Immunization of proteins from Toxascaris leonina adult worm inhibits allergic specific Th2 response

Recently, the influence of parasitic infections on the incidence of allergic diseases has become the focus of increased attention. In order to ascertain whether parasite-derived proteins could inhibit the allergic specific Th2 response, we applied excretory-secretory protein (Tl-ES) or total protein (Tl-TP) of the adult worm Toxascaris leonina to asthma model mice prior to or simultaneously with OVA challenge, after which we assessed the OVA-specific Th2 responses. The group subjected to immunization with Tl-ES and Tl-TP (immunized group) evidenced a thinning of the bronchial epithelial and muscle layer, a disruption and shedding of epithelial cells, a reduction in the number of goblet cells, and a reduction in mucus production as compared to the group treated with Tl-ES coupled with OVA challenge (challenge with OVA groups) and the OVA-induced asthma group. The administration of Tl-ES and Tl-TP, regardless of injection time, was shown to inhibit the recruitment of inflammatory cells into the airway, and in particular, macrophages, neutrophils, and lymphocytes were significantly reduced as the result of the parasite proteins. However, the total number of eosinophils was slightly reduced as the result of the administration of parasite proteins. Sensitization and OVA challenge was shown to accelerate the secretion of Th2 cytokines (IL-4 and IL-5) within the lung, but in the immunized groups, those levels were lower. The administration of Tl-TP and OVA challenge group also evidenced a significant reduction in IL-4 levels as compared to the OVA-challenged group. The concentrations of Th2 cytokines in the Tl-ES and OVA challenge group were more similar to those observed in the OVA-challenged group. The concentration of IL-10 and TGF-beta in the lung was decreased substantially in the OVA-only challenge group, but the Tl-TP immunized group exhibited significantly induced IL-10 cytokine. OVA-specific IgG2a, IgG1, and IgE levels in the immunized groups were significantly lower than those detected in the OVA-challenged group. In conclusion, parasite-derived protein is able to inhibit OVA-specific Th2 responses, and in particular, immunization with parasite proteins exerts a more profound protective effect than is seen with the treatment of allergic reactions. The results of our study are encouraging in terms of our further understanding of the molecular basis of immune evasion by nematodes.     

Disease screening profiles and colostrum management practices on 16 Irish suckler beef farms

Background

Calf output is a key element in determining the profitability of a suckler beef enterprise. Infectious agents such as Bovine Virus Diarrhoea (BVD) virus, colostrum management and parasitic challenge can all affect calf output. Prior to the national BVD eradication programme, there was little published information on either the prevalence or effect of BVD in Irish beef herds. There is little published information on colostrum management practices in Irish commercial beef herds and there have also been few studies published on the prevalence of liver fluke or rumen fluke infection in Irish beef herds. Sixteen farms participating in the Teagasc/Farmers Journal BETTER farm beef programme were used in this study. Fourteen herds were screened for the presence of BVD virus in 2010 using RT-PCR. In 13 herds, blood samples were collected from calves (2–14 days of age) in November 2011 - April 2012 to determine their passive immune status using the zinc sulphate turbidity (ZST) test, while in 12 herds, blood and faecal samples were taken in order to determine the level of exposure to gastrointestinal and hepatic helminths.

Results

The overall prevalence of BVD virus-positive cattle was 0.98% (range 0 - 3% per herd, range 0.6 - 3.0% per positive herd). Eighteen of the 82 calves (22%) sampled had ZST values less than 20 units (herd mean range 17.0 – 38.5 units) indicating a failure of passive transfer. The overall animal-level (herd-level) prevalence of liver fluke and rumen fluke infection in these herds was 40.5% (100%) and 20.8% (75%), respectively.

Conclusions

The potential costs associated with the presence of animals persistently infected with BVD virus through the increased use of antibiotics; the rate of failure of passive transfer of colostral immunoglobulins and the high prevalence of liver fluke infection in these herds highlight that some Irish suckler beef farms may not be realizing their economic potential due to a range of herd health issues. The use of farm-specific herd health plans should be further encouraged on Irish suckler beef farms.     

