Depression, anxiety and PTSD symptoms before and during the COVID-19 pandemic in the UK.

BACKGROUND: The impact of the coronavirus disease 2019 (COVID-19) pandemic on mental health is still being unravelled. It is important to identify which individuals are at greatest risk of worsening symptoms. This study aimed to examine changes in depression, anxiety and post-traumatic stress disorder (PTSD) symptoms using prospective and retrospective symptom change assessments, and to find and examine the effect of key risk factors. METHOD: Online questionnaires were administered to 34 465 individuals (aged 16 years or above) in April/May 2020 in the UK, recruited from existing cohorts or via social media. Around one-third (n = 12 718) of included participants had prior diagnoses of depression or anxiety and had completed pre-pandemic mental health assessments (between September 2018 and February 2020), allowing prospective investigation of symptom change. RESULTS: Prospective symptom analyses showed small decreases in depression (PHQ-9: -0.43 points) and anxiety [generalised anxiety disorder scale - 7 items (GAD)-7: -0.33 points] and increases in PTSD (PCL-6: 0.22 points). Conversely, retrospective symptom analyses demonstrated significant large increases (PHQ-9: 2.40; GAD-7 = 1.97), with 55% reported worsening mental health since the beginning of the pandemic on a global change rating. Across both prospective and retrospective measures of symptom change, worsening depression, anxiety and PTSD symptoms were associated with prior mental health diagnoses, female gender, young age and unemployed/student status. CONCLUSIONS: We highlight the effect of prior mental health diagnoses on worsening mental health during the pandemic and confirm previously reported sociodemographic risk factors. Discrepancies between prospective and retrospective measures of changes in mental health may be related to recall bias-related underestimation of prior symptom severity.

Examining clinical correlates, treatment outcomes and mediators in young people with comorbid obsessive-compulsive disorder and autism spectrum disorder.

Despite the high comorbidity, surprisingly little is known about the clinical features, treatment prognosis, and treatment mediators for youth with Obsessive-Compulsive Disorder (OCD) and Autism Spectrum Disorder (ASD). This study, the largest to date, compared 172 young people with OCD and ASD (OCD + ASD) to 447 without ASD (OCD) on clinical characteristics, finding those with OCD + ASD were more likely to endorse poorer insight into their OCD, have greater global functional impairment, greater levels of concurrent psychopathology, higher levels of family accommodation and to be on medication. Treatment outcomes following a course of Cognitive Behaviour Therapy with or without medication were explored for a subgroup; 100 young people with OCD + ASD and 223 with OCD. Whilst both groups benefitted from treatment, the OCD + ASD group had significantly poorer treatment outcomes. Greater global functional impairment and being on medication mediated the between-group difference in outcomes. Further research and treatment refinements are needed to improve outcomes for youth with OCD + ASD.

Evidence for distinct genetic and environmental influences on fear acquisition and extinction.

BACKGROUND: Anxiety disorders are highly prevalent with an early age of onset. Understanding the aetiology of disorder emergence and recovery is important for establishing preventative measures and optimising treatment. Experimental approaches can serve as a useful model for disorder and recovery relevant processes. One such model is fear conditioning. We conducted a remote fear conditioning paradigm in monozygotic and dizygotic twins to determine the degree and extent of overlap between genetic and environmental influences on fear acquisition and extinction. METHODS: In total, 1937 twins aged 22-25 years, including 538 complete pairs from the Twins Early Development Study took part in a fear conditioning experiment delivered remotely via the Fear Learning and Anxiety Response (FLARe) smartphone app. In the fear acquisition phase, participants were exposed to two neutral shape stimuli, one of which was repeatedly paired with a loud aversive noise, while the other was never paired with anything aversive. In the extinction phase, the shapes were repeatedly presented again, this time without the aversive noise. Outcomes were participant ratings of how much they expected the aversive noise to occur when they saw either shape, throughout each phase. RESULTS: Twin analyses indicated a significant contribution of genetic effects to the initial acquisition and consolidation of fear, and the extinction of fear (15, 30 and 15%, respectively) with the remainder of variance due to the non-shared environment. Multivariate analyses revealed that the development of fear and fear extinction show moderate genetic overlap (genetic correlations 0.4-0.5). CONCLUSIONS: Fear acquisition and extinction are heritable, and share some, but not all of the same genetic influences.

