Use of serology and real time PCR to control an outbreak of bovine brucellosis at a dairy cattle farm in the Nile Delta region,Egypt

Background

Bovine brucellosis remains one of the most prevalent zoonotic infections affecting dairy cattle in developing countries where the applied control programs often fail. We analyzed the epidemiologic pattern of bovine brucellosis in a dairy cattle herd that showed several cases of abortions after regular vaccination with RB51 (B. abortus vaccine). In 2013 thirty dairy cows, from a Holstein-Friesian dairy herd with a population of 600 cattle, aborted five months post vaccination by a regular RB51 vaccine. Blood samples were drawn from milking cows and growing heifers, as well as heifers and cows pregnant up to 6 months. These samples were collected in June 2013 (n?=?257) and May 2014 (n?=?263) and were tested by real time (rt)-PCR as well as serological tests, in particular Rose Bengal Test (RBT), Enzyme-Linked Immunosorbent Assays (ELISA) and Fluorescence Polarization Assay. Tissue specimens were also collected from an aborted fetus and cultured. Isolates were subjected to bacteriological typing tests at the genus and species levels.

Results

Five months post vaccination with RB51 vaccine, Brucella (B.) DNA was detected in blood samples of cows by rt-PCR. The serological tests also revealed the spread of Brucella field strains within the herd in 2013. Four Brucella isolates were recovered from specimens collected from the aborted fetus. These isolates were typed as follows: one B. abortus RB51 vaccine strain and three isolates of B. abortus field strain. The seropositive cows with positive rt-PCR might indicate an infection by the Brucella field strain; while the positive rt-PCR results from seronegative animals may either be due to circulating RB51 vaccine DNA in vaccinated animals or to circulating field strain in infected animals before seroconversion.

Conclusion

The results herein suggest that PCR can be a good supplementary tool in an outbreak situation, if an assay is available that can differentiate vaccine and field strains with a high analytical sensitivity. We recommend using RBT and ELISA in parallel in outbreak situations, to identify as many infected animals as possible during the initial screenings. This test procedure should be repeated for at least three successive negative tests, with one month interval.

     

Competitive and allelopathic interference between soybean crop and annual wormwood (Artemisia annua L.) under field conditions

Annual wormwood interference on soybean crop growth and yield may result from competition and allelopathy, which are modulated by crop management. Allelochemicals released by annual wormwood (e.g. artemisinin) may affect the crop directly or indirectly through the effect on the nitrogen fixing symbiont, Bradyrhizobium japonicum. The objectives were (i) to quantify the crop response (i.e. biomass production, nodulation and yield) to weed interference and (ii) to determinate the relative change of competition and allelopathy interferences, when a sublethal dose of herbicide is applied. Two split plot field experiments with three replications were used. The experiment involved a factorial combination of five weed–crop density (soybean/annual wormwood, plants m^?2) levels: D1, pure soybean, 40/0 plants m^?2; D2, 40/2 plants m^?2; D3, 40/4 plants m^?2 and D4, 40/8 plants m^?2, and D5, pure annual wormwood, 0/8 plants m^?2, two activated carbon (allelopathy) levels: C?, with activated carbon (reduced allelopathy) and C+, non activated carbon applied (with allelopathy) and two herbicide levels: H?, untreated and H+, treated with a sub-lethal dose of glyphosate. Activated carbon to adsorb allelochemicals (with and without activated carbon) and glyphosate application (with and no herbicide) were assigned to sub-plots. Increasing weed density did not affect crop biomass at flowering, but changed nodule number and soybean yield with a different pattern depending on carbon and herbicide treatment. Relative crop yield decreased with increasing relative weed biomass. This decrease was particularly drastic when allelopathy was reduced by activated carbon and without herbicide application. The maximum yield losses of 33% in 2006 and 17% in 2007 were observed with the highest weed density (8 plants m^?2). In contrast, without carbon (high allelopathy level), soybean yield remained stable within the explored range of annual wormwood biomass, despite the fact that weed biomass at high densities (D4) was high enough to generate competition. The lack of response to increasing weed density could be related to the indirect effect of allelochemicals interacting with soil microorganisms (i.e. B. japonicum) that positively affected the nodulation (e.g. larger nodules in 2006 and increased nodules biomass due to higher number of roots in 2007 at high densities). With herbicide application, soybean yield of both carbon treatments remained stable when biomass of annual wormwood increased. This research provided strong evidence in support of the existence of positive effect of allelopathic and competitive interactions between annual wormwood and soybean crop under field conditions that may be overridden under herbicide application.     

Historical processes and landscape context influence genetic structure in peripheral populations of the collared lizard (<Emphasis Type="Italic">Crotaphytus collaris</Emphasis>)

Populations at the periphery of a species’ range often show reduced genetic variability within populations and increased genetic divergence among populations compared to those at the core, but the mechanisms that give rise to this core-periphery pattern in genetic structure can be multifaceted. Peripheral population characteristics may be a product of historical processes, such as founder effects or population expansion, or due to the contemporary influence of landscape context on gene flow. We sampled collared lizards (Crotaphytus collaris) at four locations within the northern Flint Hills of Kansas, which is at the northern periphery of their range, to determine the genetic variability and extent of genetic divergence among populations for ten microsatellite loci (n = 229). We found low genetic variability (average allelic richness = 3.37 ± 0.23 SE; average heterozygosity = 0.54 ± 0.05 SE) and moderate population divergence (average F_ST = 0.08 ± 0.01 SE) among our sample sites relative to estimates reported in the literature at the core of the species’ range in Texas. We also identified differences in dispersal rates among sampling locations. Gene flow within the Flint Hills was thus greater than for other peripheral populations of collared lizards, such as the Missouri glade system where most of the mesic grasslands have been converted to forest since the last glacial retreat, which appears to have greatly impeded gene flow among populations. Our findings signify the importance of considering landscape context when evaluating core-peripheral trends in genetic diversity and population structure.     

