A method for intracerebroventricular cannulation of young broiler chicks.

The site-specific administration of neuroactive substances or pharmacological agents is a routine practice in neurological studies. To facilitate chronic treatments intercranial cannulae are often implanted in the skull. These surgical procedures are widely described and well-documented for rodents, as the most common animal model, and even refined over the years. However, precise descriptions of proper procedure for third ventricle cannulation in young broiler chicks is not readily available. This absence is surprising, as chicken models are often used in the investigation of physiological control of the metabolism and ingestion. Furthermore, as a commercial animal, there is also a particular interest in elucidating the central regulation of feed intake and metabolism to optimize feeding and living conditions for broilers. The brain nuclei that serve as the regulatory centers of feeding and metabolism are the arcuate nucleus, the ventromedial hypothalamic nucleus, and the lateral hypothalamus, which are all located near the third ventricle. Here, we provide a full protocol for intracerebroventricular (i.c.v.) cannulation of 7-day old broiler chicks, as well as a review of some of the advances made in rodent i.c.v. cannulations and whether these advances are applicable to cannulations in chickens. Using the surgical procedure described here, we were able to achieve a success rate of proper implantation of 88%, with a mortality rate of less than 1% (n = 224).•Detailed procedure for i.c.v. canula implantation in the third ventricle of 7d-old broiler chicks.•Cement cap with anchoring screws is necessary in broilers, glue does not provide sufficient stability.•Carboxylate cement and self-adhesive resin cement were tested as an alternative for methyl methacrylate cement.

Genomic Regions Associated with IgE Levels against Culicoides spp. Antigens in Three Horse Breeds.

Insect bite hypersensitivity (IBH), which is a cutaneous allergic reaction to antigens from Culicoides spp., is the most prevalent skin disorder in horses. Misdiagnosis is possible, as IBH is usually diagnosed based on clinical signs. Our study is the first to employ IgE levels against several recombinant Culicoides spp. allergens as an objective, independent, and quantitative phenotype to improve the power to detect genetic variants that underlie IBH. Genotypes of 200 Shetland ponies, 127 Icelandic horses, and 223 Belgian Warmblood horses were analyzed while using a mixed model approach. No single-nucleotide polymorphism (SNP) passed the Bonferroni corrected significance threshold, but several regions were identified within and across breeds, which confirmed previously identified regions of interest and, in addition, identifying new regions of interest. Allergen-specific IgE levels are a continuous and objective phenotype that allow for more powerful analyses when compared to a case-control set-up, as more significant associations were obtained. However, the use of a higher density array seems necessary to fully employ the use of IgE levels as a phenotype. While these results still require validation in a large independent dataset, the use of allergen-specific IgE levels showed value as an objective and continuous phenotype that can deepen our understanding of the biology underlying IBH.

Short communication: Sensitivity of estrus alerts and relationship with timing of the luteinizing hormone surge.

Both the sensitivity of an estrus detection system and the consistency of alarms relative to ovulation determine its value for a farmer. The objective of this study was to compare an activity-based system and a milk progesterone-based system for their ability to detect estrus reliably, and to investigate how their alerts are linked to the time of the LH surge preceding ovulation. The study was conducted on an experimental research farm in Flanders, Belgium. The activity alerts were generated by a commercial activity meter (ActoFIT, DeLaval, Tumba, Sweden), and milk progesterone was measured using a commercial ELISA kit. Sensitivity and positive predictive value of both systems were calculated based on 35 estrus periods over 43 d. Blood samples were taken for determination of the LH surge, and the intervals between timing of the alerts and the LH surge were investigated based on their range and standard deviation (SD). Activity alerts had a sensitivity of 80% and a positive predictive value of 65.9%. Alerts were detected from 39 h before until 8 h after the LH surge (range: 47 h, SD: 16 h). Alerts based on milk progesterone were obtained from a recently developed monitoring algorithm using a mathematical model and synergistic control. All estruses were correctly identified by this algorithm, and the LH surge followed, on average, 62 h later. Using the mathematical model, model-based indicators for the estimation of ovulation time can be calculated. Depending on which model-based indicator was used, ranges of 33 to 35 h and SD of about 11 h were obtained. Because detection of the LH surge was very labor intensive, only a limited number of potential estrus periods could be studied.

A novel system for on-farm fertility monitoring based on milk progesterone.

