Back kinematics of healthy trotting horses during treadmill versus over ground locomotion

Reasons for performing study: Treadmill locomotion is frequently used for training of sport horses, for diagnostic purposes and for research. Identification of the possible biomechanical differences and similarities between the back movement during treadmill (T) and over ground (O) locomotion is essential for the correct interpretation of research results. Objectives: To compare the kinematics of the thoracolumbar vertebral column in treadmill and over ground locomotion in healthy horses. Methods: Six sound Dutch Warmblood horses trotted on a T and O during 10 s at their own preferred velocity (mean ± s.d. 3.6 ± 0.3 m/s T and 3.6 ± 0.1 m/s O), which was the same in both conditions. Kinematics of the vertebral column was captured by infrared cameras using reflective skin markers attached over the spinous processes of selected vertebrae and other locations. Flexion‐extension and lateral bending range of motion (ROM), angular motion pattern (AMP) and intravertebral pattern symmetry (IVPS) of 5 vertebral angles (T6‐T10‐T13, T10‐T13‐T17, T13‐T17‐L1, T17‐L1‐L3 and L1‐L3‐l5) were calculated. Neck angle, linear and temporal stride parameters and protraction‐retraction angles of the limbs were also calculated. Results: The vertical ROM (flexion‐extension) was similar in both conditions, but the horizontal ROM (lateral bending) of the lumbar angles T17‐L1‐L3 and L1‐L3‐L5 was less during T locomotion (mean ± s.d. difference of 1.8 ± 0.6 and 1.7 ± 0.9°, respectively, P>0.05). During O locomotion, the symmetry pattern of the lumbar vertebral angles was diminished from 0.9 to 0.7 (1 = 100% symmetry) indicating increased irregularity of the movement (P>0.05). No differences were found in the basic linear and temporal stride parameters and neck angle. Potential relevance: Vertebral kinematics during treadmill locomotion is not identical to over ground locomotion, but the differences are minor. During treadmill locomotion lumbar motion is less, and caution should be therefore taken when interpreting lumbar kinematics.     

In vitro evaluation of metacarpophalangeal joint loading during simulated walk

Reasons for performing study: Insight into the loading pattern of the articular cartilage surface during the complete stride is important as biomechanical factors play a pivotal role in the pathogenesis of joint trauma and osteoarthritis (OA). Objectives: To determine the loading pattern in the equine MCP articulation in vitro during simulated walk. Methods: Eight cadaveric limbs from mature Dutch Warmblood horses were loaded in a pneumatic loading device in 6 different positions (A1‐A6). The pressure distribution on the articular surface of the proximal phalanx (P1) was measured at 7 sites (S1‐7) using intra‐articularly placed pressure sensitive films, which were analysed by scanning and densitometry. Results: Pressures recorded after mid‐stance (A4, 5, 6) were significantly (P<0.05) higher than those before (A1, 2, 3) and showed the biphasic loading pattern of the walk at all sites, except for the site halfway along the sagittal groove (S7). At S7, there was a linear increase in pressure during the progress of the stance phase of the stride in most horses. Medially (S4, 5, 6) the pressure was significantly higher than laterally (S1, 2, 3) (P<0.05). Conclusions and potential relevance: The heavier medial loading coincides with the location where articular cartilage degeneration in the process of OA in the equine MCP joint is known to start. The discrepancy between the loading of the central groove and the other parts of the joint may result in large stress differences at the end of the stance phase, which might be related to the pathogenesis of stress fractures in the first phalanx and distal third metacarpal bone.     

Insecticidal activity of scorpion toxin (ButaIT) and snowdrop lectin (GNA) containing fusion proteins towards pest species of different orders

