Identical 18S rRNA haplotypes of Hepatozoon canis in dogs and foxes in Brandenburg,Germany

Hepatozoon canis is a blood parasite of the suborder Adeleorina infecting wild and domestic canids. Transmission occurs by oral uptake of Rhipicephalus sanguineus sensu lato vector ticks infected with H. canis, but vertical transmission is also assumed to be possible. In German foxes, a high prevalence of H. canis has previously been reported despite the fact that R. sanguineus s.l. is not endemic. In the absence of knowledge about local transmission pathways, foxes should be considered to be possible reservoirs of H. canis and contribute to infection of domestic dogs. The present study aimed to determine how often foxes and dogs are infected in Brandenburg (Germany) and if identical or different H. canis 18S rRNA haplotypes are found in these host species. Hepatozoon spp. were detected by PCR in 46/1050 (4.4 %) of dog blood and 176/201 (77.6 %) of fox spleen samples from Brandenburg. Sequencing of 19 dog and 56 fox samples identified all as H. canis. For nine positive dogs, owners stated that they had never left Germany suggesting that autochthonous transmission occurs not only in foxes but also in dogs. Sequences for seven of these possible autochthonous cases were obtained and six were identical to the predominant haplotype found in the foxes. Haplotype network analysis confirmed that many dogs, including some without travel history, carried the same or very similar 18S rRNA haplotypes as the foxes suggesting that both hosts participate in the same epidemiological cycle.     

Seroprevalence of tick-borne encephalitis (TBE) in naturally exposed monkeys (Macaca sylvanus) and sheep and prevalence of TBE virus in ticks in a TBE endemic area in Germany

In a recently published study, a clinical case of severe tick-borne encephalitis (TBE) in a monkey (Macaca sylvanus) was described after natural exposure (tick bite) in a TBE endemic area in Germany, and from the tissue of this monkey the strain ‘Salem’, closely related to the strain ‘Neudoerfl’, was isolated and characterized.In this study, it was our aim to test all the available data for characterizing a TBE endemic area like TBE cases in humans and animals, TBE virus (TBEV) in ticks, and TBE-positive sera from animals. Sera from 283 monkeys and 100 sheep as well as 294 unfed ticks were collected at the monkey mountain Salem and its surroundings. A seroprevalence of 2.6% in monkeys and 9% in sheep were found. Furthermore, a new real-time RT-PCR method was established and used, in combination with an already published RT-qPCR, for TBEV genome detection in field-collected ticks, but no TBEV could be detected in the ticks tested.     

Outpatient antibiotic use and prevalence of antibiotic-resistant pneumococci in France and Germany: a sociocultural perspective

The prevalence of penicillin-nonsusceptible pneumococci is sharply divided between France (43%) and Germany (7%). These differences may be explained on different levels: antibiotic-prescribing practices for respiratory tract infections; patient-demand factors and health-belief differences; social determinants, including differing child-care practices; and differences in regulatory practices. Understanding these determinants is crucial for the success of possible interventions. Finally, we emphasize the overarching importance of a sociocultural approach to preventing antibiotic resistance in the community.     

GM(1,1)模型在洞庭湖区血吸虫病发病预测中应用

目的对洞庭湖区退田还湖地区中山试点血吸虫病发病进行预测。并为国家卫生机构合理分配卫生资源提供决策依据。方法应用GM(1，1)模型对中山试点血吸虫病患病率建模并进行残差修正，进行3年预测。结果中山试点GM(1，1)模型平均相对误差为13．61％，模型精度为差；残差GM(1，1)模型平均相对误差为4．85％，模型精度为优。残差修正预测模型为-↑X^(1)(k 1)=-19．373641e^-0.081742k 20．677187。连续3年预测值分别为15．71％，16．54％和17．53％。结论中山试点残差GM(1，1)模型预测效果好；血吸虫病发病在未来3年内有缓慢上升的趋势；要加强血防工作。     

Magnitude and causes of blindness and low vision in Anambra State of Nigeria (Results of 1992 point prevalence survey)

