Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

The rate of Salmonella spp. infection in zoo animals at Seoul Grand Park, Korea

Salmonellosis is an important zoonotic disease that affects both people and animals. The incidence of reptile-associated salmonellosis has increased in Western countries due to the increasing popularity of reptiles as pets. In Korea, where reptiles are not popular as pets, many zoos offer programs in which people have contact with animals, including reptiles. So, we determined the rate of Salmonella spp. infection in animals by taking anal swabs from 294 animals at Seoul Grand Park. Salmonella spp. were isolated from 14 of 46 reptiles (30.4%), 1 of 15 birds (6.7%) and 2 of 233 mammals (0.9%). These findings indicate that vigilance is required for determining the presence of zoonotic pathogen infections in zoo animals and contamination of animal facilities to prevent human infection with zoonotic diseases from zoo facilities and animal exhibitions. In addition, prevention of human infection requires proper education about personal hygiene.     

Influences of quorum‐quenching probiotic bacteria on the gut microbial community and immune function in weaning pigs

The aim of this study is to investigate the dynamic gut microbial diversity in weaning swine after administering feed supplemented with probiotic bacteria that specifically inhibit the activity of quorum molecules. Initially, the universal quorum molecule autoinducer‐2 (AI‐2) bioassay results indicated that AI‐2 activity was profoundly inhibited in enterohemorrhagic Escherichia coli (EHEC) O157:H7 in the presence of Lactobacillus acidophilus strain 30SC cell extract, although the growth of EHEC was not affected. Based on plate counting results, bacterial community analysis revealed a specific reduction in coliforms compared to the control, whereas the population of lactobacilli increased in weaning swine in in vivo trials. Supplementation with L. acidophilus strain 30SC did not affect the counts of other communities, such as total aerobes and yeast/mold. In addition, PCR‐denaturing gradient gel electrophoresis analysis showed a significant difference in the 16S rRNA gene products after administering L. acidophilus strain 30SC. Selected bands were sequenced, and most of them were identified as uncultured bacterium clones or a Lactobacillus‐ and Bifidobacterium‐specific community. Therefore, our results indicate that quorum‐quenching probiotic bacteria can significantly modulate the gut microbiota of swine and these beneficial effects can contribute to the improvement of performance and health in the gastrointestinal tract of weaning pigs.     

Controlling yellow jackets with fipronil-based protein baits in urban recreational areas

ABSTRACT

The western yellow jacket, Vespula penyslvanica (Saussare), is a serious seasonal pest of outdoor venues in the western United States. In the spring, queens and low numbers of workers were captured in heptyl-butyrate-baited traps until early July, when the number of foragers dramatically increased. Microsatellite data suggest that 18 colonies were actively foraging within the park in 2012. Foragers from 11 different colonies were collected at one trap site. In 2012 and 2013, sufficient numbers of foragers were not captured until early August when baiting trials were initiated. Baits were prepared with canned chicken mixed with fipronil for a final concentration of 0.025%. In 2012, a single baiting provided >96% reduction of foragers for at least two months. A second baiting late in the season provided >80% reductions. In 2013, one baiting trial resulted in a 74% and a 93% reduction. A bait acceptance study was conducted in 2014 to test a bait consisting of the juices from canned chicken and 0.025% fipronil incorporated into a hydrogel, and provided a much longer lasting bait in the field.     

Argentine ant control around homes: efficacy of treatments and urban runoff

ABSTRACT

Pest management professionals in California receive more customer complaints about Argentine ants than for any other urban ant pest. Fipronil, applied as a 30 × 30 cm band around the house foundation, has become the preferred treatment used to control these ants. Unfortunately, fipronil is now showing up in urban waterways at levels that are toxic to aquatic invertebrates. Our recent studies are aimed at mitigating insecticide runoff while still controlling the ant infestations. A high priority is preventing fipronil runoff from the driveway to the street, where it can flow into drains and from there to urban waterways. In this paper, two related studies address these issues. Not treating driveways with fipronil reduced by two to three orders of magnitude its runoff when compared with earlier studies. However, not treating the driveway can reduce efficacy of treatments. Granular bifenthrin, indoxacarb, botanicals, and a thiamethoxam ant bait were tested as supplemental treatments. The gel bait showed the best result as a supplement, but only after 8 weeks. We have reduced fipronil runoff while maintaining efficacy of the ant treatments.     

