Prevalence and antimicrobial resistance of Campylobacter and Salmonella in layer flocks in Honshu,Japan

Campylobacter and non-typhoidal Salmonella are the major causes of bacterial gastrointestinal infections in humans. Although antimicrobial therapy is typically not recommended in many cases of these infections, it may be life-saving in patients with severe symptoms. Since chicken eggs and meat derived from layers are destined for human consumption, we investigated the prevalence and antimicrobial resistance of these two bacterial genera in 82 layer flocks at chicken processing plants in Honshu, Japan. Campylobacter was isolated from 77 flocks (93.9%). Resistance to ampicillin, tetracycline, and ciprofloxacin was documented in 42.3 (30/71), 16.9 (12/71), and 14.1% (10/71) of Campylobacter jejuni, respectively. Multilocus-sequence typing identified ST4389 and ST5262 as the most frequent C. jejuni sequence types. In C. coli, resistance to ampicillin, tetracycline, and ciprofloxacin was found in 20.0 (7/35), 20.0 (7/35), and 25.7% (9/35), respectively. The most frequent sequence type in C. coli was ST8292. Erythromycin resistance was not observed among Campylobacter species. Salmonella was isolated from 14 flocks (17.1%). The two most frequent serovars were Salmonella Corvallis and S. Braenderup. Neither S. Enteritidis nor S. Infantis were isolated. Streptomycin resistance was observed in six isolates (26.1%), and all isolates were susceptible to cefotaxime and ciprofloxacin. Thus, chicken eggs and meat derived from layers are possible sources of these bacterial infections in humans. The antimicrobial susceptibility of these isolates was maintained, reflecting restrictions on the use of antimicrobial agents on layers.     

Novel method for improving the antioxidative properties of fish meat by direct injection of sodium l-ascorbate into the blood vessels of live fish

The color of the red muscle of pelagic fish is used as an indicator of freshness, and sustaining a bright red color is important for maintaining the commercial value of pelagic fish. Feeding fish with antioxidant compounds can delay metmyoglobin (metMb) formation, but the process requires long-term feeding. Live cultured Japanese horse mackerel Trachurus japonicus were used in this study. After anesthetization, 2 ml of blood was drawn from blood vessels, and a parenteral solution including an antioxidant compound was injected. Browning, metMb formation, and lipid oxidation of dark muscle during chilled storage were delayed by injecting sodium l-ascorbate as an antioxidant compound in the blood vessels of live fish.     

Characterization of Murine Pituitary-Derived Cell Lines Tpit/F1, Tpit/E and TtT/GF

The pituitary is an important endocrine tissue of the vertebrate that produces and secretes many hormones. Accumulating data suggest that several types of cells compose the pituitary, and there is growing interest in elucidating the origin of these cell types and their roles in pituitary organogenesis. Therein, the histogenous cell line is an extremely valuable experimental tool for investigating the function of derived tissue. In this study, we compared gene expression profiles by microarray analysis and real-time PCR for murine pituitary tumor-derived non-hormone-producing cell lines TtT/GF, Tpit/F1 and Tpit/E. Several genes are characteristically expressed in each cell line: Abcg2, Nestin, Prrx1, Prrx2, CD34, Eng, Cspg4 (Ng2), S100β and nNos in TtT/GF; Cxcl12, Raldh1, Msx1 and Twist1 in Tpit/F1; and Cxadr, Sox9, Cdh1, EpCAM and Krt8 in Tpit/E. Ultimately, we came to the following conclusions: TtT/GF cells show the most differentiated state, and may have some properties of the pituitary vascular endothelial cell and/or pericyte. Tpit/F1 cells show the epithelial and mesenchymal phenotypes with stemness still in a transiting state. Tpit/E cells have a phenotype of epithelial cells and are the most immature cells in the progression of differentiation or in the initial endothelial-mesenchymal transition (EMT). Thus, these three cell lines must be useful model cell lines for investigating pituitary stem/progenitor cells as well as organogenesis.     

Neospora caninum and complex vertebral malformation as possible causes of bovine fetal mummification

Bovine neosporosis, caused by Neospora caninum is a leading cause of abortion in cattle. We postulated that neosporosis could lead to fetal death and mummification. Fifteen mummified fetuses were tested by polymerase chain reaction (PCR) for the mutation in the bovine SLC35A3 gene that causes complex vertebral malformation (CVM) and the pNC-5 gene which identifies N. caninum infection. DNA was extracted from the mummified fetuses and the sex of the mummies was determined by PCR. The CVM mutation was not detected in the mummified fetuses, but 4 fetuses were positive for N. caninum infection. The ages of the mummies with N. caninum infection were 100, 113, 123, and 131 days. Twelve of the 15 mummified fetuses were male. To our knowledge, this is the first detection of N. caninum as a possible cause of bovine fetal mummification.     

Genetic difference between Ezo-awabi <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> and Kuro-awabi <Emphasis Type="Italic">H. discus discus</Emphasis> populations: Microsatellite-based population analysis in Japanese abalone

ABSTRACT:   This study documents the genetic population relationships between Ezo-awabi Haliotis discus hannai and Kuro-awabi H. discus discus by means of the microsatellite technique with an emphasis on the extent of genetic difference between Ezo- and Kuro-awabi populations. Eight markers were employed to screen five populations each collected from Ezo- and Kuro-awabi habitats. All eight loci showed polymorphisms in all populations (number of alleles per locus, 11.1–14.1; averaged expected heterozygosity, 0.64–0.70). The number of loci that yielded significant genetic heterogeneities (allele frequency distribution and/or pairwise θ ) between Ezo- and Kuro-awabi populations was larger than that generated in comparisons between populations within Ezo- and Kuro-awabi habitats. According to the bootstrap neighbor-joining trees constructed on the basis of two genetic distance measures ( D _A and D _ST), the 10 populations fell into two clusters of populations (Ezo-awabi and Kuro-awabi habitat groups), and the results of hierarchical amova testing also supported the neighbor-joining clustering. The outcomes presented here suggest that the microsatellite markers used in this study are potentially an efficient means to differentiate Ezo- and Kuro-awabi populations.     

