Four viral genes independently contribute to attenuation of live influenza A/Ann Arbor/6/60 (H2N2) cold-adapted reassortant virus vaccines.

Clinical studies previously demonstrated that live influenza A virus vaccines derived by genetic reassortment from the mating of influenza A/Ann Arbor/6/60 (H2N2) cold-adapted (ca) donor virus with epidemic wild-type influenza A viruses are reproducibly safe, infectious, immunogenic, and efficacious in the prevention of illness caused by challenge with virulent wild-type virus. These influenza A reassortant virus vaccines also express the ca and temperature sensitivity (ts) phenotypes in vitro, but the genes of the ca virus parent which specify the ca, ts, and attenuation (att) phenotypes have not adequately been defined. To identify the genes associated with each of these phenotypes, we isolated six single-gene substitution reassortant viruses, each of which inherited only one RNA segment from the ca parent virus and the remaining seven RNA segments from the A/Korea/1/82 (H3N2) wild-type virus parent. These were evaluated in vitro for their ca and ts phenotypes and in ferrets, hamsters, and seronegative adult volunteers for the att phenotype. We found that the polymerase PA gene of the ca parent specifies the ca phenotype and that the PB2 and PB1 genes independently specify the ts phenotype. The PA, M, PB2, and PB1 genes of the ca donor virus each contribute to the att phenotype. The finding that four genes of the ca donor virus contribute to the att phenotype provides a partial explanation for the observed phenotypic stability of ca reassortant viruses following replication in humans.     

Cloprostenol inouced luteal regression in the beef cow. I. Ovarian response and endocrine changes

Forty-two cycling, multiparous beef cows (percentage-Brahman) were given two injections of 500 ug Cloprostenol (CLP) 11 days apart. Cows were randomly allocated to be ovariectomized at 0, 12, 24, 36, 48, 60 or 72 hr after the second CLP injection. Mean CL weight declined within 36 hr after CLP. Mean concentration of P4 in luteal tissue increased between o and 60 hr, while mean P4 content per CL declined by 12 hr after CLP. There was a precipitous decrease in mean serum P4 by 12 hr following CLP injection. Serum E2 was elevated until 24 hr and then declined through 72 hr after CLP. Follicular T concentration increased from 0 to 48 hr and then decreased by 60 hr. We conclude that CLP caused rapid diminution of luteal function which was accompanied by a reduction in P4 content but not in P4 concentration. Futhermore, the concentration of E2 in large follicles decreased by 72 hr post-CLP which is consistent with an alteration of the steroidogenic pathway in the periovulatory follicle.     

Old‐growth forests buffer climate‐sensitive bird populations from warming

Aim

Habitat loss and climate change constitute two of the greatest threats to biodiversity worldwide, and theory predicts that these factors may act synergistically to affect population trajectories. Recent evidence indicates that structurally complex old‐growth forest can be cooler than other forest types during spring and summer months, thereby offering potential to buffer populations from negative effects of warming. Old growth may also have higher food and nest‐site availability for certain species, which could have disproportionate fitness benefits as species approach their thermal limits.

Location

Pacific Northwestern United States.

Methods

We predicted that negative effects of climate change on 30‐year population trends of old‐growth‐associated birds should be dampened in landscapes with high proportions of old‐growth forest. We modelled population trends from Breeding Bird Survey data for 13 species as a function of temperature change and proportion old‐growth forest.

Results

We found a significant negative effect of summer warming on only two species. However, in both of these species, this relationship between warming and population decline was not only reduced but reversed, in old‐growth‐dominated landscapes. Across all 13 species, evidence for a buffering effect of old‐growth forest increased with the degree to which species were negatively influenced by summer warming.

Main conclusions

These findings suggest that old‐growth forests may buffer the negative effects of climate change for those species that are most sensitive to temperature increases. Our study highlights a mechanism whereby management strategies to curb degradation and loss of old‐growth forests—in addition to protecting habitat—could enhance biodiversity persistence in the face of climate warming.

     

Early Neolithic agriculture (2700–2000 <Emphasis Type="SmallCaps">bc</Emphasis>) and Kushan period developments (<Emphasis Type="SmallCaps">ad</Emphasis> 100–300): macrobotanical evidence from Kanispur in Kashmir,India

Here we report the results of an archaeobotanical study carried out on Ceramic Neolithic (2700–2000 bc) and Kushan period (ad 100–300) deposits at Kanispur, Kashmir, northern India. Botanical evidence from the Ceramic Neolithic phase reflects a nascent agriculture based on cold-tolerant crops (barley, wheats, lentil, field pea and grass pea) related to farming patterns of adjacent cultures and possibly evidence for the earliest cultivation in the valley. The Kushan period is characterized by a double-cropping pattern, suggesting a change in agricultural practices associated with a population recovery following a post-Neolithic decline. Finds of Juglans regia, and Prunus cf. amygdalus shells in ceramic Neolithic levels, and Vitis cf. vinifera, Emblica officinalis and Ziziphus cf. nummularia in addition to J. regia, and P. cf. amygdalus during Kushan times suggest that horticulture and foraging played an important role in the diet of the occupants. The present dataset provides new absolute dates for the subsistence economy of the Ceramic Neolithic population in the Kashmir Valley. We also present the oldest directly dated wheat and barley in Kashmir.     

