Palms of the past: can morphometric phytolith analysis inform deep time evolution and palaeoecology of Arecaceae?

BACKGROUND AND AIMS: Palm fossils are often used as evidence for warm and wet palaeoenvironments, reflecting the affinities of most modern palms. However, several extant palm lineages tolerate cool and/or arid climates, making a clear understanding of the taxonomic composition of ancient palm communities important for reliable palaeoenvironmental inference. However, taxonomically identifiable palm fossils are rare and often confined to specific facies. Although the resolution of taxonomic information they provide remains unclear, phytoliths (microscopic silica bodies) provide a possible solution because of their high preservation potential under conditions where other plant fossils are scarce. We thus evaluate the taxonomic and palaeoenvironmental utility of palm phytoliths. METHODS: We quantified phytolith morphology of 97 modern palm and other monocot species. Using this dataset, we tested the ability of five common discriminant methods to identify nine major palm clades. We then compiled a dataset of species' climate preferences and tested if they were correlated with phytolith morphology using a phylogenetic comparative approach. Finally, we reconstructed palm communities and palaeoenvironmental conditions at six fossil sites. KEY RESULTS: Best-performing models correctly identified phytoliths to their clade of origin only 59 % of the time. Although palms were generally distinguished from non-palms, few palm clades were highly distinct, and phytolith morphology was weakly correlated with species' environmental preferences. Reconstructions at all fossil sites suggested that palm communities were dominated by Trachycarpeae and Areceae, with warm, equable climates and high, potentially seasonal rainfall. However, fossil site reconstructions had high uncertainty and often conflicted with other climate proxies. CONCLUSIONS: While phytolith morphology provides some distinction among palm clades, caution is warranted. Unlike prior spatially restricted studies, our geographically and phylogenetically broad study indicates phytolith morphology may not reliably differentiate most palm taxa in deep time. Nevertheless, it reveals distinct clades, including some likely to be palaeoenvironmentally informative.

Streptococcus caprae sp. nov., isolated from Iberian ibex (Capra pyrenaica hispanica).

Biochemical and molecular genetic studies were performed on a novel Gram-stain-positive, catalase-negative, coccus-shaped organism isolated from tonsil samples of two Iberian ibexes. The micro-organism was identified as a streptococcal species based on its cellular, morphological and biochemical characteristics. 16S rRNA gene sequence comparison studies confirmed its identification as a member of the genus Streptococcus, but the organism did not correspond to any species of this genus. The nearest phylogenetic relative of the unknown coccus from ibex was Streptococcus porci 2923-03T (96.6 % 16S rRNA gene sequence similarity). Analysis based on rpoB and sodA gene sequences revealed sequence similarity values lower than 86.0 and 83.8 %, respectively, from the type strains of recognized Streptococcus species. The novel bacterial isolate was distinguished from Streptococcus porci and other Streptococcus species using biochemical tests. Based on both phenotypic and phylogenetic findings, it is proposed that the unknown bacterium be classified as representing a novel species of the genus Streptococcus, for which the name Streptococcus caprae sp. nov. is proposed. The type strain is DICM07-02790-1CT ( = CECT 8872T = CCUG 67170T).

Nutrient and plant secondary compound composition and iron-binding capacity in leaves and green stems of commonly used plant browse (Carolina willow; Salix caroliniana) fed to zoo-managed browsing herbivores.

Plant secondary compounds are diverse structurally, and associated biological effects can vary depending on multiple factors including chemical structure and reaction conditions. Phenolic compounds such as tannins can chelate dietary iron, and supplementation of animal species sensitive to iron overload with tannins may prevent/treat iron overload disorder. We assessed the nutrient and phenolic composition and iron-binding capacity of Carolina willow (Salix caroliniana), a plant fed to zoo-managed browsing herbivores. Based on studies in other plant species and the chemical structures of phenolic compounds, we hypothesized that the concentration of condensed tannins in willow would be inversely related to the concentration of phenolic glycosides and directly related to iron-binding capacity. Our results indicated that willow nutrient composition varied by year, season, and plant part, which could be taken into consideration when formulating animal diets. We also found that the predominant plant secondary compounds were condensed tannins with minimal phenolic glycosides. Instead of binding to iron, the willow leaf extracts reduced iron from the ferric to ferrous form, which may have prooxidative effects and increase the bioavailability of iron depending on animal species, gastrointestinal conditions, and whole animal processes. We recommend identifying alternative compounds that effectively chelate iron in vitro and conducting chelation therapy trials in vivo to assess potential effects on iron balance and overall animal health.

