Comparison of three in vitro techniques to estimate benzimidazole resistance in Haemonchus contortus of sheep

Three in vitro assays to detect benzimidazole resistance, namely, the egg-hatch assay, tubulin-binding assay, and a larval-development assay, were evaluated by estimating the level of benzimidazole resistance in three field isolates of Haemonchus contortus compared with a susceptible reference strain. Comparisons were also made with estimates of benzimidazole resistance of the three field strains obtained from an in vivo controlled anthelmintic efficacy test. All three in vitro tests showed similar, consistent results which also suggested greater sensitivity than the in vivo assay. These results indicate that selection of an in vitro technique to determine benzimidazole resistance should therefore be based on considerations other than precision, such as technical expertise, availability of equipment, cost and speed in which diagnosis is required.     

Histopathology of Experimental Schistosoma bovis Infection in Goats

The inflammatory host response to Schistosoma bovis in young goats was studied at necropsy by light microscopy 34 weeks after primary exposure to 3,000 cercariae (group B, n=6), 34 weeks after primary exposure to 3,000 cercariae followed by challenge with 2,500 cercariae at week 17 (group C, n=5), and 17 weeks after primary exposure to 2,500 cercariae, given on week 17 of the experiment (group D, n=6). Three goats served as uninfected controls. The faecal egg output had been minimal for 17 weeks prior to necropsy in groups B and C and only for the last 2 weeks in group D.Histological studies were carried out on the small intestine, liver, lung and spleen, and tissue egg counts were performed. In sections of the small intestine and liver, a panel of histopathological variables were quantitated to characterize the host response and differences between groups of animals were evaluated with one way analysis of variance. The mean tissue egg count in the small intestine was slightly but not significantly higher in group C than group B and about twice as high in group D (D vs B or C p<0.01). Group means of numbers of inflammatory foci per section of gut wall corresponded well with those of tissue egg counts, suggesting that the rate of inflammatory destruction of eggs did not differ markedly between the groups. Egg material was less commonly seen in granulomas of the small intestine in group B than in group D (p<0.01), suggesting lower passage of eggs through the gut wall during the later than during the earlier phase of patent primary infection. The frequency of eosinophil-rich hepatic inflammatory foci was much higher in group D than in the other groups (D vs B p<0.05, D vs C p< 0.01), and coincided with a high degree of blood eosinophilia in this group at the time of sacrifice. Challenged goats showed a significantly higher frequency of markedly fibrotic inflammatory foci in the liver and of liver granulomas with a marked giant cell component than goats of the other groups. Hepatic portal fibrosis was least prominent in animals with 17- week- old primary infections, implying a possible relation between this change and duration of infection.     

Enzyme Immunoassays for the Determination of Ovine LH and FSH

The development of competitive enzyme immunoassays for ovine plasma LH (oLH) and FSH (oFSH) is described. Standards and plasma samples were preincubated with diluted antiserum to oLH or oFSH and the reacted solution (100 μl per well) was transferred to plates previously coated with oLH or oFSH, respectively. The second antibody used was anti‐rabbit IgG horseradish peroxidase. The measuring range was 0.39–50 ng/ml for each hormone and the 50% relative binding sensitivity was 9 ng/ml for oLH. The respective value for oFSH was 3.5 or 34 ng/ml with different hormone and antibody preparations used for the assay. The enzyme immunoassays were used to determine oLH and oFSH levels in plasma from ewes of two breeds during the oestrous cycle. The assays detected the first FSH surge coincident with the LH surge, the second FSH surge about 24 h later and the periodic fluctuations of FSH concentrations during the luteal phase of the oestrous cycle. These enzyme immunoassays are an efficient and economic alternative to the established radioimmunoassays (RIA) for oLH and oFSH.     

Optimum Soil Water Content for Chickpea Emergence in Heavy‐Textured Soils of North‐West Bangladesh

Sowing of chickpea in the heavy‐textured soils of north‐west Bangladesh with minimum tillage technology aims to increase the timely planting of large areas during a relatively short sowing window before soil water deficit limits germination and emergence. However, the seedbed conditions into which chickpea is sown need to be better quantified, so that limiting factors which affect germination and emergence can be identified. Two of the soil physical characteristics of importance are soil water and aeration. Growth cabinet studies have identified the fastest germination and emergence of chickpea on representative soils for this area at gravimetric water contents of 17–18 %, whilst soil water contents above and below this delayed germination and emergence. Emergence was recorded at soil water potentials between field capacity (?10 kPa) and wilting point (?1500 kPa). Emergence was possible at lower soil water potentials in the finer textured soil, whilst in coarser textured soil, emergence was still possible at higher soil water potentials.     

