Indoor and outdoor winter activity of Culicoides biting midges, vectors of bluetongue virus, in Italy.

Indoor and outdoor winter activity of Culicoides spp. (Diptera: Ceratopogonidae) in central Italy was investigated in order to evaluate whether indoor activity might account for the overwintering of bluetongue virus, as has been hypothesized by some authors. Weekly Culicoides collections were performed at three farms over three consecutive winter seasons. At each farm, two black-light traps were operated simultaneously, indoors and outdoors. Culicoides were identified using both morphological and molecular means. The Culicoides obsoletus group accounted for 98.2% of sampled specimens. Within this group, C. obsoletus s.s. accounted for 56.8% and Culicoides scoticus for 43.2% of samples. Nulliparous, parous and engorged females were caught throughout the entire winter, both indoors and outdoors. At times, indoor catch sizes outnumbered outdoor collections. A significant inverse correlation was found between minimum temperature and the proportion of indoor Culicoides of the total midge catch, thus indicating that lower outdoor temperatures drive Culicoides midges indoors. High rates of engorged females were recorded indoors, possibly as the result of the propensity of C. obsoletus females to feed indoors. Higher proportions of parous females were found in indoor than in outdoor catches, indicating higher survival rates indoors and, consequently, higher vectorial capacities of midges sheltering indoors compared with those remaining outdoors.

Biotic and abiotic factors influencing distribution and abundance of Culicoides obsoletus group (Diptera: Ceratopogonidae) in central Italy.

In the framework of a bluetongue surveillance program including clinical, serological, and entomological activities, Culicoides biting midges were light trapped weekly in two regions of central Italy, Lazio and Tuscany. In the period January 2002 through December 2005, 3,944 collections were carried out in 189 trap sites distributed in all the provinces of the two regions. Abundance data of C. obsoletus group were analyzed in relation to trap site altitude, distance from the sea, land use, and number of farmed animals. Species seasonality and overall temporal trend were also described. C. obsoletus was distributed over the whole study area, almost in all trapping sites and with high abundances. The species group was dominant among all captured Culicoides, with higher abundances recorded inland and in areas where land cover was partially or completely natural-wooded. Adults on the wing were caught all year round, with peaks in May-June and middle October. The observed trend through years recorded a peak during autumn 2002, in concomitance with a local epidemic of bluetongue.

Aedes albopictus and Culex pipiens implicated as natural vectors of Dirofilaria repens in central Italy.

To identify the natural vectors of Dirofilaria repens Railliet et Henry, entomological samplings were carried out in four sites within the Lazio region, foci of canine subcutaneous dirofilariasis. Collections were made in 2002-2003 by means of dog-baited and miniature Centers for Disease Control and Prevention traps as well as on humans. Microscopy identified 1576 attracted mosquito females as belonging to six species, but molecular diagnostics detected filarial DNA only in Culex pipiens L. and Aedes albopictus (Skuse, 1894). Dirofilaria immitis Leidy DNA, D. repens DNA, or both were found in the head and thorax of both mosquitoes. The simultaneous presence of vectors showing diurnal and nocturnal activity patterns is of concern for animal and human health. The finding of D. immitis DNA in mosquitoes in areas where only D. repens was been recovered in dogs also demonstrates that this filarial parasite circulates among carnivores (wild or domesticated pets).

Proliferative peritoneal and pleural cestodiasis in a cat caused by metacestodes of Mesocestoides sp. Anatomohistopathological findings and genetic identification.

A 10-year-old female cat was brought to Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana for post-mortem examination. The animal used to live, together with 26 other cats, in the big terrace of an apartment at the 8th floor in Rome; and was always fed with industrial pet food. Anamnesis referred balance troubles, vomit and convulsions, during a couple of days, followed by sudden death. At necropsy, the cat presented mucoid rhinitis, purulent tracheitis, small areas of pneumonia, dark spots in the liver, catarrhal-hemorrhagic gastritis, fibrinous enteritis and meningeal hyperemia. Thoracic and abdominal cavities were completely invaded by hundreds of larval stages of cestodes. The same parasites were also included in nodules in pancreatic, lung and kidney parenchyma. Microscopic examination of parasites allowed their identification as larval stages (metacestodes) of cestodes of the genus Mesocestoides. The molecular genotyping of the metacestodes indicates a close relationship with members of the genus Mesocestoides, although a significant variation was found with respect to the available sequences of other species of the genus.

