Role for flagella but not intimin in the persistent infection of the gastrointestinal tissues of specific-pathogen-free chicks by Shiga toxin-negative Escherichia coli O157:H7

Shiga toxin (Stx)-positive Escherichia coli O157:117 readily colonize and persist in specific-pathogen-free (SPF) chicks, and we have shown that an Stx-negative E. coli O157:117 isolate (NCTC12900) readily colonizes SPF chicks for up to 169 days after oral inoculation at 1 day of age. However, the role of intimin in the persistent colonization of poultry remains unclear. Thus, to investigate the role of intimin and flagella, which is a known factor in the persistence of non-O157 E. coli in poultry, isogenic single- and double-intimin and aflagellar mutants were constructed in E. coli O157:117 isolate NCTC12900. These mutants were used to inoculate (10(5) CFU) 1-day-old SPF chicks. In general, significant attenuation of the aflagellate and intiminaflagellate mutants, but not the intimin mutant, was noted at similar time points between 22 and 92 days after inoculation. The intimin-deficient mutant was still being shed at the end of the experiment, which was 211 days after inoculation, 84 days more than the wild type. Shedding of the aflagellar and intimin-aflagellar mutants ceased 99 and 113 days after inoculation, respectively. Histological analysis of gastrointestinal tissues from inoculated birds gave no evidence for true microcolony formation by NCTC12900 or intimin and aflagellar mutants to epithelial cells. However, NCTC12900 mutant derivatives associated with the mucosa were observed as individual cells and/or as large aggregates. Association with luminal contents was also noted. These data suggest that O157 organisms do not require intimin for the persistent colonization of chickens, whereas flagella do play a role in this process.

Neonatal environment exerts a sustained influence on the development of the intestinal microbiota and metabolic phenotype

The postnatal environment, including factors such as weaning and acquisition of the gut microbiota, has been causally linked to the development of later immunological diseases such as allergy and autoimmunity, and has also been associated with a predisposition to metabolic disorders. We show that the very early-life environment influences the development of both the gut microbiota and host metabolic phenotype in a porcine model of human infants. Farmpiglets were nursed by their mothers for 1 day, before removal to highly controlled, individual isolators where they received formula milk until weaning at 21 days. The experiment was repeated, to create two batches, which differed only in minor environmental fluctuations during the first day. At day 1 after birth, metabolic profiling of serum by 1H nuclear magnetic resonance spectroscopy demonstrated significant, systemic, inter-batch variation which persisted until weaning. However, the urinary metabolic profiles demonstrated that significant inter-batch effects on 3-hydroxyisovalerate, trimethylamine-N-oxide and mannitol persisted beyond weaning to at least 35 days. Batch effects were linked to significant differences in the composition of colonic microbiota at 35 days, determined by 16 S pyrosequencing. Different weaning diets modulated both the microbiota and metabolic phenotype independently of the persistent batch effects. We demonstrate that the environment during the first day of life influences development of the microbiota and metabolic phenotype and thus should be taken into account when interrogating experimental outcomes. In addition, we suggest that intervention at this early time could provide ‘metabolic rescue’ for at-risk infants who have undergone aberrant patterns of initial intestinal colonisation.

Aspects of designing and evaluating seasonal-to-interannual Arctic sea-ice prediction systems

Using lessons from idealised predictability experiments, we discuss some issues and perspectives on the design of operational seasonal to inter-annual Arctic sea-ice prediction systems. We first review the opportunities to use a hierarchy of different types of experiment to learn about the predictability of Arctic climate. We also examine key issues for ensemble system design, such as: measuring skill, the role of ensemble size and generation of ensemble members. When assessing the potential skill of a set of prediction experiments, using more than one metric is essential as different choices can significantly alter conclusions about the presence or lack of skill. We find that increasing both the number of hindcasts and ensemble size is important for reliably assessing the correlation and expected error in forecasts. For other metrics, such as dispersion, increasing ensemble size is most important. Probabilistic measures of skill can also provide useful information about the reliability of forecasts. In addition, various methods for generating the different ensemble members are tested. The range of techniques can produce surprisingly different ensemble spread characteristics. The lessons learnt should help inform the design of future operational prediction systems.

