Fluorogenic assay for rapid detection of Escherichia coli in chilled and frozen foods: collaborative study

A collaborative study was conducted to compare a proposed LST-MUG method with the AOAC official method for Escherichia coli detection. E. coli produces an enzyme, beta-glucuronidase, which cleaves the substrate, 4-methyl-umbelliferyl-beta-D-glucuronide (MUG), to yield a fluorescent end product. Incorporation of the MUG substrate into lauryl tryptose broth (LST) enables a rapid quantitative method for screening E. coli, which is detected by fluorescence of the medium under longwave UV light. In this collaborative study, 5 food samples, 2 frozen (entree sauce/gravy and dairy topping) and 3 chilled (hamburger, pork sausage, and cheese), were tested for E. coli detection by 17 collaborating laboratories. Results indicate that the LST-MUG method is equal to or better than the current AOAC method for detecting E. coli. The LST-MUG method has been adopted official first action.     

Comparative growth performance of two Nile tilapia (Chitralada and Red-Stirling), their crosses and the Israeli tetra hybrid ND-56

Growth performance of two Oreochromis niloticus strains, Chitralada and Red‐Stirling, their reciprocal crossbred and the Israeli tetra‐hybrid ND‐56 were assessed in net cages under on‐farming conditions. Throughout 268 days of grow‐out, the strains were weighed monthly and mortality, feed consumption and water quality were recorded. Ten rigid net cages (1.5 × 1.5 × 1.7 m) immersed in a 10 ha reservoir were linearly arranged near the reservoir outlet following a completely randomized design with two replicates for each treatment (strain). Each cage was stocked with 459 fish (120 fish m^?3) and fed twice daily to apparent satiation with a commercial tilapia diet following the recommended feeding program. The final mean weights were higher for Chitralada (557.20 g) and the reciprocal crossbreds (522.95, 496.40 g) than those of Red‐Stirling (421.90 g). All treatments outperformed the ND‐56 tetra hybrid. Daily growth showed statistical differences between Chitralada (2.04 g) and Red‐Stirling (1.52 g) but they were statistically the same when compared with the reciprocal crossbreds (1.90, 1.80 g). The relative growth ratios showed the same trend observed in the results for daily growth. The mean survival rate was 98%. The overall growth rate showed that crossbred performed as well as the parental lines. All crossbred progeny presented red colouration with variable pattern of black marks corroborating the dominant inheritance of the red trait in Red‐Stirling strain.     

Effect of Water Exchange and Mechanical Aeration on Grow‐out of the Amazon River Prawn in Ponds

Exchange and aeration of pond water are common practices in semi‐intensive freshwater prawn culture, but there is lack of scientifically based information on the results. We evaluated the effects of water flow through the ponds and mechanical aeration in semi‐intensive cultures of Macrobrachium amazonicum. A total of 40 juveniles/m² were stocked for 4 mo in 12 earthen ponds. Four randomly assigned treatments were applied: no aeration + no water exchange (NN), diurnal aeration + no water exchange (DA), nocturnal aeration + no water exchange (NA), and continuous water flow (CF). Temperature, pH, total suspended solids, and soluble orthophosphate in the water column did not differ among treatments. Dissolved oxygen (DO) concentrations in ponds with nocturnal aeration were significantly higher than in other treatments. The concentration of inorganic nitrogen was significantly higher in the CF treatment, whereas organic nitrogen was higher in treatments NN, DA, and NA. This suggests that primary production is higher in static ponds. Thermal stratification started at 0900 h, and the maximum difference between surface and bottom temperature varied from 1.5 (CF) to 2.8 C (NN). The difference between DO levels in the surface and bottom water of the ponds began to appear at 0800 h and varied from 0.50 (CF) to 5.23 mg/L (NN). Diurnal aeration and high continuous water flow were efficient in disrupting the stratification. No significant difference was found for survival, mean weight, apparent feed conversion rate, and productivity among treatments. Thus, disrupting water stratification, aerating ponds at night, or exchanging the pond water are ineffective in Amazon River prawn farming in semi‐intensive systems, at least for stocking densities lower than 40 individuals/m². About 1000 kg of Amazon River prawn can be produced in static ponds with no aeration in approximately 4 mo. This management strategy saves water and energy and reduces production costs .     