Tuberculosis, Brucellosis and Leucosis in Cattle: A Cost Description of Eradication Programmes in the Region of Lazio, Italy.

The eradication of tuberculosis, brucellosis and leucosis in cattle has not yet been achieved in the entire Italian territory. The region of Lazio, Central Italy, represents an interesting case study to evaluate the evolution of costs for these eradication programmes, as in some provinces the eradication has been officially achieved, in some others the prevalence has been close to zero for years, and in still others disease outbreaks have been continuously reported. The objectives of this study were i) to describe the costs for the eradication programmes for tuberculosis, brucellosis and leucosis in cattle carried out in Lazio between 2007 and 2011, ii) to calculate the ratio between the financial contribution of the European Union (EU) for the eradication programmes and the estimated total costs and iii) to estimate the potential savings that can be made when a province gains the certification of freedom from disease. For the i) and ii) objectives, data were collected from official sources and a costing procedure was applied from the perspective of the Regional Health Service. For the iii) objective, a Bayesian AR(1) regression was used to evaluate the average percentage reduction in costs for a province that gained the certification. The total cost for the eradication programmes adjusted for inflation to 1 January 2016 was estimated at 18 919 797 euro (5th and 95th percentiles of the distribution: 18 325 050-19 552 080 euro). When a province gained the certification of freedom from disease, costs decreased on average by (median of the posterior distribution) 47.5%, 54.5% and 54.9% for the eradication programmes of tuberculosis, brucellosis and leucosis, respectively. Information on possible savings from the reduction of control costs can help policy makers operating under budget constraints to justify the use of additional resources for the final phase of eradication.

Control and eradication of tuberculosis in cattle: a systematic review of economic evidence.

Bovine tuberculosis (TB) is a disease of zoonotic importance for which control and eradication programmes have been carried out in many countries for decades. While the impact of these programmes on public health is still uncertain, the impact on trade is significant because of movement restrictions for animals, costs of testing and culling. The objective of this systematic review was to provide a contribution to the general debate over costs against benefits for the control and eradication of bovine TB in cattle. The search strategy was performed on four electronic databases following the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. The selection process, data abstraction and quality appraisal were carried out according to the Cochrane Collaboration guidelines. The search identified 66 articles out of which eight fulfilled the inclusion criteria. The evidence gathered in this review by combining the conclusions of the most methodologically sound articles supports the idea that, when multiple cost and benefit components are taken into account, efforts to control or eradicate bovine TB may be effective in reducing disease prevalence, economically viable and worth doing.

Active circulation of bluetongue vaccine virus serotype-2 among unvaccinated cattle in central Italy.

Several seroconversions occurring in 2002 among sentinel cattle during the bluetongue-vaccination campaign in Lazio and Tuscany (central Italy) led to the suspicion of vaccine-virus circulation. Therefore in 2003, 17 seroconverting sentinel herds were investigated for the characteristics of the virus involved. From these farms, 91 unvaccinated animals and 57 Culicoides pools were tested for the presence of the bluetongue vaccine virus (serotype-2) or other strains. The presence of vaccine virus serotype-2 was confirmed by PCR followed by restriction analysis in the whole blood of 17 unvaccinated sentinel cattle and 12 pools of Culicoides imicola or C. obsoletus. Of the 17 herds, five were positive only for vaccine virus serotype-2, four were positive for other strains and two for both the vaccine and other strains; the remaining premises were virologicaly negative. The vaccine virus serotype-2 also was detected in areas not included in the vaccination campaign.

Identification of Culicoides obsoletus (Diptera: Ceratopogonidae) as a vector of bluetongue virus in central Italy.

In 2001 and 2002, 235 outbreaks of bluetongue were observed in the Lazio and Tuscany regions of central Italy. During entomological surveillance Culicoides imicola, the main vector of bluetongue virus in the Mediterranean region, was detected in only 14 of 28 municipalities affected by outbreaks; Culicoides obsoletus was the most abundant species, contributing 83 per cent of individuals in catches, whereas C. imicola contributed only 2 per cent. In affected municipalities the maximum catch of C. obsoletus was 18,000 specimens, compared with 54 of C. imicola. In October 2002 bluetongue virus serotype 2 was isolated from a single pool of wild-caught, non-blood-engorged parous C. obsoletus inoculated on to BHK-21 cells. Its identity was confirmed by reverse transcriptase-PCR.

