Seasonal Changes in Midlife Women'S Percentage Body Fat: A 1-Year Cohort Study.

Objective: The purpose of this longitudinal, observational study was to examine whether age and seasonal changes in sedentary activity (sedAct), moderate-to-vigorous physical activity (MVPA), and energy intake (EI) predict changes in body composition among midlife women. We hypothesized that reductions in MVPA and increases in sedAct and EI in winter, along with greater baseline age would predict increases in percentage body fat (%BF) across seasons. Design: This study used a longitudinal, within-subjects design. Setting: This study took place in Grand Forks, North Dakota. Participants: Participants included 52 midlife women (aged 40-60 years) who were observed over the course of one year. Measurements: Percentage body fat measures were obtained via whole body Dual Energy X-ray absorptiometry. Participants were scanned once per season. We measured EI using the ASA24®. We used a GTX3 accelerometer to measure physical activity. Each season, participants wore the monitors for 7 days, 12 hours per day. All measures began in summer. Results: Results of hierarchical multiple regression (MR) analyses showed that age increases (ß = 0.310, p = 0.021) and summer-to-fall increases in EI (ß = 0.427, p = 0.002) predicted seasonal increases in %BF (R2 = .36, F(5, 42)= 4.66, p = 0.02). Changes in MVPA and sedAct were not significant predictors. Repeated measures ANCOVA revealed that summer (M = 37.7263, 95% CI [35.8377, 39.6149]) to winter (M = 38.1463, 95% CI [36.1983, 40.0942]) increases in %BF are not reversed by spring (M = 37.8761, 95% CI [35.9365, 39.8157]). Conclusions: To minimize increases in %BF and maintain health, midlife women, particularly older women, should be encouraged to pay extra attention to their diet in the fall months.

Theileria orientalis Ikeda genotype infection associated with anaemia, abortion and death in beef cattle in Western Australia.

BACKGROUND: Theileria orientalis infection causes a clinical syndrome in cattle characterised by weakness, reluctance to walk, anaemia, jaundice and death in peri-parturient cows and young calves, referred to as bovine anaemia caused by Theileria orientalis group (BATOG). Abortions in pregnant cows are also reported. Pallor, pyrexia and elevated heart and respiratory rates are typical findings on physical examination. CASE REPORT: A syndrome of abortions, lethargy, inappetence, jaundice and deaths in beef cattle on two separate properties and a separate cluster of three properties within 15 km west of the town of Denmark in Western Australia was associated with the presence of severe regenerative anaemia and the presence of Theileria orientalis Ikeda genotype in blood samples taken from affected cattle and their cohorts. A diagnosis of bovine anaemia caused by the T. orientalis group was based on consistent clinical, haematological, biochemical and PCR findings. Conventional PCR testing detected only the T. orientalis Ikeda type. On the two properties where it was investigated, quantitative PCR testing for parasite load was suggestive of recent infections. Sequencing of T. orientalis major piroplasm surface protein gene PCR products demonstrated that they were identical to those from similar bovine cases in New South Wales. CONCLUSION: The clinical history of affected cattle and the biochemical, haematological and PCR findings were consistent with bovine anaemia caused by the T. orientalis Ikeda genotype. This clinical syndrome had not been recognised in Western Australia before this series of cases.

Proficiency testing assessments for nematode worm egg counting based on Poisson variation.

The Poisson distribution provides an appropriate model for the variation within laboratories in worm egg counting. This is demonstrated by the results from annual quality assessment exercises in which laboratories in Australia tested multiple samples from the same mixtures prepared using different worm egg densities. Confidence intervals based on simulations using the Poisson distribution are recommended in the analysis of the results to identify laboratories showing significant bias or overdispersion, hence indicating possible procedural errors.

Evaluation of a copro-antigen ELISA to detect Fasciola hepatica infection in sheep, cattle and horses.

OBJECTIVE: To determine the specificity and sensitivity of a commercial copro-antigen ELISA for the detection of Fasciola hepatica infection in cattle and sheep and to assess the suitability of the test for use in horses. METHODS: Testing was done on more than 100 negative faecal samples from each of sheep, cattle and horses and on at least 40 positive faecal samples from each species. Positive samples were selected based on a positive sedimentation test for liver fluke eggs. Faecal samples of animals from Western Australia, which is free of liver fluke infection, served as negative controls. Specificity and sensitivity were assessed for each species using the recommended kit cut-off and also custom cut-offs specific for each species based on the mean plus 3-fold standard deviation of the mean of the negative samples for each species. RESULTS: Using the cut-off recommended by the kit manufacturer, the specificity was 100% for all species and the sensitivity was 88%, 80% and 9% for sheep, cattle and horses, respectively. Using the lower custom cut-offs for each species improved the sensitivity to 100% for sheep, 87% for cattle and 28% for horses, while maintaining the specificity above 99% for all species. CONCLUSIONS: The sensitivity of the commercial copro-antigen ELISA can be improved by using custom cut-off values for each species. With this modification, it is a suitable alternative screening test to the currently used sedimentation test for border control of sheep and cattle movement. The test is not suitable for use in horses.

