Subolesin/akirin orthologs from Ornithodoros spp. soft ticks: cloning, RNAi gene silencing and protective effect of the recombinant proteins

Subolesin/akirin is a well characterized protective antigen highly conserved across vector species and thus potentially useful for the development of a broad-spectrum vaccine for the control of arthropod infestations including hard ticks, mosquitoes, sand flies and the poultry red mite Dermanyssus gallinae. Soft ticks could be also targeted by this vaccine if proved that the soft tick subolesin orthologs are conserved and induce protective immune responses too. However, to date no soft tick subolesin orthologs have been fully characterized nor tested as recombinant antigens in vaccination trials. The objectives of the present work were to clone and characterize the subolesin orthologs from two important vector species of soft ticks as Ornithodoros erraticus and O. moubata, to evaluate the effect of subolesin gene silencing by RNAi, and to test the protective value of the recombinant antigens in vaccination trials. The obtained results demonstrate that both soft tick subolesins are highly conserved showing more than 69% and 74% identity with those of hard ticks in their nucleotide and amino acid sequences, respectively. Additionally, we demonstrate that both soft ticks possess fully operative RNAi machinery, and that subolesin gene silencing by dsRNA injection inhibits oviposition indicating the involvement of subolesin in tick reproduction. Finally, vaccination with the recombinant soft tick subolesins induced a partial protective effect resulting in the reduction of the oviposition rate. These preliminary results encourage further studies on the use of recombinant subolesins as vaccines for the control of soft tick infestations, either alone or in combination with other specific molecules.     

Pathogenicity of Eimeria praecox alone or associated with Eimeria acervulina in experimentally infected broiler chickens

Eimeria praecox and Eimeria acervulina are two species of coccidia parasites infecting chickens, which develop in the duodenum. The purpose of this study was to evaluate the pathogenicity of E. praecox and to study interactions of this coccidium with E. acervulina. The results showed that the pathogenicity of E. praecox was related to the infective dose, and that its impact on individual weight and weight gain was significant from the lowest administered dose: 5000 oocysts per bird. No morbidity was observed, even with the highest infective dose, but faecal consistency alteration was higher with increasing infective doses. No consistent lesion was observed. When E. praecox was associated with E. acervulina with a low infective dose, performance deterioration seemed to be an additional effect of the two species. However, in the case of heavy infections, signs worsened along with duration of negative impact on growth, compared to a mono-infection.     

Ripening influences banana and plantain peels composition and energy content

Musa sp. peels are widely used by smallholders as complementary feeds for cattle in the tropics. A study of the influence of the variety and the maturation stage of the fruit on fermentability and metabolisable energy (ME) content of the peels was performed using banana (Yangambi Km5) and plantain (Big Ebanga) peels at three stages of maturation in an in vitro model of the rumen. Peel samples were analysed for starch, free sugars and fibre composition. Samples were incubated in the presence of rumen fluid. Kinetics of gas production were modelled, ME content was calculated using prediction equation and short-chain fatty acids production and molar ratio were measured after 72 h of fermentation. Final gas production was higher in plantain (269–339 ml g⁻¹) compared to banana (237–328 ml g⁻¹) and plantain exhibited higher ME contents (8.9–9.7 MJ/kg of dry matter, DM) compared to banana (7.7–8.8 MJ/kg of DM). Butyrate molar ratio decreased with maturity of the peels. The main influence of the variety and the stage of maturation on all fermentation parameters as well as ME contents of the peels was correlated to changes in the carbohydrate fraction of the peels, including starch and fibre.     

Dietary fat modulates dl-α-tocopheryl acetate (vitamin E) bioavailability in adult cockerels

1. A trial was designed to assess the effect of fat supplementation (amount and type of fatty acids) on vitamin E bioavailability in adult cockerels.

2. A total of 60 birds were force-fed three different diets: a semi-purified diet without added fat (Control diet) or supplemented with 3% fat as linseed (Linseed diet) or hydrogenated coconut oil (Coconut diet). The three experimental diets were also supplemented with dl-α-tocopheryl acetate to provide 40 mg vitamin E per bird.

3. After one week of depletion, blood was collected from the wing vein before (baseline) and 6, 12, 24 and 96 h after the gavage. Plasma samples were analysed for their α-tocopherol, cholesterol and triglycerides concentrations.

4. Results showed that the addition of 3% fat in the experimental diet increased post-gavage plasma α-tocopherol response by 153% for Linseed diet and by 75% for Coconut diet (P < 0.0001) compared to the Control group. Furthermore, the plasma α-tocopherol response observed with the Linseed diet was 44% greater than that observed with the Coconut diet (P < 0.0001). There was no effect of treatments on either plasma triglycerides (P = 0.91) or cholesterol (P = 0.45) responses.

5. In conclusion, this study shows that the addition of 3% fat to the diet significantly increases dl-α-tocopheryl acetate bioavailability in adult cockerels. Supplementation of fat rich in unsaturated fatty acids also leads to a higher dl-α-tocopheryl acetate bioavailability than fat rich in saturated fatty acids.