A survey to determine the prevalence and causes of blindness in Anambra State of Nigeria was conducted. The aim was to provide baseline data for the planning, implementation and evaluation of both the state's and the National Programme for Prevention of Blindness.A multistage cluster random sampling technique was used. The World Health Organization/Prevention of Blindness (WHO/PBL) Eye Examination Record Form was used. The WHOs definitions of blindness and low vision were adopted for the analysis.The prevalence of blindness in the state is estimated to be 0.33%±0.27%. Visual acuity of from 3/60 to less than 6/60 has a prevalence of 0.41%±0.30% while visual acuity of from 6/60 to less than 6/18 has a prevalence of 0.67%±0.39.There are equal numbers of blind males as females, although the prevalence among males is 0.44%±0.26% while among females it is 0.24%±0.15%. Most of the blind are above 50 y of age with prevalence of blindness in this age group being 2.62%±1.31% (3.27%±2.1% for males and 2.02%±1.58% for females).Cataract caused most of the blindness (70.59%), followed by glaucoma (17.65%). Macular degeneration is becoming important (5.88%) while obvious infective causes are rare. Errors of refraction are important public eye health care problems.Methods of tackling the cataract problem (both backlog and incident), and other eye health needs within the primary eye/health care are recommended. The need to extend refraction services to the rural areas is emphasized.     

2008-2018年云南省人间布鲁菌病流行特征及灰色GM(1,1)模型预测

目的分析云南省布鲁菌病(简称布病)流行特征,建立灰色GM(1,1)模型,预测云南省布病病情。方法收集中国疾病预防控制信息系统和云南省统计局2008-2018年云南省布病疫情数据和人口资料,分析布病流行病学特征(包括时间、地区、人群分布),并以灰色GM(1,1)模型进行建模,预测2019、2020年云南省布病发病率。结果2008-2018年云南省共报告布病病例1216例,年均发病率为0.2374/10万,呈逐年递增趋势(χ²趋势=843.34,P<0.01)。病例报告主要集中在3-9月份,占总病例数的69.41%(844/1216)。病例报告数居前5位的州市分别为红河州(289例)、曲靖市(264例)、昆明市(258例)、大理州(160例)、玉溪市(134例),占总病例数的90.87%(1105/1216)。职业以农民为主,占79.03%(961/1216)。建立灰色GM(1,1)模型预测2019、2020年云南省布病发病率分别为0.4876/10万和0.4817/10万。结论云南省布病发病较以往上升,应对重点地区、重点人群进行针对性防控,并对预测结果进行前瞻性评价,逐步完善云南省布病预测模型。     

Cost-effectiveness of heptavalent conjugate pneumococcal vaccine (Prevenar) in Germany: considering a high-risk population and herd immunity effects

In Germany, the seven-valent conjugate vaccine Prevenar is recommended for use in children at high risk of pneumococcal disease. Recent data suggest that giving conjugate vaccine to all children may lead to a decline in pneumococcal disease in unvaccinated adults, a phenomenon known as herd immunity. This analysis evaluated the cost and economic consequences in Germany of vaccinating (1) children at high risk, (2) all children when considering only benefits for vaccinated individuals and (3) all children when also considering herd immunity benefits. Costs in the model included vaccination, management of meningitis, bacteraemia, pneumonia and acute otitis media, insurance payments to parents and the costs of care for long-term disabilities. The model estimated that the cost-effectiveness of vaccination would be 38,222 euros per life year gained in children at high risk and 100,636 euros per life year gained in all children when not considering herd immunity. When considering herd immunity effects, the model estimated that offering vaccination for all children would reduce adult deaths by 3,027 per year, and vaccination would be broadly cost neutral. The findings are sensitive to the effect of conjugate vaccine on the rates of pneumonia and invasive disease in the elderly. If the herd immunity effect of conjugate vaccination in Germany is similar to that observed elsewhere, offering vaccine to all children will be more attractive than the current policy of restricting vaccination to children at high risk of pneumococcal disease.     