Reduction of faecal worm egg count, worm numbers and worm fecundity in sheep selected for worm resistance following artificial infection with Teladorsagia circumcincta and Trichostrongylus colubriformis.

We examined the changes to populations of Trichostrongylus colubriformis and Teledorsagia circumcincta in mature sheep selected for reduced faecal worm egg count (WEC). Worm resistant (n=19) and control (n=10) genotype sheep were penned and dosed with a total of 10,000 T. colubriformis and 10,000 T. circumcincta per week for 18-weeks. Sheep genotypes were from lines previously bred over 15 years for either low WEC (resistant) or non-selected (control). Weekly WEC and the proportion of larvae from each species in faecal culture were measured during weeks 3-17. Egg hatchability was assessed on a pooled faecal sample from worm resistant or control genotype sheep at weeks 7, 9, 10, 13, 14 and 17. At week 18 the number of adult and immature worms (early and late 4th, and 5th stage), and indicators of worm fecundity (female worm length and number of eggs in utero) were assessed at necropsy. Results show that resistant sheep had reduced mean WEC to only 18% of the control (P<0.05) and increased the proportion of T. circumcincta larvae in faecal culture during weeks 8-17 (P<0.10). Egg hatch assays indicated a slight reduction in the viability of eggs from worm resistant genotypes at weeks 14 (P<0.05) and 17 (P<0.10). At necropsy, resistant animals had 93% fewer adult T. colubriformis, 44% fewer adult T. circumcincta and had reduced indicators of fecundity in T. circumcincta by up to 40% (P<0.05). We observed no change in the number T. circumcincta worms but an 11% increase in the proportion of early 4th stage T. circumcincta larvae in resistant animals (P<0.05). There were different temporal patterns in WEC and different prediction equations for WEC from necropsy traits for the two sheep genotypes (P<0.05). Thus, our results suggest a changed host-parasite relationship in sheep selected for low WEC. We conclude that lower WEC is achieved through reduced number of adult worms for both species and reduced fecundity for T. circumcincta. These results support the hypotheses that worm resistant sheep with a strong immune function can regulate T. colubriformis by rejecting adult worms but that T. circumcincta is regulated through a combination of suppressed development and reduced female fecundity.

Relationships between faecal dry matter, worm burdens and inflammatory mediators and cells in parasite-resistant Merino rams.

Immune-mediated scouring due to ingested parasite larvae is a major concern for sheep producers in Mediterranean climates. We investigated immune-mediated scouring in parasite-resistant Merino sheep in Australia. Forty-adult, parasite-resistant Merino rams were judged to be either susceptible or non-susceptible to immune-mediated scouring on the basis of dag scores taken under field conditions. We hypothesised that the susceptible rams would have lower faecal dry matter during larval challenge than non-susceptible rams and that, at post-mortem examination, inflammatory mediators and granulocytes would be negatively correlated with both faecal dry matter and worm numbers. In pens, the rams received a dose of 500 Teladorsagia circumcincta L(3) and 500 Trichostrongylus colubriformis L(3) each day for 6 weeks before euthanasia. Ten rams acted as unchallenged controls. Challenging sheep with larvae reduced faecal dry matter at 2, 3 and 4 weeks after challenge began and the greatest reductions were with the sheep susceptible to scouring. The sheep showed good resistance to the parasite challenge as evidenced by low faecal worm egg counts and low total worm counts at post-mortem, with the numbers of T. colubriformis particularly low. Sheep with low faecal dry matter had significantly higher numbers of eosinophils in small intestine tissue. Sheep with low total worm counts had significantly higher levels of bradykinin in abomasum mucus. Sheep with more granulocytes in tissue and inflammatory mediators in mucus tended to have fewer numbers of T. circumcincta but there was little relationship with numbers of T. colubriformis. Our results show that dag scores are correlated to a reduction in faecal dry matter, which can be attributed to the challenge with infective parasite larvae. Inflammation during worm infection is associated with rejection of the worm challenge and may result in more fluid faeces and consequently diarrhoea. Therefore, sheep breeders should focus on breeding for both low worm egg counts and also low dag scores.

