Effects of repeated alcohol abstinence on within-subject prefrontal cortical gene expression in rhesus macaques.

Male rhesus monkeys (n = 24) had a biopsy of prefrontal cortical area 46 prior to chronic ethanol self-administration (n = 17) or caloric control (n = 7). Fourteen months of daily self-administration (water vs. 4% alcohol, 22 h access/day termed "open-access") was followed by two cycles of prolonged abstinence (5 weeks) each followed by 3 months of open-access alcohol and a final abstinence followed by necropsy. At necropsy, a biopsy of Area 46, contralateral to the original biopsy, was obtained. Gene expression data (RNA-Seq) were collected comparing biopsy/necropsy samples. Monkeys were categorized by drinking status during the final post-abstinent drinking phase as light (LD), binge (BD), heavy (HD) and very heavy (VHD drinkers). Comparing pre-ethanol to post-abstinent biopsies, four animals that converted from HD to VHD status had significant ontology enrichments in downregulated genes (necropsy minus biopsy n = 286) that included immune response (FDR < 9 × 10-7) and plasma membrane changes (FDR < 1 × 10-7). Genes in the immune response category included IL16 and 18, CCR1, B2M, TLR3, 6 and 7, SP2 and CX3CR1. Upregulated genes (N = 388) were particularly enriched in genes associated with the negative regulation of MAP kinase activity (FDR < 3 × 10-5), including DUSP 1, 4, 5, 6 and 18, SPRY 2, 3, and 4, SPRED2, BMP4 and RGS2. Overall, these data illustrate the power of the NHP model and the within-subject design of genomic changes due to alcohol and suggest new targets for treating severe escalated drinking following repeated alcohol abstinence attempts.

Six months of voluntary alcohol consumption in male cynomolgus macaques reduces intracortical bone porosity without altering mineralization or mechanical properties.

Chronic heavy alcohol consumption is a risk factor for low trauma bone fracture. Using a non-human primate model of voluntary alcohol consumption, we investigated the effects of 6 months of ethanol intake on cortical bone in cynomolgus macaques (Macaca fascicularis). Young adult (6.4 ± 0.1 years old, mean ± SE) male cynomolgus macaques (n = 17) were subjected to a 4-month graded ethanol induction period, followed by voluntary self-administration of water or ethanol (4 % w/v) for 22 h/d, 7 d/wk. for 6 months. Control animals (n = 6) consumed an isocaloric maltose-dextrin solution. Tibial response was evaluated using densitometry, microcomputed tomography, histomorphometry, biomechanical testing, and Raman spectroscopy. Global bone response was evaluated using biochemical markers of bone turnover. Monkeys in the ethanol group consumed an average of 2.3 ± 0.2 g/kg/d ethanol resulting in a blood ethanol concentration of 90 ± 12 mg/dl in longitudinal samples taken 7 h after the daily session began. Ethanol consumption had no effect on tibia length, mass, density, mechanical properties, or mineralization (p > 0.642). However, compared to controls, ethanol intake resulted in a dose-dependent reduction in intracortical bone porosity (Spearman rank correlation = -0.770; p < 0.0001) and compared to baseline, a strong tendency (p = 0.058) for lower plasma CTX, a biochemical marker of global bone resorption. These findings are important because suppressed cortical bone remodeling can result in a decrease in bone quality. In conclusion, intracortical bone porosity was reduced to subnormal values 6 months following initiation of voluntary ethanol consumption but other measures of tibia architecture, mineralization, or mechanics were not altered.

Impact of livestock hygiene education programs on mastitis in smallholder water buffalo (Bubalus bubalis) in Chitwan, Nepal.

A project implemented from 2003 to 2005 trained women in Chitwan District, Nepal, in hygienic dairy production using a process of social mobilization. The aim of this research was to assess if the prevalence of mastitis in water buffalo in the households of women who were trained was lower one year after training than in untrained households, if the training influenced knowledge and practices for the prevention or control of mastitis, and if these practices and knowledge were associated with a lower prevalence of mastitis. A total of 202 households from Eastern and Western Chitwan District were included in the study. Of these, 60 households had participated in the project and 142 had not. Milk samples were collected from 129 households (33 project households and 96 non-project households). Clinical mastitis was determined using visual inspection of udders and detection of macroscopic clots and flakes in milk. The California Mastitis Test was used to diagnose sub-clinical mastitis from milk samples, and the IDEXX SNAP test to identify the presence of tetracycline residues. The prevalence of mastitis in trained households (39.4%) was 43.78% of that in untrained households (60.4%), lower but not significantly so (p=0.08, 95% CI 0.17-1.12). Thirteen indicators of knowledge or practice for the control or prevention of mastitis were more likely to occur in trained households, four significantly so (not consuming milk from sick buffalo (p=0.001), using soap to wash hands before milking (p=0.001), discarding milk after antibiotic usage (p=0.01), and choosing appropriate flooring for their livestock (p=0.03)). Trained households that discarded milk from sick buffalo were 2.96 times more likely to have at least one animal with mastitis in the household (p=0.03, 95% CI 1.15-7.65). Trained households that knew to wash buffalos' teats after milking were less likely (OR 0.25) to have mastitis in their herd (p=0.02, 95% CI 0.08-0.80). Of the 138 buffalos tested, only one tested positive for tetracycline residues.