean intensity of T. britovi, T. spiralis and T. pseudospiralis larvae in lower forelimb muscles was 23.6, 3.5 and 13.5larvae/g, respectively. T. spiralis was detected only in the southern and eastern regions. The non-encapsulated T. pseudospiralis was recorded for the first time in Hungary. Although the overall true prevalence of Trichinella infection in foxes was only 1.8% (95% confidence interval, CI=1.5-2.1%), the spatial analysis reveals different risk regions. In the north-eastern counties bordering Slovakia and Ukraine (21% of the Hungarian territory), the true prevalence of Trichinella infection is significantly higher than that observed in other regions (6.0%, CI=4.8-7.1%). In the southern counties bordering Croatia, Serbia and Romania (41% of the Hungarian territory), the true prevalence of Trichinella infection is moderate (1.4%, CI=1.0-1.8%). In the north-western and central counties (38% of Hungarian territory), the prevalence of Trichinella infection is significantly lower (0.2%, CI=0.1-0.4%) than that of the other regions. Based on the statistical analysis and the evaluation of epidemiological data, none of the counties can be considered free of Trichinella infection. In the past decade, Trichinella infection has been detected only in few backyard pigs, and only few wild boar-related autochthonous infections in humans were described. Nevertheless, these results highlight the need of the maintenance of a strict monitoring and control programmes on Trichinella infection in farmed and hunted animals of Hungary.     

Spatial distribution of Trichinella britovi, T. spiralis and T. pseudospiralis of domestic pigs and wild boars (Sus scrofa) in Hungary

Trichinellosis is a foodborne disease caused by the consumption of raw meat and raw meat-derived products from swine, horse and some game animals infected with nematode worms of the genus Trichinella. Between June 2006 and February 2011, 16 million domestic pigs and 0.22 million wild boars (Sus scrofa) were tested for Trichinella sp. in Hungary. Trichinella infection was not found in any pigs slaughtered for public consumption. Nevertheless, Trichinella spiralis was detected in four backyard pigs when trace back was done following a family outbreak. Trichinella infection was demonstrated in 17 wild boars (0.0077%). Larvae from wild boars were identified as Trichinella britovi (64.7%), T. spiralis (29.4%) and Trichinella pseudospiralis (5.9%). Although the prevalence of Trichinella sp. infection in wild boars and domestic pigs is very low, the spatial analysis reveals that the level of risk differs by region in Hungary. Most of the T. britovi infected wild boars (63.6%) were shot in the north-eastern mountain area of Hungary; whereas domestic pigs and wild boars infected with T. spiralis were detected only in the southern counties bordering Croatia and Romania. In the north-western and central counties, the prevalence of Trichinella infection seems to be negligible.     

The genetic diversity and structure of 18 sheep breeds exposed to isolation and selection

The phylogenetic layout of the genotyped (30 microsatellite) 18 sheep breeds in this study demands and provides the opportunity to evaluate both neutral and adaptive components of genetic diversity in a naturally and artificially selected and subdivided sheep population. Seven Pramenka strains from Bosnia and Herzegovina and Croatia characterized by a very low intensity of artificial selection, preserved the highest neutral genetic variability. Eight central and north‐western European breeds under considerable artificial isolation and selection preserved the lowest genetic variability. Only combinations of various phylogenetic parameters offer a reasonable explanation for underlying evolutionary forces working in the investigated island and mainland sheep breeds under variable natural and artificial selection. More than 60% of total genetic, diversity was allocated to virtually unselected Pramenka strains, and an additional 25% to native moderately selected Graue Gehoernte Heidschnucke and intensively selected Ostfriesische Milchschafe. Some economically very important breeds and strains did not contribute to a pool with maximal genetic diversity, while they play an important role in the cultural heritage of respective countries.     

假苍耳的地理分布及潜在适生区预测

外来入侵植物假苍耳在中国东北部地区快速蔓延,威胁自然生态系统的结构并对人们的健康产生危害。本研究通过野外调查获得假苍耳在东北地区的分布范围,采用Maxent模型软件对假苍耳在我国及世界的潜在适生区进行预测,运用ArcGIS软件中的自然间断点分级法进行适生等级分类,ROC曲线评价Maxent模型预测的准确性,并结合刀切法及环境变量反应曲线判断分析影响假苍耳分布的主要环境变量。结果表明,假苍耳的适生区较广,扩散范围可能继续增大。我国的华北地区、东北地区、华东地区、华中地区、西南地区、西北地区均存在适生区,高度适生区主要分布在我国的黑龙江省、吉林省、辽宁省、内蒙古自治区、山西省、陕西省、河南省、河北省、山东省、北京市、天津市、新疆维吾尔族自治区。世界范围内除南极洲外均存在假苍耳的适生区,高度适生区主要在北美洲的加拿大和美国,非洲的摩洛哥,欧洲的西班牙、克罗地亚、匈牙利、斯洛伐克、波黑、塞尔维亚、马其顿、保加利亚、格鲁吉亚、亚美尼亚和阿塞拜疆,亚洲的土耳其、伊朗、哈萨克斯坦、吉尔吉斯斯坦、塔吉克斯坦、中国、朝鲜、韩国和日本。上述适生区预测显示出假苍耳适宜生长在年均温度较高,最冷月份最低温和最冷季度平均温度较低的中纬度地区。     

