Black soybean promotes the formation of active components with antihepatoma activity in the fermentation product of Agaricus blazei

The antihepatoma activity and related active components in the fermentation products of Agaricus blazei (AB) cultured in the medium containing soybean (S) or black soybean (BS) were investigated. AB(BS)-pE and AB(S)-pE were the ethanolic extracts from the fermentation products of AB(BS) and AB(S), respectively. According to the IC 50 values, AB(BS)-pE (161.1 and 24.0 microg/mL for Hep 3B and Hep G2 cells, respectively) exhibited stronger cytotoxicities against hepatoma cells than AB(S)-pE (>200 and 99.9 microg/mL for Hep 3B and Hep G2 cells, respectively). AB(BS)-pE was separated by silica gel column chromatography and eluted with n-hexane/ethyl acetate/methanol gradient solvent system into 21 fractions. Fraction 3 [AB(BS)-pE-F3], eluted with n-hexane/ethyl acetate (97:3 and 19:1, v/v), was the most active fraction having inhibitory activity on the proliferation of Hep 3B and Hep G2 cells (IC 50 of 3.6 and 1.9 microg/mL, respectively). Three major compounds, compounds 1- 3, were further isolated from the AB(BS)-pE-F3 fraction by reversed-phase semipreparative high-performance liquid chromatography. Compounds 2 and 3 gave better antihepatoma activity than that of compound 1. The IC 50 values of compounds 2 and 3 were 2.8 and 4.5 microg/mL for Hep 3B cells and 1.4 and 2.0 microg/mL for Hep G2 cells, respectively. The structures of compounds 2 and 3 were identified by UV, IR, electron impact mass spectrometry, and (1)H and (13)C NMR to be blazeispirols A and C, respectively. Blazeispirols A and C existed in the mycelia but not in the broth and were more in AB(BS)-pE (49.9 +/- 8.9 and 14.2 +/- 2.4 mg/g, respectively) than AB(S)-pE (15.9 +/- 1.7 and 3.9 +/- 0.6 mg/g, respectively). Additionally, the result shows that the production of blazeispirols A and C was increased after cultivation in the medium containing black soybean on day 6 and reached the maximum on day 12, and the contents of blazeispirols A and C were negatively correlated with Hep 3B and Hep G2 cell viabilities ( r = -0.84 to -0.93, P < 0.01). It suggests that blazeispirols A and C could be used as biomarkers to produce the fermentation product of A. blazei with antihepatoma activity.     

Absence of three known benzimidazole resistance mutations in Trichostrongylus tenuis, a nematode parasite of avian hosts

Benzimidazole (BZ) resistance is widespread in nematode parasites of livestock, but very little is known about the levels of BZ resistance in parasites with avian hosts. We investigated BZ resistance in Trichostrongylus tenuis, a nematode parasite of red grouse, Lagopus lagopus scotica. BZ anthelmintics had been in use in this system for up to 15 years, yet existing phenotypic evidence for resistance was inconclusive. We screened 1530 individuals from 14 populations at the principal β-tubulin locus for BZ resistance (isotype 1, residue 200) and 940 of these at two further resistance sites (isotype 1, residue 167; isotype 2, residue 200). No BZ resistant genotypes were found. Alternative mechanisms may be responsible for BZ resistance in this system, or the method and timing of treatments may reduce selection pressure for BZ resistance by creating substantial refugia for susceptible genotypes.     

Prevalence of Giardia and Cryptosporidium in dairy calves in three areas of Norway

A study was undertaken to determine the prevalences of Cryptosporidium and Giardia in dairy calves less than 6 months of age in Norway. Faecal samples were collected from a total of 1386 calves, between 3 and 183 days of age, in 136 dairy farms from three different areas of Norway. Faecal samples were examined for Cryptosporidium oocysts and Giardia cysts after concentration and immunofluorescent staining.

Giardia was found in 93% (127 out of 136) of the farms and in 49% (679 out of 1386) of the calves. Cryptosporidium was found in 53% (72 out of 136) of the farms and in 12% (167 out of 1386) of the calves. The level of Giardia and/or Cryptosporidium was low in the majority of the infected calves.