BACKGROUND: The toxicity of a fusion protein, ButalT/GNA, comprising a venom toxin (ButaIT) derived from the red scorpion, Mesobuthus tamulus (F.), and Galanthus nivalis agglutinin (GNA), was evaluated under laboratory conditions against several pest insects. Insecticidal activity was compared with SFI1/GNA, a fusion comprising a venom toxin (SFI1) derived from the European spider Segestria florentina (Rossi) and GNA, which has been previously demonstrated to be effective against lepidopteran and hemipteran pests, and to GNA itself. RESULTS: Injection assays demonstrated that both fusion proteins were toxic to lepidopteran larvae, dipteran adults, coleopteran adults and larvae and dictyopteran nymphs. ButalT/GNA was more toxic than SFI1/GNA in all cases. GNA itself made a minor contribution to toxicity. Oral toxicity of ButalT/GNA towards lepidopteran pests was confirmed against neonate Spodoptera littoralis (Boisd.), where incorporation at 2% dietary protein resulted in 50% mortality and > 85% reduction in growth compared with controls. ButaIT/GNA was orally toxic to Musca domestica L. adults, causing 75% mortality at 1 mg mL^?1 in aqueous diets and, at 2 mg g^?1 it was orally toxic to Tribolium castaneum (Herbst.), causing 60% mortality and a 90% reduction in growth. CONCLUSIONS: Toxicity of the ButaIT/GNA recombinant fusion protein towards a range of insect pests from different orders was demonstrated by injection bioassays. Feeding bioassays demonstrated the potential use of the ButaIT/GNA fusion protein as an orally active insecticide against lepidopteran, dipteran and coleopteran pests. These experiments provide further evidence that the development of fusion protein technology for the generation of new, biorational, anti‐insect molecules holds significant promise. © Crown Copyright 2009. Reproduced with permission of Her Majesty's Stationery Office. Published by John Wiley & Sons, Ltd.     

A Pressure Algometer Is a Useful Tool to Objectively Monitor the Effect of Diagnostic Palpation by a Physiotherapist in Warmblood Horses

There is a lack of scientific evidence for objective evaluation of neck and back musculoskeletal sensitivity in horses, although pressure algometry has been described as an objective tool to quantify musculoskeletal responses by mechanical nociceptive threshold (MNT) values. This study aimed to evaluate the use of pressure algometry for objectively quantifying the effect of diagnostic palpation applied by physiotherapists on the musculoskeletal function of the equine neck and back. The inter-examiner repeatability of animal physiotherapists was tested, and their subjective clinical scores for the vertebral column area were objectively compared with MNT values measured at the same locations to investigate the potential clinical implementation of the pressure algometer in daily equine rehabilitation practice. Six adult Dutch Warmblood riding school mares were randomly assigned to an experimental or a control group. The MNT of all horses was measured on 35 predefined sites on the vertebral column in the morning and in the evening of the same day. In the experimental group (n = 3), neck and back surface “temperature”, “pain”, “muscle tone”, and passive “mobility” were scored through palpation by three certified physiotherapists and related to MNT measurements at the same vertebral column locations. Agreement between the physiotherapists was determined from Spearman's rank correlation coefficients (P < .05). These correlation coefficients showed a significant agreement between the scores of individual physiotherapists and with objective MNT measurements. The three physiotherapists agreed best in their subjective gradings of “pain”, but less for “temperature” and “muscle tone”, and least for “mobility”. There was also a significant difference in MNT between individual horses. The physiotherapeutic diagnostic intervention did not significantly alter the MNT of the experimental group compared with the control group. There was a significant difference, however, between morning (7.4 kg/cm²) and evening (6.9 kg/cm²) MNT-measurements within the combined group (n = 6, P < .05). In conclusion, a pressure algometer proved to be a useful tool to objectively monitor the palpation of individual Warmbloods by individual physiotherapists. The correlation of their scores to the objective MNT measurements elucidated that there were differences on which scale (“pain”, “temperature”, “muscle tone”, “mobility”) they merely relied upon in their palpation. Significant effects of physiotherapeutic diagnostic palpation on MNT, however, were not found. The lower MNT of the horses at the second trial in the evening could be a sensitization of the measurement location because of bruising, a learning effect of the horses, or a diurnal fluctuation. The use of pressure algometry has both a potential to quantify clinical neck and back musculoskeletal sensitivity in horses possibly leading to dysfunction, as well as to objectively evaluate treatment results. Repeated measurements on the same day and on the same location along the vertebral column may influence absolute MNT values. The algometer can be used with success provided that the operator has proper and frequent training.     