Trichinella prevalence in swine in an endemic district in Serbia: Epidemiology and control

Endemic trichinellosis is re-emerging in Serbia and it is a serious problem both from the perspective of human health and animal husbandry. The widespread appearance of human trichinellosis is attributed to a high prevalence of Trichinella infection in domestic animals, especially swine. Epidemiological data presented in this paper were collected during a 12-year period (1995–2006) at small private swine farms in the region of Branicevo, Serbia where a high Trichinella prevalence in slaughter pigs (0.57%) has been detected. To further monitor Trichinella prevalence in swine, a serological survey, using ELISA, was performed in 2006. Of 916 swine tested by ELISA, Trichinella specific antibodies were detected in 15 (1.64%), while suspect results were obtained in 10 (1.09%). Positive or suspect animals originated from all parishes except one (Pozarevac). Our results point to the need for systematic monitoring in pigs to achieve a better control of trichinellosis in Serbia.     

Evaluation of ELISA and Western Blot Analysis using three antigens to detect anti-Trichinella IgG in horses

We assessed a serological method for detecting Trichinella infection in horses, specifically, an ELISA using three antigens to detect anti-Trichinella IgG (i.e. a synthetic tyvelose glycan-BSA (stg-BSA) antigen, an excretory/secretory (ES) antigen, and a crude worm extract (CWE) antigen). Serum samples were collected from 2502 horses (433 live horses from Romania and 2069 horses slaughtered in Italy and originating from Italy, Poland, Romania, and Serbia). Serum samples were also taken from horses experimentally infected with different doses of T. spiralis and T. murrelli larvae, as controls. The cut-off value of ELISA was determined on serum samples from 330 horses from Trichinella-free regions of Italy, which were also examined by artificial digestion of preferential-muscle samples. In the experimentally infected horses, the stg-BSA and ES antigens were less sensitive than the CWE antigen. Trichinella spiralis showed a higher immunogenicity than T. murrelli, and the IgG immunoresponse was dose-dependent. The kinetics of anti-Trichinella IgG were similar among all experimentally infected horses. No circulating antibodies were detected 4-5 months after experimental infection, although these horses still harbored infective larvae. Depending on the antigen used, for 4-7 of the 330 horses from Trichinella-free areas, the optical density (OD) of the serum sample was higher than the cut-off value, yet these samples were negative when subjected to Western Blot. Similar results were obtained for the 1739 horses slaughtered in Italy (originating from Italy, Poland, Romania, and Serbia) and the 433 live Romanian horses. Of the 4 horses with muscle larvae, only one was positive by ELISA and Western Blot. Because the anti-Trichinella IgG remain circulating for only a short period of time, whereas the larvae remain infective for longer periods, serology cannot be used for either diagnosing Trichinella infection in horses or estimating the prevalence of infection. Artificial digestion of at least 5 g of preferential-muscle tissue continues to be the method of choice at the slaughterhouse for preventing equine-borne trichinellosis in humans.     

Prevalence of Haemophilus parasuis serotypes in large outdoor and indoor pig units in Hungary/Romania/Serbia

Gross postmortem pathologic-anatomical and bacteriological examination, and serotyping for Haemophilus parasuis were performed in 401 weaned pigs with the clinical symptoms of Gl?sser's disease, originating from indoor (204 piglets out of 12 units) and outdoor (197 piglets out of 9 units) units in the country triangle Hungary/Romania/Serbia. The majority of the isolates in indoor units included Haemophilus parasuis serotypes 2, 5 and non typable, while in outdoor units, a high diversity (serotypes 1, 2, 4, 5, 11, 12, 14, 15, and non typable) of Haemophilus parasuis serotypes was found.     

The need for implementation of International Commission on Trichinellosis recommendations, quality assurance standards, and proficiency sample programs in meat inspection for trichinellosis in Serbia

Implementation of methods to control inspection for Trichinella in meat recommended by International Commission on Trichinellosis (ICT), particularly the introduction of the quality assurance standards and proficiency panels for certified analysts is extremely important in Serbia and other countries where Trichinellosis is endemic. In spite of existing regulations, including the inspection of 0.5 g samples of diaphragm by the compression method or by artificial digestion of 1g samples, in Serbia 280 people were diagnosed with clinical trichinellosis after consumption of inspected meat during the period 2001--2002. These outbreaks, which occurred in the municipalities of Kumane, Surcin and Bogatic, were a consequence of inadequate application of inspection methods and insufficient education of some veterinary inspectors. The problem of inadequate veterinary inspection in Serbia can be overcome by strict application of the ICT recommendations for the control of Trichinella with specific emphasis on implementing the quality assurance system (QAS) and proficiency sampling (PS/--PP/panel).     