Nutrient digestibility, rumen parameters, and (cannabinoid) residues in sheep fed a pelleted diet containing green hemp (Cannabis sativa L.) biomass.

The feeding value for ruminants of green hemp biomass, from the low Δ9-tetrahydrocannabinol (Δ9-THC) variety of Cannabis sativa L., is unknown. Twelve Merino ewes were individually penned and randomly allocated on a stratified liveweight basis to one of two pelleted dietary treatments, control (0% hemp, n = 6) or hemp (42% green hemp biomass, n = 6) that delivered a diet meeting the nutrient requirements of the animals. The experimental period consisted of 17 d dietary and housing adaptation, followed by 7 d total urine and feces collection for determination of apparent nutrient digestibility. A ruminal fluid sample was collected on day 27 and assessed for pH, ammonia, volatile fatty acid (VFA), and cannabinoid concentrations. A blood sample from the jugular vein and incisional subcutaneous fat biopsy from an area around the base of the tail were collected on day 28 with additional fat biopsies taken 35 d and 140 d post-feeding to measure cannabinoids. The dry matter (DM), organic matter (OM), and crude protein (CP) digestibilities, along with total VFA concentration did not differ (P = 0.713) between the two diets; however, acid detergent fiber (ADF) and neutral detergent fiber (NDF) digestibilities (P < 0.001), water intake (P = 0.023), and fecal water output (P < 0.001) were significantly lower for the sheep-fed Hemp. Rumen pH did not vary (P = 0.256) between diets, but ruminal ammonia concentration was significantly lower (P = 0.024) for sheep consuming Hemp. Sheep-fed Hemp had significantly greater molar proportions of butyric (P = 0.039) and hexanoic (P = 0.012) acids and lower molar proportions of propionic acid (P = 0.003). There were no differences between diets for N intake (P = 0.175), fecal N output (P = 0.253), and N balance (P = 0.695), with all sheep in positive N balance; however, there was significantly lower (P = 0.001) urinary N output for sheep-fed Hemp. Cannabidiolic acid (CBDA) and tetrahydrocannabinolic acid (THCA) were detected in plasma of all sheep-fed Hemp. ∆9-tetrahydrocannabinol was present in the subcutaneous fat of four of the six sheep on the final day of being fed Hemp, and in all (six) sheep 35 d post-feeding and one sheep 140 d post-feeding Hemp. No cannabinoids were detected in the corresponding samples taken from the sheep-fed Control. Thus, despite green hemp biomass being nutritionally a suitable feed for ruminants, under current Food Standards in Australia, the presence of these cannabinoid residues restricts its use in ruminant diets.

Enhancing performance of berseem clover genotypes with better harvesting management through farmers' participatory research at smallholder farms in Punjab.

A field study was conducted on smallholder farmer fields between 2012 to 2014 to evaluate the performance of cv. Agaitti Berseem-2002, against local landraces exchanged between farmers (LBF1) or available from local markets (LBM1). The effects of genotype and harvesting regimen on forage production, quality and seed production were evaluated. Significant differences (P < 0.05) among genotypes and cutting treatments were recorded for forage and seed yields, and forage quality across all research sites in both years. Maximum cumulative fresh forage (89.7 t/ha) and dry matter (DM; 13.4 t/ha) yields were obtained with Agaitti Berseem-2002 when harvesting occurred five times over the season. However, maximum seed yield (1048 kg/ha) with higher 1000-seed weight (3.63 g) were obtained if forage was only harvested three times and the crop then left for seed set. Agaitti Berseem-2002 also produced forage with the higher crude protein content (27%), DM digestibility (69%), digestible organic matter (DM basis; 65%) and metabolizable energy content (10%) compared to the local landraces (LBF1 and LBM1). Therefore, the harvesting regimen for greatest economic return which produced optimum fresh and DM forage yields of highest nutritive values and maximum seed yield, were comprised of taking three forage cuts (at 65, 110 and 150 days after sowing) prior to seed harvest.

