Production and in vivo testing of a recombinant bovine IL-12 as an adjuvant for Salmonella typhimurium vaccination in calves

A recombinant bovine interleukin-12 (boIL-12) that contains a histidine hexamer, rboIL-12His, was produced, purified and administered to calves. We first tried the purification of heterodimer IL-12 from a mixture of p40 homodimer, p40 monomer, and p40-p35 heterodimer with a p35 subunit tagged with a histidine hexamar at its C-terminal (p35His). A recombinant baculovirus expressing p35His was generated and used for superinfection with a recombinant baculovirus expressing p40 subunit. The expressed subunits, p40 and p35His, were assembled into a 70kDa heterodimer in insect cells, released into culture medium, and then purified using a nickel chelate column. The purified rboIL-12His was bioactive for induction of IFN-gamma in bovine peripheral blood mononuclear cells (PBMCs) in vitro.The purified rboIL-12His was then administered to calves with inactivated Salmonella Typhimurium (ST). When sera were assayed by ELISA, specific anti-ST IgG1 antibodies were detected in all ST immunized calves, but, specific anti-ST IgG2 antibodies were detected only in calves administered ST along with rboIL-12His, indicating a possible switch to a Th1 response. Administration of commercially available Salmonella vaccine did not elicit IgG2 antibodies in calves. These results suggest that co-administration of IL-12 with inactivated ST cells could induce a Th1-type response in calves.     

A new Badnavirus species detected in Bougainvillea in Brazil

Badnavirus in Bougainvillea spectabilis showing virus-like symptoms was identified by the presence of bacilliform particles, measuring 125–130 × 30–40 nm in leaf-dip preparations and by analysis of its putative open reading frame 3 sequence. The virus, tentatively named Bougainvillea bacilliform virus (BBV), had the highest identities (up to 60%) with Spiraea yellow leaf spot virus, Gooseberry vein banding associated virus, Taro bacilliform virus, and Citrus yellow mosaic virus. In phylogenetic analysis, BBV clustered with Badnavirus putative species. Attempts to transmit the virus to several hosts failed. This is the first report of a new Badnavirus detected in Bougainvillea.     

Starvation–refeeding causes cellular stress responses in the gut and liver of Masu salmon Oncorhynchus masou masou

Fisheries Science - Gene expression profiles during the transition from fasting to refeeding were investigated in the gut and liver of Masu salmon Oncorhynchus masou masou. Fish were starved for...     

Identification of enzyme genes in the liver of the Bleeker’s squid Loligo bleekeri by expressed sequence tag analysis

Expressed sequence tag (EST) analyses were performed with the aim of identifying enzyme genes in the liver of the Bleeker’s squid Loligo bleekeri. Of the 768 ESTs identified and sequenced, 669 were grouped into 324 clusters. Of these clusters, 123 comprising 245 ESTs were found to be homologous to genes reported to date. Among these, 43 clusters were annotated as enzymes according to the Enzyme Commission (EC) numbering system. Two EC groups, oxidoreductases and hydrolases, possessed a large number of ESTs. A cluster homologous to the glutathione peroxidase, an enzyme in the oxidoreductase group, contained 16 ESTs, which accounted for 2.4% of the total ESTs sequenced. There are three serine proteases, three cathepsins, two triacylgricerol lipases, and two chitinases among the clusters homologous to the enzymes in the hydrolase group. Since the squid liver functions in the digestive process, these enzymes would be involved in food digestion. Our data provide information on the various types of enzymes expressed in the squid liver and may provide a useful basis for further characterization of these enzymes.     

Gut bacterial community profile in Pacific white shrimp Litopenaeus vannamei following 5‐aminolevulinic acid supplementation

Changes in the diet have been previously reported to alter the gut bacterial profile in several organisms, including shrimp. These shifts in microbial structure either promote beneficial effects to the host or cause diseases. Supplementation of 5‐aminolevulinic acid (5‐ALA), the precursor of tetrapyrroles, has been previously reported to enhance shrimp's immune response. To know whether 5‐ALA has effects on the gut bacterial structure in Pacific white shrimp (Litopenaeus vannamei), we investigated the bacterial communities in the intestine and stomach of L. vannamei using high‐throughput Illumina sequencing of 16S rRNA gene libraries. One week of 5‐ALA supplementation altered the bacterial community structure (beta diversity) in both tissues, as shown by the results of multi‐dimensional scaling (MDS) plots and analysis of similarities (ANOSIM). DESeq2 analysis revealed enrichment of differentially abundant taxa in the 5‐ALA group (e.g. Enhydrobacter and Oceaniovalibus) and control group (e.g. Tenacibaculum and Mycobacterium). Metagenomic predictions suggested that the control group had more KEGG pathways associated with ‘metabolism’ than the 5‐ALA group. This study suggests that 5‐ALA supplementation potentially promotes the formation of a beneficial bacterial community structure in shrimp. This is the first report on the effect of 5‐ALA supplementation on the bacterial community profile in any organism.     