Postweaning multisystemic wasting syndrome (PMWS) in pigs in Croatia: detection and characterisation of porcine circovirus type 2 (PCV2)

The objective of this study was to characterise porcine circovirus type 2 (PCV2) from pigs with naturally occurring postweaning multisystemic wasting syndrome (PMWS) in Croatia, and to determine the epizootiological, clinical and pathomorphological features of the disease. During a systematic health monitoring programme conducted in the period from January 2002 to June 2003, PMWS was suspected on eight different pig-producing farms in Croatia. The diagnosis of PMWS met all three key criteria: the presence of compatible clinical signs, the presence of the characteristic microscopic lymphoid lesions, and the detection of PCV2 within the lesions by polymerase chain reaction (PCR) and by in situ hybridisation (ISH). Moreover, PCV2 DNA from swine tissues was extracted and sequenced. The phylogenetic analysis of 4 Croatian PCV2 strains showed close relationship to PCV2 strains isolated in Slovenia, France, the Netherlands, the United Kingdom, China and Hungary. PCV2 was also demonstrated by electron microscopy in the lymph node of an affected animal. This is the first demonstration of PMWS in Croatia based on all scientifically accepted diagnostic criteria.     

Post Hoc Assessment of Stand Structure Across European Wood-Pastures: Implications for Land Use Policy

Europe’s woodland and savanna rangelands, often part of silvopastoral systems known as wood-pastures, are deteriorating because of abandonment that leads to return to a forested state or lack of tree regeneration from overgrazing or tree and shrub removal. Despite numerous local studies, there has been no broader survey of the stand structure of European wood-pastures showing which systems are at risk of losing their semiopen character. This overview aims to 1) show some of the differences and similarities in wood-pastures from landscapes across Europe and 2) identify which of these wood-pastures are at risk of losing their semiopen character. We collated a dataset of 13 693 trees from 390 plots in wood-pastures from eight different European regions (western Estonia, eastern Greece, northern Germany, Hungary, northern Italy, southern Portugal, central Romania, and southern Sweden), including tree diameters at breast height, tree density, management type, and tree species composition. On the basis of their structural characteristics, we classified wood-pastures using principal component analysis (PCA) and cluster analysis. The PCA showed a gradient from dense wood-pastures with high levels of regeneration (e.g., in Estonia) to sparse wood-pastures with large trees but a lack of regeneration (e.g., in Romania). Along this gradient, we identified three main groups of wood-pastures: 1) sparse wood-pastures with mostly big trees; 2) dense wood-pastures composed of small trees, and 3) wood-pastures containing a wide range of tree ages. Our results show a large structural gradient in European wood-pastures, as well as regeneration problems varying in their severity, highlighting the importance of social-ecological context for wood-pasture conditions. To maintain the ecological and cultural integrity of European wood-pastures, we suggest 1) more comprehensively considering them in European policies such as the Common Agricultural Policy and EU Habitats Directive, while 2) taking into account their structural characteristics and social-ecological backgrounds.     

Genotyping of Bacillus anthracis Isolated from Croatia and Bosnia and Herzegovina

Anthrax is a serious disease caused by Bacillus anthracis. Humans can become infected by handling products from infected animals, by breathing spores and rarely by eating undercooked meat from infected animals. The genome of B. anthracis is highly monomorphic and thus shows very low DNA sequence variation. We analysed the molecular characteristics of 12 B. anthracis isolates from outbreaks in Croatia and Bosnia and Herzegovina, which have occurred during the past 10 years along with two vaccine strains. Genotyping system based on variable‐number tandem repeat analysis at six loci revealed that six isolates belong to genotype from the A1.a cluster whilst six isolates relate to the B2 cluster, compared to 89 previously described genotypes. The distribution of two evolutionarily distant clusters suggests an introduction of B. anthracis to this area in at least two separate events.     