Reciprocal links between anxiety sensitivity and obsessive-compulsive symptoms in youth: a longitudinal twin study.

BACKGROUND: Anxiety sensitivity, the tendency to fear the symptoms of anxiety, is a key risk factor for the development anxiety disorders. Although obsessive-compulsive disorder was previously classified as an anxiety disorder, the prospective relationship between anxiety sensitivity and obsessive-compulsive symptoms (OCS) has been largely overlooked. Furthermore, a lack of genetically informative studies means the aetiology of the link between anxiety sensitivity and OCS remains unclear. METHODS: Adolescent twins and siblings (N = 1,579) from the G1219 study completed self-report questionnaires two years apart assessing anxiety sensitivity, OCS, anxiety and depression. Linear regression models tested prospective associations between anxiety sensitivity and OCS, with and without adjustment for anxiety and depressive symptoms. A phenotypic cross-lagged model assessed bidirectional influences between anxiety sensitivity and OCS over time, and a genetic version of this model examined the aetiology of these associations. RESULTS: Anxiety sensitivity was prospectively associated with changes in OCS, even after controlling for comorbid anxiety and depressive symptoms. The longitudinal relationship between anxiety sensitivity and OCS was bidirectional, and these associations were predominantly accounted for by nonshared environmental influences. CONCLUSIONS: Our findings are consistent with the notion that anxiety sensitivity is a risk factor for OCS during adolescence, but also suggest that experiencing OCS confers risk for heightened anxiety sensitivity. The reciprocal links between OCS and anxiety sensitivity over time are likely to be largely mediated by nonshared environmental experiences, as opposed to common genes. Our findings raise the possibility that interventions aimed at ameliorating anxiety sensitivity could reduce risk for OCS, and vice versa.

Are punitive parenting and stressful life events environmental risk factors for obsessive-compulsive symptoms in youth? A longitudinal twin study.

BACKGROUND: Punitive parenting and stressful life events are associated with obsessive-compulsive symptoms (OCS). However, the lack of longitudinal, genetically-informative studies means it remains unclear whether these factors represent environmentally-mediated risks for the development of OCS. METHODS: Twins and siblings from the Genesis1219 study completed self-report questionnaires two years apart (Time 1: N = 2616, mean age = 15.0; Time 2: N = 1579, mean age = 17.0 years) assessing OCS, maternal and paternal punitive parenting, and dependent stressful life events. Multiple regression models tested cross-sectional and longitudinal associations between the putative environmental risk factors and obsessive-compulsive symptoms using: (a) individual scores; and (b) monozygotic twin difference scores. The aetiologies of significant phenotypic associations between putative risk factors and OCS were further examined using multivariate genetic models. RESULTS: At a phenotypic level, maternal and paternal punitive parenting and stressful life events were all associated with OCS both cross-sectionally and longitudinally. However, only stressful life events predicted the subsequent development of OCS, after controlling for earlier symptoms. Genetic models indicated that the association between life events and change in OCS symptoms was due to both genetic (48%) and environmental (52%) influences. Overall, life events associated with change in OCS accounted for 1.2% of variation in OCS at Time 2. CONCLUSIONS: Stressful life events, but not punitive parenting, predict OCS change during adolescence at a phenotypic level. This association exists above and beyond genetic confounding, consistent with the hypothesis that stressful life events play a causal role in the development of obsessive-compulsive symptoms.

Moderators and predictors of outcomes in telephone delivered compared to face-to-face cognitive behaviour therapy for paediatric obsessive-compulsive disorder: preliminary evidence from a non-inferiority RCT.

