Evaluation of efficacy of several disinfectants against Campylobacter jejuni strains by a suspension test

The objective of this study was to determine the efficacy of 16 active compounds and 11 commercial disinfectants against Campylobacter jejuni. Two reference strains (one of avian origin and the other isolated from bovine) and two avian field strains were tested in suspension test in the presence and absence of serum. Chloramine-T, povidone–iodine (1% available iodine), cetylpiridinium chloride, ethanol, isopropanol, chlorhexidine digluconate, formaldehyde, phenol, and 10 of the 11 commercial formulations (eight of them based on quaternary ammonium compounds) showed an excellent disinfectant capability, resulting in the highest level of reduction (>6-log₁₀) in colony-forming units of the four C. jejuni strains compared regardless of the presence or absence of organic material. These compounds might be helpful in the adoption of environmental control measures against C. jejuni.     

Single dose adenovirus vectored vaccine induces a potent and long-lasting immune response against rabbit hemorrhagic disease virus after parenteral or mucosal administration

Rabbit hemorrhagic disease virus (RHDV) is the etiological agent of a lethal and contagious disease of rabbits that remains as a serious problem worldwide. As this virus does not replicate in cell culture systems, the capsid protein gene has been expressed in heterologous hosts or inserted in replication-competent viruses in order to obtain non-conventional RHDV vaccines. However, due to technological or safety issues, current RHDV vaccines are still prepared from organs of infected rabbits. In this work, two human type 5 derived replication-defective adenoviruses encoding the rabbit hemorrhagic disease virus VP60 capsid protein were constructed. The recombinant protein was expressed as a multimer in mouse and rabbit cell lines at levels that ranged from approximately 120 to 160 mg/L of culture. Mice intravenously or subcutaneously inoculated with a single 10(8) gene transfer units (GTU) dose of the AdVP60 vector (designed for VP60 intracellular expression) seroconverted at days 7 and 14 post-immunization, respectively. This vector generated a stronger response than that obtained with a second vector (AdVP60sec) designed for VP60 secretion. Rabbits were then immunized by parenteral or mucosal routes with a single 10(9)GTU dose of the AdVP60 and the antibody response was evaluated using a competition ELISA specific for RHDV or RHDVa. Protective hemagglutination inhibition (HI) titers were also promptly detected and IgG antibodies corresponding with inhibition percentages over 85% persisted up to one year in all rabbits, independently of the immunization route employed. These levels were similar to those elicited with inactivated RHDV or with VP60 obtained from yeast or insect cells. IgA specific antibodies were only found in saliva of rabbits immunized by intranasal instillation. The feasibility of VP60 production and vaccination of rabbits with replication-defective adenoviral vectors was demonstrated.     

In vitro acaricidal effect of tannin-rich plants against the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

The objectives of this study were to evaluate the in vitro acaricidal effects of lyophilized extracts of four tannin rich plants (Acacia pennatula, Piscidia piscipula, Leucaena leucocephala and Lysiloma latisiliquum) against diverse stages of Rhipicephalus (Boophilus) microplus, and to asses whether tannins were involved in the acaricidal effect using polyethylene glycol (PEG) to block tannins. Larval immersion (LIT) and adult immersion (AIT) tests were used to evaluate the acaricidal effect of each of the lyophilized extracts against larval and adult stages of R. microplus respectively. Larvae and adult ticks were exposed to increasing concentrations of each plant extract (0, 1200, 2400, 4800, 9600 and 19,200 μg ml(-1)) for 10 min. Larval mortality was recorded at 48 h post-incubation. Adult mortality was recorded daily over 14 days, at which point their reproductive efficiency was evaluated. PEG was added to the extracts to verify whether tannins were involved in the acaricidal effect. The effect on egg laying inhibition and larval mortality was analyzed using the GLM procedure in SAS. A Kruskal-Wallis test was used to assess the effect of PEG on LIT results. Calculation of the lethal concentration 50 (LC50) was performed using a probit analysis. All extracts reduced the viability of R. microplus larval stages (P<0.001), and viability was restored with the addition of PEG suggesting an important role of tannins in the acaricidal effect (P<0.001). The LC50 values of L. latisiliquum and P. piscipula plant extracts were 6.402 and 2.466 μg ml(-1). None of the tannin-rich plant extracts affected adult mortality (P>0.05). Lysiloma latisiliquum extract inhibited egg hatching of R. microplus (P<0.01). Tannin-rich plant extracts from A. pennatula, P. piscipula, L. leucocephala and L. latisiliquum showed potential acaricidal activity. Further in vivo studies are needed to confirm this finding.     