Infection peaked in the age group 2–3 months for both Cryptosporidium and Giardia. The prevalences of both parasites were higher in samples taken during winter than in samples taken during summer, and statistically significant differences were found when prevalences in different age groups of calves were compared between the three areas. A significantly lower prevalence of Cryptosporidium was found in calves housed in shared pens that were thoroughly washed more than three times a year than in calves from pens washed less often. For Giardia there was a trend for decreasing intensity of infection with increasing age in the sampled calves. For Cryptosporidium there was a trend for increasing herd prevalence with increasing number of calves in the herd, but this trend was not statistically significant. Other parameters which were investigated such as housing, feeding or management routines were not associated with prevalence or intensity of infection with either parasite.     

A novel phenotype for Laron dwarfism in miniature Bos indicus cattle suggests that the expression of growth hormone receptor 1A in liver is required for normal growth

Mutations within the growth hormone receptor (GHR) gene that lead to an inactivated or truncated GHR protein cause abnormal growth and small adult size in a variety of species (Laron dwarfism). We studied a line of miniature Bos indicus cattle that have phenotypic (small mature size) and endocrine (increased blood growth hormone and decreased blood insulin-like growth factor-I concentrations) similarities to Laron dwarfs. Liver mRNA from miniature and control cattle was used to amplify a cDNA within the coding region of the GHR. The miniature cattle had GHR mRNA size (determined by Northern blot) and cDNA sequence that were similar to control cattle and, therefore, were unlike most Laron dwarf genotypes in which the GHR gene is mutated. Amounts of mRNA from liver as well as muscle (superficial neck and longissimus) were analyzed by ribonuclease protection assay for IGF-I, total GHR, GHR 1A (inducible, liver-specific GHR mRNA), and GHR 1B (constitutive GHR mRNA). Four control and five miniature bulls were tested. As expected, liver IGF-I mRNA was decreased in the miniature cattle (approximately 12% of control; P < 0.01). The amount of the total GHR as well as GHR 1A mRNA were also decreased in liver (17% and 19% of control, respectively; P < 0.01). Other GHR mRNA, including GHR 1B mRNA, were similar for miniature and control cattle. In muscle, there was a tendency (P < 0.10) for decreased IGF-I mRNA and increased GHR mRNA in miniature compared with control cattle. In summary, a novel phenotype for Laron dwarfism in Bos indicus cattle was associated with underexpression of GHR 1A mRNA, but not other GHR mRNA variants in liver. In addition to decreased GHR 1A mRNA, the miniature cattle had decreased liver IGF-I mRNA. Full expression of GHR 1A in liver, therefore, may be required for full liver IGF-I expression and normal growth.     

Development of an estrus synchronization protocol for beef cattle with short-term feeding of melengestrol acetate: 7-11 synch

An estrus synchronization protocol (7-11 Synch) was developed to synchronize the first follicular wave and timing of ovulation in postpartum beef cows. In Exp. 1, follicular development and timing of ovulation in response to the following protocol were evaluated. Beef heifers (n = 12) and cows (n = 6), at random stages of the estrous cycle, were fed melengestrol acetate (MGA; .5 mg x animal(-1) x d(-1)) for 7 d and injected with PGF2alpha (PG; 25 mg) on the last day of MGA. A second injection of PG was administered 11 d after cessation of MGA. After the second injection of PG, estrus was synchronized in 6/12 heifers and 3/6 cows. The interval to estrus in heifers and cows was 54 and 64 h, respectively (P > .10). All animals exhibiting estrus ovulated first-wave follicles. Animals that failed to respond to the second injection of PG were in estrus later than 6 d after cessation of MGA and had corpora lutea that were unresponsive to the injection of PG. Based on the variation in interval to estrus following the first PG injection on the last day of MGA feeding in Exp. 1, an injection of GnRH (100 microg) was added to the protocol 4 d after the cessation of MGA to ensure ovulation or luteinization of dominant follicles and synchronization of first-wave follicular development. This revised protocol was termed "7-11 Synch." In Exp. 2, two estrus synchronization protocols were compared. Multiparous beef cows were stratified by breed and postpartum interval and randomly assigned to the 7-11 Synch (n = 44) or Select Synch protocols (GnRH injection followed by PG injection 7 d later; n = 45). Timing of estrus after the last PG injection (0 h) ranged from 42 to 102 h in the 7-11 Synch group and -30 to 114 h in the Select Synch group. Eight cows (18%) in the Select Synch group exhibited estrus 30 h before to 18 h after PG. Synchronized estrus peaked between 42 and 66 h after the last PG injection, and a maximum number of cows were in estrus at 54 h for both treatment groups. Synchrony of estrus from 42 to 66 h was greater (P < .05) in 7-11 Synch (91%: 41/44) than in Select Synch cows (69%: 31/45). Artificial insemination pregnancy rate from 42 to 66 h was greater (P < .05) in the 7-11 Synch group (66%: 29/44) than in the Select Synch group (40%: 18/45). In summary, the 7-11 Synch protocol improved synchrony of estrus without reducing fertility. This protocol has potential future application for fixed-time AI in beef cattle production systems.     