Study of Health In Pomerania (SHIP): a health examination survey in an east German region: objectives and design

SummaryObjectives The reason for the Study of Health in Pomerania (SHIP) is the lack of epidemiological studies with a broad range of health indicators. Furthermore, in Germany there is a need for studies that take into account the particular situation of life after the reunification. One objective of SHIP is to provide prevalence estimates on a broad range of diseases, risk and health factors for a defined region in the former GDR.Methods A sample of 7008 women and men aged 20 to 79 years in a north-east region of Germany, 4900 expected participants. The sample was drawn in two steps: First, 32 communities in the region were selected. Second, within the communities a simple random sample was drawn from residence registries, stratified by gender and age. The data collection and instruments include four parts: oral health examination, medical examination, health-related interview, and a health- and riskfactor-related questionnaire. The oral health examination includes the teeth, periodontium, oral mucosa, craniomandibular system, and prosthodontics. The medical examination includes blood pressure measurements, electrocardiography, echocardiography, carotid, thyroid and liver ultrasounds, neurological screening, blood and urine sampling. The computer-aided health-related interview includes cardiovascular symptoms, utilisation of medical services, health-related behaviours, and socioeconomic variables. The self-administered questionnaire comprises housing conditions, social network, work conditions, subjective well-being and individual consequences from the German reunification.The research work reported is funded by grants from the German Federal Ministry for Education and Research (BMBF, grant no. 01ZZ96030), of the Ministry for Education, Research and Cultural Affairs and the Ministry for Social Affairs of the State of Mecklenburg-West Pomerania as well as the Municipal Hospital of Stralsund GmbH.     

Influenza vaccination of hospital staff in Germany: a five-year survey on vaccination coverage and policies: identified deficits in influenza immunisation campaigns for hospital employees

Summary. Objectives: Influenza vaccination of hospital staff is recommended by STIKO, the German committee for vaccination. A survey was conducted to assess compliance with this recommendation. The occupational health services of 25 hospitals participated in a survey and provided data by questionnaire on influenza vaccination and on hospital policies to promote coverage of employees.Methods: Vaccination activities were monitored by occupational health services (OHS) for five consecutive years from 1997 to 2002. The hospital sample covered a total of 17089 beds (3.23% of the hospital capacity in Germany) and a total number of 41969 employees (4.39% of hospital staff).Results: The proportion of hospitals actively offering influenza vaccination increased from 48% in 1997/98 to 92% in 2001/02. Vaccination coverage of all staff in 1997 was only 3.3% and reached 8.4% in 2001/02. Coverage of vaccinating hospitals increased from 5.0% to 10.4%. Poster campaigns and managing board commitment had significant impact.Conclusions: Considerable progress has been made to involve more hospitals and to increase coverage for vaccination of hospital employees. Nevertheless, coverage levels remain unacceptably low. Recommendations are ignored extensively.

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Zusammenfassung. Influenzaimpfung von Krankenhauspersonal in Deutschland: eine Fünfjahresuntersuchung zu Durchimpfungsraten, Impfpolitik und -defiziten in 25 deutschen KrankenhäusernFragestellung: Die Impfung von Krankenhauspersonal gegen Influenza wird von der Ständigen Impfkommission (STIKO) empfohlen. Die Umsetzung dieser Empfehlung in deutschen Krankenhäusern wurde untersucht. Die Betriebsärzte von 25 Krankenhäusern nahmen an einer Umfrage teil, für die sie Daten ihres jeweiligen Hauses zur Influenzaimpfung und Impfpolitik bereitstellten.Methode: Die Untersuchung fand in fünf aufeinander folgenden Jahren (1997–2002) statt. Die Stichprobe umfasste insgesamt 17089 Betten (3,23% der gesamten deutschen Bettenkapazität) und 41969 Angestellten (4,39% des deutschen Krankenhauspersonals).Ergebnisse: Der Anteil der impfenden Krankenhäuser stieg von 48% in der Saison 1997/98 auf 92% in 2001/02. Die Durchimpfungsrate lag 1997 bei nur 3,3% und 2001/02 erreichte sie 8,4%. Die Impfrate in impfenden Krankenhäusern stieg im gleichen Zeitraum von 5,0% auf 10,4%. Poster-Kampagnen und Einbeziehung der Krankenhausleitung haben signifikanten Einfluss.Schlussfolgerung: Es zeigt sich eine deutliche Steigerung sowohl der Krankenhäuser mit Impfangebot als auch der Durchimpfungsrate des Krankenhauspersonals. Nichtsdestotrotz bleibt diese aber auf einem nicht akzeptabel niedrigem Niveau.