Production of a novel 9,12-dihydroxy-10(E)-eicosenoic acid from eicosenoic acid by Pseudomonas aeruginosa PR3

Microbial conversions of unsaturated fatty acids often generate polyhydroxy fatty acids, giving them new properties such as higher viscosity and reactivity. A bacterial strain Pseudomonas aeruginosa (PR3) has been intensively studied to produce mono-, di-, and trihydroxy fatty acids from different 9-cis-monoenoic fatty acids such as oleic acid, ricinoleic acid, and palmitoleic acid. However, from the results and the postulated similar metabolic pathways involved in these transformations, it was assumed that the enzyme system involved in transformation of the monoenoic fatty acid by strain PR3 could utilize fatty acids with different chain lengths and locations of the double bond. In this study was used as a substrate for bioconversion by strain PR3 eicosenoic acid (C20:1, ω-9) containing a singular cis double bond at different positions from the carboxyl end as oleic acid, and it was confirmed that PR3 could produce a novel 9,12-dihydroxy-10(E)-eicosenoic acid (DED) with 6.2% yield from eicosenoic acid. The structure of DED was confirmed using GC-MS, FTIR, and NMR analyses. DED production was maximized at 72 h after the substrate was added to the 24 h culture. Some other nutritional factors were also studied for optimal production of DED.     

In vitro activity of isothiocyanates against Fusarium graminearum

Isothiocyanates are biotoxic degradation products formed as a result of enzymatic hydrolysis of glucosinolates present in Brassica species. The application of biofumigant Brassica crops, as an alternative crop protection method for soilborne pathogens and pests is increasingly gaining interest. However, little is known of the potential of biofumigation to reduce the inoculum of Fusarium species affecting cereals. The aim of this study was to evaluate the antifungal activity of five isothiocyanates, namely allyl, benzyl, ethyl, 2-phenylethyl and methyl isothiocyanates, against germination and growth of Fusarium graminearum under in vitro conditions. Aromatic isothiocyanates were more inhibitory than the aliphatic isothiocyanates against mycelial growth, whereas the reverse was observed for conidial germination. Among the tested isothiocyanates, allyl and methyl isothiocyanates were more efficient overall, showing lower ED₅₀ values (35–150 mg/L) for conidial germination and mycelial radial growth. The findings suggest that Brassica plants containing allyl and methyl glucosinolates could have a suppressive effect, reducing the inoculum of F. graminearum in soil prior to cereal production.     

Immunohistochemical Expression of Growth Factors in the Follicular Wall of Normal and Cystic Ovaries of Sows

The expression of growth factors was evaluated immunohistochemically in normal and cystic ovaries of sows. The immunohistochemically stained area (IHCSA) was quantified by image analysis to analyse the expression of these proteins in the follicular wall of secondary, tertiary and cystic follicles. IGF‐I immunoreactivity was strong in the granulosa cell layer (GC), moderate in the theca interna (TI) and mild in the theca externa (TE) of the normal follicles. There was severe reduction of the labelling to IGF‐I in the GC of the follicular and luteinized cysts. In the normal follicles, the reactivity for IGF‐II was very similar to pattern noted in IGF‐I. There was reduction of the IHCSAs in the GC of the follicular and luteinized cysts, but the decrease was not significant. The staining of the IGF‐II in the TI and TE of the cysts was increased, in comparison with normal follicles. The IHCSAs for VEGF were higher in the GC and TE of the normal follicles in contrast to TI, but this difference was noted only in the tertiary follicle. The VEGF reactivity increased in the GC of the cysts, in relation to normal follicles. The results of the current study show that the formation of ovarian cysts in sows is associated with alterations in the immunohistochemical expression of some growth factors.     

Analysis of the risk of introduction and spread of porcine reproductive and respiratory syndrome virus through importation of raw pigmeat into New Zealand

AIMS: To determine the frequency with which porcine reproductive and respiratory syndrome (PRRS) virus (PRRSv) would become established in a non-commercial pig herd in New Zealand due to illegal feeding of uncooked food waste containing virus-contaminated pigmeat. To determine the likelihood of a single incursion resulting in a multi-farm outbreak of the disease, and describe the spatio-temporal characteristics of such an outbreak.

METHODS: A Monte Carlo simulation model was constructed to determine the expected annual frequency of PRRSv infection being initiated in a non-commercial pig herd as a result of inadvertent feeding of pigmeat imported from countries endemically infected with the disease. Once the likelihood of PRRSv becoming established in a single pig herd was determined, stochastic spatially explicit infectious disease modelling software was utilised to model the temporal and spatial characteristics of the resulting epidemic.

