Disease threats to the endangered Iberian lynx (Lynx pardinus)

The Iberian lynx, (Lynx pardinus), is the most endangered felid in the world. To determine whether sympatric carnivores are reservoirs of pathogens posing a disease risk for the lynx, evidence of exposure to 17 viral, bacterial and protozoan agents was investigated in 176 carnivores comprising 26 free-living lynx, 53 domestic cats, 28 dogs, 33 red foxes (Vulpes vulpes), 24 Egyptian mongooses (Herpestes ichneumon), 10 common genets (Genetta genetta) and 2 Eurasian badgers (Meles meles) in the areas inhabited by the last two populations of Iberian lynx, both in Andalusia (South-Western Spain).The results indicated that the lynx had low rates of contact with viral pathogens, with one seropositive finding each for feline leukemia virus, parvovirus and canine adenovirus-1, whereas contact with bacteria and protozoa appeared more frequent. Active infections with parvovirus, Ehrlichia spp., Mycobacterium bovis, Leptospira interrogans and Cytauxzoon spp. were confirmed. In contrast, 53% of the domestic cats were exposed to some infectious agent (prevalence range 4.5–11.4%). Antibodies to canine distemper virus and parvovirus were frequently found in dogs (32% and 42%, respectively) and foxes (30% and 12%). Past or present infections with parvovirus, Ehrlichia spp., Chlamydophila spp., M. bovis, Salmonella enterica, L. interrogans, Toxoplasma gondii, and Neospora caninum were also detected in these and other species surveyed.Questionnaires to owners revealed that 14% of the dogs but none of the cats had been vaccinated, and no cat had been neutered. Based on the apparent absence of acquired immunity of the lynx against infectious agents, the frequent detection of agents among sympatric carnivores, and the reported lack of immunocompetence of the Iberian lynx, a disease outbreak among the local abundant carnivores may pose a serious disease risk for lynx conservation.     

Effect of Competition on Nitric Oxide, Carbon Monoxide, and Cyclic Guanosine Monophosphate Levels in Training Horses

The aim of this study was to compare the effect of exercise on nitric oxide (NO), carbon monoxide (CO), and cyclic guanosyl monophosphate (cGMP) levels in jumping and dressage horses involved in competition. Blood samples were collected from the jugular vein of 100 horses involved in jumping or dressage competition at three time points: baseline at rest, on reaching the schooling but before exercise, and over a jump or dressage course. Fourteen healthy horses not involved in competition were used as control group. Exercise increased plasma CO concentration in both jumping and dressage horses, and this effect was more apparent in dressage horses. Exercise also increased NO plasma concentration in jumping horses, whereas it did not significantly modify NO plasma concentration in dressage horses. After exercise, plasma cGMP concentration was higher in both groups. Our results show that different signaling pathways are initially activated by exercise and that this activation is specific to the different modes of exercise. This information can be used to optimize warm-up and cool-down procedures for sport horses or to optimize training programs for equine athletes.     

Evaluation of baits for oral vaccination of European wild boar piglets

The objective of this study was to develop and evaluate new baits for the oral delivery of vaccine preparations to 2–4 month-old wild boar piglets. Baits were prepared using a matrix composed of wild boar feed, wheat flour, paraffin, sacarose and cinnamon-truffle powder attractant with polyethylene capsules dipped into the matrix to introduce vaccine formulation. Physical stability studies demonstrated that baits were stable for at least three days at temperatures as high as 42 °C. Recombinant Escherichia coli expressing the membrane-displayed BM95-MSP1a fusion protein were used to test bacterial viability in the baits and the antibody response in orally immunized wild boar. The E. coli viability was not significantly affected after bait incubation at 25 and 37 °C for 96 h. Bait acceptance studies using artificial feeders in the field showed that baits were accepted by 2–3 month-old animals, the preferred age for vaccination. Orally immunized wild boar piglets excreted recombinant E. coli in the feces and developed antibody titers to recombinant BM95-MSP1a protein, thus confirming that vaccine composition was released and reached the wild boar gastrointestinal track. The results of these experiments support the use of these baits for oral delivery of vaccine formulations to 2–4 month-old wild boar piglets.     

Correlating the immune response with the clinical-pathological course of persistent mastitis experimentally induced by Mycoplasma agalactiae in dairy goats

To correlate the clinical course of mycoplasma mastitis with its immune response, right mammary glands of 15 lactating goats were inoculating with 10¹⁰ colony-forming units (cfu) of Mycoplasma agalactiae (Ma). Before sacrificing the animals at 5, 15 or 45 days post-inoculation (dpi), blood Ma antibody titres and milk mycoplasma colony and somatic cell counts were monitored. Ma colonised the mammary gland and milk counts increased to over 10¹² cfu/ml within 5 dpi. During this period, an innate immune response involving neutrophils and macrophages was observed, and Ma antigen appeared in the degenerated acinar epithelium. From 7 dpi, a specific antibody response coincided with reduced viable mycoplasmas in milk. The humoral immune response was limited; by 37 dpi, all animals scored negative for anti-Ma antibodies, and around 10⁸ cfu/ml were shed. Results indicate an early immune response to Ma inoculation unable to control mycoplasmal invasion. An ensuing humoral response, despite reducing the mycoplasma burden, leads to chronic, persistent infection.     