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Résumé. Vaccination contre la grippe du personnel hospitalier en Allemagne: une recherche de cinq ans sur les taux de vaccination et la politique appliquée en matière de vaccination dans 25 hôpitaux allemandsObjectifs: La vaccination du personnel hospitalier contre la grippe est recommandée par la Commission permanente de la vaccination du Robert Koch Institut. Cette étude a porté sur lapplication de cette recommandation dans les hôpitaux allemands. Les médecins du travail de 25 hôpitaux ont participé à une enquête dans le cadre de laquelle ils ont fourni les données suivantes: prévalence de la vaccination contre la grippe et méthodes de promotion de la vaccination.Méthodes: Lenquête a eu lieu cinq années consécutives (1997–2002). Elle a porté sur 25 hôpitaux, soit sur 41969 employés (4,39% du personnel hospitalier allemand).Résultats: La proportion dhôpitaux pratiquant la vaccination est passée de 48% pour la période 1997/98 à 92% pour 2001/02. En 1997, le taux de vaccination ne sélevait quà 3,3% pour atteindre 8,4% en 2001/02. Le taux de vaccination dans les hôpitaux pratiquant déjà la vaccination en 1997 est passé durant la même période de 5,0% à 10,4%. Limplication de la direction hospitalière ainsi que le recours à des affiches ont eu un impact significatif.Conclusions: Le nombre dhôpitaux proposant la vaccination, ainsi que le taux de vaccination ont nettement augmenté. Néanmoins, ce taux reste à un niveau inacceptablement bas.

     

Molecular identification of tick-borne pathogens (Rickettsia spp., Anaplasma phagocytophilum,Borrelia burgdorferi sensu lato,Coxiella burnetii and piroplasms) in questing and feeding hard ticks from North-Western Spain

The occurrence of tick-borne pathogens (TBPs) of human and veterinary interest was studied in questing and feeding ticks collected from wild animals in a region in North-Western Spain. A total of 529 ticks (489 questing, 40 feeding) of seven different species (386 Ixodes ricinus, 53 Haemaphysalis concinna, 27 Haemaphysalis punctata, 25 Dermacentor marginatus, 21 Haemaphysalis inermis, 15 Dermacentor reticulatus, and two Rhipicephalus bursa) were analyzed. Molecular analysis of the 16S rRNA gene in I. ricinus ticks, revealed the presence of two phylogenetic groups in the region. Most of the sequenced ticks (96%) were assigned to I. ricinus haplogroup and 4% of the ticks were phylogenetically related to I. inopinatus haplogroup. Feeding ticks were removed from 17 animals from seven wild species (seven roe deer -Capreolus capreolus-, three wolves -Canis lupus-, two Iberian red deer -Cervus elaphus hispanicus-, two European wild boar -Sus scrofa-, one Cantabrian brown bear -Ursus arctos-, one Eurasian badger -Meles meles-, and one red fox -Vulpes vulpes-). Presence of Rickettsia spp., Anaplasma phagocytophilum, piroplasms, Borrelia burgdorferi sensu lato (s.l.) and Coxiella burnetii were tested in ticks by specific PCR. A total of 92 (17.4%) of the 529 ticks analyzed were positive for at least one of the TBPs tested. Sequencing revealed the presence of the genospecies “Candidatus Rickettsia rioja”, Rickettsia raoultii, and Anaplasma phagocytophilum in both questing and feeding ticks. Rickettsia slovaca, Borrelia lusitaniae, Borrelia afzelii, Borrelia garinii, Borrelia burgdorferi sensu stricto and Babesia bigemina were only detected in questing ticks, while Babesia sp. badger type A, Theileria OT3 and Hepatozoon canis occurred only in engorged ticks. None of the ticks were positive for C. burnetii. The analysis of the 16S rRNA gene sequences of A. phagocytophilum revealed the presence of three variants (I, X and W) circulating in the region. New host-tick-pathogen interactions have been revealed, finding for the first time the human pathogen R. raoultii in D. reticulatus removed from a Cantabrian brown bear. Co-occurrence between different TBPs were detected in 4.3% of the ticks. The association B. burgdorferi s.l./Rickettsia spp. was detected in questing ticks; and Rickettsia spp./piroplasms and A. phagocytophilum/Theileria OT3 in feeding ticks. The presence of pathogenic agents constitutes a threat to human and animal health, and should be considered in the diagnosis and treatment after a tick bite. This study increases the knowledge on TBPs diversity of medical and veterinary interest circulating between ticks and their hosts in North-Western Spain.     