     

Employing a Monte Carlo algorithm in expectation maximization restricted maximum likelihood estimation of the linear mixed model

Multiple‐trait and random regression models have multiplied the number of equations needed for the estimation of variance components. To avoid inversion or decomposition of a large coefficient matrix, we propose estimation of variance components by Monte Carlo expectation maximization restricted maximum likelihood (MC EM REML) for multiple‐trait linear mixed models. Implementation is based on full‐model sampling for calculating the prediction error variances required for EM REML. Performance of the analytical and the MC EM REML algorithm was compared using a simulated and a field data set. For field data, results from both algorithms corresponded well even with one MC sample within an MC EM REML round. The magnitude of the standard errors of estimated prediction error variances depended on the formula used to calculate them and on the MC sample size within an MC EM REML round. Sampling variation in MC EM REML did not impair the convergence behaviour of the solutions compared with analytical EM REML analysis. A convergence criterion that takes into account the sampling variation was developed to monitor convergence for the MC EM REML algorithm. For the field data set, MC EM REML proved far superior to analytical EM REML both in computing time and in memory need.     

Effect of lactation on functional and morphological echocardiographic variables in adult dairy cows

ObjectivesTo study the effect of pregnancy and lactation on echocardiographic parameters in Holstein dairy cows.AnimalsNine multiparous high milk producing (HMP) dairy cows (producing more than 40 kg of milk per day in peak production) and 9 low milk producing (LMP) cows (producing less than 30 kg or milk per day in peak production).MethodsEchocardiography was performed twice; one month before calving and two months after calving.ResultsThe heart rate of HMP cows in the early lactation period was significantly higher than in the dry period. In LMP cows, there was a significant increase in left ventricular dimension in the early lactation period as compared to the dry period, and the interventricular septum in systole (IVSs) in the dry period was significantly thicker than early lactation period. In HMP cows, there was an increase in the right ventricular diameter in systole in the early lactation period as compared to the dry period. Left ventricular and aortic dimensions in the dry period of HMP were significantly higher than those of LMP cows. When the data were corrected for body weight, comparison of values of the dry period of HMP and LMP cows showed that left ventricular volume in systole in HMP was significantly higher and that IVSs, left ventricular fractional shortening and ejection fraction were significantly lower than in LMP cows.ConclusionsThis study demonstrated that lactation influences the intracardiac dimensions. The amount of milk production can influence echocardiographic parameters in dairy cows.     

Effect of breed and gender on bovine liver cytochrome P450 3A (CYP3A) expression and inter-species comparison with other domestic ruminants

The cytochrome P450 (P450) superfamily represents a group of relevant enzymes in the field of drug metabolism and several exogenous or constitutional factors contribute to regulate its expression. Cattle represent an important source of animal-derived food-products and studies concerning the P450 expression are needed for the extrapolation of pharmacotoxicological data from one species to another and for the evaluation of the consumer's risk associated with the consumption of harmful residues found in foodstuffs. In the present study, possible breed-, gender- and species-differences in P4503A (the P450 subfamily more expressed in the human liver) expression were studied in vitro in Piedmontese (PDM) and Limousin (LIM) meat cattle breeds of both sexes and in domestic Ruminants (cattle, sheep and goats). Cytochrome P450 and P4503A contents as well as CYP3A-dependent drug metabolising enzymes (DME) were measured in liver microsomes. Significant lower levels of P450 (P < 0.001) and P4503A (P < 0.05) contents were observed in PDM vs. LIM of both sexes; the P4503A-dependent DME activities were significantly (P values ranging from 0.05 up to 0.001) higher in PDM cattle, particularly in males. A gender-effect in DME activities was noticed (P < 0.05) only in PDM male cattle. With regards to the species, the expression of both P4503A apoprotein and some of the related DME activities were more pronounced in sheep (P < 0.01 vs. cattle) and in goats (P < 0.05 vs. sheep; P < 0.01 vs. cattle) than in cattle. The significant differences in P4503A expression observed in LIM and PDM cattle are consistent with previously published data on strain- and breed-differences pointed out in rats and men. As far as a possible sex-effect is concerned, no clear-cut evidence is likely to be drawn. Finally, P4503A expression was more relevant in small ruminants.     

Identification of viral pathogens in aborted fetuses and stillborn piglets from cases of swine reproductive failure in Spain

The objective of the present study was to determine the presence of recognised abortifacient viruses such as porcine reproductive and respiratory virus (PRRSV), Aujeszky's disease virus (ADV), porcine parvovirus (PPV) and porcine circovirus type 2 (PCV2), in tissues from aborted fetuses and stillborn neonates in cases of late reproductive failure in swine. A total of 293 specimens (fetuses aborted in the last third of gestation and stillborn piglets) from 100 different cases of late-term abortions and premature farrowing from 15 different Spanish provinces were studied. PRRSV was detected in 9/100 cases by RT-PCR. Only 1/100 cases analysed (corresponding to a late-term aborted fetus with a negative PRRSV RT-PCR result) was positive for PCV2 by PCR. Neither ADV (monitored by viral isolation plus antigen detection) nor PPV (monitored by ELISA antigen capture test) infection was identified. The results suggest that PRRSV is one of the most important infectious agents, if not the most relevant one, associated with fetal infection leading to abortion or premature farrowing in Spain. Moreover, other viral pathogens such as ADV, PPV and PCV2 seem to have a minor impact on reproductive disease.