Enhanced Tolerance to Mercury in a Streptomycin-Resistant Strain of Euglena gracilis

Toxicity of Hg²⁺ was determined in two strains of Euglena gracilis var. bacillaris: the wild-type (B) and a streptomycin-resistant strain (Sm^r). Cells were cultured under 12-h dark/12-h light regime or under continuous darkness. In the dark/light cultures, cellular growth was severely diminished in the B strain by Hg²⁺, whereas only a slight decrease was attained in the Sm^r strain; Hg²⁺ also affected the photosynthetic and respiratory activities of the B strain, but not those of Sm^r strain. Under continuous darkness, cellular growth of both strains was lower than under dark/light cycles, but it was inhibited by Hg²⁺ to a much lesser extent. Cell culture by 5 days under continuous dark or by 8 days of dark/light cycles resulted in a higher intracellular content of mercury in B strain than in Sm^r strain. In contrast, in both culture conditions, the fraction of mercury removed from medium by B strain was lower than that attained by Sm^r strain, whereas the ability to bio-transform (reduce) Hg²⁺ was two times higher in the mutant strain. The results suggested that Sm^r strain cells acquired an ability to remove Hg²⁺ from the medium, which was not associated to accumulation and which conferred protection against mercury.     

Genetic parameter estimation and gene association analyses for meat quality traits in open-air free-range Iberian pigs

Meat quality of Iberian pigs is defined by the combination of their genetic characteristics and the particular production system. To carry out a genetic analysis of the main meat quality traits, we estimated their heritabilities, genetic correlations and the association effects of 32 selected SNPs of 12 candidate genes. A total of ten traits were measured in longissimus dorsi samples from 1,199 Iberian pigs fattened in the traditional free-range system: water holding capacity (thawing, cooking and centrifuge force water losses), instrumental colour (lightness L*, redness a* and yellowness b*), myoglobin content, shear force on cooked meat, and shear force and maximum compression force on dry-cured loin. Estimated heritability values were low to moderate (0.01 to 0.43) being the lowest for L* and the highest for cooking loss. Strong genetic correlations between water holding capacity traits (0.93 to 0.96) and between myoglobin content and a* (0.94) were observed. The association analyses revealed 19 SNPs significantly associated with different traits. Consistent and strong effects were observed between PRKAG3 SNPs (rs319678464G > C and rs330427832C > T), MYH3_rs81437544T > C, CASP3_rs319658214G > T and CTSL_rs332171512A > G and water losses. Also for CAPN1_rs81358667G > A and CASP3_rs319658214G > T and shear force. The SNPs mapping on PRKAG3 showed the highest effects on Minolta colour traits. Genotyping of these SNPs could be useful for the selection of Iberian young boars with similar estimated breeding values for productive traits.     

Proposal for rational antibacterial use in the diagnosis and treatment of dogs with chronic diarrhoea

Chronic diarrhoea is a frequent complaint in canine practice and the diagnostic path is often characterised by numerous diagnostic tests and stepwise empirical treatments, often applied before gastrointestinal endoscopy/mucosal biopsies. These include dietary interventions (novel protein, hydrolysed protein diet), parasiticides and still, in many cases, antibacterials. Indiscriminate use of antibacterial drugs risks detrimental consequences for both the individual patient (antimicrobial resistance, long-term disruption of intestinal bacterial populations, potential worsening of gastrointestinal signs) and the general public. For that reason, in this Perspective essay we advocate use of antibacterials only after histopathologic evaluation of gastrointestinal biopsies or, for those cases in which endoscopy is not possible, after other therapeutic trials, such as diet/pre-probiotics or anti-inflammatory drugs have proven unsuccessful. They should be reserved, after appropriate dietary trials, for those canine chronic diarrhoeic patients with signs of true primary infection (i.e. signs of systemic inflammatory response syndrome or evidence of adherent-invasive bacteria) that justify antibacterial use.     

Phenylalanine ammonia‐lyase activity and ortho‐diphenolic content with regard to boron nutrition in sunflower leaves.

Homogenates were prepared from the leaves of hydroponically cultivated sunflower (Helianthus annuus, L.) under deficient, normal and toxic B conditions and the phenylalanine ammonia‐lyase activity together with the ortho‐diphenolic and micronutrient content were measured every week from one month to flowering

The restoration of enzymatic activity on the exogenous addition of borate “in vivo”; was also studied.