Prognosis for Neonatal Foals in an Intensive Care Unit

This study was conducted to develop an equation for the prediction of outcome in neonatal foals undergoing treatment in an intensive care unit (ICU). Fifty-three physical examination, historical, and clinicopathologic variables were analyzed from the records of 99 neonatal foals (<14 days of age) treated in the neonatal ICU of the Equine Medical Center. The outcome was recorded and the results were categorized into either surviving or nonsurviving groups. The mean values for the 2 groups were compared, and variables that differed significantly between the two groups were retained and used to construct a logistic regression equation. Retained variables were heart rate, temperature, and neutron phil count. The predictive equation then was tested prospectively in 2 additional groups of foals from 2 separate ICUs. The predicted outcome was compared to the actual outcome, and performance variables were calculated. Sensitivity (.83), specificity (.87), negative predictive value (.72), and positive predictive value (.93) were determined for foals from one neonatal ICU; the sensitivity (.83), specificity (.44), negative predictive value (.44), and positive predictive value (.83) were lower for foals at a second, separate ICU.     

Do early infant feeding practices vary by maternal ethnic group?

OBJECTIVE: To examine UK country and ethnic variations in infant feeding practices. DESIGN: Cohort study. SETTING: Infants enrolled in the Millennium Cohort Study, born between September 2000 and January 2002. SUBJECTS: A total of 18 150 natural mothers (11 286 (8207 white) living in England) of singleton infants. OUTCOME MEASURES: Breast-feeding initiation, breast-feeding discontinuation and introduction of solid foods before 4 months. EXPLANATORY VARIABLES: Maternal ethnic group, education and social class. RESULTS: Seventy per cent of UK mothers started to breast-feed, of whom 62% stopped before 4 months. Median age at discontinuing breast-feeding was 14, 13, 10 and 6 weeks in Scotland, England, Wales and Northern Ireland, respectively. Thirty-six per cent of UK mothers (34% in England) introduced solids before 4 months. White mothers were more likely to discontinue breast-feeding (62%) and introduce solids early (37%) than most other ethnic minority groups; those stopping before 4 months were more likely to introduce solids early compared with those continuing to breast-feed beyond this age (adjusted rate ratio (95% confidence interval): 1.3 (1.1-1.2)). Educated mothers were less likely to stop breast-feeding before 4 months (white mothers, 0.8 (0.8-0.9); non-white mothers, 0.9 (0.8-1.0)) than those with no/minimal qualifications but, among ethnic minorities, were more likely to introduce solids early (1.3 (1.0-1.6)). Socio-economic status was positively associated with breast-feeding continuation among white women, and with age at introduction of solids among non-white women. CONCLUSIONS: We have identified important geographic, ethnic and social inequalities in breast-feeding continuation and introduction of solids within the UK, many of which have not been reported previously. The factors mediating these associations are complex and merit further study to ensure that interventions proposed to promote maternal adherence to current infant feeding recommendations are appropriate and effective.     

The role of learning in fish orientation

Past research on fish orientation and migration has tended to focus on genetically fixed patterns of behaviour. However, in most aquatic environments, the physical landscape as well as biologically important locations will be subject to varying degrees of change, favouring an ability to learn. Here, we review the evidence from field and laboratory‐based experiments that fish can and do use learning and memory to orientate within their natural environments, relying on information from a diverse array of different sources. Comparisons of different species and populations suggest that fish may be predisposed to learn specific associations at specific times or places, appropriate for the particular navigational problems that they are likely to encounter.