RESULTS: Assuming the proportion of imported pigmeat remained at current levels, consumption patterns of pigmeat in households in New Zealand remained steady, and limited compliance with recently reintroduced regulations to prevent feeding of uncooked food waste, at least 4.3 pig herds per year were predicted to become infected with PRRSv. Simulation modelling of PRRSv epidemics related to initial infection of a non-commercial farm produced an estimate that 36% of these incursions would spread from the initial herd, and that these outbreaks would involve 93 herds on average in the first year. By increasing the estimated persistence of PRRSv infection in small herds, an average of 205 herds became infected in the first year.

CONCLUSIONS: Given a mean of 4.3 infected premises per year and a 36% probability of infection spreading beyond the initial infected herd, there was a 95% likelihood of a multi-farm PRRS outbreak occurring within 3 years.

CLINICAL RELEVANCE: Introduction of PRRSv through importation of virus-contaminated pigmeat presents a high risk for establishment of the disease in the pig industry in New Zealand.     

The effect of induced forelimb lameness on thoracolumbar kinematics during treadmill locomotion

REASONS FOR PERFORMING STUDY: Lameness has often been suggested to result in altered movement of the back, but there are no detailed studies describing such a relationship in quantitative terms. OBJECTIVES: To quantify the effect of induced subtle forelimb lameness on thoracolumbar kinematics in the horse. METHODS: Kinematics of 6 riding horses was measured at walk and at trot on a treadmill before and after the induction of reversible forelimb lameness grade 2 (AAEP scale 1-5). Ground reaction forces (GRF) for individual limbs were calculated from kinematics. RESULTS: The horses significantly unloaded the painful limb by 11.5% at trot, while unloading at walk was not significant. The overall flexion-extension range of back motion decreased on average by 0.2 degrees at walk and increased by 3.3 degrees at trot (P<0.05). Changes in angular motion patterns of vertebral joints were noted only at trot, with an increase in flexion of 0.9 degrees at T10 (i.e. angle between T6, T10 and T13) during the stance phase of the sound diagonal and an increase in extension of the thoracolumbar area during stance of the lame diagonal (0.7degrees at T13, 0.8 degres at T17, 0.5 degres at L1, 0.4 degrees at L3 and 0.3 degrees at L5) (P<0.05). Lameness further caused a lateral bending of the cranial thoracic vertebral column towards the lame side (1.3 degrees at T10 and 0.9 degrees at T13) (P<0.05) during stance of the lame diagonal. CONCLUSIONS: Both range of motion and vertebral angular motion patterns are affected by subtle forelimb lameness. At walk, the effect is minimal, at trot the horses increased the vertebral range of motion and changed the pattern of thoracolumbar motion in the sagittal and horizontal planes, presumably in an attempt to move the centre of gravity away from the lame side and reduce the force on the affected limb. POTENTIAL RELEVANCE: Subtle forelimb lameness affects thoracolumbar kinematics. Future studies should aim at elucidating whether the altered movement patterns lead to back and/or neck dysfunction in the case of chronic lameness.     

Comparative analysis of pathogenesis-related protein 10 (<Emphasis Type="Italic">PR10</Emphasis>) genes between fungal resistant and susceptible peppers

To elucidate the functional roles of PR10 genes from two pepper species during plant-pathogen interactions, PR10 genes were isolated from fungal-resistant (Capsicum baccatum var. PBC80) and fungal-susceptible (C. annuum var. Yeoju) pepper fruits infected with anthracnose fungus (Colletotrichum acutatum). Despite strong nucleotide sequence identity, there were significant differences in the patterns of gene expression and protein accumulation between the genes from the two host species. Induced expression of the PR10 mRNA in PBC80 (bacPR10) was highly maintained from 24 h after infection (HAI) rather than that in Yeoju (annPR10). These mRNA expression patterns were correlated with the level of respective protein that was detected as two or three bands in each species. Substantial induction of bacPR10 proteins was confirmed by 2D-gel analysis followed by immunoblotting. Immunolocalization study showed that deposition of bacPR10 was exclusively observed in the pericarp of PBC80 fruits after fungal infection, suggesting functional significance in defence. Additionally, in vitro analysis of the enzymatic properties of PR10 proteins revealed that recombinant bacPR10 had higher ribonucleolytic activity and exhibited less sensitivity to proteinase treatment than did annPR10. Taken together, these results support the idea that relative abundance and prolonged longevity of bacPR10 in PBC80 fruits may contribute to their increased resistance in response to the anthracnose fungus, as compared with Yeoju fruit.