Temporal expression analyses of pancreatic and gastric digestive enzymes during early development of the olive flounder (Paralichthys olivaceus)

The olive flounder (Paralichthys olivaceus) is one of the most important aquaculture species in Korea. Previous studies focused on the morphological changes and only analysed digestive enzyme expression patterns of certain digestive systems. A comprehensive analysis of the pancreatic and gastric digestive enzymes of the olive flounder in its early developmental stages has not been performed. In this study, we determined the mRNA expression levels of pancreas and stomach digestive enzymes by extracting total RNA from the whole body of olive flounders from hatching to post‐metamorphosis and conducting real‐time PCR analysis with gene‐specific primers for each isoform. The pancreatic digestive enzymes were initially and strongly expressed from the first feeding period to pre‐metamorphosis. The expression of trypsinogen 3 was detected from the early stage of development to metamorphosis, whereas trypsinogen 2 was significantly increased only in the post‐metamorphosis period. The mRNA expression of pepsinogen was only detected at metamorphosis and corresponded to stomach differentiation. The lipid digestive‐related enzymes had already reached a certain level at the beginning of hatching, then increased during the early developmental stage and gradually decreased before metamorphosis. These results suggest that food ingestion is exclusively digested by pancreatic digestive enzymes and trypsinogen 3 during the early developmental stage until pre‐metamorphosis, and it then fully switches to digestion by pepsinogen and trypsinogen 2 in mid‐metamorphosis. In conclusion, understanding the expression profiles of digestive enzymes in the pancreas and stomach during the early developmental stages is an important consideration when developing formulated larval diets for aquacultured olive flounder.     

A comparison of one‐slope straight broken‐line,two‐slope straight broken‐line,quadratic broken‐line and quadratic models to estimate an accurate optimum feeding rate for juvenile olive flounder (Paralichthys olivaceus)

Two feeding trials using different commercial feeds and different fish sizes were conducted to compare model performance of various regression models on the estimation of optimum feeding rates (OFR; % BW/day) for juvenile olive flounder. Six feeding rates ranging from 1% to 11% in trial I and another six feeding rates ranging from 1% to 6% in trial II were tested. Final body weight, weight gain, feed conversion ratio, condition factor, nutrient gains, and whole‐body lipid and energy contents were significantly influenced by the feeding rates in both trials. The overall responses shown in trial I were generally comparable to those observed in trial II. In trial I, the OFR was estimated to be 7.3%, 5.7%, 9.8% and 10.2% by the one‐slope straight broken‐line, two‐slope straight broken‐line, quadratic broken‐line and quadratic models, respectively. In trial II, the OFR was estimated to be 4.8%, 4.6%, 8% and 8% by the one‐slope broken‐line, two‐slope broken‐line, quadratic broken‐line and quadratic models, respectively. Based on the model selection criteria (R²_adj and AICc), the quadratic broken‐line model was selected as the best‐fit model in both trials. This model that describes the dose–response relationship best among the tested models can be recommended when determining an OFR.     

Assessing whether dairy cow welfare is “better” in pasture-based than in confinement-based management systems

ABSTRACT

Consumers perceive pasture-based systems of milk production as natural and therefore better for cow welfare than confinement systems. However both systems are heterogeneous and continually evolving, varying from total confinement to total pasture with many hybrid intermediaries. To compare the welfare of dairy cows in these various systems, we use the three spheres framework, comprising biological functioning, natural behaviour and affective states. Considering biological functioning, pasture-based cows are less at risk of subclinical and clinical mastitis, claw lesions, lameness, metritis, early embryonic mortality, culling and mortality, but at more risk of internal parasitism, malnutrition and delayed onset of oestrous activity postpartum than confined cows. Regarding natural behaviours, pasture-based cows exhibit less agonistic behaviour, better lying behaviour, more normal oestrous behaviours and better synchronicity of behaviours than confined cows. They also have the opportunity to graze, which is one of the main features of the behavioural repertoire of dairy cows, but, they may also experience long periods away from pasture in larger herds, and severe climatic stresses which will become increasingly important as the climate changes. Our current ability to assess the affective state of dairy cows is poor. For example, hunger is an important subjective state that cannot be measured directly. The growing focus on ensuring that animals have lives worth living, means that dairy cows should garner some positive emotions from their lives, and it seems clear that pasture access is essential for this. Clearly measurement of affective state is an important challenge for future dairy cow welfare research. At the extremes of management systems, there can be major differences in animal welfare but in hybrid systems, dairy cows experience elements of both confinement and pasture which may ameliorate the negative effects of each on cow welfare. Ultimately, the optimal system gives cows an element of choice between both environments. Moreover management of the system, whether it is confinement or pastured-based, may be as important as the system of management in ensuring good dairy cow welfare and addressing societal concerns.

