Estimating the probability of freedom from bovine brucellosis in the Galapagos Islands.

Bovine brucellosis is a worldwide zoonotic disease that still burdens several countries in the Mediterranean, Asia, Africa and Latin America. Although the disease is present in Ecuador, the Galapagos Islands seem to be free from the disease based on a survey conducted in 1997 where all tested animals showed negative results. This study aimed at estimating the probability of freedom from brucellosis in this Ecuadorian province in 2014. A survey was implemented on the three main cattle-producing islands of the province: Santa Cruz, Isabela and San Cristóbal. Thirty-three cattle farms and 410 cattle were tested for brucellosis using the Rose Bengal test and indirect ELISA. All animals showed negative results for both tests. Probability of freedom was estimated at 98%, 91% and 88% for Santa Cruz, Isabela and San Cristóbal, respectively, considering a herd-level design seroprevalence of 20% and animal-level design seroprevalence of 15%, and assuming a perfect specificity of the survey. The negative results found in 1997 and present surveys suggest that the Galapagos Islands are free from bovine brucellosis.

Bovine anaplasmosis and tick-borne pathogens in cattle of the Galapagos Islands.

A cross-sectional study was conducted to determine the species of Anaplasma spp. and estimate its prevalence in cattle of the three main cattle-producing Galapagos Islands (Santa Cruz, San Cristóbal and Isabela) using indirect PCR assays, genetic sequencing and ELISA. Ticks were also collected from cattle and scanned for 47 tick-borne pathogens in a 48 × 48 real-time PCR chip. A mixed effects logistic regression was performed to identify potential risk factors explaining Anaplasma infection in cattle. A. phagocytophilum was not detected in any of the tested animals. Genetic sequencing allowed detection of A. platys-like strains in 11 (36.7%) of the 30 Anaplasma spp.-positive samples analysed. A. marginale was widespread in the three islands with a global between-herd prevalence of 100% [89; 100]95% CI and a median within-herd prevalence of 93%. A significant association was found between A. marginale infection and age with higher odds of being positive for adults (OR = 3.3 [1.2; 9.9]95% Bootstrap CI ). All collected ticks were identified as Rhipicephalus microplus. A. marginale, Babesia bigemina, Borrelia theileri and Francisella-like endosymbiont were detected in tick pools. These results show that the Galapagos Islands are endemic for A. marginale.

Estimation of French cattle herd immunity against bluetongue serotype 8 at the time of its re-emergence in 2015.

BACKGROUND: From 2006 to 2010, France experienced two bluetongue epidemics caused by serotype 1 (BTV-1) and 8 (BTV-8) which were controlled by mass vaccination campaigns. After five years without any detected cases, a sick ram was confirmed in August 2015 to be infected by a BTV-8 strain almost identical to that circulating during the previous outbreak. By then, part of the French cattle population was expected to be still protected, since bluetongue antibodies are known to last for many years after natural infection or vaccination. The objective of this study was to estimate the proportion of cattle in France still immune to BTV-8 at the time of its re-emergence in 2015. RESULTS: We used BTV group-specific cELISA results from 8525 cattle born before the vaccination ban in 2013 and 15,799 cattle born after the ban. Samples were collected from January to April 2016 to estimate seroprevalence per birth cohort. The overall seroprevalence in cattle at national and local levels was extrapolated from seroprevalence results per birth cohort and their respective proportion at each level. To indirectly assess pre-immune status of birth cohorts, we computed prevalence per birth cohort on infected farms in autumn 2015 using 1377 RT-PCR results. These revealed limited BTV circulation in 2015. Seroprevalence per birth cohort was likely to be connected to past exposure to natural infection and/or vaccination with higher seroprevalence levels in older animals. A seroprevalence of 95% was observed for animals born before 2008, of which > 90% were exposed to two compulsory vaccination campaigns in 2008-2010. None of the animals born before 2008 were found to be infected, unlike 19% of the young cattle which had never been vaccinated. This suggests that most ELISA-positive animals were pre-immune to BTV-8. We estimated that 18% (from 12% to 32% per département) of the French cattle population was probably pre-immune in 2015. CONCLUSIONS: These results strongly suggest a persistence of antibodies for at least 5-6 years after natural infection or vaccination. The herd immunity of the French cattle population probably limited BTV circulation up to 2015, by which time more than 80% of cattle were naive.

Serological status for BTV-8 in French cattle prior to the 2015 re-emergence.