Implementing a low-starch biscuit-free diet in zoo gorillas: the impact on behavior.

In the wild, western lowland gorillas travel long distances while foraging and consume a diet high in fiber and low in caloric density. In contrast, gorillas in zoos typically consume a diet that is low in fiber and calorically dense. Some items commonly used in captive gorilla diets contain high levels of starch and sugars, which are present at low levels in the natural diet of gorillas. Diet items high in simple carbohydrates are associated with obesity and heart disease in humans. Typical captive gorilla diets may also encourage undesirable behaviors. In response to these issues, we tested the behavioral impact of a diet that was biscuit-free, had low caloric density, and which was higher in volume at five institutions. We hypothesized that this diet change would reduce abnormal behaviors such as regurgitation and reingestion (R/R), decrease time spent inactive, and increase time spent feeding. The biscuit-free diet significantly reduced (and in the case of one zoo eliminated) R/R and may have reduced hair-plucking behavior. However, an increase in coprophagy was observed in many individuals following the diet change. The experimental diet caused a general increase in time the gorillas spent feeding, but this increase did not occur across all institutions and varied by individual. Interestingly, the overall time gorillas spent inactive actually increased with this diet change. Future research will examine these behavioral changes in a greater number of individuals to determine if the results remain consistent with these preliminary findings. Additionally, future research will examine the physiological impact of this diet change.

Implementing a low-starch biscuit-free diet in zoo gorillas: the impact on health.

In the wild, western lowland gorillas consume a diet high in fiber and low in caloric density. In contrast, many gorillas in zoos consume a diet that is high-calorie and low in fiber. Some items commonly used in captive gorilla diets contain high levels of starch and sugars, which are minimal in the natural diet of gorillas. There is a growing concern that captive gorillas may qualify as obese. Furthermore, the leading cause of death for adult male gorillas in zoos is heart disease. In humans, a diet that is high in simple carbohydrates is associated with both obesity and the incidence of heart disease. In response to these issues, we implemented a biscuit-free diet (free of biscuits and low in fruit) and measured serum biomarkers of obesity and insulin resistance pre- and post-diet change at three institutions: North Carolina Zoological Garden, Cleveland Metroparks Zoo, and Columbus Zoo and Aquarium. We also added a resistant starch supplement to gorilla diets at two of the above institutions. We anticipated that these diet changes would positively affect biomarkers of obesity and insulin resistance. Both diet manipulations led to a reduction in insulin. Resistant starch also decreased overall serum cholesterol levels. Future research will examine these health changes in a greater number of individuals to determine if the results remain consistent with these preliminary findings.

Antimicrobial resistance in indicator Escherichia coli isolates from free-ranging livestock and sympatric wild ungulates in a natural environment (Northeastern Spain).

Antimicrobial resistance was assessed in indicator Escherichia coli isolates from free-ranging livestock and sympatric wild boar (Sus scrofa) and Iberian ibex (Capra pyrenaica) in a National Game Reserve in northeastern Spain. The frequency of antimicrobial resistance was low (0% to 7.9%). However, resistance to an extended-spectrum cephalosporin and fluoroquinolones was detected.

Streptococcus porcorum sp. nov., isolated from domestic and wild pigs.

Seven isolates of an unidentified Gram-stain-positive, catalase-negative, coccus-shaped organism isolated from domestic and wild pigs were characterized by phenotypic and molecular-genetic methods. Based on cellular morphology and biochemical criteria, the isolates were tentatively assigned to the genus Streptococcus, although the organisms did not appear to correspond to any recognized species. Comparative 16S rRNA gene sequencing showed that the unknown bacterium was phylogenetically closely related to, but distinct from, Streptococcus suis (97.5 % 16S rRNA gene sequence similarity to the type strain). rpoB and sodA sequence analysis showed minimum interspecies divergence from phylogenetically close 16S rRNA gene sequence-based relatives of 13.8 and 18.6 %, respectively. DNA-DNA hybridization of a strain of the unidentified organism demonstrated 8-18 % reassociation with S. suis NCTC 10234(T). The novel bacterium could be distinguished from S. suis and other Streptococcus species using biochemical tests. On the basis of phenotypic and phylogenetic evidence, it is proposed that the unknown isolates from domestic and wild animals be assigned to a novel species of the genus Streptococcus, Streptococcus porcorum sp. nov. The type strain is 682-03(T) ( = CCUG 58479(T)  = CECT 7593(T)).

