Receptor-specific binding of purified F4 to isolated villi.

Different procedures for preparing and purifying F4ac fimbriae of the enterotoxigenic Escherichia coli strain GIS 26 (O149:K91:F4ac LT+Sta+STb+) were performed and the purity and yield of F4ac were compared. Fimbriae were prepared by either mechanical shearing or heatshock treatment of concentrated bacterial suspensions (10(11) bacteria/ml). The mechanical shearing procedure resulted in approximately 1.7 mg fimbriae (i.e. 74.4% of the isolated protein) and 0.6 mg (25.6%) contaminating proteins per 10(12) bacteria, whereas the yield of fimbriae following heatshock treatment was lower (0.3 mg per 10(12) bacteria, i.e. 26.2%) and the relative contamination higher (1.0 mg per 10(12) bacteria, i.e. 73.8%). A further purification consisted of either anion exchange chromatography (AEC) or electro-elution from SDS-polyacrylamide gels. The electroelution procedure was performed under reducing and denaturing conditions, so that purified FaeG subunits, the major subunit of F4, were finally obtained. The binding activity of fimbriae, nonpurified as well as purified, and FaeG to F4-specific receptors on isolated intestinal villi was assessed in an inhibition adhesion assay. Native fimbriae as well as major subunits were able to bind to the receptors, and the specificity of the binding was demonstrated by blockage with F4ac-specific MAb.     

Antigenicity of cortical bone allografts in dogs and effect of ethylene oxide-sterilization.

The aims of the present study were to determine the antigenicity of cortical bone allografts and the effect of ethylene oxide-sterilization (EO-sterilization). Cortical bone allografts from one donor dog were implanted in a muscle pouch in four groups of four dogs each. The grafts were either fresh, EO-sterilized, demineralized or demineralized and EO-sterilized. The immune response against the grafts was determined by measuring the antibody response against surface antigens of donor cells and by the mixed lymphocyte reaction. Dogs receiving EO-sterilized grafts or bone matrix did not demonstrate an immune response. Only two of the four dogs with fresh cortical bone grafts showed a very weak immune response. This suggests a priming of the host by the fresh bone grafts. However, implanting skin grafts from the donor dog subdermally, in one dog of each of the groups, four months after implanting the bone grafts did not induce a secondary immune response. Macroscopic and histologic examination of the bone grafts five months after their implantation consistently revealed graft resorption (activity of osteoclasts) and vascularization of the fresh bone grafts, but not of EO-sterilized fresh grafts. For most EO-sterilized grafts, a strong inflammatory reaction was present in the tissues surrounding the graft and this was not apparent around the non-sterilized grafts. The absence of resorption and the presence of the inflammation seemed to be unwanted effects of the EO-sterilization. The EO-sterilisation did not affect osteoinduction since osteocytes were observed in the EO-sterilized demineralized grafts. Results indicate that cortical bone allografts used in the present study are very weak antigens and that the EO-sterilization procedure used has no effect on osteoinduction, but decreases bone resorption.     

Serological and parasitological survey of dourine in the Arsi–Bale highlands of Ethiopia

This study was conducted from August 2005 to January 2007 to determine prevalence and distribution of dourine in horses and to investigate the occurrence of clinical and carrier cases in donkeys and mules in the Arsi–Bale highlands. Study methodology was based on questionnaire, serological, clinical and parasitological survey. The questionnaire indicated that dourine is a major health problem of equines in the Arsi–Bale highlands. Though dourine is commonly observed throughout the year, it has a seasonal character and occurs mostly during the breeding season from June to late September. Serological screening of 646 horses showed a seroprevalence of 184 (28%), 161 (25%) and 125 (19%) for card agglutination test for trypanosomosis, LATEX and enzyme-linked immunosorbent assay, respectively. Risk factors were parity number, previous history of abortion and body condition score. No trypanosomes could be detected by Giemsa staining or by haematocrit centrifugation technique. Ten puppies inoculated with blood samples, genital washes and oedematous fluids remained parasitologically negative. Different characteristic signs of dourine were observed. During the genital stage, mares showed vaginal oedema, discharge and presence of depigmented scars over the external genitalia. In stallions, oedema of the scrotum and prepuce, prepucial and urethral discharge, and ulceration of the genital mucosae mainly of the penile were observed. In both sexes, lameness in one or both legs, partial dragging and stiffness of the hind legs and incoordination were the dominant signs observed as nervous form of the disease.     

