Molecular identification of cryptic cysticercosis: Taenia ovis krabbei in wild intermediate and domestic definitive hosts.

The complex life cycle of taeniids represents an ideal model of a multi-host system. The complexity of these parasites can therefore cover the epidemiological issues of the interface between wild and domestic animals, especially once spatial overlap between wild and domestic definitive and intermediate hosts occurs. Here we use the occurrence of Taenia ovis krabbei in two model areas as an example of this epidemiological complexity. In two contiguous areas in the Italian northern Apennines, two hunted roe deer (Capreolus capreolus) showed numerous cysticerci in the muscles of their whole body and an adult tapeworm was recorded in a semi-stray dog (Canis lupus familiaris). Through molecular typing of the mitochondrial cytochrome c oxidase I (cox1) gene, cysticerci and the adult tapeworm of T. krabbei were identified. Taenia krabbei cysticercosis was recorded for the first time in Italy. Although the role of dogs in the parasite's life cycle emerges, the overlap between wild and domestic definitive hosts and the increase of wild population densities raise concerns about the temporal (old or new) introduction and the spread of this parasite by one of these canid species (wolf (Canis lupus) or dog). Although T. krabbei is not a public health issue, economic concerns emerged for hunters and meat producers, related to the damage of carcasses by cysticerci. Therefore, there is a need to evaluate the spread of T. krabbei in the intermediate and definitive host populations, and to ensure the relevant sanitary education for hunters in order to avoid practices that could favour the spread and maintenance of its life cycle.

Characterization of Immune System Cell Subsets in Fixed Tissues from Alpine Chamois (Rupicapra rupicapra).

Immune system cell subsets in lymph nodes and spleen from alpine chamois (Rupicapra rupicapra subspecies rupicapra) living in the Italian Alps were characterized immunohistochemically. Seven primary antibodies (against human CD3, CD79αcy, CD68, or ovine CD4, CD8, CD21 and γδ T-cell receptor [TCR] epitopes) were tested on tissues fixed either in formalin or in zinc salts (ZS) and cross-reactivity with chamois immune cell epitopes was shown. ZS fixation allowed wider identification of immune cells, without the need for antigen retrieval. CD4(+) and CD21(+) cells were labelled only in ZS-fixed tissues. Reagents specific for human CD3, CD79 and CD68 antigens successfully detected chamois immune cells, both in ZS-fixed and formalin-fixed tissues. The reactivity and distribution of immune cells in lymph nodes and spleen were similar to those described in other domestic and wild ruminants. Results from this study may allow future investigation of the immune response and pathogenesis of diseases in the chamois.

Genetic variability of Haemonchus contortus (Nematoda: Trichostrongyloidea) in alpine ruminant host species.

Genetic variability of the ovine parasite Haemonchus contortus from the Alpine area was investigated using mitochondrial DNA (nd4 gene), internal transcribed spacers 1 and 2 and microsatellites, in order to assess whether cross-transmission between domestic and wild ruminants occurs. The dataset was composed of 78 individual adult male H. contortus collected from chamois (Rupicapra r. rupicapra), roe deer (Capreolus capreolus), alpine ibex (Capra ibex ibex), domestic goat (Capra hircus) and sheep (Ovis aries) from different alpine areas. The data obtained show low host specificity and high genetic variation within H. contortus populations. The analyses indicate the presence of two mitochondrial haplotype clusters among host species and the absence of cryptic parasite species, confirming H. contortus as a generalist nematode and suggesting that parasite transmission between populations of domestic and wild ruminants normally occurs.

Experimental ELISA for diagnosis of ovine dicrocoeliosis and application in a field survey.