Block of pinprick and cold sensation poorly correlate with relief of postoperative pain during epidural analgesia.

OBJECTIVE: To test the following hypotheses: there is a correlation between spread of epidural analgesia as assessed postoperatively by pinprick/cold test and postoperative pain intensity; block of pinprick/cold sensation is associated with absence of postoperative pain. DESIGN: Correlation analysis on prospectively collected data. SETTING: University hospital. PATIENTS: One hundred patients undergoing major surgery. Consecutive sample. INTERVENTIONS: Patients received an epidural infusion of bupivacaine 1 mg/ml, fentanyl 2 microg/ml, and epinephrine 2 microg/ml for at least 48 hours postoperatively. The infusion rate was adjusted according to pain intensity, occurrence of hypotension, or motor block. OUTCOME MEASURES: Assessments were made on three time points: 20-24 hours, 32-36 hours, and 4248 hours after extubation. Assessments included pinprick and cold sensitivity from C2 to S5, pain intensity (visual analogue scale, VAS) at rest, after cough, and after mobilization. Data were analyzed by multiple regression. RESULTS: VAS significantly decreased with increasing spread (number of dermatomes for which hyposensitivity to pinprick or cold was observed). Spread could explain only 2-5% of the variability of VAS. Absence of both pinprick and cold sensation at all dermatomes corresponding to the surgical wound was frequently associated with pain. A high proportion of patients manifesting an upper level of block above T5 had pain after abdominal surgery. CONCLUSIONS: Spread and efficacy of epidural analgesia as assessed by pinprick and cold stimulation correlate poorly with postoperative pain. These methods are of limited value both as clinical indicators of the efficacy of postoperative pain control and for investigating the effect of epidural drugs and techniques.

Isolation of an adenovirus from an ostrich (Struthio camelus) causing pancreatitis in experimentally infected guinea fowl (Numida meleagris).

An adenovirus was isolated from the pancreas, kidney, and lung of a dead 4-month-old ostrich (Struthio camelus). Experimental infection of week-old guinea fowl (Numida meleagris) was carried out in order to study transmission. Of the infected keets, six out of 15 died; these exhibited pancreatitis, presence of a hemorrhagic liquid in the abdominal cavity, kidney degeneration, and pulmonary lesions. The remaining birds, euthanatized on day 21 postinoculation, showed chronic pancreatitis. The virus was reisolated from the pancreas, lung, and kidney of the infected keets.

Tick-borne diseases in ruminants of Central and Southern Italy: epidemiology and case reports.

Sera and blood from cattle and sheep were examined for the presence of Babesia and Theileria spp by microscopy and serology at the Parasitology Department of the Istituto Zooprofilattico Sperimentale of Abruzzo and Molise (IZSAM). Of the 47 bovine herds (323 animals) tested, 15 were found positive for Babesia bigemina and 1 for Babesia bovis. Two outbreaks occurred, one caused by B. bigemina and one by B. bovis. The B. bigemina outbreak occurred in Abruzzo and has been followed for two years. The isolate of B. bigemina was very pathogenic leading to the death of two cows out of 57. The vector responsible of the transmission appeared to be Rhipicephalus bursa. Parasites were observed in the erythrocytes for 30 days whereas sera were positive to indirect immunofluorescence (IFA) for at least one year. The B. bovis outbreak occurred in the province of Mantova (Northern Italy) in a group of 70 beef cattle imported from France. The infection resulted in the death of 5 animals and severe illness in another 6. In contrast with what occurred for Babesia infection, no clinical cases were recorded in cattle when species of Theileria were detected by microscopy. Of the 24 bovine herds (252 animals) tested for Theileria, 21 were found positive for the T. "sergenti"/buffeli/orientalis group. Single and mixed infection of T. "sergenti" and T. buffeli/orientalis were detected in herds of cross-bred cattle from Abruzzo and Marche. The parasites were identified by using a polymerase chain reaction which amplified DNA encoding p32/34. Most of the collected ticks (90%) were adults of R. bursa whereas the others were adults of Hyalomma detritum. During the period the animals have been observed (18 months), no clinical cases have been recorded and no associations have been found between blood abnormalities and animals found infected with Theileria. Prevalences of subclinically infected carriers increased from February till December (95.4%) even if the animals were indoors and no ticks were present. The prevalence then dropped dramatically six months later (76.7%). In calves less than 1 year old, the prevalence of infection significantly (p<0.05) increased with age, however intraerythrocytic stages of Theileria were found in the blood of three newborn calves (<7 days of age). Of the 18 ovine flocks tested for Babesia spp. (150 animals examined), 1 was positive for B. ovis and 2 for B. motasi. B. motasi infection was not associated with symptoms, while an outbreak of babesiosis caused by B. ovis occurred in Abruzzo. The infection resulted in the death of 3 animals (0.75% of the flock), two rams (20% of the total number) and a ewe, and severe illness in another 5 ewes (2% of the flock). Specimens of R. bursa and R. turanicus were collected from the infected animals. Of the 18 flocks (150 animals) examined, 12 were microscopically positive for Theileria spp. No clinical cases were recorded and identification at species level was not possible on the basis of morphological criteria. The prevalence distribution of infected herds and infected animals within herds and flocks have been calculated by a Monte Carlo simulation model, running 10,000 iterations. The most likely levels of prevalence of infected herds and infected animals within herds found for the species observed were as follows: 20% for B. bigemina with a prevalence within herd of 27%, 11% for B. bovis (18% within herd), 10% for Babesia ovis (19% within herd), 10% for B. motasi (17.5% within herd), 63% for Theileria in cattle (66% within herd) and 51% for Theileria in sheep (55% within herd).