Greenland sledge dogs (Canis familiaris) develop liver lesions when exposed to a chronic and dietary low dose of an environmental organohalogen cocktail

We assessed the relationship between exposure to organohalogen polluted minke whale (Balaenoptera acutorostrata) blubber and liver morphology and function in a generational controlled study of 28 Greenland sledge dogs (Canis familiaris). The prevalence of portal fibrosis, mild bile duct hyperplasia, and vascular leukocyte infiltrations was significantly higher in the exposed group (all Chi-square: p<0.05). In case of granulomas, the frequency was significantly highest in the bitches (P generation) while the prevalence of portal fibrosis was highest in the F generation (pups) (both Chi-square: p<0.05). No significant difference between exposed and controls was found for bile acid, ALAT, and ALKP, while ASAT and LDH were significantly highest in the control group (both ANOVA: p<0.05). We therefore suggest that a daily intake of 50-200g environmentally organohalogen polluted minke whale blubber can cause liver lesions in Greenland sledge dogs. It is reasonable to infer that other apex predators such as polar bears (Ursus maritimus) and humans may suffer from similar impacts.     

Comprehensive screening of tick-borne microorganisms indicates that a great variety of pathogens are circulating between hard ticks (Ixodoidea: Ixodidae) and domestic ruminants in natural foci of Anatolia

Grazing domestic ruminants serve as important reservoirs and/or amplificatory hosts in the ecology of tick-borne pathogens (TBPs) and tick vectors in the natural foci; however, many enzootic life cycles including ruminants and ticks are still unknown. This study investigated a wide range of TBPs circulating among ticks and grazing ruminants in the natural foci of Anatolia, Turkey. Tick specimens (n = 1815) were collected from cattle, sheep, and goats in three ecologically distinct areas (wooded, transitional, and semi-arid zones) of Anatolia and identified by species: Dermacentor marginatus, Dermacentor reticulatus, Hyalomma anatolicum, Hyalomma excavatum, Hyalomma marginatum, Hyalomma scupense, Haemaphysalis inermis, Haemaphysalis parva, Haemaphysalis punctata, Haemaphysalis sulcata, Ixodes ricinus, Rhipicephalus bursa, and Rhipicephalus turanicus. PCR-sequencing analyses revealed TBPs of great diversity, with 32 different agents identified in the ticks: six Babesia spp. (Babesia occultans, Babesia crassa, Babesia microti, Babesia rossi, Babesia sp. tavsan1, and Babesia sp. Ucbas); four Theileria spp., including one putative novel species (Theileria annulata, Theileria orientalis, Theileria ovis, and Theileria sp.); one Hepatozoon sp.; four Anaplasma spp., including one novel genotype (Anaplasma phagocytophilum, Anaplasma marginale, Anaplasma ovis, and Anaplasma sp.); six unnamed Ehrlichia spp. genotypes; Neoehrlichia mikurensis; nine spotted fever group rickettsiae, including one putative novel species (Rickettsia aeschlimannii, Rickettsia slovaca, Rickettsia hoogstraalii, Rickettsia monacensis with strain IRS3, Rickettsia mongolitimonae, Rickettsia raoultii, Candidatus Rickettsia goldwasserii, Candidatus Rickettsia barbariae, and Rickettsia sp.); and Borrelia valaisiana. Detailed phylogenetic analyses showed that some of the detected pathogens represent more than one haplotype, potentially relating to the tick species or the host. Additionally, the presence of Neoehrlichia mikurensis, an emerging pathogen for humans, was reported for the first time in Turkey, expanding its geographical distribution. Consequently, this study describes some previously unknown tick-borne protozoan and bacterial species/genotypes and provides informative epidemiological data on TBPs, which are related to animal and human health, serving the one health concept.     