Leukotoxin family genes in Staphylococcus aureus isolated from domestic animals and prevalence of lukM-lukF-PV genes by bacteriophages in bovine isolates

Leukotoxin family genes in Staphylococcus aureus isolated from domestic animals were examined by polymerase chain reaction. LukS and lukF genes were detected in all 48 avian and 72 porcine isolates of S. aureus. LukE and lukD genes, located in a putative staphylococcal pathogenicity island (Sapln3/Saplm3), were recognized in 44 (91.7%) of 48 avian isolates, but these genes were not detected in porcine isolates. In 297 bovine isolates collected from mastitic cow's milk and bulk milk from dairy farms in two regions, lukM and lukF-PV(P83) genes in addition to lukS-lukF and lukE-lukD genes were detected in 100 (62.5%) of the 160 isolates from Ishikawa and in118 (86.1%) of the 137 isolates from Hokkaido. When the lysogeny of S. aureus bovine isolates was examined by treatment with mitomycin C, clearing of the culture due to cell lysis was observed in 34 (91.9%) of 37 lukM-lukF-PV(P83) genes--positive isolates. In addition, we isolated a novel lukM-lukF-PV(P83)-carrying (designated phiLukM), and revealed that the lukM-lukF-PV(P83) genes were located very close to an amidase gene on the temperate phage genomes. These results suggest horizontal transmission of lukM-lukF-PV(P83) genes by temperate bacteriophages in S. aureus of bovine origin.     

Surveillance of avian influenza viruses in Northern pintails (Anas acuta) in Tohoku District, Japan

Among winter migratory waterfowl, Northern pintails (Anas acuta), in one of the largest flocks in Tohoku district, northeast Japan, were surveyed for influenza A viruses at five wintering sites in three prefectures, viz., Aomori, Akita, and Miyagi. A total of 38 influenza A viruses were isolated from 2066 fecal samples collected during November 2006 through March 2007. The overall isolation rate was 1.84%. Eleven different subtypes were isolated, including nine H5N2, seven H6N8, seven H10N1, four H4N6, three H6N1, three H11N9, and one each of H1N1, H6N2, H6N5, H10N9, H11N1. Only the H4N6 subtype was detected during two successive months, November and December, from Lake Ogawara of Aomori prefecture. One wintering site, Lake Izunuma of Miyagi prefecture, was negative for virus isolation throughout the study period. During the sampling period, the highest virus isolation rate was in December (4.90%) followed by November (2.18%), January (0.91%), and February (0.30%). Virus isolation was negative for samples collected in March 2007. These results suggest that influenza viruses are introduced by Northern pintail when they migrate into Japan, but the viruses are not maintained in the flocks, most likely because the birds are not breeding during the winter. We believe that this relatively large data set creates a strong foundation for future studies of avian influenza virus (AIV) prevalence, evolution, and ecology in wintering sites, along with the role of Northern pintails in the spread of AIV during their migration from northern Russia and Asia to Japan.     

Distribution,body length,and abundance of blue shark and shortfin mako offshore of northeastern Japan,as determined from observed pelagic longline data, 2000–2014

Longline surveys have been conducted in the Northwest Pacific Ocean from 2000 to 2014 using chartered commercial longline vessels. Each year, two cruises were conducted offshore of northeastern Japan from mid‐April to mid‐June. For each longline set during the surveys, onboard scientists collected detailed biological information about the species caught, such as the size and sex, and recorded the catch numbers for all species. Blue shark (Prionace glauca) and shortfin mako (Isurus oxyrinchus) have eurythermal distributions, but the application of a generalized additive model (GAM) showed that the sea surface temperatures (SSTs) at catch sites positive for shortfin mako were warmer than those for blue shark. On the basis of the GAM, the probabilities of occurrence of both sharks differed by size category: small sharks had a narrower SST range than that of large sharks. Most catches of both sharks were juveniles, and the nominal catch rate of blue shark was more than 10 times that of shortfin mako. The standardized catch per unit effort (CPUE) for both species was calculated using a generalized linear model (GLM) with negative binomial errors, or a delta‐lognormal GLM. The standardized CPUE for blue shark in the second quarter of the year peaked in the mid‐2000s and then decreased, but it has been increasing since 2012. The CPUE for shortfin mako in the second quarter generally increased, with fluctuations.     

Tumor necrosis factor-alpha induces apoptosis in cultured porcine luteal cells

Some studies in mammalian species recently demonstrated that tumor necrosis factor (TNF)-alpha plays an important role for corpus luteum (CL) function by way of apoptosis during the estrous cycle. The objectives of this study were to clarify the induction of apoptosis in cultured porcine luteal cells by TNF-alpha treatment. Luteal cells prepared from porcine ovaries collected from crossbred mature gilts on Days 10-14 of the estrous cycle were isolated and examined as follows: 1) Flow cytometric analysis was carried out to determine apoptosis in cultured luteal cells. 2) Single-cell gel electrophoresis (comet assay) was performed to investigate apoptotic DNA fragmentation in luteal cells. The results of the flow cytometric analysis and comet assay demonstrated coincidentally that TNF-alpha induces DNA fragmentation in luteal cells causing apoptosis. These results revealed that TNF-alpha is an inducing factor of apoptosis in luteal cells.