This study provides a preliminary exploration of factors which differentially predict treatment response to telephone-delivered cognitive behavioural therapy (TCBT) compared to face-to-face CBT (CBT) in a randomised non-inferiority controlled trial of 72 children (aged 11-18 years) with obsessive-compulsive disorder (OCD). Potential moderator variables, their interaction with treatment group (CBT, TCBT) and baseline levels of OCD severity were entered into separate regression models where the primary outcome measure was the post-intervention Children's Yale-Brown Obsessive-Compulsive Scale total score (CYBOCS). Separate regressions were also used to test associations between predictors and outcome controlling for pretreatment CYBOCS. Only pretreatment level of parent-rated child peer problems moderated the effects of the two interventions on CYBOCS severity at post-treatment. After controlling for baseline CYBOCS, only family accommodation rated by mothers predicted poorer outcomes in both groups. While CBT and TCBT may be equally effective for adolescents with OCD, the current results tentatively suggest that higher baseline level of peer problems strengthened the response to therapy for youth receiving TCBT and the predictor analyses reinforce the importance of directly addressing family accommodation during CBT for paediatric OCD regardless of delivery mode. Limitations of the current findings and directions for future work are discussed.

Prevalence and effect of Theileria orientalis infection in homebred calves in the Gloucester region of New South Wales, Australia.

OBJECTIVES: To examine the prevalence of Theileria orientalis in the Gloucester area of New South Wales and its effect on individual animals. METHODS: Blood samples (EDTA and clotted blood) were collected from a total of 55 calves and their dams from 6 properties over a 16-week period. A total of 202 and 190 blood samples were collected from the calves and dams, respectively, and were examined via blood film for the presence of intraerythrocytic T. orientalis piroplasms. Packed cell volume (PCV) was measured to determine infection resulting in anaemia. The presence of antibodies against the T. orientalis major piroplasm surface protein (MPSP) was tested using ELISA. RESULTS: The overall prevalence of T. orientalis infection in both dams and calves from all herds examined was 95%. Mean peak parasitaemia was observed in calves between 6 and 9 weeks of age (P = 0.051), coinciding with a decline in mean PCV. Only 3 (6%) of the blood samples collected from the dams were positive for Theileria-associated antibodies and no significant relationship (P > 0.05) was found between the presence of antibodies in the dams and PCV levels in the calves. There was no evidence that passive transfer of antibodies from dams to calves protected the calves against a decline in PCV. CONCLUSION: This study confirmed a high prevalence of low-level Theileria infection, but low MPSP seroconversion rates, in dams and calves in an area where the disease has been endemic for a number of years.

Differential rumination, intake, and enteric methane production of dairy cows in a pasture-based automatic milking system.

Proper performance monitoring of cows on pasture-based diets is crucial to inform nutritional recommendations that minimize undesirable effects of high ruminant CH4 emissions into the environment. The prediction of linkages between rumination patterns, methane emissions, and correlated production traits of cows in a pasture-based automatic milking system was tested. A previous 10-d baseline measurement of rumination activity by acoustic methodology of 156 Holstein-Friesian cows was used for frequency analysis of rumination time and identification of 2 treatment groups (n = 37 cows/group) represented by cows with consistently high (HR; 75th rumination percentile = 617.55 ± 81.37 min/d) or low (LR; 25th rumination percentile = 356.65 ± 72.67 min/d) rumination. The HR and LR cows were paired by nearest parity, days in milk, body weight (BW), and previous 10-d milk production, and within pairs randomly assigned to 1 of 2 experimental groups managed on a voluntary milking system with diets consisting of at least 75% pasture, plus concentrates. Animal traits, including rumination time, mass flux of CH4 (QCH4) and carbon dioxide (QCO2), milk production, and estimated dry matter intake according to individual QCO2 fluxes over a 22-d period were analyzed with repeated measure mixed models for a completely randomized design, structural equation modeling, and nonlinear regression. High rumination and methane was seen in older and heavier cows that had greater estimated dry matter intake and milk production. A consistent difference in rumination time and QCH4 across days was detected between HR and LR, even after adjustment for metabolic BW. Estimated dry matter intake had direct positive effects on rumination and QCH4, but no independent direct effect of rumination on QCH4 was detected. The LR cows produced more QCH4/milk, associated with lower milk, BW, concentrate intake, and greater activity at pasture. A typical dilution of maintenance effect on QCH4/milk was detected as a consequence of increasing milk yield and similar significant reduction of QCO2/milk. The results raise challenging questions regarding the rumination patterning of grazing dairy cows and alternatives to reduce ruminant methane emissions in grazing dairy cows.

Acute renal failure associated with Amaranthus species ingestion by lambs.