Both B toxicity and deficiency resulted in increased phenylalanine ammonia‐lyase activity in the homogenates

The toxicity effect was early and sustained and the deficiency effect late and transitory. “In vitro”; borate addition to the reaction mixture did not significantly change the phenylalanine‐ammonia‐lyase activity in the homogenates.

There was no statistical correlation between the phenylalanine‐ammonia‐lyase activity and any micronutrient content in leaves.

A high accumulation of ortho‐diphenolics appeared in B‐deficient leaves only after 28 days of differential culture and may be responsible for the drop in phenylalanine‐ammonia‐lyase activity noted at this time.     

Polyphenol oxidase activity and ortho‐diphenolic content in sunflower leaves in relation to boron nutrition.

Homogenates were prepared from the leaves of hydroponically cultivated sunflowers (Helianthus annuus, L.) under deficient, normal and toxic B conditions and the polyphenol oxidase activity together with the ortho‐diphenolic and micronutrient content was measured every week from one month to flowering.

B deficiency slightly depressed polyphenol oxidase activity and B toxicity decreased it significantly at all times, the response fitting a line versus the logarithm of the B concentration in solution. We found a positive correlation between polyphenol oxidase activity and the B content in leaves.

B action on enzymatic activity seemed not to be caused by direct interaction with substrates as the infiltration of B into deficient leaves did not restore the normal activity

According to our results we suggest that polyphenol oxidase activity may be a valid functional parameter to indicate the nutritional status of B in sunflower plants, if its specificity for B can be demonstrated.

The ortho‐diphenolic content did not change with the B levels used in our experiments. Neither did we find any correlation between enzymatic activity and the o‐diphenolic content of the leaves.     

Microbial response of soils with organic and conventional management history to the cultivation of Bacillus thuringiensis (Bt)-maize under climate chamber conditions

An experiment was carried out in a climate chamber to analyse if Bt-maize may cause particular changes in soils with different levels of microbial biomass and activity due to long-term management history. Among the soils selected, the ones managed organically for 30 years exhibited twice the microbial biomass and 2.6 times the dehydrogenase activity (DHA) of the soil from a field with long-term conventional maize monoculture. Soils were cultivated twice in a row with Bt-maize, its near-isogenic line and a conventional breeding line. We tested the hypotheses that (a) soil microbial biomass and activity are affected by the cultivation of Bt-maize and that (b) the influence of Bt-maize depends on the level of soil microbial biomass and activity. Shoot and root yield and shoot C-content of Bt-maize were higher than the ones of the near-isogenic line. DHA under Bt-maize was 6 % higher, and the metabolic quotient for CO₂ (qCO₂) was 9 % lower than under its near-isogenic line, giving some support to hypothesis (a). No significant interactions of the soils and the varieties used were found in this study, thus hypothesis (b) was not confirmed, and soils with different microbial biomass and activity appear to react in a similar way to the cultivation of Bt-maize.     

Unravelling the relationships among Verticillium wilt,irrigation, and susceptible and tolerant olive cultivars

Survival, germination, olive colonization, and water-use efficiency (WUE) impairments by Verticillium dahliae could be influenced by cultivar susceptibility or irrigation, and this could modify the irrigation–pathogen–disease relationship. In this study, the combined effects of irrigation and cultivar susceptibility on Verticillium wilt (VW) development were modelled by the temporary assessment of V. dahliae propagules (total inoculum density, density of micropropagules, and sclerotia in wet and air-dried soil; ID, MpD, S_wD, and S_dD, respectively), root (RCI) and shoot (SCI) colonization indexes, and WUE. The relationship of disease severity to the measured parameters was then explored. Under controlled conditions, plants of cultivars ‘Picual’ and ‘Frantoio’ were irrigated to a high and low rate by varying drip-irrigation frequencies: daily, twice weekly, and a combination of daily for 11 days and then twice weekly. Disease severity and colonization parameters were higher in ‘Picual’, while WUE was higher in ‘Frantoio’. However, high rate and twice weekly and combination treatments significantly increased disease incidence and reduced time-to-symptoms-onset only in ‘Picual’, while high rate reduced WUE and increased relative ID, MpD, and S_wD in both cultivars. Irrigation did not affect SCI, but a higher RCI was found at high rate during the development of symptoms in ‘Picual’. By using classification trees to examine parameters—disease severity relationships, it was possible to determine the degree to which VW was affected by irrigation and/or cultivar susceptibility. MpD was the best indicator for VW detection at any time, WUE was best before symptoms developed, and RCI, total ID, and S_dD after symptoms developed.