Dicrocoeliosis is a parasitic disease caused by trematodes belonging to the genus Dicrocoelium. It usually produces mild symptoms, and for this reason, dicrocoeliosis often remains undetected. Its diagnosis is mostly based on postmortem examination of the liver or on coprological assays for in vivo diagnosis. However, the latter method has scant sensitivity and because of the long pre-patency of Dicrocoelium spp. only permits late diagnosis. In the present study, an enzyme-linked immunosorbent assay (ELISA) was developed based on excretory/secretory antigen of Dicrocoelium dendriticum. The ELISA detected antibodies (IgG) in experimental infected sheep starting from day 30 post-infection (d.p.i.), whereas coprological samples were positive from 58 d.p.i. The ELISA was used in a field study in order to assess dicrocoeliosis seroprevalence in ovine flocks from the province of Trento (northeastern Italy), and this was the first sero-epidemiological study of ovine dicrocoeliosis in northern Italy. Altogether, 842 sheep sera were tested. In accordance with previous surveys carried out in other regions of Italy, a high prevalence of 80% to 100% was found.

The effect of gender on autonomic and respiratory responses during sleep among both young and middle-aged subjects.

BACKGROUND AND OBJECTIVES: Sleep affects the control of circulation and respiratory function. Gender and age are also known to have a profound impact on the neural control of circulation. We investigated whether gender affects sleep-related cardiovascular and respiratory responses and whether these vary according to healthy subjects being young or middle-aged. METHODS: We studied 32 subjects: 8 women and 8 men aged 20-30 years (young), and 8 women and 8 men aged 50-60 years (middle-aged). Young women were under oral contraceptive therapy and middle-aged women were postmenopausal and not receiving hormonal replacement therapy. One-night polysomnography was used to assess RR variability during non-rapid eye movement (NREM) (stage 2) and rapid eye movement (REM) sleep. Low-frequency (LF) and high-frequency (HF) components, in normalized units (LFnu and HFnu) and LF/HF ratio were calculated on five-minute segments selected across the night and averaged for each sleep stage. The respiration frequency in NREM and REM sleep was also measured. Interaction between gender, age and sleep on autonomic and respiration variables was assessed by 2 x 2 x 2 analysis of variance (ANOVA). RESULTS: Compared to men, women had a greater NREM-to-REM increment in LFnu (gender-by-state interaction, p<0.01), a greater decrement in HFnu (interaction, p<0.01) and a greater increment in LF/HF (interaction, p<0.05). Women also showed a more pronounced increase in respiratory frequency during REM sleep compared to men in both groups of age (gender-by-state interaction, F=7.1, p<0.05). No gender-by-age-by-state interaction was observed to affect autonomic and respiration variables. CONCLUSION: NREM-to-REM excitatory cardiac and respiratory responses are more marked among women compared to men, regardless of their hormonal status and whether they are young or middle-aged.

Nocturnal blood pressure changes in patients with restless legs syndrome.

OBJECTIVES: To assess heart rate (HR) and blood pressure (BP) changes associated with periodic leg movements during sleep (PLMS) with or without EEG signs of arousal in subjects with primary restless legs syndrome (RLS). METHODS: Ten patients with RLS (4 women, aged 47.3 +/- 13.5 years) underwent one night of polysomnography along with noninvasive beat-to-beat BP monitoring. Ten PLMS with microarousals (PLMS-MA) and 10 PLMS without microarousals (PLMS-noMA) were analyzed in each subject. Systolic and diastolic BP (SBP, DBP) were measured within a 25-beat temporal window comprising 10 beats before and 15 beats after onset of each movement. PLMS-related BP changes were assessed by repeated measures one-way analysis of variance. BP changes associated with PLMS-MA and PLMS-noMA were compared by paired t-tests. Pearson correlation coefficients were used to assess the relationship between cardiovascular changes and clinical and polysomnographic variables. RESULTS: BP increased significantly in association with all PLMS (on average, SBP 22 mm Hg, DBP 11 mm Hg). BP changes associated with PLMS-MA were greater vs those associated with PLMS-noMA (p < 0.05). SBP and DBP changes increased with age and the duration of illness. CONCLUSIONS: Periodic leg movements-related repetitive nocturnal blood pressure fluctuations could contribute to the risk of cardiovascular diseases in patients with restless legs syndrome, especially in the elderly.

Dynamics of parasite communities and interactions between wild and domestic ruminants.

Dynamics of parasite communities and host-parasite-environment interactions can be influenced by different factors. The present note, by discussing some field experiences in wild and domestic ruminant populations, approaches eco-epidemiology of abomasal nematodes in relation with host health and dynamics. Factors possibly playing a role in the host-parasite relationship are discussed, as well as possible use of macroparasites as ecological and sanitary indicators. The above topics are approached in a management perspective, in particular regarding interactions between domestic and wild ruminant populations.