Multi-criteria spatial identification of carnivore conservation areas under data scarcity and conflict: a jaguar case study in Sierra Nevada de Santa Marta,Colombia

Human–wildlife conflict, habitat loss, and prey hunting are the main threats to carnivore species worldwide. Forest conversion as consequence of deforestation and agricultural expansion increases the proximity between carnivores and humans, thereby escalating conflicts. Knowledge about carnivore species in data-poor countries, such as Colombia, is scarce which has the potential to result in poor landscape planning decisions. For many species, the only existing spatial information resides in expert-driven approaches which result in coarse-resolution ‘extent-of-occurrence’ maps. There is an increasing need for the development of methodologies to identify conservation and management areas at appropriate scales. Multi-criteria approaches will allow the inclusion of diverse species attributes enabling environmental institutions to address complex landscape decisions that result in conservation and management of carnivore habitat. We present a multi-criteria spatial identification tool for conservation and management areas, focused on Jaguars (Panthera onca) in the Sierra Nevada de Santa Marta, in northern Colombia. Our approach identifies areas based on the relationship between three spatial criteria: (1) suitable habitat patches, (2) habitat connectivity, and (3) zones of higher likelihood of human–jaguar conflict. We identified areas with the presence of at least one spatial criteria in 32% of the study area. Only 16.28% of these occur within protected areas (PAs) and the remaining fall on private lands (83.72%), either within (35.68%) or outside (48.04%) buffer zones of PAs. Our results highlight the need for multi-stakeholder collaborative approaches given that most proposed conservation areas fall on private rather than public lands.     

Evaluation of methods for the isolation and detection of Escherichia coli O157 in minced beef

This study has evaluated enrichment and detection procedures for the isolation and detection of Escherichia coli O157 inoculated into minced beef. The use of a 24 h enrichment in modified EC broth containing novobiocin allowed low numbers of contaminating cells to multiply to levels detectable on culture media and by ELISA test kits. Total analysis time was reduced by the use of the Dynabead^TM immunomagnetic separation system. The use of the Petrifilm^TM Test Kit-HEC for E. coli O157: H7 and Organon Teknika EHEC-TEK system detected low numbers of contaminating cells following enrichment and reduced analysis time by 1 d. The incorporation of cefixime and tellurite into Sorbitol MacConkey Agar increased the rate and ease of isolation of E. coli O157 and its use is therefore recommended.     

Cross-clade human immunodeficiency virus (HIV)-specific cytotoxic T-lymphocyte responses in HIV-infected Zambians.

We have examined cross-clade HIV-specific cytotoxic T-lymphocyte (CTL) activity in peripheral blood of eight Zambian individuals infected with non-B-clade human immunodeficiency virus type 1 (HIV-1). Heteroduplex mobility assay and partial sequence analysis of env and gag genes strongly suggests that all the HIV-infected subjects were infected with clade C HIV-1. Six of eight C-clade HIV-infected individuals elicited CTL activity specific for recombinant vaccinia virus-infected autologous targets expressing HIV gag-pol-env derived from B-clade HIV-1 (IIIB). Recognition of individual recombinant HIV-1 B-clade vaccinia virus-infected targets expressing gag, pol, or env was variable among the patients tested, indicating that cross-clade CTL activity is not limited to a single HIV protein. These data demonstrate that HIV clade C-infected individuals can mount vigorous HIV clade B-reactive CTL responses.     

Butterflies of the Cambrian benthos? Shield position in bradoriid arthropods

Mode of preservation and method of recovery strongly influences our understanding of the life habits of extinct organisms. Bradoriid arthropods were abundant, and diverse members of early Cambrian ecosystems and most life reconstructions display these animals with the two shields of the carapace open in a ‘butterfly’ configuration. This favoured reconstruction is largely based on the abundance of ‘crack‐out’ specimens preserved in this position (e.g. Kunmingella from the early Cambrian of China). In contrast, large collections of acid processed bradoriids from the Arrowie Basin of South Australia (Cambrian Stage 3) are preserved with a narrow gape at the ventral margin or completely closed with the carapace folded along the dorsal midline. The relative abundance of conjoined, closed (or partially closed) specimens from the lower Cambrian Hawker Group succession suggests that at least some bradoriid taxa were capable of withdrawing appendages and tightly closing the shields, challenging the common view that the majority of bradoriids usually held their carapaces open in a ‘butterfly’ configuration during life. New data show that layers of the bradoriid carapace are continuous through the dorsal fold with no evidence for complex articulating structures as in ostracod hinges. The relatively pliable, sclerotized or lightly mineralized calcium phosphate composition of the carapace and the simple, flexible dorsal fold facilitated opening and closing of the shields. Despite not being closely related, ostracods share close biomechanical and ecological similarities with bradoriids. The evolution of more complex articulating hinge structures – together with well‐developed musculature – in ostracods during the Early Ordovician, may have provided more efficient means for shield articulation and movement, thus promoting the ecological success of ostracods throughout the Phanerozoic.     

The kinematics of Heteroptera in free flight

The kinematics of six species of Heteroptera in free flight are analysed and compared.

• (1) 

  Using nested analysis of variance techniques, statistically significant variation was detected between species for several of the flight parameters measured: mean angular velocity; pronation/supination ratio; upstroke/downstroke ratio; and wing beat frequency. In each case this is discussed in terms of variation in flight behaviour.
• (2) 

  Beneficial aerodynamic forces are generated during the upstroke and the downstroke, in both fast forward and rising flight.
• (3). 

  When the insects change from level, forward flight to near vertical, rising flight, the following parameters are altered in most of the sequences analysed:

• (a). 

  the stroke plane angle becomes steeply, negatively inclined, associated with an increase in body angle;
• (b). 

  the stroke amplitude is reduced;
• (c). 

  wing beat frequency is lowered, associated with a drop in mean angular velocity;
• (d). 

  the speed of stroke reversal (rotational velocity) is increased. This may be associated with increased wing torsion and tip flexion which in turn could improve any beneficial unsteady aerodynamic effects generated at stroke reversal.

The reasons for this change in flight performance and the deviations from that seen in other insects are discussed.
It is shown that Heteroptera may make use of wing drag in flight, particularly during rising flight.