Streptococcus rupicaprae sp. nov., isolated from a Pyrenean chamois (Rupicapra pyrenaica).

Biochemical and molecular genetic studies were performed on an unknown Gram-stain-positive, catalase-negative, coccus-shaped organism isolated from clinical samples of a Pyrenean chamois. The micro-organism was identified as a streptococcal species based on its cellular morphological and biochemical tests. 16S rRNA gene sequence comparison studies confirmed its identification as a member of the genus Streptococcus, but the organism did not correspond to any species of this genus. The nearest phylogenetic relative of the unknown coccus from chamois was Streptococcus ovis (95.9 % 16S rRNA gene sequence similarity). The rpoB and sodA sequence analysis showed sequence similarity values of less than 85.7 % and 83.0 %, respectively, with the currently recognized species of the genus Streptococcus. The novel bacterial isolate was distinguished from S. ovis and other species of the genus Streptococcus using biochemical tests. Based on both phenotypic and phylogenetic findings, it is proposed that the unknown bacterium be classified as a novel species of the genus Streptococcus, Streptococcus rupicaprae sp. nov., with the type strain 2777-2-07(T) ( = CECT 7718(T)  = CCUG 59652(T)).

Decreasing prevalence of brucellosis in red deer through efforts to control disease in livestock.

When a pathogen infects a number of different hosts, the process of determining the relative importance of each host species to the persistence of the pathogen is often complex. Removal of a host species is a potential but rarely possible way of discovering the importance of that species to the dynamics of the disease. This study presents the results of a 12-year programme aimed at controlling brucellosis in cattle, sheep and goats and the cascading impacts on brucellosis in a sympatric population of red deer (Cervus elaphus) in the Boumort National Game Reserve (BNGR; NE Spain). From February 1998 to December 2009, local veterinary agencies tested over 36 180 individual blood samples from cattle, 296 482 from sheep and goats and 1047 from red deer in the study area. All seropositive livestock were removed annually. From 2006 to 2009 brucellosis was not detected in cattle and in 2009 only one of 97 red deer tested was found to be positive. The surveillance and removal of positive domestic animals coincided with a significant decrease in the prevalence of brucellosis in red deer. Our results suggest that red deer may not be able to maintain brucellosis in this region independently of cattle, sheep or goats, and that continued efforts to control disease in livestock may lead to the eventual eradication of brucellosis in red deer in the area.

The impact of public transport on the health of work commuters: a systematic review.

Although the public transport (PT) commute can form a substantial part of the working day, there is a significant gap in our understanding of how it influences health of those who engage in it. The purpose of this systematic review was to therefore generate evidence from 1972 about the extent to which the PT commute (involving train, bus, subway, tram, or metro) impacts on the mental health, physical health and well-being of the working people. We identified 47 studies in English worldwide involving an empirical quantitative focus which met the inclusion criteria. Of these, 23 studies involved over 500 participants. Although initial multi-modal comparisons showed impact on sickness rate, self-rated health complaints, perceived stress level and reduction in sleep, a more homogeneous analysis of rail commuters showed elevation in salivary cortisol, perceived stress, and affective reactions to crowding. Findings also revealed a bias towards use of endogenous self-report measures. On this basis, we argue that it would be of benefit to test theoretical models to account for more objective measures of job and commuting stress. Recommendations were made for flexible working agendas.

High seroprevalence of Neospora caninum in the red fox (Vulpes vulpes) in the Pyrenees (NE Spain).

