Peripubertal stress-induced behavioral changes are associated with altered expression of genes involved in excitation and inhibition in the amygdala.

Early-life stress is a critical risk factor for developing psychopathological alterations later in life. This early adverse environment has been modeled in rats by exposure to stress during the peripubertal period-that is, corresponding to childhood and puberty-and has been shown to lead to increased emotionality, decreased sociability and pathological aggression. The amygdala, particularly its central nucleus (CeA), is hyperactivated in this model, consistent with evidence implicating this nucleus in the regulation of social and aggressive behaviors. Here, we investigated potential changes in the gene expression of molecular markers of excitatory and inhibitory neurotransmission in the CeA. We found that peripubertal stress led to an increase in the expression of mRNA encoding NR1 (the obligatory subunit of the N-methyl D-aspartate (NMDA) receptor) but to a reduction in the level of mRNA encoding glutamic acid decarboxylase 67 (GAD67), an enzyme that is critically involved in the activity-dependent synthesis of GABA, and to an increase in the vesicular glutamate transporter 1 (VGLUT1)/vesicular GABA transporter (VGAT) ratio in the CeA. These molecular alterations were present in addition to increased novelty reactivity, sociability deficits and increased aggression. Our results also showed that the full extent of the peripubertal protocol was required for the observed behavioral and neurobiological effects because exposure during only the childhood/prepubertal period (Juvenile Stress) or the male pubertal period (Puberty Stress) was insufficient to elicit the same effects. These findings highlight peripuberty as a period in which stress can lead to long-term programming of the genes involved in excitatory and inhibitory neurotransmission in the CeA.

Detection of bovine herpesvirus type 4 antibodies and bovine lymphotropic herpesvirus in New Zealand dairy cows.

AIM: To detect the presence of bovine herpesvirus (BoHV) type 4 in New Zealand dairy cows with clinical metritis. METHODS: Serum samples taken from 92 dairy cows with clinical metritis, each from a different farm, were tested for the presence of antibodies against BoHV-4 using a commercially available, indirect ELISA. Peripheral blood mononuclear cells (PBMC) were collected from 10 BoHV-4 seropositive cows, and PBMC were examined by a pan-herpesvirus nested PCR to detect herpesvirus. PCR products were sequenced directly and a proportion of the PCR products were cloned and sequenced to identify the virus present. RESULTS: Antibodies to BoHV-4 were detected in 23/92 (25%) serum samples. The pan-herpesvirus PCR was positive in 8/10 PBMC samples. Cloning and sequencing identified that all of the eight PCR-positive PBMC contained bovine lymphotropic herpesvirus (BLHV); no BoHV-4 DNA was detected. CONCLUSIONS: This study reports the finding of the presence of apparent antibodies to BoHV-4, and BLHV DNA in New Zealand dairy cows affected by metritis. CLINICAL RELEVANCE: Bovine herpesvirus type 4 and BLHV are reported to have the potential to cause reproduction failure in cows. This is the first report of apparent BoHV-4 antibodies, and BLHV in New Zealand. The importance and epidemiology of these viruses in cattle in New Zealand requires further investigation.

Invited review: Systematic review of diagnostic tests for reproductive-tract infection and inflammation in dairy cows.

The objective of this study was to conduct a systematic and critical appraisal of the quality of previous publications and describe diagnostic methods, diagnostic criteria and definitions, repeatability, and agreement among methods for diagnosis of vaginitis, cervicitis, endometritis, salpingitis, and oophoritis in dairy cows. Publications (n=1,600) that included the words "dairy," "cows," and at least one disease of interest were located with online search engines. In total, 51 papers were selected for comprehensive review by pairs of the authors. Only 61% (n=31) of the 51 reviewed papers provided a definition or citation for the disease or diagnostic methods studied, and only 49% (n=25) of the papers provided the data or a citation to support the test cut point used for diagnosing disease. Furthermore, a large proportion of the papers did not provide sufficient detail to allow critical assessment of the quality of design or reporting. Of 11 described diagnostic methods, only one complete methodology, i.e., vaginoscopy, was assessed for both within- and between-operator repeatability (κ=0.55-0.60 and 0.44, respectively). In the absence of a gold standard, comparisons between different tests have been undertaken. Agreement between the various diagnostic methods is at a low level. These discrepancies may indicate that these diagnostic methods assess different aspects of reproductive health and underline the importance of tying diagnostic criteria to objective measures of reproductive performance. Those studies that used a reproductive outcome to select cut points and tests have the greatest clinical utility. This approach has demonstrated, for example, that presence of (muco)purulent discharge in the vagina and an increased proportion of leukocytes in cytological preparations following uterine lavage or cytobrush sampling are associated with poorer reproductive outcomes. The lack of validated, consistent definitions and outcome variables makes comparisons of the different tests difficult. The quality of design and reporting in future publications could be improved by using checklists as a guideline. Further high-quality research based on published standards to improve study design and reporting should improve cow-side diagnostic tests. Specifically, more data on intra- and interobserver agreement are needed to evaluate test variability. Also, more studies are necessary to determine optimal cut points and time postpartum of examination.