CASE REPORT: This report documents an outbreak of acute renal failure in a flock of 9-month-old White Suffolk-cross ewe lambs. The mortality rate was moderate but the morbidity was high. The lambs had grazed a wheat stubble paddock in which there were several weed species, including Amaranthus spp. CLINICAL SIGNIFICANCE: Renal failure in livestock has been reported internationally following the ingestion of Amaranthus spp. and although it has previously been suspected in Australia no cases have been documented. It is important for Australian veterinarians and farmers to realise the toxic potential of this group of plants.

Transformation obsessions in paediatric obsessive-compulsive disorder: Clinical characteristics and treatment response to cognitive behaviour therapy.

BACKGROUND AND OBJECTIVES: Transformation obsessions denote an under-reported symptom of Obsessive Compulsive Disorder (OCD), characterised by an excessive fear of turning into another person/object or acquiring unwanted characteristics. Relative to other OCD symptoms, little is known about the clinical presentation of transformation obsessions. The study aims to examine the clinical correlates and treatment prognosis of transformation obsessions in a paediatric OCD sample. METHODS: The sample consisted of 346 youths with a primary diagnosis of OCD. Patients with and without transformation obsessions were compared in terms of demographic and clinical characteristics, and CBT outcomes. RESULTS: 10% of the sample endorsed transformation obsessions. Patients with transformation obsessions were more likely to be boys, to be on augmented medication regimes, and to present with more severe obsessions at assessment. A factor analysis identified four major OCD symptom clusters, with transformation obsessions loading on a 'forbidden thoughts' factor alongside aggressive, sexual, and religious obsessions. No group differences in treatment outcomes were observed. LIMITATIONS: Limitations include the cross-sectional and retrospective nature of the study, the representativeness of our sample, and use of concomitant medication, among others. CONCLUSIONS: The study provides the first empirical evidence regarding phenomenological similarities and differences between paediatric OCD patients presenting with and without transformation obsessions. The findings suggest that transformation obsessions are best conceptualised as related to 'forbidden' obsessions and respond to exposure-based CBT.

Evaluation of the Brix refractometer as an on-farm tool for the detection of passive transfer of immunity in dairy calves.

OBJECTIVES: To investigate the accuracy of both the optical and digital Brix refractometers compared with radial immunodiffusion (RID) for determining the immunoglobulin G (IgG) concentrations in dairy calf serum. DESIGN: The experiment design was a cross-sectional survey of four dairy farms. Serum was sampled from 12 calves from each farm at approximately 48 hours of age. METHODS: Serum IgG concentrations of 48 calves were measured using RID and both types of Brix refractometer. RESULTS: IgG concentrations measured by Brix refractometer scores correlated with RID results: 0.74 and 0.71 for the digital and optical devices, respectively. The minimum Brix score that identified calf serum with success of passive immunity (>1000 mg/dL RID IgG) with 100% accuracy was 10% for both devices. The optical and digital devices performed similarly at identifying IgG concentrations in calf serum, with a concordance of 87%. CONCLUSION: Brix refractometer score ≥10% can be used to classify calves with successful transfer of passive immunity and the devices are sufficiently accurate for use as a simple, inexpensive on-farm tool for the monitoring of neonatal dairy calf immunity levels.

Paediatric Obsessive-Compulsive Disorder and Depressive Symptoms: Clinical Correlates and CBT Treatment Outcomes.

