Susceptibility of sheep to European bat lyssavirus type-1 and -2 infection: a clinical pathogenesis study

European bat lyssaviruses (EBLVs) have been known to cross the species barrier from their native bat host to other terrestrial mammals. In this study, we have confirmed EBLV-1 and EBLV-2 susceptibility in sheep (Ovis ammon) following intracranial and peripheral (intramuscular) inoculation. Notably, mild clinical disease was observed in those exposed to virus via the intramuscular route. Following the intramuscular challenge, 75% of the animals infected with EBLV-1 and 100% of those that were challenged with EBLV-2 developed clinical signs of rabies and then recovered during the 94-day observation period. Disease pathogenesis also varied substantially between the two viruses. Infection with EBLV-1 resulted in peracute clinical signs, which are suggestive of motor neuron involvement. Antibody induction was observed and substantial inflammatrory infiltrate in the brain. In contrast, more antigen was detected in the EBLV-2-infected sheep brains but less inflammatory infiltrate and no virus neutralising antibody was evident. The latter involved a more protracted disease that was behaviour orientated. A high infectious dose was required to establish EBLV infection under experimental conditions (> or =5.0 logs/ml) but the infectious dose in field cases remains unknown. These data confirm that sheep are susceptible to infection with EBLV but that there is variability in pathogenesis including neuroinvasiveness that varies with the route of infection. This study suggests that inter-species animal-to-animal transmission of a bat variant of rabies virus to a terrestrial mammal host may be limited, and may not always result in fatal encephalitis.     

Prevalence and clonal diversity of Campylobacter jejuni from dairy farms and urban sources

AIM: To investigate the role of free-living animals such as spar- rows, rodents and flies as potential reservoirs of Campylobacter spp on a dairy farm, and to assess the genetic diversity among Campylobacter isolates from the farm and an urban source.

METHODS: A total of 290 samples (bovine, passerine and ro- dent faeces, and whole flies) were collected from a large com- mercial dairy farm in the Manawatu district in New Zealand, and from faeces from urban sparrows in a nearby city. Other samples collected from the dairy farm included five from silage, two from aprons worn by workers during milking, two from workers' boots and two from water in troughs in a paddock. Isolates of thermophilic Campylobacter spp were identified mor- phologically and phenotypically and further characterised mo- lecularly using pulsed-field gel electrophoresis (PFGE) and the restriction enzyme SmaI.

RESULTS: Campylobacter jejuni was the only Campylobacter species isolated from all samples. The highest prevalence was found in faeces from dairy cows (54%), followed by faeces from sparrows from the urban area (40%) and the farm (38%), and from rodents (11%) and whole flies (9%). Other samples from the farm environment such as silage, trough water, and work- ers' aprons and boots were also positive for C. jejuni. Of the 22 restriction patterns obtained, seven were common to more than one source.

CONCLUSIONS: Cattle, sparrows, rodents and flies are po- tential reservoirs of C. jejuni on dairy farms. Identical clones of C. jejuni carried by cattle, sparrows, flies and rodents prob- ably indicate a common source of infection. The high level of asymptomatic carriage of C. jejuni by healthy dairy cows could be sufficient to maintain infections within the dairy farm sur- roundings via environmental contamination.     

Effect of setting a maximum milking time,from peak lactation,on production,milking time and udder health

Abstract

AIM: To examine the effect of setting a maximum milking time, from peak lactation until drying-off, on production, duration of milking, and udder health of dairy cows.

METHODS: Forty cows were assigned in twin-pairs to be either milked until cups were removed at a milk flow-rate threshold of 0.35 kg/minute (Control), or until cups were removed at a milk flow-rate threshold of 0.35 kg/minute, or maximum time, whichever came first (MaxT). The maximum time was set by determining the milking time of the 70th percentile cow when ranked from fastest to slowest, irrespective of yield. The milking routine was typical of that practised on dairy farms in New Zealand, and involved no pre-milking preparation. The study began at peak lactation (68 (SD 7) days in milk; DIM) and continued for 26 weeks. Duration of milking and milk yield were measured for each milking. Composition of milk was determined from weekly herd tests, and milk quality from fortnightly somatic cell counts (SCC). Completeness of milking and teat condition were assessed during the study. The bacterial status of quarter milk samples was determined at the beginning and end of the study, and all treated cases of clinical mastitis recorded. ANOVA was used to examine the effect of treatment group on variables of interest.