Interacting controls on innate sources of CO2 efflux from a calcareous arid zone soil under experimental acidification and wetting

More than half of global soil carbon is stored as carbonates, primarily in arid and semi-arid zones. Climate change models predict more frequent and severe rainfall events in some parts of the globe, many of which are dominated by calcareous soils. Such events trigger substantial increases in soil CO2 efflux. We hypothesised that the primary source of CO2 emissions from calcareous, arid zone soil during a single wetting event is abiotic and that soil acidification and wetting have a positive, potentially interacting, effect. We manipulated soil pH, soil moisture, and controlled soil respiration by gamma irradiating half of an 11 day incubation experiment. All manipulated experimental treatments had a rapid and enormous effect on CO2 emission. Respiration contributed ca. 5% of total CO2 efflux; the major source (carbonate buffering) varied depending on the extent of acidification and wetting. Maximum CO2 efflux occurred when pH was lowest and at intermediate matric potential. CO2 efflux was lowest at native pH when soil was air dry. Our data suggest that there may be an underestimate of soil-atmosphere carbon fluxes in arid ecosystems with calcareous soils. There is also a clear potential that these soils may become net carbon sources depending on changes in rainfall patterns, rainfall acidity, and future land management. Our findings have major implications for carbon cycling in arid zone soil and further study of carbon dynamics in these terrestrial systems at a landscape level will be required if we are to improve global climate and carbon cycling models.

Profile of cortisol, glycaemia, and blood parameters of American Bullfrog tadpoles Lithobates catesbeianus exposed to density and hypoxia stressors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of timing of estradiol benzoate administration upon synchronization of ovulation in suckling Nelore cows (Bos indicus) treated with a progesterone-releasing intravaginal device

The present study investigated how the timing of the administration of estradiol benzoate (EB) impacted the synchronization of ovulation in fixed-time artificial insemination protocols of cattle. To accomplish this, two experiments were conducted, with EB injection occurring at different times: at withdrawal of the progesterone-releasing (N) intravaginal device or 24 h later. The effectiveness of these times was compared by examining ovarian follicular dynamics (Experiment 1, n = 30) and conception rates (Experiment 2, n = 504). In Experiment 1, follicular dynamics was performed in 30 Nelore cows (Bos indicus) allocated into two groups. on a random day of the estrous cycle (Day 0), both groups received 2 mg of EB i.m. and a P4-releasing intravaginal device, which was removed on Day 8, when 400 IU of eCG and 150 mu g of PGF were administered. The control group (G-EB9; n = 15) received 1 mg of EB on Day 9, while Group EB8 (G-EB8; n = 15) received the same dose a day earlier. Ovarian ultrasonographic evaluations were performed every 8 h after device removal until ovulation. The timing of EB administration (Day 8 compared with Day 9) did affect the interval between P4 device removal to ovulation (59.4 +/- 2.0 h compared with 69.3 +/- 1.7 h) and maximum diameter of dominant (1.54 +/- 0.06 a cm compared with 1.71 +/- 0.05 b cm, P = 0.03) and ovulatory (1.46 +/- 0.05 a cm compared with 1.58 +/- 0.04 b cm, P < 0.01) follicles. In Experiment 2,504 suckling cows received the same treatment described in Experiment 1, but insemination was performed as follows: Group EB8-AI48h (G-EB8-AI48h; n = 119) and Group EB8-AI54h (G-EB8-AI54h; n = 134) received 1 mg of EB on Day 8 and FrAI was performed, respectively, 48 or 54 h after P4 device removal. Group EB9-AI48h (G-EB9-AI48h; n = 126) and Group EB9-AI54h (G-EB9-AI54h n = 125) received the same treatments and underwent the same FTAI protocols as G-EB8-AI48h and G-EB8-AI54h, respectively; however, EB was administered on Day 9. Conception rates were greater (P < 0.05) in G-EB9-AI54h 163.2% (79/125) a], G-EB9-AI48h [58.7% (74/126) a] and G-EB8-AI48h [58.8% (70/119) a] than in G-EB8-AI54h [34.3% (46/134) b]. We concluded that when EB administration occurred at device withdrawal (D8), the interval to ovulation shortened and dominant and ovulatory follicle diameters decreased. Furthermore, when EB treatment was performed 24 h after device removal, FTAI conducted at either 48 or 54 h resulted in similar conception rates. However, EB treatment on the same day as device withdrawal resulted in a lesser conception rate when FTAI was conducted 54 h after device removal. (C) 2007 Elsevier B.V. All rights reserved.