Sleep bruxism is associated to micro-arousals and an increase in cardiac sympathetic activity.

Sleep bruxism (SB) subjects show a higher incidence of rhythmic masticatory muscle activity (RMMA) than control subjects. RMMA is associated with sleep micro-arousals. This study aims to: (i) assess RMMA/SB episodes in relation to sleep cycles; (ii) establish if RMMA/SB and micro-arousals occur in relation to the slow wave activity (SWA) dynamics; (iii) analyze the association between RMMA/SB and autonomic cardiac activity across sleep cycles. Two nights of polygraphic recordings were made in three study groups (20 subjects each): moderate to high SB, low SB and control. RMMA episodes were considered to occur in clusters when several groups of RMMA or non-specific oromotor episodes were separated by less than 100 s. Correlations between sleep, RMMA/SB index and heart rate variability variables were assessed for the first four sleep cycles of each study group. Statistical analyses were done with SYSTAT and SPSS. It was observed that 75.8% of all RMMA/SB episodes occurred in clusters. Micro-arousal and SB indexes were highest during sleep cycles 2 and 3 (P < 0.001). Within each cycle, micro-arousal and RMMA/SB indexes showed an increase before each REM sleep (P

[Increase in right-to-left intracardiac shunt with non-invasive ventilation]. / Majoration d'un shunt intracardiaque droit-gauche sous ventilation non invasive.

INTRODUCTION: Hypoxia caused by an increase in right-to-left shunt has been reported in patients with patent foramen ovale treated with levels of positive end-expiratory pressures (PEEP) greater than 10 cmH2O. This phenomenon has not previously been described with non-invasive ventilation (NIV). CASE REPORT: A 23 year-old man with tetralogy of Fallot and a severe kyphoscoliosis was admitted at the hospital for chronic dyspnoea. Arterial blood gases on room air: pH 7.43, PaCO2 39 mmHg, PaO2 67 mmHg, HCO3- 25 mmol/l, SaO2 95%. Nocturnal oxymetry showed severe hypoxaemia resistant to oxygen. NIV with PEEP of 3 cm H2O was commenced. With ventilation, his oxygenation worsened. An echocardiogram performed during NIV showed an increase in the right-to-left interventricular gradient from 22 to 37 cmH2O, and of the right ventriculo-auricular gradient from 76 to 142 mmHg. Furthermore, his oxygen saturation decreased progressively from 95 to 85%. Following removal of NIV, the patient recovered in 15 minutes. DISCUSSION: We report an increased right-to-left intracardiac shunt in a patient with tetralogy of Fallot. Compression of pulmonary vessels and cardiac cavities induced by NIV may have been enhanced by a reduction in thoracic compliance related to kyphoscoliosis. Right-to-left shunt in patients with kyphoscoliosis may be a contra-indication to NIV.

Seasonal 4-year investigation into the role of the alpine marmot (Marmota marmota) as a carrier of zoophilic dermatophytes.