Subpopulations of equine blood lymphocytes expressing regulatory T cell markers

Several distinct T lymphocyte subpopulations with immunoregulatory activity have been described in a number of mammalian species. This study performed a phenotypic analysis of cells expressing regulatory T cell (Treg) markers in the peripheral blood of a cohort of 18 horses aged 6 months to 23 years, using antibodies to both intracellular and cell surface markers, including Forkhead box P3 (FOXP3), CD4, CD8, CD25, interferon gamma (IFNγ) and interleukin 10 (IL-10). In peripheral blood, a mean of 2.2 ± 0.2% CD4+ and 0.5 ± 0.1% CD8+ lymphocytes expressed FOXP3. The mean percentage of CD4+FOXP3+ cells was found to be significantly decreased in horses 15 years and older (1.5%) as compared to horses 6 years and younger (2.7%), but did not differ between females and males and ponies and horses. Activation of peripheral blood mononuclear cells by pokeweed mitogen resulted in induction of CD25 and FOXP3 expression by CD4+ cells, with peak expression noted after 48 and 72 h in culture respectively. Activated CD4+FOXP3+ cells expressed IFNγ (35% of FOXP3+ cells) or IL-10 (9% FOXP3+ cells). Cell sorting was performed to determine FOXP3 expression by CD4(+)CD25(-), CD4(+)CD25(dim) and CD4(+)CD25(high) subpopulations. Immediately following sorting, the percentage of CD4+FOXP3+ cells was higher within the CD4(+)CD25(high) population (22.7-26.3%) compared with the CD4(+)CD25(dim) (17% cells) but was similar within the CD4(+)CD25(dim) and CD4(+)CD25(high) cells after resting in IL-2 (9-14%). Fewer than 2% of cells in the CD4(+)CD25(-) population expressed FOXP3. These results demonstrate heterogeneity in equine lymphocyte subsets that express molecules associated with regulatory T cells. CD4+FOXP3+ cells are likely to represent natural Tregs, with CD4+FOXP3+IL-10+ cells representing either activated natural Tregs or inducible Tregs, and CD4+FOXP3+IFNγ+ cells likely to represent activated Th1 cells.     

Sequential medium with GH and IGF‐1 improved in vitro development of bovine preantral follicles enclosed in ovarian tissue

The growth hormone (GH) and growth insulin‐like factor‐1 (IGF‐1) act directly upon the regulation and growth in the different phases of preantral follicles. Thus, it is necessary to define their sequentiality until the in vitro preovulatory development. Therefore, the study aimed to assess the effects of a sequential medium containing GH and/or IGF‐1 in the long‐duration in vitro culture of preantral ovarian follicles. Ovarian fragments were cultivated: first half (days 1–7), second half (days 7–14) or during 14 culture days. Treatments were identified as: αMEM+; GH → IGF‐1; IGF‐1 → GH and GH + IGF‐1. The culture was designed in 24‐well plates, in an incubator at 37°C and 5% CO₂. The parameters of normality, viability, follicles (primordial/in developing) and follicle diameter were evaluated. In addition, the ultrastructure was confirmed with electron transmission microscopy. The results showed that the culture treated with GH → IGF‐1 kept the follicular normality and the viability until the 14th day of culture and increased both in the follicular development until 7th day and in the follicular diameter until 14th day, when compared to the control. The treatments IGF‐1 → GH and GH + IGF‐1 were not effective in the developing and follicular diameter after 7 days of culture, and also reduced the percentage of viability. It is concluded that the bovine preantral follicles cultured in the sequential medium treated with GH → IGF‐1 improved the follicular development until the first half of the culture and kept these parameters with normality, viability and ultrastructure until the second half of the in vitro culture.     

Effect of sorghum grain variety and reconstitution on site and extent of starch and protein digestion in steers

Hetero-yellow (HY), red (RED) and brown (BR, high tannin) sorghums were fed dry-rolled or reconstituted (RED and BR only) to evaluate the effect of variety and reconstitution on the site and extent of starch and protein digestion in steers fitted with ruminal, duodenal and ileal cannulae. Processed grains were incorporated into 88% sorghum (DM basis) diets fed at 2% of body weight in a 5 X 5 Latin square. Ruminal fermentation of organic matter, starch and protein tended to be lower for the dry-rolled RED than for either the dry-rolled HY or BR sorghum. Digestion of organic matter (OM) and starch in the small intestine was very low for dry-rolled sorghums. Total tract starch digestibility was lower for the dry-rolled RED sorghum (86.9%) than the BR (90.8%) and HY (91.4%). Nitrogen (N) digestibility ranged from 53.1% for the dry-rolled BR to 64.5% for the HY. Tannins were extensively (95.2%) degraded in the rumen, which may have enhanced fermentation of the BR sorghum. Reconstitution increased (P less than .05) total-tract starch digestion of the RED and tended to increase starch digestion of the BR as well. Total N flow to the duodenum tended to increase with reconstitution, with most of the increase being due to greater (P less than .05) microbial-N. Reconstitution also increased (P less than .05) total-tract N digestibility of the RED. The response to reconstitution for the RED sorghum appeared to be due primarily to an increase (P less than .10) in the extent of fermentation of organic matter and starch in the rumen. Reconstitution of BR, however, enhanced disappearance of starch from the small intestine. In both cases, most (97.3%) of the digestible starch of the reconstituted sorghums had disappeared before the terminal ileum. In contrast, 14.5% (621 g) of the digestible starch of dry-rolled RED disappeared in the large intestine. Sorghum grain variety and reconstitution appear to alter site and extent of starch and protein digestion, which may result in variable performance of cattle fed sorghum grain diets.     