Saccharomyces Cerevisiae cell wall dietary supplementation on the performance and intestinal mucosa development and integrity of broiler chickens vaccinated against coccidiosis

This study was carried out to verify if Saccharomyces cerevisiae cell wall (SCCW) dietary supplementation (0.2%) was capable of protecting the intestinal mucosa of broiler chickens vaccinated against coccidiosis. Body weight gain, feed intake, feed conversion and intestinal mucosa morphometric parameters and epithelial loss were evaluated. In the experiment,400 day-old male chicks were distributed according to a completely randomized design in a 2x2 factorial arrangement. The following treatments were applied: T1 - no vaccination/ no SCCW supplementation; T2 - no vaccination/SCCW supplementation; T3 - vaccination/no SCCW supplementation; and T4 - vaccination/SCCW supplementation to four replicates of 25 birds each. Birds were vaccinated on the first day of age using a spray vaccine (Coccivac B®, Coopers), containing E. acervulina, E. maxima, E. mivati and E. tenella. S. cerevisiae cell wall was supplied from the first day of age. Live performance, intestinal morphometric parameters and epithelial loss were evaluated at 14, 21 and 28 days of age. Performance was affected by vaccination only at 21-days of age, when body weight gain was reduced in the vaccinated birds, but no body weight difference was observed on day 28. Vaccine also increased the crypt depth (p<0.05) in the duodenum and jejunum, suggesting a high cell activity in the crypt:villus transition area to maintain the epithelial cell turnover. Villi number/area (103,269 µm²) was not affected (p>0.05) by vaccine or cell wall supplementation, and epithelial loss was more pronounced in the duodenum and jejunum. In conclusion, the findings of this study suggest that S. cerevisiae cell wall supplementation may be an useful management tool to maintain the intestinal integrity of broilers vaccinated against coccidiosis.

Effect of oils sources on blood lipid parameters of commercial laying hens

The experiment was carried out to verify if total cholesterol, HDL-cholesterol and triacylglicerol plasma levels are affected when laying hens are fed rations containing different dietary oil sources. One hundred sixty 50 week-old hens, assigned to four treatments with five replicates using 8 hens per replicate were used. The experimental period was of 84 days divided in 3 cycles of 28 days each. In the last day of each cycle, blood samples of 2 hens per replicate were randomly choose and blood samples were collected. on the other hand, blood was also collected at 7 am, 11 am and 3 pm aiming to study the daily changes of these lipids. Blood lipid parameters were not affected by different dietary oil sources (p > 0.05); however, HDL-cholesterol did change during the day, giving evidence that this lipid is indeed involved in the egg yolk formation.

Effect of environmental temperature on immune response of broilers

Several studies demonstrate that environmental temperature can influence the immune response of poultry. The objective of this research was to determine at which stage in the life of a bird this effect is greatest. In experiment 1, broiler breeder eggs were incubated at three different temperatures (36.8+/-0.2, 37.8+/-0.2, and 38.8+/-0.2degreesC from the 13th day of incubation to hatching. After hatching, birds were raised in thermoneutral temperature. In experiment 2, 144 1-d-old broiler chicks were distributed into three environmental chambers with different temperatures (18+/-2, 24+/-2, and 32+/-2degreesC). In both experiments, the humoral immune responses to Newcastle disease virus (NDV) and infectious bursal disease (IBDV) were evaluated. NDV and IBDV antibody titers were not significantly different (P > 0.05) among treatments.