RESULTS: Total milk, fat and protein yields during the study period did not differ between treatments. On average, 30.3% of the morning and 27.6% of the afternoon milkings of MaxT cows reached the maximum time at which cups were removed, and were therefore shortened. While the average milking time of the slowest-milking cow was longer for the Control compared with MaxT group in Weeks 1–18, the average milking time did not differ between treatments. There was no difference in overall SCC, and the incidence of clinical mastitis, or the percentage of infected quarters at drying-off, was similar for the MaxT and Control cows.

CONCLUSION: The results show that setting a maximum milking time can reduce the milking time of slower-milking cows in a herd without compromising overall herd production and udder health.

CLINICAL RELEVANCE: Although the numbers of cows in the study were small there was no evidence of a major increase in SCC, or subclinical or clinical mastitis when a maximum milking time was set for slower-milking cows.     

The adventive flora of New Zealand

The ecological grouping (Decreaser and Increaser groups) of individual species was determined on two topographical units in the Highland Sourveld of the Orange Free State. For this purpose gradients were selected, giving special attention to fence‐line comparisons, vegetation at different distances from watering points and ungrazed vegetation. The ecological grouping (Decreaser and Increaser groups) of species on a grazing gradient differed not only for different topographical units, but also for sub‐habitats within a certain unit. Broad standardized groupings of species into Decreaser and Increaser groups are therefore scientifically wrong and of little practical use.     

Effect of meloxicam and butorphanol on minimum alveolar concentration of isoflurane in rabbits

OBJECTIVE: To determine the effects of meloxicam and butorphanol on minimum alveolar concentration of isoflurane (MAC(ISO)) in rabbits. ANIMALS: 10 healthy young adult female rabbits. PROCEDURE: Rabbits were anesthetized with isoflurane on 3 occasions in a blinded, randomized complete block design to determine the MAC(ISO) associated with administration of meloxicam (0, 0.3, or 1.5 mg/kg, PO) and butorphanol (0.4 mg/kg, IV). The MAC(ISO) was determined by use of a paw clamp technique as the end-tidal concentration of isoflurane halfway between the values that allowed or inhibited purposeful movement. Rectal temperature, end-tidal CO2 concentration, heart rate, oxygen saturation, and arterial blood pressure were measured to evaluate cardiopulmonary function. RESULTS: Mean +/- SE MAC(ISO) in saline (0.9% NaCl) solution-treated rabbits was 2.49 +/- 0.07% and was not significantly different from that associated with administration of meloxicam at 0.3 mg/kg (2.56 +/- 0.07%) or 1.5 mg/kg (2.66 +/- 0.07%). Butorphanol significantly reduced the MAC(ISO) to 2.30 +/- 0.07% when administered with saline solution alone, 2.27 +/- 0.07% when administered with 0.3 mg of meloxicam/kg, and 2.33 +/- 0.07% when administered with 1.5 mg of meloxicam/kg. The percentage reduction in MAC(ISO) was significantly greater for rabbits that received butorphanol and meloxicam at either dose, compared with butorphanol and saline solution. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that meloxicam does not have a direct isoflurane-sparing effect and does not interfere with the anesthetic-sparing effect of butorphanol in rabbits.     

Rapid vacuolar sequestration: the horseweed glyphosate resistance mechanism

BACKGROUND: Glyphosate‐resistant (GR) weed species are now found with increasing frequency and threaten the critically important GR weed management system. RESULTS: The reported ³¹P NMR experiments on glyphosate‐sensitive (S) and glyphosate‐resistant (R) horseweed, Conyza canadensis (L.) Cronq., show significantly more accumulation of glyphosate within the R biotype vacuole. CONCLUSIONS: Selective sequestration of glyphosate into the vacuole confers the observed horseweed resistance to glyphosate. This observation represents the first clear evidence for the glyphosate resistance mechanism in C. canadensis. Copyright © 2010 Society of Chemical Industry     

High-Tc superconductors in the two-dimensional limit:

The free modulation of interlayer distance in a layered high-transition temperature (high-Tc) superconductor is of crucial importance not only for the study of the superconducting mechanism but also for the practical application of high-Tc superconducting materials. Two-dimensional (2D) superconductors were achieved by intercalating a long-chain organic compound into bismuth-based high-Tc cuprates. Although the intercalation of the organic chain increased the interlayer distance remarkably, to tens of angstroms, the superconducting transition temperature of the intercalate was nearly the same as that of the pristine material, suggesting the 2D nature of the high-Tc superconductivity.     