Antibodies to Neospora caninum were determined in the red fox (Vulpes vulpes) in the Pyrenees, north-eastern Spain. Sera from 53 red foxes (29 male and 24 female) were tested using a Neospora agglutination test (NAT). Seroprevalence at dilutions of 1/40, 1/80 and 1/160 was 69.8%, 47.2% and 7.5%, respectively. Sex differences were significant only at a dilution of 1:40. The high seroprevalence observed in red fox suggests that this species is highly exposed to N. caninum in this area.

Ingesta passage and gastric emptying times in loggerhead sea turtles (Caretta caretta).

Ingesta passage times of soft flat foam dishes and gastric emptying time of barium-impregnated polyethylene spheres (BIPS) were measured in 22 and 8 loggerhead sea turtles (Caretta caretta), respectively. Transit time (T(1)) was considered as the time between ingestion and first elimination, and retention time (T(50)) and total transit time (T(85)) the expulsion time of 50% and 85% of the markers, respectively. The experiments were carried out at different times of the year and water temperature was recorded. A set of dorso-ventral radiographs was taken to locate the BIPS, and the gastrointestinal anatomy of 5 dead turtles was studied to help with interpretation of the radiographs. No significant correlation was observed between T(1), T(50), T(85) and minimum straight carapace length (SCLmin) or body mass and no statistical difference was found in ingesta passage transit times between juvenile (n = 6) and sub-adult turtles (n = 16). Mean passage times of the dishes (in days) were: T(1) = 9.05, T(50) = 12.00 and T(85) = 13.19. Gastric emptying time using BIPS was 24-48 h. The transit time (T(1)) for the BIPS was longer (13.25 +/- 4.86 days) than the foam markers (8.5 +/- 2.73 days) in 8 turtles studied simultaneously. Although the total transit time tended to be faster in turtles submitted to water temperatures between 20 degrees C and 23.6 degrees C no significant correlation was observed between T(1), T(50) and T(85) and the temperature.

First isolation of Haemophilus parasuis and other NAD-dependent Pasteurellaceae of swine from European wild boars.

Haemophilus parasuis is a colonizer of the upper respiratory tract of pigs and the etiological agent of Glässer's disease, which is characterized by a fibrinous polyserositis, meningitis and arthritis. Glässer's disease has never been reported in wild boar (Sus scrofa), although antibodies against H. parasuis have been detected. The goal of this study was to confirm the presence of this bacterium in wild boar by bacterial isolation and to compare the strains to H. parasuis from domesticated pigs. Therefore, nasal swabs from 42 hunted wild boars were processed for bacterial isolation and subsequent H. parasuis identification by specific PCR, biochemical tests and 16S rRNA gene sequencing. Two different strains of H. parasuis from two wild boars were isolated. These strains belonged to serotype 2 and were included by 16S rRNA gene sequencing and MLST analysis in a cluster with other H. parasuis strains of nasal origin from domestic pigs. During this study, Actinobacillus minor and Actinobacillus indolicus, which are NAD-dependent Pasteurellaceae closely related to H. parasuis, were also isolated. Our results indicate similarities in the respiratory microbiota of wild boars and domestic pigs, and although H. parasuis was isolated from wild boars, more studies are needed to determine if this could be a source of H. parasuis infection for domestic pigs.

Seroprevalence of Neospora caninum in non-carnivorous wildlife from Spain.

Serum samples from 1034 non-carnivorous wildlife from Spain were tested for antibodies to Neospora caninum by competitive screening enzyme linked immunosorbent assay (ELISA) and confirmed by an indirect fluorescent antibody test (IFAT). High agreement was observed between results in both techniques (kappa value higher than 0.9). Prevalences of N. caninum antibodies positive by both techniques were 11.8% of 237 red deer (Cervus elaphus), 7.7% of 13 barbary sheep (Ammotragus lervia), 6.1% of 33 roe deer (Capreolus capreolus) and 0.3% of 298 wild boar (Sus scrofa). In one of 53 hares (Lepus granatensis), antibodies were found in the ELISA but could not be confirmed by IFAT due to lack of sample. Antibodies to N. caninum were not found in any of 251 wild rabbits (Oryctolagus cuniculus), 79 fallow deer (Dama dama), 27 mouflon (Ovis ammon), 40 chamois (Rupicapra pyrenaica) and three Spanish ibex (Capra pyrenaica). Statistically significant differences were observed between N. caninum seroprevalence in red deer and management of hunting estates (open versus fenced) with higher prevalence in fenced estates, and among sampling sites. Seroprevalence was particularly high in some areas (MO estate in South-Central Spain or some estates of Catalonia, North-East Spain), while no contact with N. caninum was observed in others. Results indicate that in certain areas of Spain, N. caninum is present in wildlife, especially in red deer. These results have important implications in both sylvatic cycles and may influence the prevalence of infection in cattle farms in those areas. To our knowledge, this is the first report of antibodies to N. caninum in wildlife from Spain and the first report of N. caninum antibodies in barbary sheep and wild boar.