Systemic antibody response in colostrum-deprived pigs experimentally infected with Haemophilus parasuis

The serum antibody response to an experimental infection by Haemophilus parasuis, the etiological agent of Glässer’s disease in pigs, was characterized by ELISA measuring IgM and IgGt levels against whole-cells and outer-membrane-proteins (OMPs) as antigens. Five groups of pigs were studied, four of those were previously immunized with different formulations, and the fifth was maintained as non-immunized control. All groups were challenged with 5 × 10⁹ CFU of H. parasuis. The non-commercial bacterin induced a full protection against disease, the OMP-vaccine and the exposure to a sublethal dose of 10⁵ CFU protected only partially, and the recombinant TbpB-vaccine conferred no protection. The humoral response in the pigs that died after infection (all controls, all those vaccinated with the recombinant TbpB, and two of both those inoculated with OMPs and those exposed to the sublethal dose) could be only measured before it, but it was irrelevant in all cases. However, a specific IgM and IgGt production was observed before challenge in all the surviving pigs, irrespective of the type of immunization received. This antibody response was even greater after H. parasuis infection, especially in those survivors receiving the sublethal dose. These results suggest a role of the antibodies developed after the different immunization protocols in preventing infection and death; therefore, the humoral immunity is protective against experimental Glässer’s disease.     

Organic‐carbon and nitrogen stocks and organic‐carbon fractions in soil under mixed pine and oak forest stands of different ages in NE Germany

The objective of this work was to evaluate the C and N stocks and organic‐C fractions in soil under mixed forest stands of Scots pine (Pinus sylvestris L.) and Sessile oak (Quercus petraea [Matt.] Liebl.) of different ages in NE Germany. Treatments consisted of pure pine (age 102 y), and pine (age 90–102 y) mixed with 10‐, 35‐, 106‐, and 124‐y‐old oak trees. After sampling O layers, soils in the mineral layer were taken at two different depths (0–10 and 10–20 cm). Oak admixture did not affect total organic‐C (TOC) and N stocks considering the different layers separately. However, when the sum of TOC stocks in the organic and mineral layers was considered, TOC stocks decreased with increasing in oak age (r² = 0.58, p < 0.10). The microbial C (C_MB) was not directly correlated with increase of oak age, however, it was positively related with presence of oak species. There was an increase in the percentage of the C_MB‐to‐TOC ratio with increase of oak‐tree ages. On average, light‐fraction C (C_LF) comprised 68% of the soil TOC in upper layer corresponding to the highest C pool in the upper layer. C_LF and heavy‐fraction C (C_HF) were not directly affected by the admixture of oak trees in both layers. The C_HF accounted on average for 30% and 59% of the TOC at 0–10 and 10–20 cm depths, respectively. Despite low clay contents in the studied soils, the differences in the DCB‐extractable Fe and Al affected the concentrations of the C_HF and TOC in the 10–20 cm layers (p < 0.05). Admixture of oak in pine stands contributed to reduce topsoil C stocks, probably due to higher soil organic matter turnover promoted by higher quality of oak litter.     

Organic vs conventionally grown Rio Red whole grapefruit and juice: comparison of production inputs, market quality, consumer acceptance, and human health-bioactive compounds

Most claims that organic produce is better tasting and more nutritious than nonorganic (conventional) produce are largely unsubstantiated. This is due mainly to a lack of rigor in research studies matching common production variables of both production systems, such as microclimate, soil type, fertilizer elemental concentration, previous crop, irrigation source and application, plant age, and cultivar. The aforementioned production variables common to both production systems were matched for comparison of Texas commercially grown conventional and certified organic Rio Red red-fruited grapefruit. Whole grapefruits from each production system were harvested between 800 and 1000 h at commercial early (November), mid- (January), and late season (March) harvest periods for three consecutive years. Within each harvest season, conventional and organic whole fruits were compared for marketable qualities (fruit weight, specific gravity, peel thickness, and peel color), and juices were compared for marketable qualities (specific gravity, % juice, and color), human health-bioactive compounds (minerals, ascorbic acid, lycopene, sugars, pectin, phenols, and nitrates), and consumer taste intensity and overall acceptance. Conventional fruit was better colored and higher in lycopene, and the juice was less tart, lower in the bitter principle naringin, and better accepted by the consumer panel than the organic fruit. Organic fruit had a commercially preferred thinner peel, and the juice was higher in ascorbic acid and sugars and lower in nitrate and the drug interactive furanocoumarins.     