Two hundred and six samples of alpine marmot (Mamota marmota) hair (148 from adults and 58 from young subjects), 102 soil samples from the entrances to the burrows of the above individuals and 20 control specimens (obtained from adjoining areas away from the burrow systems where the rodents are not usually present) were examined from May 1994 to September 1997. Seventy-five isolates belonging to six species of dermatophytes were found in 69 of the 206 hair samples examined (33.5%). Two species were zoophilic, Microsporum canis (7.8%) and Trichophyton mentagrophytes (11.2.%), and four geophilic, Microsporum cookei (2%), M. gypseum (5.8%), Trichophyton ajelloi (3.9%) and T. terrestre (5.8%). The prevalence of each species in the hair samples did not change significantly according to year, season (chi-squared test [limit significance: P <0.05] gives no significant values [P>0.05] both in year and in season comparison) or age/sex (adult versus juvenile: P=0.1; male versus female: P=0.8) of the marmot. Twenty-three of the 102 soil samples (22.5%) were positive for dermatophytes found in the hair of marmots from the same burrow systems. Five of the 20 control soil samples (25%) were positive for dermatophytes. One isolate of M. gypseum, three of T. terrestre and one of T. mentagrophytes were obtained. Compared with other free-ranging rodent hosts studied in Europe, this mycoflora is characterized by the presence and relatively high prevalence of M. canis, frequently reported in symptomatic and asymptomatic cats, dogs and fur animals. M. canis has not been isolated in other rodents in the wild. However, it has recently been reported in asymptomatic foxes (Vulpes vulpes) from northern Italy. The close link between V vulpes and M. marmota, with the former representing the most important mammal predator of the latter in the Alps (only a fraction of the predator's attacks result in the death of the rodent) may have favoured the adaptation of M. canis to this rodent host. The stable character of the M. canis/M. marmota relationship (no seasonally or annually related difference in the prevalence of this dermatophyte has been found) suggests the inclusion of the alpine marmot in the reservoir of this zoophilic pathogenic agent. In this situation, hibernation in labyrinthine burrow systems, where temperature and moisture ranges are quite uniform the whole year round, may favour the viability of M. canis arthroconidia, whose survival in mountain habitat might otherwise be compromised. This seems to be confirmed by the fact that the fungus has never been found in the control samples collected at a distance of 300 m from the outer edge of the sampled burrow systems.

Seasonal changes in serum metabolites in free-ranging alpine marmots (Marmota marmota).

Circannual changes in serum parameters and body mass were studied in free-ranging Marmota marmota that had been shot in Switzerland (Grisons) for management reasons in May, July, and September of 1995, 1996, and 1997; and in April 1996. Markers of lipid (triglycerides, cholesterol), protein (total protein, urea-nitrogen) and mineral (calcium, inorganic phosphate) metabolism were evaluated in 111 haemolysis-free serum samples; the effects of sex, age, reproductive status, season and year were tested. Mean body mass was higher in adult males than in adult females in July and September, and serum cholesterol concentrations were lower in adult males in May. Pregnant females had lower concentrations of total protein than non-pregnant females, and triglyceride concentrations were negatively correlated with the number of uterine ampullae. Inorganic phosphate decreased and total protein increased with age. In adults, triglycerides, cholesterol and urea-nitrogen increased mainly from May to July and decreased between September and May. Total protein, calcium and phosphate did not change throughout the year. This suggests that, during the active season, ingested lipids and amino acids were utilised for metabolic needs or lipid storage, while in winter, lipids were catabolised and protein was spared. Quantitative differences between years, observed for triglycerides and total protein during the active season, were probably due to different climatic conditions in each year.

Echinococcus granulosus in the wolf in Italy.

During the period 1987-1999, 119 wolf cadavers were examined and checked for the presence of Echinococcus granulosus. All the animals were retrieved along the whole Apennines range of distribution of the species in Italy and most of them were illegally killed. Eighteen wolves resulted positive (15%). The mean intensity was 697.5. The force of infection for prevalence was 8.2 year(-1). The prevalence of the parasite was significantly and positively influenced by the local prevalence of cystic echinococcosis (CE) in sheep. Mean intensity was significantly and positively influenced by both the age of the wolf and the prevalence of CE in sheep. A deterministic model was used in order to simulate a purely theoretical sylvatic cycle of the parasites having the wolf as the only definitive host with 15% of prevalence. The expected prevalence of CE in wild intermediate species ranges between 10% and 25%. This prevalence overlaps the one observed in sheep. Even if both the wolf and the wild ungulate populations are increasing, the wolf still acts as a part of the main dog-sheep cycle of the parasite.

Serological study of a population of alpine chamois (Rupicapra r rupicapra) affected by an outbreak of respiratory disease.