Computed tomography of the vertebral column and coelomic structures in the normal loggerhead sea turtle (Caretta caretta).

The aim of this study was to determine the normal computed tomography (CT) appearance of the vertebral column and coelomic structures of the loggerhead sea turtle (Caretta caretta) and to use three-dimensional (3D) and multiplanar reconstructions to indicate the position of each organ in relation to the vertebrae and carapace. Transverse sections of 1mm thickness were performed in seven clinically healthy and in five dead loggerhead sea turtles using multi-detector CT equipment. A computer workstation was used for multiplanar and 3D reconstructions. Dead turtles were frozen and sectioned in the transverse, dorsal and sagittal planes to compare the anatomical structures' appearance with CT images. Clinically relevant organs including the oesophagus, stomach, trachea, bronchi, lungs, liver, gallbladder, heart, spleen, kidneys and vertebral canal were identified in CT images. Computed tomography provides detailed information on the respiratory system and skeleton; the location of the coelomic structures with respect to the carapace and the vertebrae that is provided in this work will facilitate the use of other ancillary diagnostic techniques such as ultrasound, radiography and biopsy, thereby improving safety of access in surgical procedures.

Ultrasonographic imaging of loggerhead sea turtles (Caretta caretta).

Twenty live and five dead juvenile and subadult loggerhead sea turtles were examined ultrasonographically. Ten soft tissue areas of the integument were used as acoustic windows: cervical-dorsal and cervical-ventral, left and right cervicobrachial, left and right axillary, left and right prefemoral and left and right postfemoral windows. Anatomical cross-sections were performed on the dead turtles to provide reference data. The fourth and fifth cervical vertebrae, the spinal cord, and the venous sinuses of the external jugular vein were clearly visible through the cervical-dorsal acoustic window, and the oesophagus and the heart were imaged through the cervical-ventral acoustic window. The stomach was more frequently visible through the left axillary acoustic window. The liver could be imaged through both sides, but the right axillary acoustic window was better for visualising the gall bladder. The large and small intestines and the kidneys were visible through the right and left prefemoral acoustic windows; the kidneys were easily identified by their intense vasculature.

Effect of acepromazine on the signs of capture stress in captive and free-ranging roe deer (Capreolus capreolus).

The differences between the capture stress responses of captive and free-ranging roe deer (Capreolus capreolus) and their modulation by acepromazine, during a period of three hours' physical restraint after capture in drive-nets, were examined in 16 free-ranging and 16 captive roe deer. Eight of the free-ranging and eight of the captive animals received acepromazine intramuscularly, and the other eight free-ranging and eight captive deer received the same volume of saline. Heart rate, body temperature and haematological and serum biochemical parameters were analysed. In the groups treated with acepromazine, the heart rate stabilised sooner, and the red blood cell (RBC) count, haemoglobin concentration, packed-cell volume, the serum activities of creatine kinase (CK), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) and the concentrations of creatinine and lactate were significantly lower, and serum glucose started to decrease earlier, than in the untreated groups. Serum potassium levels decreased over time only in the untreated groups. The body temperature stabilised earlier, and the RBC count, haemoglobin concentration, serum CK, AST, ALT and LDH activities, and serum creatinine, lactate, cholesterol and glucose concentrations were significantly lower in the free-ranging roe deer than in the captive deer.

Serological Follow-up of Tuberculosis in a Wild Boar Population in Contact with Infected Cattle.