Levels and predictors of urinary nickel concentrations of children in Germany: Results from the German Environmental Survey on children (GerES IV)

Human biomonitoring of nickel has gained interest in environmental medicine due to its wide distribution in the environment and its allergenic potential. There are indications that the prevalence of nickel sensitization in children is increased by nickel exposure and that oral uptake of nickel can exacerbate nickel dermatitis in nickel-sensitive individuals. Urinary nickel measurement is a good indicator of exposure. However, data on nickel levels in urine of children are rare. For the first time, the German Environmental Survey on children (GerES IV) 2003–2006 provided representative data to describe the internal nickel exposure of children aged 3–14 years in Germany. Nickel was measured after enrichment in the organic phase of urine by graphite furnace atomic absorption spectrometry with Zeeman background correction. Nickel levels (n = 1576) ranged from <0.5 to 15 μg/l. Geometric mean was 1.26 μg/l. Multivariate regression analysis showed that gender, age, socio-economic status, being overweighted, consumption of hazelnut spread, nuts, cereals, chocolate and urinary creatinine were significant predictors for urinary nickel excretion of children who do not smoke. 20.2% of the variance could be explained by these variables. With a contribution of 13.8% the urinary creatinine concentration was the most important predictor. No influence of nickel intake via drinking water and second hand smoke exposure was observed.     

Survey on viral pathogens in wild red foxes (Vulpes vulpes) in Germany with emphasis on parvoviruses and analysis of a DNA sequence from a red fox parvovirus.

The seroprevalence of canine parvovirus (CPV), canine distemper virus (CDV), canine adenovirus (CAV) and canine herpesvirus (CHV) infections in red foxes (Vulpes vulpes) was determined in fox sera collected between 1991 and 1995. A total of 500 sera were selected and the seroprevalences were estimated to be 13% (65 of 500 sera) for CPV, 4.4% (17 of 383 sera) for CDV, 35% (17 of 485 sera) for CAV, and 0.4% (2 of 485 sera) for CHV, respectively. No statistically significant differences were observed between the two (rural and suburban) areas under study. Parvovirus DNA sequences were amplified from tissues of free-ranging foxes and compared to those of prototype viruses from dogs and cats. We report here a parvovirus sequence indicative of a true intermediate between the feline panleukopenia virus-like viruses and the canine parvovirus-like viruses. The red fox parvoviral sequence, therefore, appears to represent a link between those viral groups. The DNA sequence together with a significant seroprevalence of parvovirus infections in foxes supports the hypothesis that the sudden emergence of canine parvovirus in the domestic dog population may have involved the interspecies transmission between wild and domestic carnivores.     

Molecular characterization of tick-borne bacteria and protozoans in yaks (Bos grunniens), Tibetan sheep (Ovis aries) and Bactrian camels (Camelus bactrianus) in the Qinghai-Tibetan Plateau Area,China

Due to the specific plateau climate, a variety of unique animals live in the Qinghai-Tibetan Plateau Area (QTPA) including yaks (Bos grunniens), Tibetan sheep (Ovis aries) and Bactrian camels (Camelus bactrianus). However, information on tick-borne diseases (TBDs) in the QTPA and on the molecular characteristics of tick-borne pathogens (TBPs) in the area is limited. Therefore, the aim of this study was to investigate Anaplasma spp., Babesia spp., Theileria spp., Borrelia burgdorferi sensu lato and Rickettsia spp. infecting yaks, Tibetan sheep and camels in this area. A total of 276 animals were screened. Overall, 84.5% (164/194) of yaks, 58% (23/40) of Tibetan sheep, and 38% (16/42) of camels tested positive for at least one pathogen. Theileria spp., Anaplasma ovis and spotted fever group (SFG) Rickettsia spp. were detected as TBPs in the current study with overall infection rates of 10.9% (30/276), 8.3% (23/276) and 62.9% (171/276), respectively. Further study revealed that 1.5% (3/194) of the yaks were infected with Theileria sp. OT3, 1.5% (3/194) with T. luwenshuni, 6.2% (12/194) with T. uilenbergi, 1.1% (2/194) with T. ovis and 82% (159/194) with SFG Rickettsia spp. It was also shown that 58% (23/40) of the Tibetan sheep were infected with A. ovis and 15% (6/40) with T. ovis. Among the camels, 10% (4/42) were infected with T. equi, while 29% (12/42) were positive for Rickettsia spp. Sequence analysis revealed that the Rickettsia spp. infecting yaks and camels were Rickettsia raoultii and Rickettsia slovaca. To the best of our knowledge, this study reports the first detection and characterization of these pathogens in yaks, Tibetan sheep and camels in the country, except for T. luwenshuni infections in yaks.     