Hydrophilic chromatographic determination of carnosine, anserine, balenine, creatine, and creatinine

A new HPLC procedure based on hydrophilic interaction chromatography (HILIC) has been developed for the simultaneous determination of carnosine, anserine, balenine, creatine, and creatinine in meat. This is the first time that HILIC has been directly applied to the study of meat components, having the advantage of not requiring complex cleanup and/or sample derivatization procedures. The chromatographic separation has been developed using a silica column (4.6 x 150 mm, 3 microm), and the proposed methodology is simple, reliable, and fast (<13 min per sample). The method has been validated in terms of linearity, repeatability, reproducibility, and recovery and represents an interesting alternative to methods currently in use for determining the mentioned compounds and other polar substances. The detection limits are 5.64, 8.23, 3.66, 3.99, and 0.06 microg/mL for carnosine, anserine, balenine, creatine, and creatinine, respectively.     

Transient etiolation: protochlorophyll(ide) and chlorophyll forms in differentiating plastids of closed and breaking leaf buds of horse chestnut (Aesculus hippocastanum)

An accompanying paper reports the accumulation of photoactive protochlorophyllide (Pchlide) in the innermost leaf primordia of buds of many tree species. In this paper, we describe plastid differentiation, changes in pigment concentrations and spectral properties of bud scales and leaf primordia of horse chestnut (Aesculus hippocastanum L.) from January until the end of bud break in April. The bud scales contained plastids with grana, stroma thylakoids characteristic of chloroplasts and large dense bodies within the stroma. In January, proplastids and young chloroplasts were present in the leaf primordia, and the fluorescence spectra of the primordia were similar to those of green leaves except for a minor band at 630 nm, indicative of a protochlorophyll(ide). During bud break, the pigment concentrations of the green bud scales and the outermost leaf primordia increased, and Pchlide forms with emission maxima at 633, 644 and 655 nm accumulated in the middle and innermost leaf primordia. Depending on the position of the leaf primordia within the bud, their plastids and their pigment concentrations varied. Etio-chloroplasts with prolamellar bodies (PLBs) and prothylakoids with developing grana were observed in the innermost leaves. Besides the above-mentioned Pchlide forms, the middle and innnermost leaf primordia contained only a Chl band with an emission maximum at 686 nm. The outermost leaf primordia contained etio-chloroplasts with well-developed grana and small, narrow-type PLBs. These outermost leaves contained only chlorophyll forms like the mature green leaves. No Pchlide accumulation was observed after bud break, indicating that etiolation of the innermost and middle leaves is transient. The Pchlide forms and the plastid types of the primordia in buds grown in nature were similar to those of leaves of dark-germinated seedlings and to those of the leaf primordia of dark-forced buds. We conclude that transient etiolation occurs under natural conditions. The formation of PLBs and etio-chloroplasts and the accumulation of the light-dependent NADPH:protochlorophyllide oxidoreductase are involved in the natural greening process and ontogenesis of young leaf primordia of horse chestnut buds.     

Optical properties of bud scales and protochlorophyll(ide) forms in leaf primordia of closed and opened buds

The transmission spectra of bud scales of 14 woody species and the 77 K fluorescence emission spectra of the innermost leaf primordia of closed and opened buds of 37 woody species were studied. Pigment concentrations were determined in some species. Bud scales had low transmittance between 400 and 680 nm with a local minimum around 680 nm. Transmittance increased steeply above 680 nm and was > 80% in the 700-800 nm spectral region. Significant protochlorophyllide (Pchlide) accumulation was observed in leaf primordia of tightly packed, closed buds with relatively thick, dark bud scales. In common ash (Fraxinus excelsior L.) and Hungarian ash (Fraxinus angustifolia Vahl.), the innermost leaf primordia of the closed buds contained protochlorophyll (Pchl) and Pchlide (abbreviated as Pchl(ide)), but no chlorophyll. We observed Pchl(ide) forms with emission maxima at 633, 643 and 655 nm in these leaves. Complete transformation of Pchlide(655) (protochlorophyllide form with maximum emission at 655 nm) into Chlide(692) (chlorophyllide form with maximum emission at 692 nm) occurred after irradiation for 10 s. The innermost leaf primordia of the buds of four species (flowering ash (Fraxinus ornus L.), horse chestnut (Aesculus hippocastanum L.), tree of heaven (Ailanthus altissima P. Mill.) and common walnut (Juglans regia L.)) contained Pchl(ide)(633), Pchl(ide)(643) and Pchlide(655) as well as an emission band at 688 nm corresponding to a chlorophyll form. The Pchlide(655) was fully photoactive in these species. The outermost leaf primordia of these four species and the innermost leaf primordia of 28 other species contained all of the above described Pchl(ide) forms in various ratios but in small amounts. In addition, Chl forms were present and the main bands in the fluorescence emission spectra were at 690 or 740 nm, or both. The results indicate that Pchl(ide) accumulation occurs in leaf primordia in near darkness inside the tightly closed buds, where the bud scales and the external leaf primordia function as optical filters.