There is an increasing concern in several European countries over the role that tuberculosis (TB)-infected wild boar may play in the progress of bovine TB eradication campaigns. In 2004, as a consequence of the detection of a TB focus in wild boar from a National Game Reserve (NGR) located in southern Catalonia, a surveillance programme based on post-mortem inspection for detection of macroscopic TB-like lesions (TBLL) was initiated in the affected area. The source of infection for wild boar was linked to a tuberculous cattle herd located in the same area. Besides, the results of the surveillance programme in wild boar were used for the validation of an indirect enzyme-linked immunosorbent assay (ELISA) specific for Mycobacterium tuberculosis complex (MTBC) IgG antibodies. Using this ELISA, a seven-year serological study of MTBC in wild boar from the NGR was conducted in 173 animals (93 adults, 44 juveniles-yearlings and 36 piglets) culled between 2004 and 2010. ELISA results and presence of TBLL showed excellent agreement for adult and juvenile wild boar (Kappa index = 0.85; 95% CI: 0.76-0.95). Of the thirty-eight adults, yearlings and juveniles classified as positives by the ELISA, 34 (89%) showed TBLL at necropsy. In contrast, none of the ELISA-positive wild boar piglets (n = 20) showed TBLL, suggesting the detection of early antibody responses to the infection. Overall, this study contributes to the knowledge of wild boar humoral responses to MTBC. The results also highlight the usefulness of this serological test for wild boar TB surveillance.

Perfluorinated alkyl substances (PFASs) in northern Spain municipal solid waste landfill leachates.

Landfill leachates have been recognized as significant secondary sources of poly- and perfluoroalkyl substances (PFASs). This study presents data on the occurrence and concentration of 11 perfluoroalkyl carboxylates (PFCAs) and 5 perfluoroalkyl sulfonates (PFSAs) in leachates from 4 municipal solid waste landfill sites located across northern Spain. To the best of our knowledge, this is the first report of the presence of PFASs in Spanish landfill leachates. Two of the landfill sites applied on-site treatment using membrane bioreactors (MBR), and its effect on PFASs occurrence is also reported. Total PFASs (∑PFASs) in raw leachates reached 1378.9 ng/L, while in treated samples ∑PFASs was approximately two-fold (3162.3 ng/L). PFCAs accounted for the majority of the detected PFASs and perfluorooctanoic acid (PFOA) was the dominant compound in raw leachates (42.6%), followed by shorter chain PFHxA (30.1%), PFPeA and PFBA. The age of the sites might explain the PFASs pattern found in raw leachates as all of them were stabilized leachates. However, PFASs profile was different in treated samples where the most abundant compound was PFHxA (26.5%), followed by linear perfluorobutane sulfonate (L-PFBS) (18.7%) and PFOA (17.7%). The overall increase of the PFASs content as well as the change in the PFASs profile after the MBR treatment, could be explained by the possible degradation of PFASs precursors such as fluorotelomer alcohols or fluorotelomer sulfonates. Using the volume of leachates generated in the landfill sites, that served 1.8 million people, the discharge of 16 ∑PFASs contained in the landfill leachates was estimated as 1209 g/year.

Post-Natal Persistent Infection With Classical Swine Fever Virus in Wild Boar: A Strategy for Viral Maintenance?

In this study, fifteen wild boar piglets were intranasally inoculated <10 h after birth with the moderately virulent classical swine fever virus (CSFV) strain Catalonia 01. At 5 days post-inoculation, seven other animals within 48 h of birth were put in contact with them. Viral replication and innate and specific immune responses were evaluated. Of the inoculated animals, 46.67% remained post-natally persistently infected and were apparently healthy with neither humoral nor cellular immunological responses specific to CSFV and with high viral loads in their blood, organs and body secretions. Moreover, the present data extend the time period to 48 h after birth when a moderately virulent CSFV strain could lead to post-natal persistent infection given the generation of persistently infected wild boars in the contact group (33.33%). The innate immune response to the virus, as measured by type I IFN-α in serum, was mostly not impaired in the persistently infected wild boars. Interestingly, a decrease and lack of IFN-γ-producing cells against CSFV and PHA was observed. In endemic countries where wild swine species are increasing and low and moderate virulence CSFV strains are prevalent, the possible generation of this form of disease cannot be ruled out.