Cytokine mRNA profiles in pigs exposed prenatally and postnatally to Schistosoma japonicum

The pig is a natural host for Schistosoma japonicum and a useful animal model of human infection. The aim of the present study was to assess the differences between the cytokine profiles in prenatally or postnatally S. japonicum exposed pigs. Seven prenatally exposed pigs, 7 postnatally exposed pigs and 4 uninfected control pigs were compared 27 weeks post exposure. Variables included worm burdens, tissue egg counts, liver pathology and mRNA levels of IL-2, IL-4, IL-10, IL-12, TNF-alpha, TGF-beta1 and IFN-gamma in the liver and the caecum, assessed by RT-PCR. Infection intensity and level of septal fibrosis were significantly higher in the postnatal group compared to the prenatal group (P < 0.05). A significant increase of IL-4 (P < 0.01), IL-10 (P < 0.01), IL-12 (P < 0.01) and TNF-alpha (P < 0.05) mRNA level was also observed in the caecum of prenatally infected animals compared to the control group (P < 0.01). The prenatal group showed higher levels of TGF-beta1 in the liver compared with the postnatally infected group (P< 0.05) and the control group (P< 0.01). This suppressive immune response correlated with previously reported low hepatic pathogenesis in prenatally exposed pigs.     

Response of the pigeonpea-Rhizobium symbiosis to salinity stress: variation among Rhizobium strains in symbiotic ability

Summary There were significant differences among pigeonpea [Cajanus cajan (L.) Millsp] Rhizobium sp. strains (IC 3506, IC 3484, IC 3195, and IC 3087) in their ability to nodulate and fix N₂ under saline conditions. Pigeonpea plants inoculated with IC 3087 and IC 3506 were less affected in growth by salinity levels of 6 and 8 dS m^-1 than plants inoculated with the other strains. For IC 3506, IC 3484, and IC 3195, there was a decrease in the number of nodules with increasing salinity, while the average nodule dry weight and the specific nitrogenase activity remained unaffected. However, in IC 3087, the number of nodules increased slightly with increasing salinity. Leaf-P concentrations increased with salinity in the inoculated plants irrespective of the Rhizobium sp. strain, and leaf-N concentrations decreased with increasing salinity in IC 3484 and IC 3195 only. Shoot-Na and-Cl levels were further increased in these salt-sensitive strains only at 8 dS m^-1. Therefore there may be scope for selecting pigeonpea Rhizobium sp. symbioses better adapted to saline conditions. The Rhizobium sp. strains best able to form effective symbioses at high salinity levels are not necessarily derived from saline soils.Submitted as JA No. 919 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

Comparison of image and rapid field assessments of riparian zone condition in Australian tropical savannas

Suitable methods for measuring and monitoring the condition of riparian environments are being investigated by government agencies responsible for maintaining these environments in Australia. The objective of this work was to compare two riparian condition assessment approaches, the Tropical Rapid Appraisal of Riparian Condition (TRARC) method developed for rapid on-ground assessment of the environmental condition of savanna riparian zones and an image based riparian condition monitoring scheme. Measurements derived from these two approaches were compared and correlated. The sample representativeness of the TRARC method was evaluated and the cost-effectiveness and suitability for multi-temporal analysis of the two approaches were assessed. Two high spatial resolution multi-spectral QuickBird satellite images captured in 2004 and 2005 and coincident field data covering sections of the Daly River in the Northern Territory, Australia were used in this work. Both field and image data were processed to map indicators of riparian zone condition including percentage canopy cover, organic litter on the ground, canopy continuity, tree clearing, bank stability, and flood damage. Spectral vegetation indices, image segmentation, and supervised classification were used to produce riparian health indicator maps. QuickBird image data were used to examine if the spatial distribution of TRARC transects provided a representative sample of ground based estimates of riparian health indicators. Covering approximately 3% of the study area, the sample mean of the TRARC estimates of individual indicators of riparian zone condition were in most cases within 20% of the global mean derived from the whole imaged riparian area. The cost-effectiveness of the image based approach was compared to that of the ground based TRARC method. Results showed that the TRARC method was more cost-effective at spatial scales from 1 km to 200 km of river in relatively homogeneous riparian zones along rivers with only one channel, while image based assessment becomes more feasible at regional scales (200–2000 km of river). A change detection analysis demonstrated that image data can provide detailed information on gradual change, while the TRARC method is less suited for multi-temporal analysis due to the ranked data format, which inhibits precise detection of change. However, results from both methods were considered to complement each other for single date assessment of riparian zones if used at appropriate spatial scales.     

