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.     

A Fluted Point from the Uptar Site, Northeastern Siberia

Lanceolate bifacial points, including one fluted specimen, have been collected from beneath an early Holocene tephra at the Uptar site, northeastern Siberia. Thus, the technology associated with the well-known Paleoindian tradition was not confined to the Americas. The Uptar collection does not compare readily with other Beringian complexes and demonstrates that there is greater diversity in the archaeological record of northeastern Siberia than traditional colonization models imply.     

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.     

Hunting behavior of a marine mammal beneath the antarctic fast Ice

The hunting behavior of a marine mammal was studied beneath the Antarctic fast ice with an animal-borne video system and data recorder. Weddell seals stalked large Antarctic cod and the smaller subice fish Pagothenia borchgrevinki, often with the under-ice surface for backlighting, which implies that vision is important for hunting. They approached to within centimeters of cod without startling the fish. Seals flushed P. borchgrevinki by blowing air into subice crevices or pursued them into the platelet ice. These observations highlight the broad range of insights that are possible with simultaneous recordings of video, audio, three-dimensional dive paths, and locomotor effort.     

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.     

Effect of Early Postpartum Ovulation on Fertility in Dairy Cows

Our objectives were to determine the effects of early ovulation on fertility and uterine health of dairy cows. Four hundred and forty‐five Holstein cows (185 primiparous and 260 multiparous) from five herds were used. Blood samples were collected at 21, 35 and 49 days in milk (DIM) and cows were considered to be cyclic at 21 DIM (Cyc21) if serum progesterone (P4) concentration was above 1 ng/ml, cyclic by 49 DIM (Cyc49) if P4 concentration was above 1 ng/ml at 35 or 49 DIM, or not cyclic (NotCyc) if P4 concentration was below 1 ng/ml at all sample times. Endometrial cytology for diagnosis of subclinical endometritis was examined at 49 DIM in a subset of 414 cows. Cows in the group Cyc21 had increased hazard of insemination, for the first service, compared with cows in Cyc49 [hazard ratio (HR) = 1.40; 95% CI = 1.10–1.79; p = 0.006] and NotCyc (HR = 2.07; 95% CI = 1.52–2.82; p < 0.001). Cows in the Cyc49 group also had increased hazard of insemination compared with cows in the NotCyc group (HR = 1.47; 95% CI = 1.13–1.93; p = 0.005). Median days to insemination were, respectively, 71, 76 and 96 for cows in Cyc21, Cyc49 and NotCyc groups. Cows in Cyc21 had greater first service pregnancy per AI than Cyc49 [38.6 vs 28.1%; adjusted odds ratio (AOR) = 1.67; 95% CI = 1.01–2.75; p = 0.04] and NotCyc (38.6 vs 23.6%; AOR = 2.08; 95% CI = 1.08–4.00; p = 0.03). Pregnancy per AI was similar in Cyc49 and NotCyc cows (28.1 vs 23.6%; AOR = 1.25; 95% CI = 0.70–2.24; p = 0.45). Cows in Cyc21 had increased hazard of pregnancy up to 300 DIM compared with Cyc49 (HR = 1.52; 95% CI = 1.17–1.96; p = 0.002) and NotCyc (HR = 1.98; 95% CI = 1.41–2.78; p < 0.001). Cows in Cyc49 tended to have increased hazard of pregnancy compared with NotCyc (HR = 1.31; 95% CI = 0.96–1.77; p = 0.09). Median days to pregnancy were, respectively, 103, 147 and 173 for cows in Cyc21, Cyc49 and NotCyc groups. Cows in the Cyc21 group had decreased prevalence of subclinical endometritis compared with cows in the NotCyc group (29.9 vs 43.7%; AOR = 0.53; 95% CI = 0.29–0.97; p = 0.04); however, the prevalence did not differ from the Cyc49 group (29.9 vs 39.1%; AOR = 0.68; 95% CI = 0.41–1.14; p = 0.15). Cyc49 cows had similar prevalence of subclinical endometritis compared with NotCyc cows (AOR = 0.77; 95% CI = 0.46–1.29; p = 0.32). Early postpartum ovulation was associated with improved uterine health and fertility.     

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.     

Vaginal isolation of beta‐haemolytic Streptococcus from bitches with and without neonatal deaths in the litters

The aim of the study was to identify beta‐haemolytic streptococci in the vagina of bitches who had delivered healthy litters and bitches who had delivered litters in which neonatal deaths occurred. Fifty‐one bitches divided into two groups were used. Group 1 (G1) included 28 bitches that had delivered healthy litters and group 2 (G2) included 23 bitches that had delivered puppies who died in the neonatal period. Two vaginal samples were taken, one in proestrus and the other at the end of gestation (EG). Beta‐haemolytic Streptococcus (BS) was isolated from 16 bitches (57%) in G1 and from 21 bitches (91%) in G2. The bacteriological cultures, serological tests (Streptex^®) and PCR assay allowed identification of Streptococcus canis and Streptococcus dysgalactiae in G1 and G2. Ultramicroscopic studies allowed the observation of M Protein and capsules in strains of S. dysgalactiae and S. canis in G1 and G2. The S. canis strains isolated from G2 showed thicker capsules than S. canis strains isolated from G1 (234 ± 24.2 vs 151.23 ± 28.93 nm; p < .001.). No differences were observed in capsule thickness between strains of S. dysgalactiae isolated from G1 and G2 (210 ± 13.54 vs 211.66 ± 19.67 nm; p > .70). All strains of beta‐haemolytic Streptococcus isolated were penicillin sensitive. Penicillin was administered from EG to 5 days post‐partum in 10 G2 females with isolation of BS (G2A). Saline solution was administered in eleven G2 females with isolation of BS (G2B). Ninety per cent of the puppies survived in G2A and 25% survived in G2B. Our results suggest BS is involved in canine neonatal deaths.