Control of Salmonella enterica serovar Enteritidis in laying hens by inactivated Salmonella Enteritidis vaccines

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Performance of supplemented heifers on Marandu grass pastures in the wet-to-dry transition and dry seasons

The objective of the present study was to evaluate the characteristics of the sward canopy of Marandu grass during the rainy season, the wet-to-dry transition and the dry seasons, between March and September 2004, under intermittent grazing, and to correlate those characteristics with the performance of crossbred heifers receiving mineral supplements ad libitum or protein supplements. The experiment consisted of a randomized block design with three blocks (set of 13 paddocks), each containing five crossbred heifers per experimental unit, totaling 15 replicates. The heifers were given protein supplements daily in individual stalls and received an average 4 g/kg/day of the supplement during the rainy season and 5 g/kg/day during the dry season. Their weight gain was assessed monthly. The pasture structure was assessed through destructive sampling, and the bromatological composition of esophageal extrusa samples was also assessed. Analysis of variance was used to assess performance, and regression analysis was used to evaluate the sward canopy characteristics in relation to the months of the year. A cluster procedure was used to determine the similarity between the months of the year under assessment. Two different groups were formed for pasture evaluation: one group including the months of March to July and another group including the months of August and September. The first group exhibited a better canopy structure than the second group. This fact was corroborated by the animal performance, which was lower during the months of the second group. Low-intake protein supplementation was effective in increasing the performance of the grazing heifers. Pasture structure is critical for animal performance in a grazing environment, regardless of the type of supplementation.

Effects of beta-aminopropionitrile fumarate and exercise on equine tendon healing: Gross and histological aspects

In the present study, the effects of intralesional injections of beta-aminopropionitrile fumarate (BAPN-F) was assessed in equine experimental tendinitis. BAPN-F is a lathyrogen which inhibits crosslinking of collagen, permitting more time for parallel reorientation of the repair tissue. Sixteen two-year-old Arabian horses without health problems were used in this experiment. The animals were divided into two groups: group one was left in box rest, and group two was submitted to controlled exercise during the experiment. Tendinitis was induced with collagenase in the superficial flexor tendon of both forelimbs. Twenty days after the induction of tendinitis, intralesional treatment with BAPN-F was performed and the contralateral limbs received saline. A biopsy was obtained and gross and histopathological analysis was performed on the 150th day of the experiment. The collagen fibrillar alignment pattern in the heating area was better in the BAPN-F group submitted to controlled exercise than in the other group, as observed by sonographic and histopathologic examination. The present results indicate that BAPN-F in combination with controlled loading improved sear remodeling and tendon wound collagen maturation.

Strategic supplementation of calcium salts of polyunsaturated fatty acids to enhance reproductive performance of Bos indicus beef cows