Undetected in Europe since 2010, bluetongue virus serotype 8 (BTV-8) re-emerged in August 2015 in Central France. To gain insight into the re-emergence on the French territory, we estimated the seroprevalence in cattle before the detection of BTV-8 in 2015, in areas differentially affected by the current outbreak. A retrospective survey based on the analysis of stored sera was thus conducted in the winter preceding the re-emergence in seven French departments including the one where the virus was first detected. A total of 10,066 sera were retrieved from animals sampled in 444 different herds in winter 2014/15. Between-herd seroprevalence revealed the presence of seropositive animals in almost all herds sampled (97.4%). The animal-level seroprevalence averaged at 44%, with a strong age pattern reflecting the cumulative exposure to both natural infection and to vaccination. A multivariable analysis allowed separating the respective effects of both exposures. A higher proportion of seropositivity risk was attributed to vaccination (67.4%) than to exposure to natural infection (24.2%). The evolution of seroprevalence induced by the two main risk factors in 74 mainland departments was reconstructed between the vaccination ban (2013) and the re-emergence (2015). We showed a striking decrease in seroprevalence with time after the vaccination ban, due to population renewal, which could have facilitated virus transmission leading to the current outbreak situation.

Circulation of bluetongue virus 8 in French cattle, before and after the re-emergence in 2015.

Bluetongue virus serotype 8 (BTV-8) re-emerged in Central France in August 2015. The viral strain identified is nearly identical to the one that circulated during the 2006/2009 massive outbreak throughout Europe. To address the question of an undetected BTV-8 circulation on the French territory, a serological study was conducted on young cattle along a transect of seven departments, three of them located in areas where the virus presence had been confirmed by RT-PCR by winter 2015/2016. Sera from 2,565 animals were collected during the winters preceding and following the re-emergence, with 414 animals being sampled in each of the two consecutive years. All samples were tested by competitive ELISA (IDVet) and, when enough serum was available, ELISA-positive samples were confirmed by seroneutralization tests. In areas with infected holdings, seropositive animals were found before the re-emergence (N = 14 of 511), significantly more on the following year (N = 17 of 257), and eight animals (N = 158) seroconverted over 2015. Seropositive animals were also detected as early as winter 2014/2015 in one department without known infected holdings (N = 12 of 150), and in winter 2015/2016 in three of them (N = 21 of 555), where seven animals (N = 154) seroconverted over 2015. These results suggest that BTV-8 may have spread at low levels before the re-emergence, even in areas considered virus-free. Unfortunately, whole blood from the seropositive animals was not available to definitely confirm the virus presence by RT-PCR.

Likely Introduction Date of Schmallenberg Virus into France According to Monthly Serological Surveys in Cattle.

To estimate the date of introduction of Schmallenberg virus (SBV) into France, the prevalence of antibodies against the virus was determined monthly in cattle from two northern departments from August 2011 to April 2012. Seropositive cattle were detected from October 2011 in both departments with a prevalence of 55.6% in the westernmost department (Meurthe-et-Moselle) and of 12.7% in the easternmost department (Manche). Schmallenberg virus seroprevalence then increased rapidly to high levels.

A one-year follow-up of antibody response in cattle and sheep after vaccination with serotype 8- and serotype 1-inactivated BT vaccines.

Sixteen sheep and 18 cattle were followed up during 1 year to estimate the duration of immunity induced by inactivated bluetongue virus serotype 8 (BTV-8) vaccines (sheep and cattle) and a bluetongue virus serotype 1 (BTV-1) vaccine (cattle) under field conditions using cELISA and seroneutralization test (SNT). Four sheep never seroconverted. Those that seroconverted were all seronegative by BTV-8 SNT at the date of last sampling [378 days post-vaccination (dpv)]. Eight sheep were still positive by competitive ELISA (cELISA) 378 dpv. All the cattle seroconverted. At the end of the study, eight and 11 cattle were still positive by BTV-8 SNT and cELISA, respectively (335 dpv); and nine were still positive by BTV-1 SNT (301 dpv).

Clinical pattern characterization of cattle naturally infected by BTV-8.

Forty-one cattle from seven Belgian farms and two French farms confirmed as infected with bluetongue virus serotype 8 (BTV-8) were monitored from the onset of clinical signs to describe the disease pattern and estimate the duration of blood RT-qPCR and competitiveELISA positivity under field conditions. On each visit, blood samples were taken, and a standardized clinical form was filled in for each animal. A clinical score was calculated for every week until the end of clinical signs. A classification and regression tree (CART) analysis was conducted to determine the most important clinical signs every week for the first 7 weeks. The highest scores were recorded within 2 weeks of clinical onset. The first recorded clinical signs were quite obviously visible (lethargy, conjunctivitis, lesions of nasal mucosa, nasal discharge). Skin lesions, a drop in milk production and weight loss appeared later in the course of the disease. A biphasic pattern regarding nasal lesions was noticed: the first peak concerned mainly congestive and ulcerative lesions, whereas the second peak mainly concerned crusty lesions. The median time estimated by survival analysis to obtain negative RT-qPCR results from the onset of clinical signs was 195 days (range 166-213 days) in the 23 cattle included in the analysis. Serological results remained strongly positive until the end of the study. These results should ensure more accurate detection of an emerging infectious disease and are of prime importance in improving the modelling of BTV-8 persistence in Europe.

