Quantification of bioavailable chlortetracycline in pig feces using a bacterial whole-cell biosensor

Bacterial whole-cell biosensors were used to measure the concentration of chlortetracycline (CTC) in the feces of pigs. In this study, the Escherichia coli biosensor used has a detection limit of 0.03 mg/kg CTC in pig feces. The tetracycline concentration was correlated with the appearance and maintenance of fecal coliform bacteria resistant to tetracycline. Initially, large quantities of water-extractable CTC were excreted from the pigs and measurable amounts were detected even at 30 days after treatment cessation. This led to a sharp rise in the number of tetracycline resistant coliform bacteria in the feces, to within the same order of magnitude as the total coliform count. The high level of tetracycline resistance was maintained in spite of the declining concentration of tetracycline.     

Matrix metalloproteinase activity in tumor, stromal tissue, and serum from cats with malignancies

Matrix metalloproteinases (MMPs) are enzymes that play key roles in angiogenesis, tumor invasion, and metastasis in a wide variety of species. The purpose of this study was to evaluate pro and active MMP 2 and 9 concentrations in tumor, normal stromal tissue, and serum from tumor-bearing cats. We hypothesized that serum concentrations of pro and active forms of MMPs 2 and 9 would be predictive of MMP concentrations in tumor tissue and that these MMP concentrations would correlate with the histopathologic grade of the malignancies. Pro and active forms of MMPs 2 and 9 were determined by gelatin zymography and subsequent computerized densitometry from tumor and nearby stromal tissue and serum from 49 cats with various malignancies. The serum concentrations of MMPs from these tumor-bearing cats were compared with serum concentrations of MMPs from 44 normal cats of similar age and gender. Measurable concentrations of MMPs 2 and 9 were found within tumor, stromal, and serum samples. Mean concentrations of total pro and active MMPs 2 and 9 within tumor tissue were significantly higher (P values <.0001, .0031, <.001, and .0064, respectively) when compared with stromal tissue from the same animals. Serum MMP concentrations from tumor-bearing cats were higher than those from normal cats. Poor correlation was found between serum MMP concentrations and tissue MMP concentrations of increasing histologic grades of malignancies.     

Simultaneous one-tube quantification of host and pathogen DNA with real-time polymerase chain reaction

ABSTRACT Phaeocryptopus gaeumannii is a widespread foliar parasite of Douglas-fir. Although normally innocuous, the fungus also causes the defoliating disease Swiss needle cast in heavily infected needles. The extent of P. gaeumannii colonization in Douglas-fir foliage was estimated with real-time quantitative polymerase chain reaction (PCR) using TaqMan chemistry. In order to derive a normalized expression of colonization, both pathogen and host DNA were simultaneously amplified but individually detected by species-specific primers and TaqMan probes labeled with different fluorescent dyes. Detection of host DNA additionally provided an endogenous reference that served as both an internal positive control and adjusted for variation introduced by sample-to-sample differences in DNA extraction and PCR efficiencies. The genes employed for designing the TaqMan probes and primers were beta-tubulin for the pathogen and a LEAFY/FLORICAULA-like gene involved in floral development for the tree host. Both probe/primer sets exhibited high precision and reproducibility over a linear range of 4 orders of magnitude. This eliminated the need to analyze samples in multiple dilutions when comparing lightly with heavily infected needles. Quantification of the fungus within needles was successful as early as 1 month after initial infection. Real-time PCR is the only method currently available to quantify P. gaeumannii colonization early in the first year of the colonization process.     

Effect of nematode burden as assessed by means of faecal egg counts on body condition in goats farmed under resource-poor conditions in South Africa

A longitudinal study was conducted of the nematode faecal egg counts (FECs) and body condition scores (BCSs) of goats of resource-poor farmers at Rust de Winter, Gauteng Province, Impendle, KwaZulu-Natal Province, and Kraaipan, North-West Province, South Africa. Periods of higher FECs occurred from December/January to March/April at Rust de Winter and at Impendle and from January to March at Kraaipan. Seasonal variations in body condition were evident in the goats at Impendle with the animals showing lower BCSs from June to October. The goats at Rust de Winter and at Kraaipan did not show clear seasonal variations, although the goats at Rust de Winter showed lower BCSs from mid-July to early December. The BCSs for Rust de Winter where the animals were grazed on a private farm were generally higher than those of the other sites, where communal grazing is practised.     

Revealing genetic relationships between compounds affecting boar taint and reproduction in pigs

