Effect of endocytic and metabolic inhibitors on the internalization and intracellular growth of Brucella abortus in Vero cells.

Uptake, transfer to rough endoplasmic reticulum, and intracellular growth of Brucella abortus were studied in Vero cells treated with endocytic and metabolic inhibitors. Infection of Vero cells was suppressed when inhibitors of energy metabolism (iodoacetate, dinitrophenol), receptor-mediated endocytosis (monodansylcadaverine, amantadine, methylamine), or endosomal acidification (chloroquine, ammonium chloride, monensin) were added to the inoculum. Inhibition was not observed when these drugs were added after the inoculation period. Infection of Vero cells by B abortus was inhibited by dibutyryl-cyclic adenosine monophosphate and Vibrio cholerae enterotoxin, but was stimulated by dibutyryl-cyclic guanosine monophosphate and escherichia coli heat-stable enterotoxin a. Uptake of B abortus by Vero cells was not prevented by colchicine, but was abolished by cytochalasin B. Uptake of heat-killed B abortus and noninvasive E coli was similar to that of viable brucellae. Intracellular growth of B abortus was not affected by cycloheximide. Results indicate that: B abortus may be internalized by a receptor-mediated phagocytic process; transfer of B abortus from phagosomes to rough endoplasmic reticulum may require endosomal acidification; and replication of B abortus within the rough endoplasmic reticulum may not depend on protein synthesis by the host cell.     

Cell Proliferation in the Atretic Follicles of Buffalo and Cattle Ovary

The present study was carried out to describe the proliferative activity of granulosa and theca cells in healthy antral and atretic follicles of Philippine buffaloes (BU) and Holstein-Friesian (HF) cows. Paraffin sections of ovary were immunostained with mouse monoclonal antibody to proliferating cell nuclear antigen (PCNA). Then the follicles were classified into healthy and various stages of atretic follicles. The granulosa layer of healthy follicles had a significantly higher frequency of PCNA-positive cells than the early and advanced atretic follicles in both breeds. In the theca interna, significantly reduced populations of the PCNA-positive cells were found in both breeds as atresia progressed. Moreover, HF had significantly higher PCNA-positive cells in the theca interna of healthy, early atretic and advanced atretic follicles than BU. A reduction of PCNA-positive cells during atresia was also noted in the theca externa in both animals although differences were not significant. The results of the present work suggest that the proliferative activity of granulosa and theca cells decreases in association with follicular atresia in the BU similar to HF. Furthermore, a significantly deficient cell proliferative activity of theca interna was found in BU compared with HF.     

Development of a marker assisted selection program for cacao

ABSTRACT Production of cacao in tropical America has been severely affected by fungal pathogens causing diseases known as witches' broom (WB, caused by Moniliophthora perniciosa), frosty pod (FP, caused by M. roreri) and black pod (BP, caused by Phytophthora spp.). BP is pan-tropical and causes losses in all producing areas. WB is found in South America and parts of the Caribbean, while FP is found in Central America and parts of South America. Together, these diseases were responsible for over 700 million US dollars in losses in 2001 (4). Commercial cacao production in West Africa and South Asia are not yet affected by WB and FP, but cacao grown in these regions is susceptible to both. With the goal of providing new disease resistant cultivars the USDA-ARS and Mars, Inc. have developed a marker assisted selection (MAS) program. Quantitative trait loci have been identified for resistance to WB, FP, and BP. The potential usefulness of these markers in identifying resistant individuals has been confirmed in an experimental F(1) family in Ecuador.     

Effects of postweaning dietary energy source on reproductive traits in primiparous sows

An experiment was conducted to study the effects of major dietary energy source fed from weaning to ovulation or from ovulation to d 35 of pregnancy on reproductive traits in primiparous sows. Dietary energy sources were used to manipulate the plasma insulin concentration. One hundred thirteen sows were used in a split-plot design. From weaning to ovulation sows were fed at two times maintenance either a diet with tallow (Fat) or maize starch plus dextrose (Starch) as the major energy source. From ovulation onward, sows within each dietary group were alternately reassigned to either the Fat or the Starch diet and were fed at 1.25 times maintenance. Estrus detection was performed three times a day from d 3 to 9 after weaning and sows were inseminated each day of standing estrus. On d 35 of pregnancy, the sows were slaughtered and their reproductive tracts were removed. Plasma insulin concentration was higher in sows fed the Starch-rich diet than in sows fed the Fat-rich diet on d 4 after weaning (1.30 vs 0.97 ng/mL, P = 0.08) and on d 32 of pregnancy (1.20 vs 0.51 ng/mL, P < 0.001). Plasma glucose and IGF-I concentration on d 4 after weaning and d 32 of pregnancy did not differ between sows on the two dietary energy sources. The percentage of sows exhibiting estrus within 9 d after weaning was 52 and 67% for the Fat and Starch diet before ovulation, respectively (P = 0.11), whereas the weaning-to-estrus interval was 134 vs 123 h, respectively (P = 0.12). Survival analysis showed that sows fed the Fat-rich diet had a 1.6 times higher risk to remain anestrous until d 9 after weaning than sows fed the Starch-rich diet (P = 0.04). No effect of dietary energy source, either before or after ovulation, on uterine, placental, or embryonal development on d 35 of pregnancy was found. It can be concluded that the dietary energy source provided after weaning can affect the risk of sows to remain anestrous but does not affect uterine, placental, or embryonic traits.     

