Role of wild ruminants in the epidemiology of bluetongue virus serotypes 1, 4 and 8 in Spain

ABSTRACT: Although the importance of wild ruminants as potential reservoirs of bluetongue virus (BTV) has been suggested, the role played by these species in the epidemiology of BT in Europe is still unclear. We carried out a serologic and virologic survey to assess the role of wild ruminants in the transmission and maintenance of BTV in Andalusia (southern Spain) between 2006 and 2010.A total of 473 out of 1339 (35.3%) wild ruminants analyzed showed antibodies against BTV by both ELISA and serum neutralization test (SNT). The presence of neutralizing antibodies to BTV-1 and BTV-4 were detected in the four species analyzed (red deer, roe deer, fallow deer and mouflon), while seropositivity against BTV-8 was found in red deer, fallow deer and mouflon but not in roe deer. Statistically significant differences were found among species, ages and sampling regions. BTV RNA was detected in twenty-one out of 1013 wild ruminants (2.1%) tested. BTV-1 and BTV-4 RNA were confirmed in red deer and mouflon by specific rRT-PCR.BTV-1 and BTV-4 seropositive and RNA positive wild ruminants, including juveniles and sub-adults, were detected years after the last outbreak was reported in livestock. In addition, between the 2008/2009 and the 2010/2011 hunting seasons, the seroprevalence against BTV-1, BTV-4 and BTV-8 increased in the majority of provinces, and these serotypes were detected in many areas where BTV outbreaks were not reported in domestic ruminants. The results indicate that wild ruminants seem to be implicated in the dissemination and persistence of BTV in Spain.     

Peripheral white blood cell counts throughout pregnancy in non-aborting Neospora caninum-seronegative and seropositive high-producing dairy cows in a Holstein Friesian herd

Pregnancy is characterized by transient changes in the maternal immune system, also evident at peripheral level. The present study analyzes the kinetics and possible factors affecting peripheral white blood cell populations throughout pregnancy in a herd of high-producing dairy cows chronically infected or not with Neospora caninum. We examined 54 pregnant parous cows: 29 Neospora-seronegative and 25 Neospora-seropositive cows. Blood samples were collected on Days 90, 120, 150, 180 and 210 of gestation. General Linear Model (GLM) repeated measures analysis of variance showed that the interaction Neospora-seropositivity × parity significantly affected total leukocyte, neutrophil and monocyte counts with lower levels of total leukocytes, lower neutrophil and higher monocyte counts recorded in primiparous Neospora-seropositive cows. In addition, N. caninum-seropositive cows had significantly increased monocyte counts on Day 180 of gestation compared to seronegative ones. Other factors significantly associated with changes in total and/or differential leukocyte profiles were period of pregnancy, season, twin pregnancy and milk production. In conclusion, a parity-associated effect of chronic N. caninum infection was observed on peripheral blood cell profiles in dairy cattle during gestation.     

New Bruce-ladder multiplex PCR assay for the biovar typing of Brucella suis and the discrimination of Brucella suis and Brucella canis

Rapid and specific identification of Brucella suis at the biovar level is necessary because some of the biovars that infect animals are pathogenic for humans. None of the molecular typing methods described so far are able to discriminate B. suis biovars in a single test and differentiation of B. suis from Brucella canis by molecular approaches can be difficult. This article describes a new multiplex PCR assay, Suis-ladder, for fast and accurate identification of B. suis at the biovar level and the differentiation of B. suis, B. canis and Brucella microti. An advancement of the original Bruce-ladder PCR protocol which allows the correct discrimination of all known Brucella species is also described.     

Lipophilic Toxins in Wild Bivalves from the Southern Gulf of California,Mexico

Most of the shellfish fisheries of Mexico occur in the Gulf of California. In this region, known for its high primary productivity, blooms of diatoms and dinoflagellates are common, occurring mainly during upwelling events. Dinoflagellates that produce lipophilic toxins are present, where some outbreaks related to okadaic acid and dinophisystoxins have been recorded. From January 2015 to November 2017 samples of three species of wild bivalve mollusks were collected monthly in five sites in the southern region of Bahía de La Paz. Pooled tissue extracts were analyzed using LC-MS/MS to detect lipophilic toxins. Eighteen analogs of seven toxin groups, including cyclic imines were identified, fortunately individual toxins did not exceed regulatory levels and also the total toxin concentration for each bivalve species was lower than the maximum permitted level for human consumption. Interspecific differences in toxin number and concentration were observed in three species of bivalves even when the samples were collected at the same site. Okadaic acid was detected in low concentrations, while yessotoxins and gymnodimines had the highest concentrations in bivalve tissues. Although in low quantities, the presence of cyclic imines and other lipophilic toxins in bivalves from the southern Gulf of California was constant.     