A new species of Culicoides (Diptera, Ceratopogonidae) from southern Italy.

Culicoides paolae, a new species of Ceratopogonidae from southern Italy, and which superficially resembles an important vector of animal diseases, is described in both sexes. Its potential veterinary importance is discussed briefly.

Investigation on Opisthorchis felineus occurrence and life cycle in Italy.

Opisthorchiasis is a fish borne parasitic infection caused by helminths of the genus Opisthorchis (Digenea, Opisthorchiidae), affecting humans and other fish-eating mammals. Despite Opisthorchis felineus was first described in Italy in 1884, no cases of human opisthorchiasis were reported in this country until 2004; from then on, 4 outbreaks due to this species have been recorded in Central Italy. Following the more relevant of these outbreaks, involving 34 people in August 2007, snails, fishes and fecal samples collected from the Bolsena and Bracciano lakes (Central Italy) were analyzed in order to define the cycle of O. felineus in the area and investigate its prevalence in the different hosts. Pools of 20-40 snails each (4983 specimens altogether) of the genus Bithynia were analyzed by PCR for parasite DNA detection. Eight hundred and ninety-four fish belonging to 12 species were collected from the two lakes and tested for metacercariae both by muscle compression and digestion techniques. Eighty-seven fecal samples of 5 putative definitive host species were collected very close to the two lakes and tested for parasite eggs detection by formalin-ethyl acetate concentration technique. Identification at the species level of metacercariae and eggs, respectively, from fish and stool was confirmed by PCR analysis and sequencing. O. felineus DNA was detected in 0.08% (overall minimum infection rate) of snails of the genus Bithynia from the two lakes. The tench, Tinca tinca, was the only fish found infested in both lakes (prevalence 88.5%). O. felineus eggs were found only in cat feces (prevalence 46.4%). The tench represents the only threat for the human consumption in the study area while Coregonus sp., the most economically important species for the local fishery and frequently consumed raw marinated, resulted to be not infected. The high prevalence recorded both in fish and in definitive host suggests a widespread and massive presence of the parasite in the area. Further studies are needed to better investigate the possible role of some cyprinids species as intermediate hosts, in order to check their safety for human consumption.

Risk factors associated with Cryptosporidium parvum infection in cattle.

A 2-year, cross-sectional study was conducted to identify risk factors for Cryptosporidium sp. infection in bovine farms in central Italy. Faecal samples were collected on 248 farms, from 2024 calves and analysed using ELISA and immunofluorescent assay (IFA) commercial kits. In all 101 samples confirmed to be positive with IFA, the aetiological agent was identified as Cryptosporidium parvumand a large genetic variability was detected by subtype analysis. The prevalence of farm infection ranged from 3.4% to 35.6%. Univariate analysis showed a number of putative risk factors, including the type of farm, stalling of calves, late supply of colostrum, number of heads and contact between calves and adults. However, multivariate analysis confirmed that the higher risk for calves was associated with housing calves separately from their dams, a characteristic practice of dairy herd, whereas calves being nursed by their dams, a characteristic of cow-calf herd resulted as a protective factor.