Modelling transmission of bovine tuberculosis in red deer and wild boar in Normandy, France.

In early 2001, Mycobacterium bovis infection was confirmed in red deer (RD) (Cervus elaphus) shot in Normandy region, France. An epidemiological survey conducted during the following hunting season in two connected forests confirmed the occurrence of the disease in both free-ranging RD and wild boar (WB) (Sus scrofa). This was the first detected bovine tuberculosis outbreak in wildlife in France. We present a simple deterministic age-structured model of the within- and between-species M. bovis transmission in RD and WB populations that distinguishes direct transmission (horizontal and pseudo-vertical) and indirect transmission through contaminated offal left behind by hunters. Results issued from the epidemiological surveys conducted in Normandy forests were used to estimate transmission parameters. Because data for RD and WB populations were not available, population sizes at demographic equilibrium were estimated and used to run the model. We qualitatively tested different control measure scenarios with our model, considering different mortality rates and offal harvesting, to determine which ones affect the success of infection control. The most realistic control scenario would combine the total depopulation of RD and good compliance with offal harvesting, because the model suggests that infected offal left by hunters represents the main transmission source of M. bovis in the field.

Bluetongue virus serotype 8: abortion and transplacental transmission in cattle in the Burgundy region, France, 2008-2009.

During the incursion of bluetongue virus (BTV) serotype 8 in France in 2007, an increase in the number of abortions in cattle was observed, but the cause was not clearly established. A survey of all the reported cases of abortion in cattle from November 2008 to April 2009 was conducted in the Nièvre district (Burgundy region) to determine the percentage of abortions as a result of BTV-8 and to study factors that could have played a role in BTV-8 transplacental transmission. BTV-8 was present in 16% of the fetuses or newborn calves that died within 48 h, from 780 dams. Dams inseminated before the BTV epizootic peak recorded from July to September 2008 were more likely to have BTV-positive abortions (OR=5.7, P<0.001) and those vaccinated in May or June 2008 were less likely to have BTV-positive abortions (OR=0.3, P=0.01 and OR=0.4, P=0.001, respectively). The gestational month was not a predictor of BTV abortion. In blood or spleen, fetuses/calves from RT-PCR-positive dams had significantly higher RNA concentrations than fetuses/calves from RT-PCR-negative dams. Of the 128 dams that had BTV-positive fetuses or calves, 60% were RT-PCR-negative. BTV-8-positive fetuses/calves were significantly more frequent (n=42 vs n=21, P=0.082) amongst those showing clinical signs or lesions suggestive of cerebral damage.

Occurrence and antibiotic resistance of coliform bacteria and antimicrobial residues in pasteurized cow's milk from Brazil.

In this study we examined the hygienic and sanitary quality of pasteurized cow's milk in the state of Paraná, Brazil, by determining the presence of coliforms and occurrence of antimicrobial residues. A total of 260 milk samples were collected from commercial establishments in different regions of the state. Coliform populations were estimated by the multiple-tube test, and antimicrobial residues were detected by enzyme-linked immunosorbent assay. Overall, 105 samples (40.4%) were unsuitable for consumption according to Brazilian legal standards. Among the coliforms, Escherichia coli and Klebsiella pneumoniae were respectively identified in 77.05 and 36.07% of the samples. The highest rates of resistance to antimicrobial agents were observed for ampicillin (19.2%), cephalothin (18.9%), and tetracycline (17.1%). Antimicrobial residues were detected in 80 samples (30.8%). Forty-eight samples (18.5%) were positive for tetracycline, 29 (17.4%) for neomycin, 9 (3.5%) for beta-lactams, 6 (2.3%) for gentamicin, 4 (1.5%) for chloramphenicol, and 1 (0.4%) for streptomycin-dihydrostreptomycin. The results demonstrate a high prevalence of coliforms and also a high occurrence of antimicrobial residues in pasteurized cow's milk from Paraná, Brazil.

Patterns of lesions of bovine tuberculosis in wild red deer and wild boar.