Schizochytrium as a replacement for fish oil in a fishmeal free diet for jade perch,Scortum barcoo (McCulloch & Waite)

A 10‐week trial was conducted to determine the response of juvenile jade perch Scortum barcoo on the replacement of dietary fish oil (FO) in a fishmeal free diet. Three iso‐nitrogenous, isocaloric and isolipidic diets were formulated, each containing a different primary fat source: FO, linseed oil (LO), and a mixture of Schizochytrium and LO. The substitution of FO with the mixture of Schizochytrium and LO did not cause a difference in growth. However, there was an 8% reduction in weight gain in fish fed dietary LO, indicating that juvenile jade perch do require a minimal concentration of dietary n‐3 highly unsaturated fatty acids (HUFA). Fish fed the Schizochytrium diet stored more efficient n‐3 HUFA and in particular DHA in their flesh, and retained a higher fillet recovery compared to fish fed FO. In addition, we demonstrated that jade perch are able to produce both n‐3 HUFA and n‐6 HUFA when dietary PUFA are present. Fish fed the LO diet for 10 weeks contained the lowest amount of n‐3 HUFA in fillets among dietary treatment groups. However, feeding these fish the Schizochytrium diet for an additional 4 weeks increased the n‐3 HUFA content towards the same concentration of n‐3 HUFA found in the flesh of fish fed FO, without affecting the sensory properties of the fillets. In contrary, feeding the Schizochytrium diet for a continuous period of 14 weeks lowered overall sensory property scores.     

Supplementation with sunflower seeds in beef cattle did not impact on oocyte and in vitro embryo production

Supplementation with compounds rich in linoleic acid, including sunflower seed supplementation, promotes increase in conception rates in cows. We aimed to evaluate whether the sunflower seed (linoleic acid source) supplementation in beef donor females alters the plasma concentrations of cholesterol, triglycerides, HDL and LDL, increases the number and quality of oocytes, increases the cleavage rates and determines an improvement in number and quality of in vitro produced blastocysts. Thus, Nelore females were divided into two groups of 15 animals to receive supplementation with or without sunflower seed for 57 days. Females underwent follicular aspiration and the oocytes were subjected to in vitro embryo production. There was no difference (p > .1) between control group and group supplemented with sunflower seed on the number of displayed follicles; number of aspired oocytes; recovery rate; cleavage rate; number of embryos; number of blastocysts; embryos number of grades I and II; plasma concentrations of total cholesterol, triglycerides; HDL and LDL. Therefore, sunflower seed supplementation in oocyte donors did not increase the number and quality of oocytes, cleavage rates and the number and quality of blastocysts produced in vitro.     

Effect of body condition score and reuse of progesterone‐releasing intravaginal devices on conception rate following timed artificial insemination in Nelore cows

This study had the aim of investigating the efficiency of timed artificial insemination (TAI) through the progesterone‐releasing intravaginal device (PRID), used in new condition and for the second and third times in Nelore cows. The effects of device reuse and body condition score (BCS) on the conception rate (CR) were evaluated in 1,122 multiparous Nelore cows (mean BCS of 2.7 ± 0.4), which were randomly distributed into three groups that received new (n = 330), once (n = 439) and twice used (n = 353) PRID. Among the 1,122 females that underwent TAI, 573 became pregnant, thus representing an overall CR of 51.06%. Cows with BCS between 2.75 and 4.0 had greater (p < .0001) CR (69.75%) than cows with BSC of 2.0–2.5 (32.98%). It was observed that the CR through using PRID was 60.00%, 51.71% and 41.93% for new, once and twice used PRID, respectively, with difference between all groups (p < .0001). Under tropical conditions, animals with BCS greater than 2.5 had a higher CR, and the CR decreased proportionally with the number of times that the PRID had been used.     

The food contaminant fumonisin B1 reduces the maturation of porcine CD11R1+ intestinal antigen presenting cells and antigen-specific immune responses, leading to a prolonged intestinal ETEC infection

Consumption of food or feed contaminated with fumonisin B₁ (FB₁), a mycotoxin produced by Fusarium verticillioides, can lead to disease in humans and animals. The present study was conducted to examine the effect of FB₁ intake on the intestinal immune system. Piglets were used as a target and as a model species for humans since their gastro-intestinal tract is very similar. The animals were orally exposed to a low dose of FB₁ (1 mg/kg body weight FB₁) for 10 days which did not result in clinical signs. However, when compared to non-exposed animals, FB₁-exposed animals showed a longer shedding of F4⁺ enterotoxigenic Escherichia coli (ETEC) following infection and a lower induction of the antigen-specific immune response following oral immunization. Further analyses to elucidate the mechanisms behind these observations revealed a reduced intestinal expression of IL-12p40, an impaired function of intestinal antigen presenting cells (APC), with decreased upregulation of Major Histocompatibility Complex Class II molecule (MHC-II) and reduced T cell stimulatory capacity upon stimulation. Taken together, these results indicate an FB₁-mediated reduction of in vivo APC maturation.