Activity against Ostertagia ostertagi of low doses of oxfendazole continuously released from intraruminal capsules in cattle.

Slow release from intraruminal capsules of 0.48 mg/kg oxfendazole per day for 5.5 days gave percentage efficiencies of 99 +/- 0.6, 86.6 +/- 6.3 and 93.1 +/- 4.5 for the removal of adult worms, developing 4th stages and inhibited larvae of Ostertagia ostertagi respectively. These efficiencies were not significantly different from those obtained from a single oral dose of 2.5 mg/kg. Peak plasma levels of oxfendazole were similar for the 2 types of administration but high levels were maintained longer in cattle given capsules.     

Association between floor type and behaviour, skin lesions, and claw dimensions in group-housed fattening bulls

Rubber mats covering concrete slatted flooring are a developing market in dairy barns but remain rare in beef cattle facilities. The aim of this study was to investigate the effects of covering slatted concrete floor with perforated rubber mats on behaviour and occurrence of skin and claw lesions in fattening bulls. The groups of six bulls each with a total average age of 9.8 months were kept over 1 year on either slatted concrete (CONCRETE PEN) or on slatted concrete covered completely (RUBBER PEN) or partially (CHOICE PEN) with perforated rubber mats. Every quarter-year, behaviour (preference of flooring, lying, aggression, mounting) was recorded. In two-weekly intervals the incidence of skin lesions was examined. At 12 and 18 months of age the rising time of the bulls was measured. At the beginning of the study and after slaughter claw dimensions were recorded. Bulls in the CHOICE PEN preferred (P<0.01) the rubber coated area throughout the experiment. Animals in the RUBBER and the CHOICE PENS showed more lying periods (P<0.01) and had a lesser incidence of skin lesions (P<0.01) compared to bulls in the CONCRETE PEN. Bulls in the CHOICE PEN needed less time for rising (2.7+/-0.3s) than bulls in the CONCRETE PEN (4.4+/-0.5s, P<0.01). Net claw growth differed significantly between all pens (RUBBER>CHOICE>CONCRETE; P<0.05). In conclusion, the results of this study indicate that rubber coated slatted flooring has a positive influence on the housing conditions of beef cattle.     

Molecular characterization and genetic relationship among almond cultivars assessed by RAPD and SSR markers

RAPDs and SSRs were used to study the genetic diversity of Iranian almond cultivars and their relationship to important foreign cultivars and three related species. Eight unidentified almond Shahrodi cultivars and three wild almonds (Prunus communis, Prunus orientalis and Prunus scoparia) were also included. Of the primers tested, 42 (out of 80) RAPD and 18 (out of 26) SSR primers were selected for their reproducibility and high polymorphism. A total of 664 polymorphic RAPD bands were detected out of 729 bands. The number of presumed alleles revealed by the SSR analysis ranged from 3 to 10 alleles per locus with a mean value of 6.64 alleles per locus. Both techniques discriminated the genotypes very effectively, but only RAPDs were able to discriminate the cultivars Monagha and Sefied. Results demonstrated an extensive genetic variability within the tested cultivars as well as the value of SSR markers developed in peach for characterization of almond and related species of Prunus. Dice similarity coefficient was calculated for all pair wise comparisons and was used to construct a UPGMA dendrogram. For both markers a high similarity in dendrogram topologies was obtained although some differences were observed. All dendrograms, including that obtained by the combined use of both the marker data, depicted the phenetic relationships among the cultivars and species, depending upon their geographic region and/or pedigree information. Almond cultivars clustered with accession of P. communis showing their close relationship. P. orientalis and P. scoparia were clustered out of the rest of P. dulcis.     

Human–Dromedary Camel Interactions and the Risk of Acquiring Zoonotic Middle East Respiratory Syndrome Coronavirus Infection

Middle East respiratory syndrome coronavirus (MERS‐CoV) cases without documented contact with another human MERS‐CoV case make up 61% (517/853) of all reported cases. These primary cases are of particular interest for understanding the source(s) and route(s) of transmission and for designing long‐term disease control measures. Dromedary camels are the only animal species for which there is convincing evidence that it is a host species for MERS‐CoV and hence a potential source of human infections. However, only a small proportion of the primary cases have reported contact with camels. Other possible sources and vehicles of infection include food‐borne transmission through consumption of unpasteurized camel milk and raw meat, medicinal use of camel urine and zoonotic transmission from other species. There are critical knowledge gaps around this new disease which can only be closed through traditional field epidemiological investigations and studies designed to test hypothesis regarding sources of infection and risk factors for disease. Since the 1960s, there has been a radical change in dromedary camel farming practices in the Arabian Peninsula with an intensification of the production and a concentration of the production around cities. It is possible that the recent intensification of camel herding in the Arabian Peninsula has increased the virus' reproductive number and attack rate in camel herds while the ‘urbanization’ of camel herding increased the frequency of zoonotic ‘spillover’ infections from camels to humans. It is reasonable to assume, although difficult to measure, that the sensitivity of public health surveillance to detect previously unknown diseases is lower in East Africa than in Saudi Arabia and that sporadic human cases may have gone undetected there.