Length–mass allometries in amphibians

Body mass is rarely recorded in amphibians, and other body measurements (e.g. snout to vent length, SVL) are generally collected instead. However, length measurements, when used as proxies of body mass in comparative analyses, are problematic if different taxa and morphotypes are included. We developed allometric relationships to derive body mass from SVL measurements. We fitted phylogenetic generalized least square models for frogs (Anura) and salamanders (Caudata) and for several families separately. We tested whether allometric relationships differed between species with different habitat preferences and between morphs in salamanders. Models were fitted with SVL–mass measurements for 88 frog and 42 salamander species. We assessed the predictive performance of the models by cross‐validation. Overall, the models showed high explained variance and low forecasting errors. Models differed among semi‐aquatic, terrestrial and arboreal frogs, and between paedomorphic and non‐paedomorphic salamanders. Body mass estimates derived from our models allow for comparability of studies on multiple taxa and can be used for testing theories built upon evolutionary and ecological processes which are directly related to body mass.     

Attribution of human infections with Shiga toxin‐producing Escherichia coli (STEC) to livestock sources and identification of source‐specific risk factors,The Netherlands (2010–2014)

Shiga toxin‐producing Escherichia coli (STEC) is a zoonotic pathogen of public health concern whose sources and transmission routes are difficult to trace. Using a combined source attribution and case–control analysis, we determined the relative contributions of four putative livestock sources (cattle, small ruminants, pigs, poultry) to human STEC infections and their associated dietary, animal contact, temporal and socio‐econo‐demographic risk factors in the Netherlands in 2010/2011–2014. Dutch source data were supplemented with those from other European countries with similar STEC epidemiology. Human STEC infections were attributed to sources using both the modified Dutch model (mDM) and the modified Hald model (mHM) supplied with the same O‐serotyping data. Cattle accounted for 48.6% (mDM) and 53.1% (mHM) of the 1,183 human cases attributed, followed by small ruminants (mDM: 23.5%; mHM: 25.4%), pigs (mDM: 12.5%; mHM: 5.7%) and poultry (mDM: 2.7%; mHM: 3.1%), whereas the sources of the remaining 12.8% of cases could not be attributed. Of the top five O‐serotypes infecting humans, O157, O26, O91 and O103 were mainly attributed to cattle (61%–75%) and O146 to small ruminants (71%–77%). Significant risk factors for human STEC infection as a whole were the consumption of beef, raw/undercooked meat or cured meat/cold cuts. For cattle‐attributed STEC infections, specific risk factors were consuming raw meat spreads and beef. Consuming raw/undercooked or minced meat were risk factors for STEC infections attributed to small ruminants. For STEC infections attributed to pigs, only consuming raw/undercooked meat was significant. Consuming minced meat, raw/undercooked meat or cured meat/cold cuts were associated with poultry‐attributed STEC infections. Consuming raw vegetables was protective for all STEC infections. We concluded that domestic ruminants account for approximately three‐quarters of reported human STEC infections, whereas pigs and poultry play a minor role and that risk factors for human STEC infection vary according to the attributed source.     

Recreational sandboxes for children and dogs can be a source of epidemic ribotypes of Clostridium difficile

Different studies have suggested that the sand of public playgrounds could have a role in the transmission of infections, particularly in children. Furthermore, free access of pets and other animals to the playgrounds might increase such a risk. We studied the presence of Clostridium difficile in 20 pairs of sandboxes for children and dogs located in different playgrounds within the Madrid region (Spain). Clostridium difficile isolation was performed by enrichment and selective culture procedures. The genetic (ribotype and amplified fragment length polymorphism [AFLP]) diversity and antibiotic susceptibility of isolates was also studied. Overall, 52.5% (21/40) of samples were positive for the presence of C. difficile. Eight of the 20 available isolates belonged to the toxigenic ribotypes 014 (n = 5) and 106 (n = 2), both regarded as epidemic, and CD047 (n = 1). The other 12 isolates were non‐toxigenic, and belonged to ribotypes 009 (n = 5), 039 (n = 4), and 067, 151 and CD048 (one isolate each). Nevertheless, all isolates (even those of a same ribotype) were classified into different AFLP genotypes indicating non‐relatedness. In conclusion, our results revealed the presence of epidemic ribotypes of C. difficile in children's and dog's sandboxes located nearby, which constitutes a major health risk.     