Definition of the mutation responsible for maple syrup urine disease in Poll Shorthorns and genotyping Poll Shorthorns and Poll Herefords for maple syrup urine disease alleles.

The organisation of the E1alpha subunit of bovine branched-chain alpha-keto acid dehydrogenase gene was established. c DNA was cloned from Poll Shorthorn x Poll Hereford calves affected with Maple Syrup Urine Disease to identify the mutation responsible for the disease in Poll Shorthorns. Clones containing the c DNA sequences inherited from the Poll Shorthorn sire of the affected calves were identified. Paternal clones were sequenced and a cytidine to thymidine transition was found at nucleotide 1380. The mutation is predicted to substitute leucine in place of a highly conserved proline at codon 372. A polymerase chain reaction procedure was developed for detection of the 1380C-->T mutation in genomic DNA. Three Poll Shorthorn parents of affected calves and three affected Poll Shorthorn x Poll Hereford calves were heterozygous and an affected Poll Shorthorn calf was homozygous for this mutation. An improved polymerase chain reaction procedure was also devised to genotype Poll Herefords for the 248C-->T mutation. The procedures will facilitate disease prevention programs and assist in differential diagnosis of conditions in new-born calves that present with a rapid onset of progressive neurological disease and are characterised histologically by 'status spongiosus'. Maple Syrup Urine Disease (MSUD) is an autosomal recessive defect reported in humans (Danner and Elsas 1989), and in Poll Hereford (PH) and Poll Shorthorn (PS) calves (Harper et al 1986, Healy et al 1992). The clinical, biochemical and pathological manifestations of the disease are identical in the two breeds of cattle, and are characterised by the rapid onset of progressive neurological disease, leading to death within a few days of birth. The disease is caused by a deficiency of activity of the mitochondrial enzyme branched-chain alpha-keto acid dehydrogenase (BCKADH). This deficiency leads to elevated concentrations, in blood and tissues, of branched chain alpha-keto acids and their precursors, the branched chain amino acids, valine, leucine and isoleucine. BCKADH consists of four subunits E1alpha, E1beta, E2 and E3 that are encoded by separate genes, and MSUD may result from deficiency of any of the subunits. In PH s, the disease in caused by premature termination of translation, of the E1alpha subunit, that is induced by a cytidine to thymidine transition exon 2 (248C-->T), that converts the glutamine codon -6 to a stop codon (Q-6ST; Zhang et al 1990). We have shown that MSUD -affected PSxPH calves are heterozygous at the PH locus, illustrating molecular heterogeneity exists for bovine MSUD (Healy and Dennis 1994a). The fact that these crossbred calves are affected, indicates the PS, like the PH mutation, resides in the E1alpha subunit.     

Context-Dependent Viral Transgenerational Immune Priming in Honey Bees (Hymenoptera: Apidae)

Transgenerational immune priming is the process of increased resistance to infection in offspring due to parental pathogen exposure. Honey bees (Apis mellifera L. (Hymenoptera: Apidae)) are hosts to multiple pathogens, and this complex immune function could help protect against overwhelming infection. Honey bees have demonstrated transgenerational immune priming for the bacterial pathogen Paenibacillus larvae; however, evidence for viral transgenerational immune priming is lacking across insects in general. Here we test for the presence of transgenerational immune priming in honey bees with Deformed wing virus (DWV) by injecting pupae from DWV-exposed queens and measuring virus titer and immune gene expression. Our data suggest that there is evidence for viral transgenerational immune priming in honey bees, but it is highly context-dependent based on route of maternal exposure and potentially host genetics or epigenetic factors.