Structure and biological activities of lipochitooligosaccharide nodulation signals produced by Bradyrhizobium japonicum USDA 138 under saline and osmotic stress

The establishment of a symbiotic interaction involves a signal exchange between the host legume (flavonoids) and the nitrogen-fixing rhizobia (nodulation factors (NFs)). Likewise, abiotic stress conditions, such as salinity and drought, strongly reduce the nodulation process, possibly affecting also the signal exchange. In this work we characterized the structure and biological activity of NFs produced by Bradyrhizobium japonicum USDA 138 under control, salt, and osmotic stress conditions. This strain is the most widely used in Argentine soybean culture; under control conditions, it produces a mixture of four types of NFs (V(C_16:0,MeFuc), V(C_18:1,MeFuc), IV(C_18:1), and V(C_18:1,Ac,MeFuc)). Interestingly, under stress conditions, this strain produces new types of NFs, one common for both stress conditions (V(C_16:1,MeFuc)) and another one only present under salt stress (IV(C_18:1,MeFuc)). All mixtures of NFs, extracted from control, salt, and osmotic stress conditions, showed biological activity in soybean plants, such as root hair deformation, and the radical application of purified NFs induced systemic differences in dry matter accumulation. The inoculation of soybean with genistein-induced bacteria cultured under both control and stress conditions had a positive effect on the number of nodules formed and in some cases on dry matter accumulation. These responses are not related to changes in chlorophyll fluorescence or greenness index.     

Native maize landraces from Los Tuxtlas, Mexico show varying mycorrhizal dependency for P uptake

Different degrees of dependency on the activity of arbuscular mycorrhizal fungi (AMF) exist between native maize landraces and hybrids. In Los Tuxtlas, Mexico, the Popoluca people maintain a traditional polycultural land management with more than 15 native landraces of maize; however, it is not known whether the recent substitution of local maize for improved hybrids and fertilization has affected the integrity of the mycorrhizal symbiosis in these naturally phosphorus-poor systems. A greenhouse experiment was conducted to evaluate the response of four Popoluca maize landraces and the hybrid Texcoco to the presence of native AMF in conditions of low and medium P input (5 and 65 mg kg^?1, respectively). After 120 days in both P treatments, the native landraces Black and Yellow presented higher colonization and had acquired more P in their shoot biomass than the hybrid. The moderate fertilization did not appear to have affected the integrity of the mycorrhizal symbiosis, since all of the maize types presented a positive mycorrhizal dependency (2–14 %). Under low P conditions, the Texcoco hybrid maize presented one of the highest mycorrhizal dependencies; however, unlike the local landraces, this was not reflected in a higher tissue P concentration. The results obtained indicate that the native maize Black was the best at capturing symbiotic and direct P, which makes this landrace an important genetic and cultural heritage for the Popoluca and for the world.     

Exploitation of Exotic, Cultivated Solanum Germplasm for Breeding and Commercial Purposes

Native potato species (NPS) are cultivated potatoes which do not belong to S. tuberosum ssp. tuberosum. They are maintained by the farmers of the Andes for subsistence under harsh environmental conditions where other potatoes cannot compete. Within the huge phenotypic variability between and within these species certain accessions possess resistances to different pests and diseases, remarkable culinary and processing quality and favourable nutritional properties. We present first evaluation results of a set of NPS for these characters and discuss prospects for exploiting this valuable germplasm for breeding and for developing novel products for niche markets.     

A new lupene triterpenetriol and anticholinesterase activity of Salvia sclareoides

A new lupene triterpenetriol was isolated from the acetone extract of the aerial parts of Salvia sclareoides and characterised as (1beta,3beta)-lup-20(29)-ene-1,3,30-triol (1). In addition, nepetidin (2), nepeticin (3), lupendiol (4), (1beta,11alpha)-dihydroxy-lup-20(29)-en-3-one (5), ursolic acid (6), sumaresinolic acid (7) and hederagenin (8), were identified in this Salvia sp. To the best of our knowledge, the compounds 2 and 7 are new constituents in Salvia spp. The acetone, ethanol, butanol and water extracts of the plant were screened for the in vitro inhibitory activity of acetylcholinesterase (AChE) and butyrilcholinesterase (BChE), enzymes which play a role in the Alzheimer disease. All extracts inhibited acetylcholinesterase activity at 10 microg/ml, a remarkable activity since the standard drug rivastigmine does not inhibit acetylcholinesterase at the same concentration. Regarding the butyrilcholinesterase, the acetone extract at 1000 microg/ml was able to inhibit completely the enzyme activity and the butanol and ethanol extracts, at this concentration, produced a potent inhibition of BchE.     

Dynamics of violaxanthin and lutein epoxide xanthophyll cycles in Lauraceae tree species under field conditions

Two xanthophyll cycles have been described in higher plants: the violaxanthin xanthophyll (V or VAZ) cycle, which is present in all species, and the taxonomically restricted lutein epoxide xanthophyll (Lx) cycle, which involves the light-induced de-epoxidation of Lx to lutein (L) and its epoxidation back to Lx in low light. Laboratory experiments indicate that the first reaction occurs quickly, but the second reaction is much slower. We investigated the Lx cycle under field conditions in several tree species of the Lauraceae family to determine its relationship with the ubiquitous V cycle. The field study was conducted in two natural laurel forests: one in the Canary Islands, where Laurus azorica (Seub.) Franco, Ocotea foetens (Aiton.) Benth, Apollonias barbujana (Cav.) Bornm. and Persea indica (L.) Spreng were studied; and one in the Basque Atlantic coast where Laurus nobilis L. was studied. The results were complemented by a taxonomic study. The presence of Lx was widespread among Lauraceae species, but its concentration varied even among closely related species. The V pool size correlated positively with growth irradiance, whereas the relationship between Lx pool size and growth irradiance varied with species. A functional Lx cycle was confirmed under field conditions only in O. foetens and L. nobilis. Furthermore, in O. foetens, a correlation between Lx de-epoxidation and photoinhibition suggested a protective role for this cycle. We conclude that, unlike the V cycle, which is normally correlated with irradiance, the operation and light dependence of the Lx cycle is species-dependent.