Timely identification of a cow's reproduction status is essential to minimize fertility-related losses on dairy farms. This includes optimal estrus detection, pregnancy diagnosis, and the timely recognition of early embryonic death and ovarian problems. On-farm milk progesterone (P4) analysis can indicate all of these fertility events simultaneously. However, milk P4 measurements are subject to a large variability both in terms of measurement errors and absolute values between cycles. The objective of this paper is to present a newly developed methodology for detecting luteolysis preceding estrus and give an indication of its on-farm use. The innovative monitoring system presented is based on milk P4 using the principles of synergistic control. Instead of using filtering techniques and fixed thresholds, the present system employs an individually on-line updated model to describe the P4 profile, combined with a statistical process control chart to identify the cow's fertility status. The inputs for the latter are the residuals of the on-line updated model, corrected for the concentration-dependent variability that is typical for milk P4 measurements. To show its possible use, the system was validated on the P4 profiles of 38 dairy cows. The positive predictive value for luteolysis followed by estrus was 100%, meaning that the monitoring system picked up all estrous periods identified by the experts. Pregnancy or embryonic mortality was characterized by the absence or detection of luteolysis following an insemination, respectively. For 13 cows, no luteolysis was detected by the system within the 25 to 32 d after insemination, indicating pregnancy, which was confirmed later by rectal palpation. It was also shown that the system is able to cope with deviating P4 profiles having prolonged follicular or luteal phases, which may suggest the occurrence of cysts. Future research is recommended for optimizing sampling frequency, predicting the optimal insemination window, and establishing rules to detect problems based on deviating P4 patterns.

Evaluation of the antibacterial and antibiofilm activities of novel CRAMP-vancomycin conjugates with diverse linkers.

We report the design, synthesis and antibacterial activity analysis of conjugates of vancomycin and cathelicidin-related antimicrobial peptides (CRAMP). Vancomycin inhibits the nascent peptidoglycan synthesis and is highly active against Gram-positive bacteria, whereas Gram-negative bacteria are generally insensitive due to a protective outer membrane. CRAMP is known to translocate across the Gram-negative outer membrane by a self-promoted uptake mechanism. Vancomycin-CRAMP conjugates were synthesized using click chemistry with diverse hydrophilic and hydrophobic linkers, with CRAMP functioning as a carrier peptide for the transfer of vancomycin through the outer membrane. Small hydrophobic linkers with an aromatic group result in the most active conjugates against planktonic Gram-negative bacteria, while maintaining the high activity of vancomycin against Gram-positive bacteria. These conjugates thus show a broad-spectrum activity, which is absent in CRAMP or vancomycin alone, and which is strongly improved compared to an equimolar mixture of CRAMP and vancomycin. In addition, these conjugates also show a strong inhibitory activity against S. Typhimurium biofilm formation.

A PKS/NRPS/FAS hybrid gene cluster from Serratia plymuthica RVH1 encoding the biosynthesis of three broad spectrum, zeamine-related antibiotics.

Serratia plymuthica strain RVH1, initially isolated from an industrial food processing environment, displays potent antimicrobial activity towards a broad spectrum of Gram-positive and Gram-negative bacterial pathogens. Isolation and subsequent structure determination of bioactive molecules led to the identification of two polyamino antibiotics with the same molecular structure as zeamine and zeamine II as well as a third, closely related analogue, designated zeamine I. The gene cluster encoding the biosynthesis of the zeamine antibiotics was cloned and sequenced and shown to encode FAS, PKS as well as NRPS related enzymes in addition to putative tailoring and export enzymes. Interestingly, several genes show strong homology to the pfa cluster of genes involved in the biosynthesis of long chain polyunsaturated fatty acids in marine bacteria. We postulate that a mixed FAS/PKS and a hybrid NRPS/PKS assembly line each synthesize parts of the backbone that are linked together post-assembly in the case of zeamine and zeamine I. This interaction reflects a unique interplay between secondary lipid and secondary metabolite biosynthesis. Most likely, the zeamine antibiotics are produced as prodrugs that undergo activation in which a nonribosomal peptide sequence is cleaved off.

Total synthesis of Septocylindrin B and C-terminus modified analogues.

The total synthesis is reported of the peptaibol Septocylindrin B which is related to the well documented channel forming peptaibol antibiotic Alamethicin. Several analogues were synthesized with a modified C-terminus, to investigate the SAR of the terminal residue Phaol. All these peptides were tested for their membrane perturbation properties by fluorescent dye leakage assay and for their antibacterial activity.