A serological survey of respiratory virus infections was carried out from 1998 to 2001 in Lecco province, Italy, as part of a health monitoring programme in a population of alpine chamois, many of which died of pneumonia in autumn and winter 2000 to 2001; 194 carcases of all age classes were found over a short period and in a small area. Eighteen of them, which were examined postmortem, consistently showed signs of severe fibrinous lobar pneumonia or catarrhal bronchopneumonia. Samples of serum from 145 chamois collected from hunted animals and carcases were tested by a virus neutralisation test against bovine respiratory syncytial virus (BRSV), bovine viral diarrhoea virus, bovine herpesvirus type 1 and parainfluenzavirus type 3. Positive results were detected only for BRSV. The area was divided into two subunits on the basis of the distribution of deaths; in the areas where fatalities were observed there was a significant increase of BRSV titres at the beginning of the outbreak. Furthermore, during the 2000 and 2001 hunting seasons antibody titres to BRSV were significantly higher in the areas where mortality occurred. The roe deer living in the same area were not affected by pneumonia and had a low prevalence of titres to BRSV which did not vary during the period of the study.

Wildlife veterinarian, conservation and public health.

In recent years wildlife diseases (infectious and non-infectious) have played a relevant role in both wildlife conservation and public health. Global environmental changes have determined a bimodal evolution of wildlife. On one side a huge loss of biodiversity has been observed leading to the increasing of threatened or endangered species. In contrast few opportunistic taxa increased their aboundances and ranges. The above scenarios claim the intervention of wildlife veterinarians. In conservation the understanding of the ecological role of the host parasite relationship and the perturbations on the host population dynamics have to be assessed and eventually modified. In public health the increased overlapping among wildlife, livestock, pets and human beings represents a risk for diseases spread (no matter in which directions). Serious limits are, still now, observed in the acceptance of this 'new world' by veterinary academics. As a consequence curricula often fail in providing adequate skill at both undergraduate and graduate levels. An addressed approach towards wildlife diseases should be promoted as an essential component of environmental management.

Obstructive sleep apnea and vascular disease.

There is emerging evidence linking obstructive sleep apnea (OSA) to vascular disease, including hypertension. This relationship may be independent of co-morbidity, such as obesity. Even apparently healthy OSA patients have evidence of subtle functional vascular abnormalities that are known to occur in patients with hypertension and atherosclerosis. Untreated OSA may possibly contribute to the initiation and/or progression of pathophysiologic mechanisms involved in hypertension, heart failure, cardiac ischemia and stroke. This brief commentary will examine the evidence and mechanisms linking OSA to vascular disease.

Restrictive left ventricular filling pattern as a strong predictor of depressed baroreflex sensitivity in heart failure.

BACKGROUND: The aim of this study was to test the hypothesis that a restrictive left ventricular diastolic filling pattern, as an index of elevated pulmonary wedge pressure, would predict a depressed baroreflex sensitivity (BRS) in patients with chronic heart failure. METHODS: A total of 189 consecutive patients with an ejection fraction < or = 40% at echocardiography, in sinus rhythm and clinically stable for at least 1 month in oral therapy, underwent clinical examination, echo-Doppler study and the phenylephrine test. RESULTS: The correlations between the NYHA functional class, echo-Doppler variables and BRS were weak, although significant (r ranging from -0.15 to 0.40). However, patients with a deceleration time < 140 ms as an expression of restrictive filling, compared to those with a deceleration time > or = 140 ms, had a lower BRS (3 +/- 4 vs 6 +/- 4 ms/mmHg, p < 0.00001), a lower ejection fraction (20 +/- 6 vs 28 +/- 7%, p < 0.00001), greater left ventricular (end-diastolic volume index 137 +/- 43 vs 113 +/- 45 ml/m2, p < 0.00001) and left atrial dimensions (25 +/- 6 vs 20 +/- 5 cm2, p < 0.00001), more severe mitral regurgitation (3 +/- 1 vs 2.3 +/- 1, p < 0.00001) and were in a higher NYHA class (2.3 +/- 0.6 vs 1.8 +/- 0.5, p < 0.00001). Medications at the time of the study were similar in the two groups. At stepwise regression analysis, the deceleration time emerged as the most powerful independent predictor of a depressed BRS (< 3 ms/mmHg), followed by mitral regurgitation, age, and NYHA class (all data p = 0.0001). CONCLUSIONS: In patients with chronic heart failure, the presence of a restrictive left ventricular filling pattern is highly predictive of autonomic derangement as expressed by low values of BRS.

Blood pressure and heart rate during periodic breathing while asleep at high altitude.