Randomized control trial to test the effect of a feed additive on Campylobacter contamination in commercial broiler flocks up to slaughter

A randomized controlled trial (RCT) was carried to evaluate the effect of a feed additive on Campylobacter contamination of broilers reared in commercial conditions. Twenty‐four broiler flocks naturally contaminated with Campylobacter were enrolled in the RCT: 12 were assigned to a control group (C) fed with a conventional finishing feed from 4 weeks of age to slaughter (around 35 days), and the other group of 12 flocks (S) was fed with a finishing feed supplemented with 250 ppm of a patented feed additive (an ion‐exchanged clay compound) previously proven to reduce Campylobacter contamination in broiler caeca under experimental conditions. Enumeration of Campylobacter colonies in caeca (8 per flock) was carried out following ISO standards before feed distribution and at slaughter. Before treatment, the caecal Campylobacter load tended to be lower in C flocks (7.1 ± 1.9 log CFU/g, CI_95% [6.6–7.5]) than in S flocks (7.7 ± 1.0 log UFC/g, CI_95% [7.5–7.9]) (p = .05). At slaughter, the bacterial load was similar in the S (7.7 ± 1.0 log CFU/g, CI_95% [7.5–7.9]) and C groups (7.5 ± 1.2 log CFU/g, CI_95% [7.2–7.8]) (p = .73). Therefore, the feed additive had no significant effect on the caecal Campylobacter load at slaughter under the tested conditions. The logistical constraints inherent in field trials and the natural variability of Campylobacter contamination in naturally infected broiler flocks make it difficult to reproduce experimental results in in situ farm conditions. RCT testing of an intervention strategy in commercial situation is therefore a key step in evaluating pre‐harvest interventions against food‐borne pathogens.     

Genetic structure of the Amur tiger (Panthera tigris altaica) population: Are tigers in Sikhote‐Alin and southwest Primorye truly isolated?

We used molecular genetic analyses to noninvasively identify individual Amur tigers and define subpopulations of tigers in the Russian Far East. We identified 63 individuals after genotyping 256 feces, 7 hair and 11 blood samples collected within southern, central and northern Sikhote‐Alin, as well as Southwest Primorye. Analysis of nuclear DNA at 9 microsatellite loci demonstrated greater genetic similarity between animals from southern and northern Sikhote‐Alin (some 500 km apart) than between animals from Ussuriskii State Nature Reserve and Southwest Primorye (less than 10 km apart at their nearest point), suggesting that a true barrier exists preventing movements of tigers between Southwest Primorye and the southern Sikhote‐Alin Mountains.     

Modelling the Phenological Relationships of Questing Immature Ixodes Ricinus (Ixodidae) Using Temperature and NDVI Data

All active stages of the tick Ixodes ricinus were collected monthly at two sites in northern Spain between the years 2000 and 2007. We used percentile accumulation of the active stage in the environment to evaluate simple and coherent correlations between accumulation of the active stages of larvae and nymphs and medium‐resolution MODIS satellite‐derived information on the climate, including monthly and accumulated temperature and the Normalized Difference Vegetation Index (NDVI). This framework is not intended to predict the actual abundance of ticks in the field as a measure of the hazard to humans, but to provide a basic structure for addressing the phenology of the tick in its geographic range. We demonstrated that the accumulation of larval ticks in the active stage is a sigmoid function of the accumulated temperature from the beginning of the calendar year. We also demonstrated that the accumulated temperature necessary to recruit nymphs from the questing larval stage is a function of the changes in accumulated larvae and nymphs and the accumulated temperature and NDVI recorded by the Aqua sensor. The low p‐values obtained in the regressions confirmed that such recruitment can be calculated using time intervals to estimate, for example, the beginning of the questing period or the time of the year when a population peak can be expected. The comparison among predicted and actual accumulated temperatures between larvae and nymph recruitment had an averaged error of ±20 days in one complete year. The use of accumulated temperature and NDVI proposed in this study opens up the re‐evaluation of reports on the phenology of the tick in Europe. This framework is intended to evaluate the same correlations along the tick's range and predict its phenological patterns in areas of pathogen transmission risk for humans.     

Seroepidemiology of Hepatitis E in Selected Population Groups in Croatia: A Prospective Pilot Study

Hepatitis E has become an emerging infection in many European countries. We analysed the prevalence of hepatitis E virus (HEV) infection in selected population groups in Croatia. Overall HEV IgG seropositivity was 5.6%, while 1.9% participants showed IgM antibodies suggestive of recent infection. No IgM‐positive sample was positive for HEV RNA. HEV IgG antibodies were most prevalent in alcohol abusers (8.9%) and war veterans (8.6%), compared with 6.1% among injecting drug users and 2.7% in healthcare professionals. No individual with high‐risk sexual behaviour tested HEV seropositive. HEV IgG positivity increased significantly with age from 1.8% to 2.3% in individuals younger than 40 years to 11.3% in individuals older than 50 years (P = 0.023). The mean age of HEV‐positive participants was significantly higher than that of HEV‐negative participants (50.9 ± 11.8 years versus 41.2 ± 11.8 years, P = 0.008). Seroprevalence rates were significantly higher in residents of suburban and rural areas compared with residents of urban areas (14.5% versus 2.5%, P = 0.003). Additionally, an increasing prevalence of HEV IgG antibodies was observed from 1.8% in participants living in families with two household members to 12.1% in those living with more than four members (P = 0.046). Gender, marital status, educational level, sexual orientation, source of drinking water, history of blood transfusions, surgical procedures, tattooing and travelling were not associated with HEV seroprevalence. Logistic regression showed that living in suburban/rural areas was the main risk factor for HEV seropositivity (OR = 6.67; 95%CI = 1.89–25.0; AOR = 7.14, 95%CI = 1.89–25.0).     