Are epidemiological and sociological surveys a proper instrument for detecting true problem drinkers? (The low sensitivity of an alcohol survey in Rotterdam)

The sensitivity of an alcohol survey in Rotterdam is estimated at only 16%. Taking account of sampling fluctuations, an upper confidence limit of 47% for the sensitivity can be computed. The statistical procedure is not based upon interviewing true problem drinkers to be identified from registers, but upon an anonymous verification of positively scoring individuals and a sample of negatively scoring individuals in the survey with registers of true problem drinkers. The above results may have implications for the interpretation of survey based prevalence figures.     

Aids in sicily: Prevalence of antibodies to human immunodeficiency virus (HIV) in low and high risk groups

A seroepidemiological survey, carried out to evaluate the prevalence of antibodies to HIV in patients with ARC and in healthy individuals at risk for AIDS, showed the infection to be widespread in the groups at risk, namely in drug abusers and hemophiliacs.However, remarkable difference existed between the prevalence of antibodies to HIV in drug abusers of the city of Palermo and those of other Sicilian provinces. Spread of the virus among Sicilian thalassemics, however, was very low and quite similar in all geographic areas. Antibodies were found very rarely (0.06)% in unpaid voluntary blood donors. The spread of the virus is still confined in high risk groups, and the major part of the seropositive blood donors were identified a posteriori as drug abusers.     

Rapid identification of Babesia canis and Babesia gibsoni (Asian genotype) in canine blood samples using a customized portable real-time PCR analyzer and TaqMan-based assay

Canine babesiosis is a serious infectious disease in subtropical and tropical regions. Typically, clinical detection of canine babesiosis is performed by blood smear observation or the traditional polymerase chain reaction (PCR). Herein, we developed a new TaqMan-based real-time PCR assay combined with a customized portable real-time PCR platform for a rapid and accurate detection of canine babesiosis. Two new primer/probe pairs (B18S and BITS1) were designed based on 18S ribosomal RNA and an internal transcribed spacer 1 (ITS1) sequence to differentiate Babesia canis and B. gibsoni (Asian genotype) DNAs from canine blood samples. Additionally, a corresponding customized compact real-time PCR platform with low 6-carboxyfluorescein fluorescence detection (≤5 nM), including a fast and accurate thermal cycling ability with a user-friendly interface for thermal control and data analysis, was designed for the limited space use. Both assays (B18S and BITS1) demonstrated a sensitivity of 100 copies/reaction based on the 95 % confidence interval evaluation method. The self-developed customized portable real-time PCR analyzer presented high repeatability and reproducibility with the TaqMan-based assay. Moreover, 501 clinical specimens were collected for evaluating the performance of the proposed PCR. The positive and negative predictive values were 90 % (18 of 20) and 100 % (226 of 226), respectively, for samples suspected with B. canis infection and 98 % (55 of 56) and 100 % (199 of 199), respectively, for samples suspected with B. gibsoni infection.     

Molecular cloning of translocation breakpoints in a case of constitutional translocation t(11;22)(q23;q11) and preparation of probes for preimplantation genetic diagnosis

In vitro fertilization (IVF) centres with preimplantation genetic diagnosis (PGD) programmes are often confronted with the problem of identifying chromosomal abnormalities in interphase cells biopsied from preimplantation embryos of carriers of a reciprocal translocation. The present authors have developed a DNA testing based approach to analyse embryos from translocation carriers, and this report describes breakpoint-spanning probes to detect abnormalities in cases of the most common human translocation (i.e. the t(11;22)(q23;q11)). Screening a yeast artificial chromosome (YAC) library for probes covering the respective breakpoint regions in the patient lead to probes for the breakpoint on chromosome 11q23. The physically mapped YAC and bacterial artificial chromosome (BAC) clones from chromosome 22 were then integrated with the cytogenetic map, which allowed localization of the breakpoint on chromosome 22q11 to an interval of less than 84 kb between markers D22S184 and KI457 and to prepare probes suitable for interphase cell analysis. In summary, breakpoint localization could be accomplished in about 4 weeks with additional time needed to optimize probes for use in PGD.