Five experiments evaluated the effects of supplemental Ca salts of PUFA on reproductive function of Bos indicus beef cows. In Exp. 1, nonlactating and multiparous grazing cows (n = 51) were assigned to receive (as-fed basis) 0.1 kg of a protein-mineral mix + 0.1 kg of ground corn per cow/d, in addition to 0.1 kg per cow/d of 1) Ca salts of PUFA (PF), 2) Ca salts of SFA (SF), or 3) kaolin (control). Treatments were offered from d 0 to 20 of the estrous cycle. No treatment effects were detected on serum progesterone concentrations (P = 0.83), day of luteolysis (P = 0.86), or incidence of short cycles (P = 0.84). In Exp. 2, nonlactating and multiparous grazing cows (n = 43) were assigned to receive PF, SF, or control from d 0 to 8 of the estrous cycle. on d 6, all cows received (intramuscularly) 25 mg of PGF(2 alpha). No treatment effects were detected on serum progesterone concentrations on d 6 (P = 0.37), and incidence (P = 0.67) or estimated time of luteolysis (P = 0.44). In Exp. 3, twenty-seven lactating and multiparous grazing cows, approximately 30 to 40 d postpartum, were assigned to receive PF or control for 10 d beginning at the first postpartum ovulation. No treatment effects were detected (P = 0.85) on incidence of short cycles. In Exp. 4, lactating and multiparous grazing cows (n = 1,454), approximately 40 to 60 d postpartum, were assigned to receive 1 of the 7 treatments for 28 d after timed AI (TAI; d 0): 1) control from d 0 to 28, 2) SF from d 0 to 14 and then control, 3) PF from d 0 to 14 and then control, 4) SF from d 0 to 21 and then control, 5) PF from d 0 to 21 and then control, 6) SF from d 0 to 28, and 7) PF from d 0 to 28. Cows receiving PF for more than 21 d after TAI had greater (P < 0.01) pregnancy to TAI compared with all other treatments combined (50.4 vs. 42.4%, respectively). In Exp. 5, lactating and multiparous grazing cows (n = 501), approximately 40 to 60 d postpartum, were assigned to receive 1 of the 4 treatments for 21 d after TAI (d 0): 1) PF from d 0 to 14 and then control, 2) control from d 0 to 6 and then PF, 3) control from d 0 to 13 and then PF, and 4) PF from d 0 to 21. Cows receiving PF after d 14 of the experiment had greater (P = 0.02) pregnancy to TAI compared with cows not receiving PF during the same period (46.8 vs. 33.1%, respectively). In summary, supplemental Ca salts of PUFA during the expected time of luteolysis increased pregnancy to TAI in beef cows.

Concentrations of Progesterone and Insulin in Serum of Nonlactating Dairy Cows in Response to Carbohydrate Source and processing

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of calcium salts of polyunsaturated fatty acids on productive and reproductive parameters of lactating Holstein cows

Two experiments evaluated milk production, serum progesterone and insulin, and reproductive performance of lactating Holstein cows receiving or not receiving Ca salts of polyunsaturated fatty acids (PUFA), or receiving Ca salts of PUFA at different daily frequencies. In experiment 1, 1,125 cows randomly distributed in 10 freestall barns were enrolled. Barns were assigned randomly to receive a high-concentrate diet containing (PI?) or not containing (control, CON) 1.1% (dry matter basis) Ca salts of PUPA. Diets were offered 6 times daily, whereas the Ca salts of PUFA were included in the PF treatment in the first feeding of the day. In experiment 2, 1,572 cows were randomly distributed in 10 freestall barns, which were assigned randomly to receive a diet similar to PF, but with Ca salts of PUFA included only in the first feeding of the day (PF1X), or equally distributed across all 6 feedings (PF6X). During both experiments, cows were artificially inseminated 12 h after the onset of estrus. Once per month, cows that did not conceive to artificial insemination were assigned to a fixed-time embryo transfer protocol. Pregnancy was determined via transrectal ultrasonography 28 and 60 d after expected ovulation. Pregnancy loss was considered in cows that were pregnant on d 28 but nonpregnant on d 60. During both experiments, feed intake, milk yield, and milk protein and fat content were recorded weekly. Blood samples were collected concurrently with embryo transfer. During experiment 1, feed intake was similar between treatments. Compared with CON, PF cows had greater milk yield (37.8 vs. 35.3 kg/d), and reduced milk fat content (3.41 vs. 3.55%). However, PF cows had reduced pregnancy losses per service compared with CON (12.6 vs. 18.3%). Serum progesterone was greater and serum insulin tended to be greater in primiparous cows receiving PF compared with CON cohorts (4.50 vs. 3.67 ng of progesterone/mL, and 10.4 vs. 7.5 mu UI of insulin/mL). During experiment 2, no treatment effects were detected for feed intake, milk yield, or milk fat, whereas PF1X cows tended to have reduced pregnancy losses per service compared with PF6X (14.4 vs. 18.4%). In summary, feeding Ca salts of PUFA to dairy cows increased milk production, did not alter feed intake, and reduced pregnancy losses per service. Further, the total daily amount of Ca salts of RITA should be fed during the first feeding of the day to optimize its benefits on pregnancy maintenance of dairy cows.