Faecal dry matter, inflammatory cells and antibodies in parasite-resistant sheep challenged with either Trichostrongylus colubriformis or Teladorsagia circumcincta.

Sheep that are highly resistant to parasitic nematodes can suffer bad diarrhoea due to the inflammation associated with rejection of ingested larvae from pasture. We hypothesised that challenging parasite-resistant sheep indoors with nematode larvae would result in reduced faecal dry matter, and that faecal dry matter would be lowest in sheep challenged with a mixture of Trichostrongylus colubriformis and Teladorsagia circumcincta compared to those challenged with either species alone. We also hypothesised that inflammatory cells and serum antibodies and interleukin-5 would be highest in those sheep that received the mixed larval challenge. We found that faecal dry matter was reduced (P<0.05) in challenged sheep compared to unchallenged sheep, with the fastest reduction being in those sheep challenged with only Tric. colubriformis. At 14 and 23 days after challenge began, there were no differences in faecal dry matter between the three challenged groups. Within the abomasum, there were no differences in inflammatory cell numbers between unchallenged sheep and those challenged only with Tric. colubriformis. Cell numbers in sheep challenged with Tela. circumcincta were higher (P<0.05) than those in unchallenged sheep, but there were no differences between sheep challenged only with Tela. circumcincta or as a mixed challenge. In the small intestine, inflammatory cell numbers were higher (P<0.05) in sheep that received the mixed challenge compared to controls. Cell numbers in sheep challenged with either Tela. circumcincta or Tric. colubriformis were also slightly higher than those in controls. Larval challenge increased (P<0.05) levels of IgA and IgE in serum, but there were no differences between the three challenged groups. Larval challenge also increased (P<0.05) levels of IL-5, with the greatest increase being in those sheep challenged with both species. We concluded that both Tela. circumcincta and Tric. colubriformis can cause immune-mediated diarrhoea in sheep, and that a mixed challenge will not necessarily lead to worse diarrhoea or higher concentrations of antibodies in serum. We also concluded that challenge with Tric. colubriformis leads to no inflammation in the abomasum, but challenge with Tela. circumcincta may lead to some inflammation in the small intestine.

In vitro isolation and characterisation of the first canine Neospora caninum isolate in Australia.

Neospora caninum was isolated and established in vitro from the skin lesion of a naturally infected dog. The identity of the parasite was evaluated by immunofluorescent antibody test (IFAT), microscopy, Western blotting and polymerase chain reaction (PCR). N. caninum DNA was detected in the whole blood, serum, skin lesion, rectal scrapings and faeces of the infected dog utilising a nested PCR targeting the Nc-5 gene of N. caninum. Antigenic and genetic characterisation of the isolate, designated WA-K9, at a number of loci including the Nc-5 gene, heat shock protein 70 (HSP-70) gene, alpha-tubulin and beta-tubulin genes revealed no variation between this isolate and two N. caninum isolates from different geographic areas. Clinical aspects of this case, which included cutaneous and neurological disease, are also discussed.

Molecular epidemiology of Giardia and Cryptosporidium infections in dairy calves originating from three sources in Western Australia.

A longitudinal study was undertaken to determine the spread of naturally acquired Giardia and Cryptosporidium in dairy cattle in Western Australia. Samples were collected weekly from birth to weaning, and only calves that were sampled four or more times were included in the analysis. It was found that calves rapidly acquire infections with Cryptosporidium parvum and Giardia, with Cryptosporidium being the first to appear within 1-3 weeks after birth whereas Giardia was most prevalent in calves 4-7 weeks of age. Cryptosporidium and Giardia were detected in 48 and 89% of sampled calves, respectively. No significant association was observed in the present study between parasite occurrence and season or management practices (housing), and calf-to-calf contact appears to be the most likely source of transmission. Molecular characterization of isolates of Giardia and C. parvum recovered from calves demonstrated that only the occurrence Cryptosporidium represents a public health risk in terms of the potential for zoonotic transmission.

Intrahepatic cholestasis of pregnancy: a critical review.