Boar taint is characterized by an unpleasant taste or odor in intact male pigs and is primarily attributed to increased concentrations of androstenone and skatole and to a lesser extent by increased indole. The boar taint compounds skatole and indole are produced by gut bacteria, metabolized in the liver, and stored in the fat tissue. Androstenone, on the other hand, is synthesized in the testis along with testosterone and estrogens, which are known to be important factors affecting fertility. The main goal of this study was to investigate the relationship between genetic factors involved in the primary boar taint compounds in an attempt to discover ways to reduce boar taint without decreasing fertility-related compounds. Heritabilities and genetic correlations between traits were estimated for compounds related to boar taint (androstenone, skatole, indole) and reproduction (testosterone, 17β-estradiol, and estrone sulfate). Heritabilities in the range of 0.47 to 0.67 were detected for androstenone concentrations in both fat and plasma, whereas those for skatole and indole were slightly less (0.27 to 0.41). The genetic correlations between androstenone in plasma and fat were extremely high (0.91 to 0.98) in Duroc and Landrace. In addition, genetic correlations between androstenone (both plasma and fat) and the other sex steroids (estrone sulfate, 17β-estradiol, and testosterone) were very high, in the range of 0.80 to 0.95. Furthermore, a genome-wide association study (GWA) and a combined linkage disequilibrium and linkage analysis (LDLA) were conducted on 1,533 purebred Landrace and 1,027 purebred Duroc to find genome regions involved in genetic control of the boar taint compounds androstenone, skatole, and indole, and sex hormones related to fertility traits. Up to 3,297 informative SNP markers were included for both breeds, including SNP from several boar taint candidate genes. From the GWA study, we found that altogether 27 regions were significant at a genome-wide level (P < 0.05) and an additional 7 regions were significant at a chromosomal level. From the LDLA study, 7 regions were significant on a genome-wide level and an additional 7 regions were significant at a chromosomal level. The most convincing associations were obtained in 6 regions affecting skatole and indole in fat on chromosomes 1, 2, 3, 7, 13, and 14, 1 region on chromosome 6 affecting androstenone in plasma only, and 5 regions on chromosomes 3, 4, 13, and 15 affecting androstenone, testosterone, and estrogens.     

Effects of egg disinfection and incubation temperature on early life stages of spotted wolffish

Eggs of spotted wolffish (Anarhichas minor Olafsen)were incubated at constant 4, 6 and 8 °C, and disinfected withglutaric dialdehyde (150 p.p.m. for 5 min) once ortwice a month during two thirds of the incubation period, to prevent growth ofmicroorganisms. Hatching of apparently normal larvae started earlier when eggswere disinfected twice a month compared to once a month at all incubationtemperature regimes. The time to 50% hatch was 900 and 920 day-degrees (16 and16,5 weeks) at 8 °C, 835 and 880 day-degrees (20 and 21 weeks)at 6 °C and 725 and 800 day-degrees (26 and 28,5 weeks) at 4°C, in the egg groups disinfected twice or once a month,respectively. The best survival until hatching was noted when eggs weredisinfected twice a month and incubated at 6 and 8 °C.Survivalwas very low at 4 °C. Prematurely hatched larvae wereregistered in all egg groups disinfected twice a month and the highestfrequencywas noted in the 8 °C groups. The larval weight at normalhatching in the 6 and 8 °C groups was negatively correlatedwith incubation temperature and intervals of disinfection during the incubationperiod, but after 42 days feeding with live feed (unenrichedArtemia) the weights of the larvae were not significantlydifferent. The specific growth rates of the larvae from the eggs incubated at 6°C and 8 °C were 3.0% and 3.2%, respectively.The mean survival of larvae was between 88% and 96% at 42 days post-hatching.Young wolffish originating from the 6 °C incubation groupsshowed lowest mortality.     

Identification and fine mapping of quantitative trait loci for backfat on bovine chromosomes 2, 5, 6, 19, 21, and 23 in a commercial line of Bos taurus

Backfat thickness is one of the major quantitative traits that affects carcass quality in beef cattle. In this study, we identified and fine-mapped QTL for backfat EBV on bovine chromosomes 2, 5, 6, 19, 21, and 23 using an identical-by-descent haplotype-sharing analysis in a commercial line of Bos taurus. Eleven haplotypes were found to have significant associations with backfat EBV at the comparison-wise P-value threshold, and one at the chromosome-wise P-value threshold on bovine chromosomes 5, 6, 19, 21, and 23. On average, the 12 significant haplotypes had an effect of 0.62 SD on backfat EBV, ranging from 0.38 SD to 1.33 SD. The 12 significant haplotypes spanned nine chromosomal regions, one on chromosome 5 (65.4 to 70.0 cM), three on 6 (8.2 to 11.8 cM, 63.6 to 68.1 cM, and 81.5 to 83.0 cM), three on 19 (4.8 to 15.9 cM, 39.4 to 46.5 cM, and 65.7 to 99.5 cM), one on 21 (46.1 to 53.1 cM), and one on 23 (45.1 to 50.9 cM). Among the nine chromosomal regions, six were new QTL regions and three showed remarkable agreement with QTL regions that were previously reported. Eight of the nine QTL regions were localized to less than or close to 10 cM in genetic distance. The results provide a useful reference for further positional candidate gene research and marker-assisted selection for backfat.     

A two-year comparison on nutritive value and yield of eight lucerne cultivars and one red clover cultivar

Red clover (Trifolium pratense) and lucerne (Medicago sativa) are among the most common legume species used for ruminant feeding systems. Besides the greater nutritive quality and first-year ley yield compared to lucerne, red clover decreases its yield over the years. However, under unfavourable soil conditions the persistency of lucerne can be compromised, not performing as expected. This study compared yield and chemical composition of eight lucerne cultivars and one tetraploid red clover cultivar, with similar growth capacity as lucerne, replicated over two years. In the second year, four lucerne cultivars and the red clover cultivar were analysed for crude protein (CP) fractions, dry matter and neutral detergent fibre (NDF) digestibility. A randomized block design was used with three field blocks per cultivar for the multiple comparisons within each year. There were differences in DM yield, concentrations of CP and NDF and CP fractions among the lucerne cultivars, but the differences were inconsistent between years and between growth cycles within year. The red clover cultivar had lower CP and NDF concentrations than a majority of the lucerne cultivars, and greater in vitro organic matter digestibility compared to all of the lucerne cultivars. Also, the red clover cultivar had greater rumen undegradable-to-degradable CP ratio but confirmed its lower yield persistency compared to the lucerne cultivars.