Isolation of the pigeon PMV-1 variant of Newcastle disease virus from imported pheasants (Phasianus colchicus).

Isolation of a virulent pigeon paramyxovirus 1 (PPMV-1) strain of Newcastle disease virus (NDV) from pheasants imported for release is reported. Although no clinical signs or postmortem lesions suggesting NDV infection were detected, the intestine was processed for attempted virus isolation as part of a surveillance programme on wild-bird populations. The potential role of pheasants as carriers of the disease, and the importance of developing surveillance programmes for wild bird populations is discussed.

A multifactorial analysis of the spread of epidural analgesia.

The controversies about the factors determining the spread of epidural analgesia are partly due to inappropriate methodology or sample size of previous studies. We performed a multivariate regression analysis on 803 ASA class 1-2 non-atherosclerotic adults, undergoing lumbar epidural anaesthesia according to a predefined standardised procedure. The spread of epidural analgesia is more accurately studied by analysing dose/segment (R2 = 0.671) instead of spread (R2 = 0.271) as dependent variable. The impact of local anaesthetic (2% lidocaine CO2 or 0.5% bupivacaine) and addition of adrenaline is not significant. Spread significantly increases with increasing age, weight, body-mass index, dose of local anaesthetic, addition of fentanyl, higher site of injection, and decreasing body height. The impact of age and dose is higher under the age of 40 and at doses lower than 20 ml. Increasing the total dose increases the dose needed to block one spinal segment. Unknown idiosyncratic factors still determine a certain proportion of the sample variance. The addition of adrenaline to lidocaine and the use of bupivacaine improve the predictability of spread. In conclusion, we found clinically significant correlations between a group of factors and epidural spread. Alternative anaesthetic solutions lead to different degrees of predictability.

A multifactorial analysis to explain inadequate surgical analgesia after extradural block.

A multivariate analysis of inadequate extradural analgesia was carried out prospectively on 1051 patients undergoing lumbar extradural anaesthesia for surgery performed on structures innervated by T10-S5. Ninety-six patients (9%) experienced pain during surgery. Age, extradural fentanyl, diazepam sedation and duration of surgery had no significant influence. We found some weak evidence that the type of surgery affects the risk of feeling pain. The probability of pain increased with increasing weight, except in overweight women, and was significantly greater for both shorter and taller patients, relative to patients of average height. The probability of pain decreased with increasing dose of local anaesthetic, increasing spread of extradural analgesia, addition of adrenaline, and fentanyl or thiopentone sedation. In conclusion, patient-, surgery- and anaesthesia-related factors influence the risk of inadequate extradural analgesia. If such factors are taken into account, an increase in the success rate may be anticipated.

Factors associated with hypotension and bradycardia after epidural blockade.

In order to identify patient-, anesthesia-, and surgery-related factors influencing the probability of hypotension and bradycardia after epidural blockade, an observational study was conducted on 1050 nonpregnant patients. Backward stepwise logistic regression was performed on the variables hypotension (systolic blood pressure < 90 mm Hg) and bradycardia (heart rate < or = 45 bpm). Hypotension and bradycardia occurred in 158 and 24 patients, respectively. The probability of hypotension increased when epidural fentanyl was administered (odds ratio [OR] = 2.18; 95% confidence interval [CI] = 1.16-4.11), with body weight and spread of epidural analgesia, and decreased when a tourniquet was used (OR = 0.01, CI = 0.01-0.02) and bupivacaine instead of carbonated lidocaine was administered (OR = 0.28, CI = 0.14-0.60). Sensitivity and specificity of the model were 89% and 88%, respectively. The probability of bradycardia was less in women (OR = 0.05, CI = 0.01-0.41) and when a tourniquet was used (OR = 0.04, CI = 0.02-0.09). Sensitivity and specificity were 50% and 97%, respectively. In conclusion, our analysis can contribute to identification of patients at high risk to develop hypotension and bradycardia after epidural blockade. If bupivacaine instead of carbonated lidocaine is used and epidural fentanyl is not administered a decrease in the incidence of hypotension may be anticipated.