Relationship between endogenous protein disulfide isomerase family proteins and glutenin macropolymer

The effects of endogenous protein disulfide isomerase (PDI) family proteins on the properties of gluten proteins in dough during breadmaking were determined using bacitracin, an inhibitor of PDI. Bread loaf volume in the presence of bacitracin was increased to 118% of that in the absence of bacitracin. The addition of bacitracin caused a decrease in the extension tolerance of the dough. The amount of sodium dodecyl sulfate (SDS)-insoluble glutenin macropolymer (GMP) in dough decreased to approximately 70% of that in flour during the 20 min of mixing for doughmaking. The addition of bacitracin to dough caused a dramatic GMP decrease, corresponding to ～20-30% of that in flour during the 20 min of mixing. The decrease in GMP was compensated by an increase in SDS-soluble glutenin polymer. Taken together, these results suggest that the endogenous PDI family proteins in flour suppress the depolymerization of GMP during dough mixing.     

Estimating the annual supply potential and availability of timber and logging residue using forest management records of the Tochigi prefecture, Japan

We used forest management records to estimate the annual supply potential and availability of timber and logging residue from profitable subcompartments for all the cities and towns in the Tochigi prefecture. Five log markets and three factories in the Tochigi prefecture were assumed to be the destination of timber and logging residue, and the forest operation systems were set on the basis of interviews with forestry cooperative officials. The results showed that the annual supply potential of timber and logging residue was 450,304 m³ and 549,957 tons, of which 6 % (26,304 m³) and 37 % (204,122 tons) were from precommercial thinning operations, 61 % (276,180 m³) and 50 % (276,276 tons) were from commercial thinning operations, and 33 % (147,820 m³) and 13 % (69,559 tons) were from final-felling operations, respectively. When the unit price of the logging residue was 10,000 yen/ton, the annual logging residue availability from profitable subcompartments could almost cover the annual demand of the three facilities that we considered in this study. Introduction of feed-in tariffs had a significant impact. However, the ratios of the availability to supply potential with unit prices of logging residue of 3,000, 6,000, and 10,000 yen/ton were estimated to be only 1.67, 2.06, and 4.09 %, respectively. Considering the subsidies, the ratios respectively increased to 5.79, 7.35, and 13.09 %. Furthermore, the annual logging residue availability with subsidy could meet 70 % of the annual woody biomass demand of the large-scale factory in Sano city, which is 100,000 tons.     

Midbrain control of three-dimensional head orientation

Little is known about the neural mechanisms controlling head posture and why they fail in clinical syndromes like torticollis. It is well established, however, that the brain controls eye position by integrating eye velocity commands. By electrically stimulating and reversibly inactivating midbrain sites in the head-free (nonimmobilized) monkey, we found that the interstitial nucleus of Cajal functions as a neural integrator for head posture. We suggest that a bilateral imbalance in this structure, through either direct damage or inappropriate input, could be one of the mechanisms underlying torticollis.     

Atypical ocular Chelonoid herpesvirus manifestations in a captive Loggerhead turtle (Caretta caretta)

A captive loggerhead turtle (Caretta caretta) of unknown sex, 3 years of age, presented with bilateral mucoid secretions, severe chemosis, conjunctival hyperemia, and globe retraction. The animal was evaluated ophthalmologically and systemically, and hematological, microbiological, and conjunctival cytological and biopsy samples were collected for complementary diagnosis. The histopathological examination showed amphophilic intranuclear inclusions associated with severe inflammatory infiltrate. The diagnosis of Chelonid alphaherpesvirus 5 (ChAHV 5) was confirmed with end point PCR. Following systemic treatment with L‐lysine, acyclovir and vitamin A, the ocular signs resolved. No amphophilic intranuclear inclusions were seen in a follow‐up biopsy 5 months later, and there has been no recurrence of clinical ophthalmic signs during a 4‐year follow‐up. It is suggested that ChAHV 5 be considered as a differential diagnosis in captive marine turtles that present for conjunctival disease other than fibropapillomatosis.     

Effects of cooking on concentrations of polychlorinated dibenzo-p-dioxins and related compounds in fish and meat

We investigated the cooking-induced changes in concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (PCBs) (dioxins) using mackerel and beef. The concentrations of dioxins (29 congeners) were determined by isomer specific analyses and were compared between uncooked and cooked samples. The cooking procedures examined in this study included grilling as a fillet, boiling as a fillet, and boiling as tsumire (small, hand-rolled balls) for mackerel and boiling as a slice, broiling as a slice, and broiling as a hamburger for beef. Three trials were carried out for each cooking method. Generally, concentrations of dioxins were reduced in every cooking trial. When nondetected congener concentrations were assumed to be half the limit of detection for mackerel, the maximum percentage reductions of total concentrations given as 2,3,7,8-tetraCDD equivalents (TEQ) were 31% in grilling as a slice, 14% in boiling as a slice, and 21% in boiling as tsumire under the conditions of this study. In contrast, for beef, the reductions were 42% in boiling as a slice, 42% in broiling as a slice, and 44% in broiling as a hamburger. These results suggest that ordinary cooking processes with heating undoubtedly reduce the dioxin content in animal products, and the reductions estimated should be considered when dioxin intake is evaluated using contamination data for individual food items.