Modelling forest recolonization by an epiphytic lichen using a landscape genetic approach

The process of recolonization after disturbance is crucial for the persistence and dynamics of patch-tracking metapopulations. We developed a model to compare the spatial distribution and spatial genetic structure of the epiphytic lichen Lobaria pulmonaria within the perimeter of two reconstructed 19th century disturbances with a nearby reference area without stand-level disturbance. Population genetic data suggested that after stand-replacing disturbance, each plot was colonized by one or a few genotypes only, which subsequently spread clonally within a local neighborhood. The model (cellular automaton) aimed at testing the validity of this interpretation and at assessing the relative importance of local dispersal of clonal propagules vs. long-distance dispersal of clonal and/or sexual diaspores. A reasonable model fit was reached for the empirical data on host tree distribution, lichen distribution, and tree- and plot-level genotype diversity of the lichen in the reference area. Although model calibration suggested a predominance of local dispersal of clonal propagules, a substantial contribution of immigration of non-local genotypes by long-distance dispersal was needed to reach the observed levels of genotype diversity. The model could not fully explain the high degree of clonality after stand-replacing disturbance, suggesting that the dispersal process itself may not be stationary but depend on the ecological conditions related to disturbance.     

Differential association of MUC5AC and CLCA1 expression in small cartilaginous airways of RAO‐affected and control horses

Reasons for performing study: Airway mucus accumulation is associated with indoor irritant and allergen exposure in horses with recurrent airway obstruction (RAO). Epidermal growth factor receptor (EGFR) and a chloride channel (calcium activated, family member 1; CLCA1) are key signalling molecules involved in mucin gene expression. Objectives: We hypothesised that exposure to irritants and aeroallergens would lead to increased expression of the mucin gene eqMUC5AC and increased stored mucosubstance in the airways of RAO‐affected horses, associated with increased neutrophils and CLCA1 and EGFR mRNA levels. Methods: We performed quantitative RT‐PCR of eqMUC5AC, CLCA1 and EGFR; volume density measurements of intraepithelial mucosubstances; and cytological differentiation of intraluminal inflammatory cells in small cartilaginous airways from cranial left and right and caudal left and right lung lobes of 5 clinically healthy and 5 RAO‐affected horses that had been exposed to indoor stable environment for 5 days before euthanasia. Results: Neutrophils were increased in RAO‐affected horses compared to clinically healthy controls. EqMUC5AC mRNA levels were positively correlated with both CLCA1 and EGFR mRNA levels in RAO‐affected horses but only with CLCA1 in controls. The relationship between eqMUC5AC and CLCA1 differed in the 2 groups of horses with RAO‐affected animals overexpressing CLCA1 in relation to eqMUC5AC. Conclusions: These data implicate CLCA1 as a signalling molecule in the expression of eqMUC5AC in horses but also suggest differential regulation by CLCA1 and EGFR between horses with RAO and those with milder degrees of airway inflammation.     

Technical and economic feasibility of integrating seahorse culture in shrimp/oyster farms

Integrated multi‐trophic aquaculture systems (IMTA) are building ecosystems designed to produce aquatic organisms with less environmental impact. We have developed a simple system and a management strategy for introducing seahorse (Hippocampus spp.) culture in shrimp/oyster farms to increase economic sustainability. The system includes broodstock, nursery and grow‐out phases of Hippocampus reidi G. culture. We performed a test in a 42.4‐ha marine shrimp/oyster farm, which showed that this system is technically feasible. Then, we performed a partial budget analysis including cost‐return, cash flow and financial feasibility analysis. More than 12 000 marketable size seahorses may be produced in 402 m² annually. Initial investment was only US$ 21 103.00. The benefit–cost ratio was ~US$ 20.00 for each US$ 1.00 spent, internal rate of return was 131% and payback period <2 years. In conclusion, the introduction of seahorse in shrimp/oyster IMTA system is technically feasible, profitable, resilient and shows high liquidity. This system may be easily established in developing countries and has potential to provide seahorses to supply different markets (live specimens for the marine aquarium trade and dried specimens for traditional Chinese medicine and the curio trade). In addition, it can indirectly contribute to preserve natural populations.     

Cardiovascular effects of xylazine and detomidine in horses

The cardiovascular effects of xylazine and detomidine in horses were studied. Six horses were given each of the following 5 treatments, at 1-week intervals: xylazine, 1.1 mg/kg, IV; xylazine, 2.2 mg/kg, IM; detomidine, 0.01 mg/kg, IV; detomidine, 0.02 mg/kg, IV; and detomidine, 0.04 mg/kg, IM. All treatments resulted in significantly decreased heart rate, increased incidence of atrioventricular block, and decreased cardiac output and cardiac index; cardiac output and cardiac index were lowest following IV administration of 0.02 mg of detomidine/kg. Mean arterial pressure was significantly reduced for various periods with all treatments; however, IV administration of 0.02 mg of detomidine/kg caused hypertension initially. Systemic vascular resistance was increased by all treatments. Indices of ventricular contractility and relaxation, +dP/dt and -dP/dt, were significantly depressed by all treatments. Significant changes were not detected in stroke volume or ejection fraction. The PCV was significantly reduced by all treatments. Respiratory rate was significantly decreased with all treatments, but arterial carbon dioxide tension did not change. Arterial oxygen tension was significantly decreased briefly with the 3 IV treatments only.