Prevalence and Characterization of Salmonella Isolated from Feral Pigs Throughout Texas

Feral pigs are one of the most abundant free‐roaming ungulates in the United States, yet their role in the ecology and transmission of foodborne pathogens is poorly understood. Our objectives were to estimate the prevalence of Salmonella shedding among feral pigs throughout Texas, to identify risk factors for infection, and to characterize the isolates. Faecal samples were collected from feral pigs in Texas from June 2013 through May 2015. Standard bacteriologic culture methods were used to isolate Salmonella from samples, and isolates were characterized via serotyping and anti‐microbial susceptibility testing. The prevalence of faecal Salmonella shedding among sampled pigs was 43.9% (194/442), with positive pigs originating from 50 counties. Pigs sampled during fall and summer were significantly more likely to be shedding Salmonella than pigs sampled during winter. High serovar diversity was evident among the isolates, and many of the detected serovars are leading causes of human salmonellosis. The most common serovars were Montevideo (10.0%), Newport (9.1%), and Give (8.2%). Resistance to anti‐microbial agents was rare. The burgeoning feral pig population in the United States may represent an emerging threat to food safety.     

High‐Level Ciprofloxacin‐Resistant Campylobacter jejuni Isolates Circulating in Humans and Animals in Incheon,Republic of Korea

Campylobacter jejuni is one of the major causative pathogens of outbreaks or sporadic cases of diarrhoeal diseases worldwide. In this study, we compared the phenotypic and genetic characteristics of C. jejuni isolates of human and food‐producing animal origins in Korea and examined the genetic relatedness between these two groups of isolates. Regardless of isolation source, all C. jejuni isolates harboured four virulence genes, cadF, cdtB, ciaB and racR, whereas the wlaN and virB11 genes were more frequently observed in human isolates. Antimicrobial susceptibility testing showed that the majority of C. jejuni isolates displayed high‐level resistance to fluoroquinolone (95.2%) or tetracycline (76.2%) antibiotics, and 12.4% of isolates exhibited multidrug resistance (more than three classes of antibiotics tested). Pulsed‐field gel electrophoresis (PFGE) of all Campylobacter isolates revealed 51 different SmaI‐PFGE patterns and six major clusters containing both human and animal isolates. These results indicate that genetically diverse strains of C. jejuni with antimicrobial drug‐resistance and virulence properties have prevailed in Incheon. Nevertheless, some particular populations continue to circulate within the community, providing the evidence for an epidemiological link of C. jejuni infections between humans and food‐producing animals. Therefore, the continued monitoring and surveillance of C. jejuni isolates of human and food‐producing animal origins are required for public health and food safety.     

Influenza A Viruses Detected in Swine in Southern Germany after the H1N1 Pandemic in 2009

Infections with influenza A viruses (IAV) are highly prevalent in swine populations, and stable cocirculation of at least three lineages has been well documented in European swine – till 2009. However, since the emergence of the human pandemic pdmH1N1 virus in 2009, which has been (re)introduced into individual swine herds worldwide, the situation has been changing. These variations in the respective IAV pools within pig populations are of major interest, and the zoonotic potential of putative emerging viruses needs to be evaluated. As data on recent IAV in swine from southern Germany were relatively sparse, the purpose of this study was to determine the major IAV subtypes actually present in this region. To this aim, from 2010 to 2013, 1417 nasal swabs or lung tissue samples from pigs with respiratory disease were screened for IAV genomes. Overall, in 130 holdings IAV genomes were detected by real‐time RT‐PCR targeting the matrix protein gene. For further analyses, several PCR protocols were adapted to quickly subtype between H1, pdmH1, H3, N1 and N2 sequences. Taken together, cocirculation of the three stable European lineages of IAV was confirmed for Bavaria. H1N1 sequences were identified in 59, whereas H1N2 genomes were only diagnosed in 14, and H3N2 in 9 of the holdings analysed. However, pdmH1 in combination with N1 was detected in 2010, 2012 and 2013 confirming a presence, albeit in low prevalence, likewise pdmH1N2 reassortant viruses. Interestingly, individual cases of coinfections with more than one subtype were diagnosed. Partial genome sequences were determined and phylogenetic analyses performed. Clearly other than in the human population classically circulating IAV have not been displaced by pdmH1N1 in Bavarian swine. However, some interesting viruses were detected. Further surveillance of these viruses in the Bavarian pig population will be of major importance, to monitor future developments.