Depression frequently co-occurs with paediatric obsessive-compulsive disorder (OCD), yet the clinical correlates and impact of depression on CBT outcomes remain unclear. The prevalence and clinical correlates of depression were examined in a paediatric specialist OCD-clinic sample (N = 295; Mean = 15 [7 - 18] years, 42 % female), using both dimensional (Beck Depression Inventory-youth; n = 261) and diagnostic (Development and Wellbeing Assessment; n = 127) measures of depression. The impact of depressive symptoms and suspected disorders on post-treatment OCD severity was examined in a sub-sample who received CBT, with or without SSRI medication (N = 100). Fifty-one per-cent of patients reported moderately or extremely elevated depressive symptoms and 26 % (95 % CI: 18 - 34) met criteria for a suspected depressive disorder. Depressive symptoms and depressive disorders were associated with worse OCD symptom severity and global functioning prior to CBT. Individuals with depression were more likely to be female, have had a psychiatric inpatient admission and less likely to be attending school (ps < 0.01). OCD and depressive symptom severity significantly decreased after CBT. Depressive symptoms and depressive disorders predicted worse post-treatment OCD severity (ßs = 0.19 and 0.26, ps < 0.05) but became non-significant when controlling for pre-treatment OCD severity (ßs = 0.05 and 0.13, ns). Depression is common in paediatric OCD and is associated with more severe OCD and poorer functioning. However, depression severity decreases over the course of CBT for OCD and is not independently associated with worse outcomes, supporting the recommendation for treatment as usual in the presence of depressive symptoms.

Genetic and environmental influences on obsessive-compulsive behaviour across development: a longitudinal twin study.

BACKGROUND: Little is known about the factors influencing the stability of obsessive-compulsive behaviour (OCB) from childhood to adolescence. The current study aimed to investigate: (1) the stability of paediatric OCB over a 12-year period; (2) the extent to which genetic and environmental factors influence stability; and (3) the extent to which these influences are stable or dynamic across development. METHOD: The sample included 14 743 twins from a population-based study. Parental ratings of severity of OCB were collected at ages 4, 7, 9 and 16 years. RESULTS: OCB was found to be moderately stable over time. The genetic influence on OCB at each age was moderate, with significant effects also of non-shared environment. Genetic factors exerted a substantial influence on OCB persistence, explaining 59-80% of the stability over time. The results indicated genetic continuity, whereby genetic influences at each age continue to affect the expression of OCB at subsequent ages. However, we also found evidence for genetic attenuation in that genetic influences at one age decline in their influence over time, and genetic innovation whereby new genes 'come on line' at each age. Non-shared environment influenced stability of OCB to a lesser extent and effects were largely unique to each age and displayed negligible influences on OCB at later time points. CONCLUSIONS: OCB appears to be moderately stable across development, and stability is largely driven by genetic factors. However, the genetic effects are not entirely constant, but rather the genetic influence on OCB appears to be a developmentally dynamic process.

[Renal denervation for resistant hypertension without general anesthesia: advantage of a MEOPA morphine protocol. Preliminary experience]. / Dénervation rénale pour HTA réfractaire sans anesthésie générale : intérêt d'un protocole MEOPA Morphine. Expérience préliminaire.

UNLABELLED: Renal denervation using the technique of radiofrequency is used only recently for the treatment of resistant hypertension. Normally, it is done under general anesthesia because the ablation point technique is painful. We suggest an alternative to general anesthesia comprising an association of morphin 0.1mg/kg IV to MEOPA (gas combining oxygen and azot protoxyd) delivered through an oxygen mask. Our series includes 12 consecutive patients treated between October 2011 and June 2013, the first five patients (group 1) have received only an hydroxizin and morphin sedation. Every five have felt the ablation painful, in two cases bearable pain (EVA<5), in three cases intense (EVA>5) pain leading to increasing doses of morphin, (total dose of 0.25mg/kg in two cases, 0.17mg in one case). For the seven following patients, a protocol including hydroxyzin, morphin and MEOPA given through a mask has been set up. Only one patient has felt a mild pain (EVA 5) leading to an increasing dose of morphin (total dose 0.17mg/kg). None of the six other patients has felt any pain during the procedure. The average dose of morphin is 0.17mg/kg in group 1, 0.11mg/kg in group 2. This is a preliminary study; if confirmed, it will allow a lot of hospitals without on-site possibilities of general anesthesia, to realize such procedures. CONCLUSION: regarding pain, the procedure of renal ablation was well tolerated for six among seven patients receiving the association MEOPA and IV morphin. In contrast, in the five patients treated only with IV morphin, we observed a less good tolerance to pain and the need to increase the doses of IV morphin.

Using faecal DNA to determine consumption by kangaroos of plants considered palatable to sheep.