Intrahepatic cholestasis of pregnancy (ICP) is a disease predominantly of the third trimester of pregnancy, characterized primarily by pruritus, biochemical disturbances in liver enzymes, and less frequently jaundice. Although maternal pruritus can be severe, overall maternal morbidity and mortality associated with ICP is low. However, fetal morbidity and mortality are significant with associated risks for meconium-stained amniotic fluid, acute onset of fetal compromise, spontaneous preterm labor, and intrauterine fetal demise. Current literature recommends obstetric management that includes frequent fetal surveillance with delivery when fetal lung maturity has been established.

Modulation of p53 expression in cultured colonic adenoma cell lines by the naturally occurring lumenal factors butyrate and deoxycholate.

The high incidence of colorectal cancer in Western society is believed to be strongly related to diet. Mutation of the p53 gene is a late event in colorectal carcinogenesis, and thus, the majority of pre-malignant adenomas express wild-type p53. As loss of p53 protein function is an important step in colorectal carcinogenesis, we investigated whether naturally occurring lumenal factors can modulate the expression of p53 in non-tumorigenic human colonic adenoma cell lines. Levels of p53 protein and mRNA were measured in adherent cells which had been incubated with growth-inhibitory concentrations of sodium butyrate (a by-product of dietary fibre fermentation) or sodium deoxycholate (a bile acid) for up to 48 hr. We report that both butyrate and deoxycholate can down-regulate the expression of wild-type and mutant p53. In contrast, incubation for 48 hr with the endogenous inhibitory growth factor TGFbeta1 did not alter p53 protein expression. Thus, in addition to cellular mechanisms which regulate p53 function, such as post-translational stabilisation, nuclear exclusion, negative feedback inhibition of p53 mRNA translation or binding of p53 by cellular proteins, p53 protein levels also may be regulated by changes in the level of p53 gene transcription. Furthermore, we show that lumenal factors are able to affect directly the expression of p53 protein in colonic epithelial cells.

Lectin staining of trichostrongylid nematode eggs of sheep: rapid identification of Haemonchus contortus eggs with peanut agglutinin.

The binding of 10 different lectins to the surface of trichostrongylid nematode eggs of sheep were investigated. Osage orange seed agglutinin bound to eggs from Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis, jack bean agglutinin to H. contortus and O. circumcincta eggs and peanut agglutinin to H. contortus eggs only. Staining of egg mixtures with fluorescein-labelled peanut agglutinin showed that the lectin correctly identified the percentage of H. contortus eggs present. Identification of eggs by lectin staining correlated well with results from larval differentiation. The findings suggest that peanut agglutinin can be used in routine diagnostic parasitology for the rapid identification of H. contortus eggs.

Changes in the phenotype of monocytes/macrophages and expression of cytokine mRNA in peripheral blood and synovial fluid of patients with rheumatoid arthritis.

Data from a previous study suggested that peripheral blood monocytes in patients with rheumatoid arthritis (RA) may be activated. Therefore, in this study we sought further evidence of 'presynovial' activation of monocytes. Our results show that phenotypic changes are demonstrable in peripheral blood monocytes in patients with RA, including increased expression of CR3 (CD11b/CD18) and FcRI (CD64). However, changes are most extensive in synovial monocytes/macrophages and especially for HLA-DR and intercellular adhesion molecule-1 (ICAM-1) (CD54). We conclude that monocyte/macrophage activation is most evident within the joint, and that 'presynovial' changes occur but are of limited extent.

Measurement of an eosinophil differentiation factor in sheep before and after infection with Trichostrongylus colubriformis.

A bioassay for the measurement of an ovine eosinophil differentiation factor (EDF) was developed. Recombinant murine interleukin-5 (rmIL-5) and supernatant from concanavalin A stimulated lymphocytes induced eosinophil differentiation and proliferation in ovine bone marrow cell culture, which was determined by an eosinophil peroxidase assay (EPO). The assay was used to measure the production of an ovine eosinophil differentiation factor by lymphocytes following experimental infection of sheep with Trichostrongylus colubriformis. Sixteen sheep were immunised and challenged with T. colubriformis and grouped into responders (n = 10) and non-responders (n = 6) based on the lower 99% confidence limit of the worm burden of the unimmunised control group (n = 10). Total worm burdens of all animals were established 10 days after initial challenge. Peripheral blood lymphocytes at Day 0 (pre-challenge values) and Day 9 (post-challenge values) from all sheep and lymphocytes of mesenteric lymph nodes from eight sheep were stimulated with third stage larval antigen of T. colubriformis. The resulting lymphocyte conditioned medium (LyCM) was tested for EDF by its ability to induce growth and differentiation of eosinophils in liquid bone marrow cell cultures. No significant difference was found between the animal groups. No correlation was found between EDF activity and total worm numbers.