The Control of Reactive Oxygen Species Influences Porcine Oocyte In Vitro Maturation

The aim of this study was to examine the effect of varying intracellular reactive oxygen species (ROS) levels during oocyte in vitro maturation with enzymatic ROS production systems (xanthine + xanthine oxidase or xanthine + xanthine oxidase + catalase), scavenger systems (catalase or superoxide dismutase + catalase) or cysteine on porcine oocyte maturation. Oocyte ROS levels showed an increase when H₂O₂ or O₂?^‐ production systems were added to the culture medium (p < 0.05). On the other hand, the presence of ROS scavengers in the maturation medium did not modify oocyte ROS levels compared with the control after 48 h of maturation, but the addition of cysteine induced a decrease in oocyte ROS levels (p < 0.05). The ROS production systems used in this work did not modified the percentage of oocyte nuclear maturation, but increased the decondensation of sperm head (p < 0.05) and decreased the pronuclear formation (p < 0.05). In turn, the addition of O₂?^‐ and H₂O₂ scavenging systems during in vitro maturation did not modify the percentage of oocytes reaching metaphase II nor the oocytes with decondensed sperm head or pronuclei after fertilization. However, both parameters increased in the presence of cysteine (p < 0.05). The exogenous generation of O₂?^‐ and H₂O₂ during oocyte in vitro maturation would not affect nuclear maturation or later sperm penetration, but most of the spermatozoa cannot progress to form the pronuclei after fusion with the oocyte. The decrease in endogenous ROS levels by the addition of cysteine would improve pronuclear formation after sperm penetration.     

Herd diagnosis of low pathogen diarrhoea in growing pigs – a pilot study

Background

The major indication for antibiotic use in Danish pigs is treatment of intestinal diseases post weaning. Clinical decisions on antibiotic batch medication are often based on inspection of diarrhoeic pools on the pen floor. In some of these treated diarrhoea outbreaks, intestinal pathogens can only be demonstrated in a small number of pigs within the treated group (low pathogen diarrhoea). Termination of antibiotic batch medication in herds suffering from such diarrhoea could potentially reduce the consumption of antibiotics in the pig industry. The objective of the present pilot study was to suggest criteria for herd diagnosis of low pathogen diarrhoea in growing pigs.Data previously collected from 20 Danish herds were used to create a case series of clinical diarrhoea outbreaks normally subjected to antibiotic treatment. In the present study, these diarrhoea outbreaks were classified as low pathogen (<15% of the pigs having bacterial intestinal disease) (n =5 outbreaks) or high pathogen (≥15% of the pigs having bacterial intestinal disease) (n =15 outbreaks). Based on the case series, different diagnostic procedures were explored, and criteria for herd diagnosis of low pathogen diarrhoea were suggested. The effect of sampling variation was explored by simulation.

Results

The diagnostic procedure with the highest combined herd-level sensitivity and specificity was qPCR testing of a pooled sample containing 20 randomly selected faecal samples. The criteria for a positive test result (high pathogen diarrhoea outbreak) were an average of 1.5 diarrhoeic faecal pools on the floor of each pen in the room under investigation and a pathogenic bacterial load ≥35,000 per gram in the faecal pool tested by qPCR. The bacterial load was the sum of Lawsonia intracellularis, Brachyspira pilosicoli and Escherichia coli F4 and F18 bacteria per gram faeces. The herd-diagnostic performance was (herd-level) diagnostic sensitivity =0.99, diagnostic specificity =0.80, positive predictive value =0.94 and negative predictive value =0.96.

Conclusions

The pilot study suggests criteria for herd diagnosis of low pathogen diarrhoea in growing pigs. The suggested criteria should now be evaluated, and the effect of terminating antibiotic batch medication in herds identified as suffering from low pathogen diarrhoea should be explored.

Electronic supplementary material

The online version of this article (doi:10.1186/2046-0481-67-24) contains supplementary material, which is available to authorized users.