Prevalence of antimicrobial residues in raw table eggs from farms and retail outlets in Enugu State,Nigeria

The use of antimicrobial agents in poultry production results in their accumulation in the body tissues and products such as milk and egg. The subsequent accumulation of these drugs and their metabolites in body cells is known as drug residue. This study was designed to determine the prevalence of antimicrobial residues in eggs from poultry farms and retail outlets in Enugu State, Nigeria. Eggs from 25 selected commercial farms and ten retail outlets were screened for the prevalence of antimicrobial residue. Also, structured questionnaires were administered to 25 commercial farms in the state to determine the management practices and the most widely used antimicrobial drugs in farms and possible association between the management practices and the occurrence of antimicrobial residues in eggs from these farms. All the 25 farms surveyed use oxytetracycline. Eggs from nine of the surveyed farms tested positive for antimicrobial residue and three of the ten surveyed farms also tested positive for antimicrobial residue. No association was observed (p 0.05; Fisher’s exact test) between the occurrence of antibiotic residues in eggs and farm size, feed source and housing systems. This study was able to demonstrate the presence of antimicrobial residues in eggs destined for human consumption. Drugs like nitrofurans which has been banned for use in food animals are still very much in use in Enugu State, Nigeria. Antibiotics given as feed additives may give rise to drug residues in food animals.     

Morphological characterization of the IVIA persimmon (Diospyros kaki Thunb.) germplasm collection by multivariate analysis

Diospyros kaki Thunb. originated in Eastern Asia, as evidenced of its culture in China as early as several centuries B.C. In the seventh century, persimmon was introduced to Japan and later, in the fourteenth century to Korea. There is no information about persimmon culture in Europe until the seventeenth century, with the spread over the world occurring in the eighteenth century. The genus Diospyros contains more than 400 species, with levels of ploidy ranging from diploid (2n = 2x = 30) up to nonaploid (2n = 9x = 135). The primary economic crop species is Diospyros kaki Thunb., which is mainly hexaploid (2n = 6x = 90) and includes hundreds of cultivars. Although a relatively recent introduction in Europe, the species has adapted well, and the genetic diversity have been expanded with culture and selection for the past 200 years in the Mediterranean basin. These locally adapted cultivars were evaluated with cultivars from Asian origin in a germplasm collection established at IVIA in Valencia, Spain. In this paper 27 cultivars from the IVIA collection were studied by multivariate analysis, and 37 variables were analyzed using a Principal Components Analysis and cluster analysis following the method UPGMA. Studies on correlations and significance among variables identified the most relevant ones, and thus provided information for a future core collection.     

Application to soil of disinfectants through irrigation reduces Verticillium dahliae in the soil and verticillium wilt of olive

The application of disinfectants through drip irrigation could be a feasible practice against verticillium wilt (Verticillium dahliae) of olive. OX-VIRIN (activated peroxide) and OX-AGUA AL25 (quaternary ammonium compounds) are two disinfectants that have shown efficacy against V. dahliae in irrigation water and potential for reducing the disease in young olive plants. In this work, various post-planting application strategies incorporating OX-VIRIN (once a month, or twice a month on alternate or successive weeks) or OX-AGUA AL25 (once a month, or twice a month on alternate weeks) were assessed for their effect on V. dahliae in soil, disease in olive trees, and olive yield, in a 2-year pot-experiment under natural environmental conditions. The disinfectants were injected via metering pumps into a drip irrigation system that irrigated olive trees planted in V. dahliae-inoculated soil. All the application strategies significantly reduced the total inoculum density in soil compared to controls with no disinfectants and noninoculated soil. The microsclerotia density was also significantly reduced in disinfested soils by 73.6–86.8%, depending on the strategy. The symptoms and infection incidence were always lower in treatments subjected to disinfestation. The treatment with OX-AGUA AL25 applied twice a month on alternate weeks most reduced the symptoms (by 53.0%) and colonization index (by 70.8%) with respect to untreated water control. This soil disinfestation also significantly strengthened the symptom remission. Tree growth and production were negatively affected by soil inoculation (reduced by 45.6% and 88.7%, respectively), but not so by disinfectants, which even relieved the reduction in inoculated soils, especially when OX-AGUA AL25 was applied.     

Cobalt reduces nitrate inhibition of nodulation in mungbean (Vigna radiata)

 A cobalt-mediated decrease in ethylene production reduced the inhibition of nodulation by nitrate in Vigna radiata (mungbean). Nitrate increased the ethylene production in 5-day-old seedlings, while it caused a reduction in the nodulation status (nodule number and nodule weight) and nodule efficiency (acetylene reduction activity) in mungbean plants. The application of cobalt chloride inhibited nitrate-affected ethylene production and also decreased the inhibitory effect of nitrate on nodulation. The effect of cobalt was most marked on nodule number. Received: 6 August 1999     

A putative RNA silencing component protein FoQde-2 is involved in virulence of the tomato wilt fungus <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis>

RNA silencing pathways in filamentous fungi are composed of multiple component proteins and known to be involved in vegetative growth, virulence or sexual reproduction. We found that the tomato wilt fungus, Fusarium oxysporum f. sp. lycopersici (Fol), carries four homologues genes of Qde-2, an argonaute protein gene and one of the main component protein genes in Neurospora crassa. Gene targeting revealed that FoQde-2, one of the Qde-2 homologues in Fol, is involved in virulence to tomato but not in vegetative growth.