The ventilatory and arterial blood pressure (ABP) responses to isocapnic hypoxia during wakefulness progressively increased in normal subjects staying 4 wk at 5,050 m (Insalaco G, Romano S, Salvaggio A, Braghiroli A, Lanfranchi P, Patruno V, Donner CF, and Bonsignore G; J Appl Physiol 80: 1724-1730, 1996). In the same subjects (n = 5, age 28-34 yr) and expedition, nocturnal polysomnography with ABP and heart rate (HR) recordings were obtained during the 1st and 4th week to study the cardiovascular effects of phasic (i.e., periodic breathing-dependent) vs. tonic (i. e., acclimatization-dependent) hypoxia during sleep. Both ABP and HR fluctuated during non-rapid eye movement sleep periodic breathing. None of the subjects exhibited an ABP increase during the ventilatory phases that correlated with the lowest arterial oxygen saturation of the preceding pauses. Despite attenuation of hypoxemia, ABP and HR behaviors during sleep in the 4th wk were similar to those in the 1st wk. Because ABP during periodic breathing in the ventilatory phase increased similarly to the ABP response to progressive hypoxia during wakefulness, ABP variations during ventilatory phases may reflect ABP responsiveness to peripheral chemoreflex sensitivity rather than the absolute value of hypoxemia, suggesting a major tonic effect of hypoxia on cardiorespiratory control at high altitude.

[The assessment of breathing during sleep: a curiosity or clinical necessity?]. / La valutazione del respiro durante il sonno: curiosità o necessità clinica?

The study of sleep, which initially focused on the neurophysiological mechanisms and cardiorespiratory function during the night, has shown the presence of sleep-related breathing disorders that epidemiological, pathophysiological and clinical data have indicated to be associated with increased cardiovascular morbidity and mortality: the obstructive sleep apnea syndrome (OSAS) and the central sleep apnea syndrome (CSAS). OSAS is a condition characterized by repetitive respiratory pauses due to the pharynx wall collapse, with a subsequent obstruction to the airflow. The hemodynamic consequences due to the markedly increased negative intrathoracic pressure (induced by the respiratory muscle effort towards the closed upper airways), the progressive hypercapnic hypoxemia and the arousal terminating the apneas, are the pathophysiological keys of the cardiovascular effects of OSAS and may explain the association between OSAS and the documented increase of cardiovascular morbidity and mortality. CSAS is a breathing disorder characterized by recurrent episodes of central hypopneas or apneas and hyperventilation which, is the classical form described by Cheyne and Stokes, show a crescendo-decrescendo pattern of respiration. Pathophysiological and epidemiological data clearly indicate the link between CSAS and heart failure, also showing a correlation between respiratory disorders and the severity of hemodynamic impairment. However, other mechanisms are involved in the genesis of CSAS in explaining the variable presence of CSAS independent of cardiac function and, more importantly, the impact of CSAS on poor prognosis in heart failure. In conclusion, the data available indicate the need to include screening for sleep-related breathing disorders in the evaluation of cardiac patients who are at risk for OSAS and, particularly, in patients with heart failure, who could really benefit from treatment of the respiratory disorder.

Host specificity of abomasal nematodes in free ranging alpine ruminants.

Abomasums from 641 alpine wild ruminants representing five different species (Cervus elaphus, Capreolus capreolus, Rupicapra rupicapra, Capra ibex, Ovis musimon) and from 19 domestic sheep (Ovis aries) from alpine areas were examined in order to investigate the host-specificity of abomasal helminths. Nine out of 20 helminth species were found in at least five different host species. A discriminant analysis was able to significantly discriminate the hosts on the basis of their helminth community composition with the exception of O. musimon and O. aries. Based on the correlation between each variable represented by helminth species with the most explanatory discriminant axis, it was possible to classify helminths into specialists and generalists. Specialists are represented by the dominant species in a community of an host species or family while generalists appear in the communities of many different hosts as intermediate species. Due to the pathogenic potential of some of these generalist species (i.e. Haemonchus contortus, Trichostrongylus axei) and their ability to adapt easily to the conditions found in several different hosts, they appear to be the most important from a sanitary point of view.