Cellular Composition and Viability of Cloned Bovine Embryos Using Exogene-Transfected Somatic Cells

The present study compared the efficiency of transgenic (TG) cloned embryo production by somatic cell nuclear transfer (SCNT) with fetal-derived fibroblast cells (FFCs) which were transfected with pEGFP-N1 to in vitro-fertilized (IVF), parthenogenetic and SCNT counterparts by evaluating the rates of cleavage and blastocyst formation, apoptosis rate at different developmental stages, cell number, ploidy and gene expression in blastocysts. In SCNT and TG embryos, the rates of cleavage and blastocyst formation were significantly lower (p < 0.05) than those of IVF controls, but it did not differ between SCNT and TG embryos. In IVF control, 86.7% embryos displayed diploid chromosomal complements and the rates were significantly (p < 0.05) higher than those of SCNT and TG embryos. Most TG embryos (79%) with FFCs expressed the gene by both PCR and under fluorescence microscopy. The expression of apoptosis by TUNEL was first detected at six to eight cell stages in all embryos of IVF, SCNT and TG groups, but the expression rate at each developmental stages was significantly higher (p < 0.05) in SCNT and TG embryos than in IVF counterparts. The expression rate in inner cell mass (ICM) of TG embryos was significantly higher (p < 0.05) than in SCNT and IVF embryos. These results indicate that the high occurrence of apoptosis observed in SCNT and TG embryos compared with IVF counterparts might influence the developmental competence. Moreover, the SCNT embryos derived using non-transfected donor cells exhibited a lower apoptosis expression in ICM cells than in TG embryos derived using pEGP-N1-transfected donor cells suggesting a possible role of negative gene effect in TG embryos.     

Computational analysis of mutated AHAS in response to sulfonylurea herbicides

Sulfonylurea (SU) herbicides that target acetohydroxyacid synthase (AHAS, also called acetolactate synthase (ALS)) are regarded as one of the most important classes of herbicides, due to their extremely low toxicity towards mammals, ultra‐low dosage application and high selectivity. However, mutations of AHAS to a herbicide‐insensitive form appear to be a worldwide problem for acquired resistance. In this study, three mutated AHAS sequences were used. In the mutated AHASs, the proline (Pro) was replaced by one of threonine (Thr), alanine (Ala) or serine (Ser) in AHAS amino acid (AA) position 197Pro. To understand the herbicide resistance mechanism, we built dimeric AHAS complex models based on 1YBH of protein data bank (PDB) with SU herbicide, chlorimuron ethyl (CIE), using a homology. The three mutated AHASs in complex with CIE were analysed, and the resistance mechanism to SU herbicides was studied by molecular dynamics (MD) simulation performed using GROMACS 4.5.5. MD simulation analysis of wild‐type and the three mutated dimeric AHAS–CIE complex systems revealed the conformational stability and changes in herbicide binding to dimeric AHAS system due to the mutation. Additionally, it showed that conformational change of amino acid residue 200Met (methionine) was associated with the imidazolinone herbicide resistance mechanism. According to our computation, the detailed mechanism of herbicide resistance was variable, depending on the type of mutated amino acid, providing new insights for designing herbicides specific to each mutant AHAS.     

Comparison of the bovine blood gas parameters produced with three types of portable blood gas analyzers

BackgroundA definite diagnosis should be made in the bovine practice field, however, it was difficult to perform laboratory analysis immediately. Currently, three types of portable blood gas analyzers are available in Korea.ObjectivesThis study aimed to evaluate the correlations among these three analyzers.MethodsSeventy-two plasma samples from Holstein-Friesian cows were used for blood gas analysis, and three instruments (EDAN i15 Vet, VETSCAN i-STAT, and EPOC) were operated simultaneously. Moreover, plasma calcium levels were compared between these portable analyzers and blood chemistry device, which is usually used in a laboratory environment. Pearson analysis was performed to confirm the correlation of each parameter produced with the three instruments and blood chemistry analyzer.ResultsAs results, high correlation was observed in parameters of pH, pO₂, potassium ion, ionized calcium, and glucose (p < 0.001, r > 0.7). In addition, pCO₂ showed a moderate correlation among the three analyzers (p < 0.001, r > 0.5), and there was no correlation among all instruments for sodium ions. There was also a high correlation between ionized calcium from the three portable devices and total calcium from the biochemistry analyzer (p < 0.001, r > 0.9).ConclusionsIn conclusion, there was a high correlation between results from the three different blood gas analyzers used in the bovine clinical field in Korea. Thus, a consistent diagnosis can be made even with different equipment if the operator is aware of the strengths and weaknesses of each piece of equipment and operates it properly.