Disagreement exists within the scientific community with regards to the level of competition for feed between sheep and kangaroos in the Australian rangelands. The greatest challenge to solving this debate is finding effective means of determining the composition of the diets of these potential grazing competitors. An option is to adopt a non-invasive approach that combines faecal collection and molecular techniques that focus on faecal DNA as the primary source of dietary information. As proof-of-concept, we show that a DNA reference data bank on plant species can be established. This DNA reference data bank was then used as a library to identify plant species in kangaroo faeces collected in the southern rangelands of Western Australia. To enhance the method development and to begin the investigation of competitive grazing between sheep and kangaroos, 16 plant species known to be palatable to sheep were initially targeted for collection. To ensure that only plant sequences were studied, PCR amplification was performed using a universal primer pair previously shown to be specific to the chloroplast transfer RNA leucine (trnL) UAA gene intron. Overall, genus-specific, single and differently sized amplicons were reliably and reproducibly generated; enabling the differentiation of reference plants by PCR product length heterogeneity. However, there were a few plants that could not be clearly differentiated on the basis of size alone. This prompted the adoption of a post-PCR step that enabled further differentiation according to base sequence variation. Restriction endonucleases make sequence-specific cleavages on DNA to produce discrete and reproducible fragments having unique sizes and base compositions. Their availability, affordability and simplicity-of-use put restriction enzyme sequence (RES) profiling as a logical post-PCR step for confirming plant species identity. We demonstrate that PCR-RES profiling of plant and faecal matter is useful for the identification of plants included in the diet of kangaroos. The limitations, potential and the opportunities created for researchers interested in investigating the diet of competing herbivores in the rangelands are discussed.

Feeding grape seed extract to horses: effects on health, intake and digestion.

A feeding trial involving four Thoroughbred race horses was undertaken to establish whether inclusion of grape seed extract (GSE) in the diet of horses undergoing mild exercise had any effects on their general health, intake and digestion. Supplementation with GSE had no effect on either feed or water intake of the horses and the supplement was readily palatable to the horses at all levels of inclusion. Feeding GSE caused no adverse effects in terms of animal health (temperature, pulse and respirations rates), and there were some positive effects related to a presumed alteration in fermentation in the hindgut. Feeding GSE increased faecal pH, changing from acid faeces (pH 6.6) when no GSE was fed to neutral faeces (pH 7.0) when 150 mg GSE/kg body weight (BW) was fed. In addition, blood glucose concentrations were significantly (P < 0.05) decreased when GSE was fed at 100 and 150 mg/kg BW (5.50 ± 0.26 and 5.32 ± 0.72 mmol/l, respectively) compared with the control diet (5.77 ± 0.31 mmol/l). The actual mechanisms causing these alterations are yet to be elucidated, but could have important implications for the prevention of acidosis.

Effects of Lewisite on cell membrane integrity and energy metabolism in human keratinocytes and SCL II cells.

Lewisite is a highly toxic arsenic compound which can cause skin damage. In the present study effects of Lewisite on cell membrane integrity and energy metabolism as well as antidotal effects of DL-2,3-dimercaptopropanesulfonate (DMPS), and meso-2,3-dimercaptosuccinic acid (m-DMSA) were investigated in a keratinocyte derived cell line (SCL II) and primary human keratinocytes (HK). Cells were incubated in Lewisite (60 microM) containing medium for 5 min. During the following 6 h lactate dehydrogenase (LDH) activity in the supernatant, intracellular ATP content, tetrazolium reduction, glucose consumption and lactate formation were measured. Glucose consumption and lactate production were decreased in both cell lines after Lewisite exposure. In SCL II cells an increase of LDH activity in the supernatant, a decrease of ATP content, and an impaired ability to reduce tetrazolium was found 3 h after Lewisite exposure. In HK cultures tetrazolium reduction was significantly decreased already after 2 h, whereas LDH increase in the supernatant and ATP content decrease occurred only at 6 h after Lewisite exposure. When DMPS or m-DMSA was added directly after Lewisite exposure to SCL II cells, glucose consumption and lactate formation were restored and LDH leakage was prevented. SCL II cells might be more prone to membrane damage whereas in keratinocytes mitochondrial impairment seems to be the predominant effect of Lewisite.