猪Kobu病毒感染概况

猪Kobu病毒是小核糖核酸病毒科Kobu病毒属成员,是2008年从匈牙利健康猪群中分离到的新病毒,目前在一些亚洲和欧洲的国家均报道了本病毒感染的发生。论文重点对猪Kobu病毒在猪群中的感染状况进行了综述,该病毒在临床表现正常的猪群或表现腹泻病症的猪群均出现较高的阳性率,可能在猪的胃肠炎致病作用中充当了一定的角色。     

Epidemiological and diagnostic findings during a 16-year-long trichinellosis surveillance in Timis County,Romania

The main source of trichinellosis in Romania is insufficiently cooked pork infected with Trichinella spiralis. Timis, the largest Romanian county, is located in the western part of the country where pork and traditional food prepared from pork are extensively consumed. The objective of the present study was to conduct a retrospective investigation of the incidence of human trichinellosis in Timis County over a period of 16 years (1990–2005). The medical records of 521 patients were used as the source of data. Patients were hospitalized at “Victor Babes” Hospital of Infectious Diseases from Timisoara during the period 1990–2005. The highest number of cases was recorded in 1994 (16.90%) and most of the patients were in the age group of 20–29 (23.22%). Males and females were almost equally affected by trichinellosis. The clinical pattern was mainly characterised by myalgia (72.55%) and edema (54.12%). Laboratory investigations included mainly elevated eosinophil and leucocyte values. In 52.02% of the patients, the hospitalization period ranged between 8 and 14 days. Most of the patients were from urban areas, but often the source of infection was found in rural areas. Hospitalization of the affected patients required considerable health care resources. The decreasing incidence of trichinellosis in Romania over the last decade has been due to sanitary education programmes for both swine breeders and consumers.     

Fighting Rabies in Eastern Europe,the Middle East and Central Asia ‐ Experts Call for a Regional Initiative for Rabies Elimination

MEEREB is an informal network of rabies experts from the Middle East, Eastern Europe and Central Asia, seeking to eliminate rabies from the region. They met for the second time to review the current rabies situation, both globally and in their respective countries, highlighting current rabies control problems and potential solutions. Success stories in Latin America, in Western Europe, in some Asian countries, as well as in Croatia and Serbia prove that elimination of human rabies is achievable in the MEEREB region. It requires political willingness and cooperation of all stakeholders, including Ministries of Health and of Agriculture; adequate management of animal bites through post‐exposure prophylaxis; pre‐exposure prophylaxis for populations at high risk of rabies exposure, animal vaccination and humane control of stray dog populations. MEEREB members called for a regional initiative for rabies elimination in Eastern Europe and the Middle East. They are confident that the elimination of human rabies of canine origin can be achieved in the region through adopting a One Health approach, and that campaigns for rabies elimination will have significant benefit for public health, including strengthening the structure for control of other zoonoses.     

Spatial distribution of Dermacentor reticulatus and Ixodes ricinus in Hungary: evidence for change?

A survey was conducted to investigate the spatial distribution of Ixodes ricinus and Dermacentor reticulatus in Hungary and to compare these data with the results of a previous large-scale survey. In the survey conducted in the 1950s, D. reticulatus adults were detected in two isolated areas of two counties, and the presence of these ticks in the collection was explained by accidental introduction. In the present survey, D. reticulatus became the second most common species occurring in all 16 counties involved in the monitoring and showed high prevalence. The change in the spatial distribution of this tick species, the increase of incidence of Babesia canis infection in Hungary, and the increasing number of canine babesiosis case reports from other Central and Central Eastern European countries since the 1970s suggest an expansion of the geographic range of D. reticulatus and intensification of the transmission rate of B. canis and probably other D. reticulatus-borne diseases (e.g. tularemia and tick-borne lymphadenopathy) in the region. The spatial distribution of I. ricinus was roughly in line with the results of the earlier survey. I. ricinus was the most common tick species being present in all 16 counties with the highest prevalence. Nevertheless, the comparison of the data of the previous and current survey cannot be used for fine-scale analysis; thus, it cannot be dismissed that the spatial distribution of I. ricinus also changed during the past decades. The spatial distribution patterns of tick-borne encephalitis in Hungary and other Central Eastern European countries may indicate such a change.     

Trichinella in horses: a low frequency infection with high human risk

After the initial report in 1976 of a trichinellosis epidemic caused by the consumption of infected horsemeat, 12 other outbreaks have been described in Europe. Since the first serious human outbreak several experiments have confirmed the susceptibility of horses to Trichinella species and the rapid disappearance of specific antibodies in this host that prevents the use of serological methods for routine screening. A review of the distribution of parasite burdens in muscles of naturally or experimentally infected horses indicates that the tongue is the most likely sample to contain detectable numbers of Trichinella larvae in low level infections. Requirements for testing of horsemeat are specified in legislation of the European Union, and other recommendations are published elsewhere. The EEC directives have evolved into very specific requirements which specify the testing of at least 5g of tongue, masseter or diaphragm per horse using a pooled digestion assay. More recently, France has revised the requirement for sample size to 10g for horsemeat originating from countries with high prevalence of Trichinella. To address the continuing outbreaks of human trichinellosis due to infected horsemeat, the development and implementation of a quality assurance system for testing is being considered.