Abbreviations: BCS: Body condition score; TMR: Total mixed ration     

High-throughput detection of antioxidants in mulberry fruit using correlations between high-resolution mass and activity profiles of chromatographic fractions

Background

Plant extracts contain a huge variety of pharmacologically active substances. Conventionally, various chromatographic methods must be applied several times to purify functional compounds to measure their functional activity. However, conventional purification methods are time-consuming and expensive due to the laborious purification process. Recently, a high-throughput discovery method that replaces such time-consuming purification processes was introduced; this method uses 15 T ultra-high-resolution Fourier transform ion cyclotron resonance mass spectrometry (15 T FT-ICR MS) and a high-throughput screening method. This 15 T FT-ICR MS provides unparalleled resolution and sub-ppm accuracy in mass measurements, while simultaneously detecting multiple compounds without separation. The high-throughput, simultaneous multi-component discovery method known as Scaling of Correlations between Activity and Mass Profiles (SCAMP) was used to detect functional compounds in a plant extract. We validated the performance of SCAMP using 33 fractions from antioxidant-rich mulberry ethyl acetate extract and known standard antioxidants.

Results

The mulberry fruit was first separated into 33 fractions by LC and analyzed using high-resolution mass spectrometry. The antioxidative strength of the 33 fractions and standard antioxidants was measured. To validate the efficiency of this antioxidant discovery method, correlations between the antioxidation activity profile and changes in mass intensity of components within the 33 fractions were calculated to provide relative scores for the antioxidant candidate list. Enrichment curves and area under the curve (AUC) values were then calculated to compare the performance of the methods. Using this improved scoring method, five strong antioxidants, chlorogenic acid (14.2 ng), dihydoxy quercetin (46.2 ng), rutin (154.0 ng), quercetin (71.7 ng) and luteolin (3.5 ng) in 2 kg mulberry fruit, were found within the top 20 candidates.

Conclusions

We calculated AUCs in order to compare scoring methods quantitatively. Scoring systems were compared and calculated AUCs, where the AUCs for new scoring systems (0.98 and 0.99) were higher than the previously used correlation coefficient (AUC = 0.89). Using the new scoring algorithms, we successfully enriched thirteen unknown strong antioxidant candidates in addition to known antioxidants, methyl syringin and naringenin (3.5 ng) in mulberry extract. Targeted purification of these unknown candidates will significantly reduce purification time and labor.

     

The role of rpoS,hmp, and ssrAB in Salmonella enterica Gallinarum and evaluation of a triple-deletion mutant as a live vaccine candidate in Lohmann layer chickens

Salmonella enterica Gallinarum (SG) causes fowl typhoid (FT), a septicemic disease in avian species. We constructed deletion mutants lacking the stress sigma factor RpoS, the nitric oxide (NO)-detoxifying flavohemoglobin Hmp, and the SsrA/SsrB regulator to confirm the functions of these factors in SG. All gene products were fully functional in wild-type (WT) SG whereas mutants harboring single mutations or a combination of rpoS, hmp, and ssrAB mutations showed hypersusceptibility to H₂O₂, loss of NO metabolism, and absence of Salmonella pathogenicity island (SPI)-2 expression, respectively. A triple-deletion mutant, SGΔ3 (SGΔrpoSΔhmpΔssrAB), was evaluated for attenuated virulence and protection efficacy in two-week-old Lohmann layer chickens. The SGΔ3 mutant did not cause any mortality after inoculation with either 1 × 10⁶ or 1 × 10⁸ colony-forming units (CFUs) of bacteria. Significantly lower numbers of salmonellae were recovered from the liver and spleen of chickens inoculated with the SGΔ3 mutant compared to chickens inoculated with WT SG. Vaccination with the SGΔ3 mutant conferred complete protection against challenge with virulent SG on the chickens comparable to the group vaccinated with a conventional vaccine strain, SG9R. Overall, these results indicate that SGΔ3 could be a promising candidate for a live Salmonella vaccine against FT.     

Calf health from birth to weaning. I. General aspects of disease prevention

Calfhood diseases have a major impact on the economic viability of cattle operations. This is the first in a three part review series on calf health from birth to weaning, focusing on preventive measures. The review considers both pre- and periparturient management factors influencing calf health, colostrum management in beef and dairy calves and further nutrition and weaning in dairy calves.