The data obtained from a survey of Mycobacterium bovis infection in wild red deer (Cervus elaphus) and wild boar (Sus scrofa) conducted in France in the 2005/06 hunting season were used to describe and quantify the pathological findings in the two species. The red deer had caseous abscessed lesions in their organs and lymph nodes, whereas in the wild boar the lesions were predominantly caseocalcareous and occurred mainly in the lymph nodes. The severity of the gross tuberculosis-like lesions was estimated on the basis of a numerical score. The significant difference between the distribution of the scores in the two species indicated that the disease was more serious in the red deer than in the wild boar. Unlike the red deer, the wild boar did not show a generalised pattern of disease. Among the lymph nodes examined systematically, gross lesions were most frequently observed in the mesenteric lymph nodes in the red deer and in the retropharyngeal lymph nodes in the wild boar. In both species, the presence of gross lesions showed the closest agreement with the isolation of M bovis from the same lymph nodes. The different patterns of the lesions of tuberculosis in the two species suggest that red deer might play an important role in the intraspecies and interspecies dissemination of the infection, whereas in wild boar the spread of the infection would be more likely to be restricted to other wild boar.

Piperacillin versus cefotetan as single-dose prophylaxis in abdominal hysterectomy: a prospective, randomized, multicenter study.

Five hundred and sixty-six women, consecutively admitted to nine different hospitals for elective abdominal hysterectomy, were randomized to receive a single 2 g i.v. dose of either cefotetan or piperacillin at induction of anesthesia. Five patients were excluded and 561 (287 given cefotetan and 274 given piperacillin) could be evaluated. In 10 patients (5 for each drug) the concentrations of either cefotetan or piperacillin in serum and subcutaneous tissue at the beginning and at the end of surgery and in uterus, salpinx and ovary samples, were microbiologically assessed. No significant differences could be found between cefotetan and piperacillin groups for any of the considered infectious complications (bacteremia, wound or vaginal cuff infection, pelvic cellulitis and febrile morbidity) nor for the pooled data (cefotetan group: 4.9%, piperacillin group: 5.8%, p:NS). As for the concentrations of prophylactic drugs, serum levels throughout surgery were found to be adequate.

Expression of doxorubicin-daunorubicin resistance genes in different anthracycline-producing mutants of Streptomyces peucetius.

Two DNA fragments, ric1 and ric2, were isolated from the Streptomyces peucetius 7600 mutant, which produces daunorubicin and doxorubicin, on the basis of their abilities to confer doxorubicin and daunorubicin resistance to Streptomyces lividans. These two fragments are unrelated by restriction mapping and do not show any homology by Southern analysis, yet both of them increase the level of resistance 10-fold in transformed S. lividans. Functional analysis revealed that ric1 also contains two genes of daunorubicin biosynthesis: one coding for the aklavinone C-11 hydroxylase and the other corresponding to the putative dnrR2 regulatory gene of wild-type S. peucetius ATCC 29050 (K. J. Stutzman-Engwall, S. L. Otten, and C. R. Hutchinson, J. Bacteriol. 174:144-154, 1992). Northern (RNA) blot experiments, performed with a ric1 fragment containing daunorubicin-doxorubicin resistance gene(s), revealed a transcript of about 2,100 nucleotides that is present only during the phase of anthracycline metabolite production. The amount of this transcript is higher in strain 7600 than in strain 7900, a mutant which produces 5-fold more daunorubicin and 10-fold less doxorubicin than 7600. Furthermore, two 7900-derived blocked mutants, 8600 and 9700, do not express the 2,100-nucleotide transcript in spite of the absence of gross rearrangements in the ric1 region such as occur with the 7900 parental strain.

[Study of pollen of allergologic significance in the urban atmosphere of Parma in the years 1983-1984]. / Studio di alcuni pollini di interesse allergologico nell'atmosfera urbana di Parma negli anni 1983-1984.

From 1983 to 1984 pollen air samples from Parma's urban atmosphere were collected weekly by means of a Burkard recording volumetric spore trap. Data regarding a few of the meteorologic variables were also collected at the same time: temperature, insulation, water precipitation, humidity and wind speed. Some of the more allergenic pollens and their concentration/m3 air were determined from the collected samples: Graminea, Urticaceae, Fagaceae, Salicaceae and Betulaceae. Looking at the results from the period of study, it can be seen that Graminea and Urticaceae were the most represented in comparison with the other families. Graminea was most represented in May 1983, in June 1984 and during the first week of September 1984; Urticaceae in September 1983, May 1984 and September 1984. These pollen concentrations were also compared to the meteorologic data.