In vivo and in vitro genetic recombination between conventional and gene-deleted vaccine strains of pseudorabies virus

Pseudorabies virus (PRV), an alpha-herpesvirus, causes substantial economic losses in the swine industry and is currently the focus of eradication and control programs. Some of these programs rely on the ability of veterinarians to differentiate animals exposed to virulent strains of PRV from animals exposed to avirulent vaccine strains of PRV on the basis of a serologic response to nonessential glycoproteins that are deleted in some vaccine strains of PRV. Genetic recombination resulting in the creation of virulent strains of PRV with the same negative immunologic markers as vaccine strains could disrupt these programs. Two strains of PRV were coinoculated either into tissue culture or into sheep to facilitate recombination. Progeny viruses were selected to detect a specific recombinant phenotype. We were able to detect genetic recombination between vaccine strains of PRV following in vitro or in vivo coinoculation of 2 strains of PRV. The selected recombinants had marker-deleted phenotypes in strains with restored virulence genes. Increased virulence was observed in sheep after coinoculation of 2 avirulent vaccine strains of PRV.     

Endocrine systems and behavior

Estradiol, either directly from the ovary of females or as a metabolite of testosterone from the testes of males, seems to be the necessary and probably sufficient endocrine signal for sexual differentiation and adult sexual expression. Progesterone, in females, may play an important priming or facilitative role prior to estrus, especially following pregnancy, when blood estrogen levels are high for a prolonged period. At high doses, progesterone clearly inhibits the expression of sexual behavior.     

The Source and Fate of Massive Carbon Input During the Latest Paleocene Thermal Maximum

Lithologic, faunal, seismic, and isotopic evidence from the Blake Nose (subtropical western North Atlantic) links a massive release of biogenic methane approximately 55.5 million years ago to a warming of deep-ocean and high-latitude surface waters, a large perturbation in the combined ocean-atmosphere carbon cycle (the largest of the past 90 million years), a mass extinction event in benthic faunas, and a radiation of mammalian orders. The deposition of a mud clast interval and seismic evidence for slope disturbance are associated with intermediate water warming, massive carbon input to the global exogenic carbon cycle, pelagic carbonate dissolution, a decrease in dissolved oxygen, and a benthic foraminiferal extinction event. These events provide evidence to confirm the gas hydrate dissociation hypothesis and identify the Blake Nose as a site of methane release.     

Cloning and heterologous expression of the epothilone gene cluster

The polyketide epothilone is a potential anticancer agent that stabilizes microtubules in a similar manner to Taxol. The gene cluster responsible for epothilone biosynthesis in the myxobacterium Sorangium cellulosum was cloned and completely sequenced. It encodes six multifunctional proteins composed of a loading module, one nonribosomal peptide synthetase module, eight polyketide synthase modules, and a P450 epoxidase that converts desoxyepothilone into epothilone. Concomitant expression of these genes in the actinomycete Streptomyces coelicolor produced epothilones A and B. Streptomyces coelicolor is more amenable to strain improvement and grows about 10-fold as rapidly as the natural producer, so this heterologous expression system portends a plentiful supply of this important agent.     

An evaluation of anthelmintic properties,assessed using faecal nematode egg counts,of New Zealand native flax (Phormium tenax)

AIM: To investigate the anthelmintic properties of New Zealand native flax (Phormium tenax) for cattle, using a faecal egg count reduction test (FECRT).

METHODS: Twenty-six heifer calves with high (>300 epg) faecal nematode egg counts (FEC) were paired into two groups and fed either chopped flax leaves ad libitum, or barley straw at an equivalent dry matter (DM) rate, from Day 0–7; all were supplemented with 1.5 kg/head/day cereal feed supplement. On Days 8–10, all heifers were fed a common diet of 3.5 kg DM/ head/day barley straw and 1.5 kg/head/day cereal feed supplement. Heifers were weighed and faecal samples were collected on Days —6, 0, 7, and 10; samples were analysed in triplicate for FEC. The nutritive value of the flax and straw was analysed.

RESULTS: Both the flax and straw had low dry matter digestibility (DMD) and protein content. Although the flax-fed claves grew more than the controls, this may have been related to gut fill. Total daily faecal egg output was similar for flax and straw groups on Day 7 (14.7 vs 15.0 x 10⁶ eggs/day, respectively) and Day 10 (14.9 vs 15.1 x 10⁶ eggs/day, respectively). There was no difference in the change in FEC with time between the calves fed flax or straw diets.

CONCLUSION: Consumption of flax leaves did not reduce FEC in calves with a mixed nematode infection.     

GM2 Gangliosidosis in Shiba Inu Dogs with an In‐Frame Deletion in HEXB

Consistent with a tentative diagnosis of neuronal ceroid lipofuscinosis (NCL), autofluorescent cytoplasmic storage bodies were found in neurons from the brains of 2 related Shiba Inu dogs with a young‐adult onset, progressive neurodegenerative disease. Unexpectedly, no potentially causal NCL‐related variants were identified in a whole‐genome sequence generated with DNA from 1 of the affected dogs. Instead, the whole‐genome sequence contained a homozygous 3 base pair (bp) deletion in a coding region of HEXB. The other affected dog also was homozygous for this 3‐bp deletion. Mutations in the human HEXB ortholog cause Sandhoff disease, a type of GM2 gangliosidosis. Thin‐layer chromatography confirmed that GM2 ganglioside had accumulated in an affected Shiba Inu brain. Enzymatic analysis confirmed that the GM2 gangliosidosis resulted from a deficiency in the HEXB encoded protein and not from a deficiency in products from HEXA or GM2A, which are known alternative causes of GM2 gangliosidosis. We conclude that the homozygous 3‐bp deletion in HEXB is the likely cause of the Shiba Inu neurodegenerative disease and that whole‐genome sequencing can lead to the early identification of potentially disease‐causing DNA variants thereby refocusing subsequent diagnostic analyses toward confirming or refuting candidate variant causality.     

Growth,morphometrics and nutrient content of farmed eastern oysters,Crassostrea virginica (Gmelin), in New Hampshire,USA

When harvested, oysters represent a removal from the ecosystem of nutrients such as nitrogen (N) and carbon (C). A number of factors potentially affect nutrient content, but a quantitative understanding across the geographical range of the eastern oysters is lacking. This study was designed to quantify the relationships among various metrics of farmed eastern oysters near its northern geographical range focusing on nutrient content. Hatchery‐reared oysters were deployed in polyethylene bags at six sites, and were measured on multiple occasions from 2010 to 2012. A quadratic polynomial fit to the combined datasets for shell height indicated that on average a ‘cocktail’ size oyster (63 mm shell height) would be reached after 2 year, and ‘regular’ size (76 mm) would require 3 year. There were significant differences in growth rates and oyster nutrient content among the sites; means for %N in soft tissue ranged from 6.9 to 8.6, and 0.07 to 0.18 in shell. Per cent N in soft tissue and shell were highest at two sites at the mouths of rivers with elevated dissolved inorganic N concentrations in the water. Grand means (all sites, seasons and years combined) of soft tissue N and C for regular size oysters were 7.3% and 38.5%, respectively; and for shell N and C were 0.13% and 12.0% respectively. Our study extends the range of data on nutrient content of the eastern oyster to northern New England, and indicates that oyster size, seasonality, and nutrient concentration in ambient water potentially affect %N and %C content of oysters.