The positive aspects of correct negative reinforcement

Abstract

In the scheme of contemporary animal training, horse training is virtually unique because it relies on negative reinforcement (NR) rather than positive reinforcement (PR). Furthermore, horse trainers are largely unaware that they are using NR in training. Instead, they believe in the benevolent nature of the horse and see their task in training as one of improving the balance and gymnastic ability of the horse—outcomes that emerge when the rider is similarly properly balanced. Under these conditions, it is claimed the willing horse will perform its required maneuvers. These beliefs may be associated with several welfare issues and indicate areas requiring future research: 1. The absence of release of pressure, the release of pressure at the wrong times, the use of opposing pressures simultaneously and the absence of shaping procedures are central to the development of acute and chronic stress responses in horses.

2. Resultant conflict behaviors contribute to equine wastage statistics and include behaviors that are dangerous to horses and humans.

3. There is a need for research into the mechanics of NR because it is poorly researched compared to PR.

4. When NR responses are installed correctly, only mild pressures need to be used, and results are obtained in few trials.

5. Many qualified animal trainers misunderstand NR and confuse it with punishment. They believe that PR has positive welfare implications and thus NR being “negative,” has negative welfare implications. So there is a need for horse trainers to understand learning theory and the principles that surround NR.

6. Horse trainers are isolated from advances in animal training. Therefore they increasingly seek knowledge and solutions from the growing number of “horse whisperers” and unqualified “horse psychologists.” This is potentially detrimental for the welfare of the horse and the need is urgent for universities throughout the world to become the knowledge bases for equitation science.

     

A mini‐STR typing system for degraded equine DNA

Degraded biological samples are a challenge for testing laboratories. Genotyping success can be improved through the use of mini‐STRs, by which primers are placed adjacent to the repeat motifs to reduce amplicon size. Here, we present a genetic profiling system comprising 13 autosomal and one X‐linked dinucleotide‐repeat markers and the SRY gene based on the internationally accepted equine parentage panel. The markers are divided into two panels with all alleles falling at or below 182 bp. The application of this method significantly increases the ability to profile difficult samples and to provide discriminating results to clients.     

Genetic risk for squamous cell carcinoma of the nictitating membrane parallels that of the limbus in Haflinger horses

Squamous cell carcinoma (SCC) is the most common cancer affecting the equine eye, with a higher incidence documented in Haflinger horses. Recently, a missense variant in the gene damage specific DNA binding protein 2 (DDB2, p.Thr338Met) on ECA12 was identified as a risk factor for the development of limbal SCC in Haflinger horses. SCC also occurs on the nictitating membrane; therefore, investigating the role of this missense variant in nictitating membrane SCC is warranted. In this study, a common ancestor was identified among Haflinger horses affected with limbal SCC or with nictitating membrane SCC, thus supporting a recessive risk factor for the development of cancer at both ocular locations. Analysis of genotype data from Haflinger horses with and without nictitating membrane SCC revealed that the same region on ECA12 associated with limbal SCC was also associated with nictitating membrane SCC (P < 2.04 × 10^?5). Fine mapping of this locus using 25 cases and 49 controls supported the hypothesis that DDB2:c.1013C>T, p.Thr338Met, is a risk factor for nictitating membrane SCC, as 88% of our cases were homozygous for this variant and no other polymorphism was more strongly associated (P = 4.13 × 10^?14). These data indicate that the genetic risk is the same for the development of both limbal and nictitating membrane SCC in Haflinger horses and validates utilization of genetic testing of the DDB2 variant for both clinical management and the guidance of mating decisions.     

Characterization of a red blood cell antigen in donkeys and mules associated with neonatal isoerythrolysis

A red cell antigen of donkeys and mules was identified using antibodies in serum from a mare which produced a mule foal affected with neonatal isoerythrolysis (NI). Subsequently antibodies with similar activity were identified in the sera of other mares which had produced mule foals and were produced by immunization of horses with blood from donkeys. The antigen detected by these antibodies does not correspond to any recognized horse red cell alloantigen. This may be a xenoantigen since all donkeys (and mules) tested have shared this antigen and all horses tested have lacked the antigen. The results suggest that all mule pregnancies (donkey sire x horse dam) are incompatible with regard to this factor and a potential for neonatal isoerythrolysis exists in all cases.     

Dissemination of western equine encephalomyelitis virus in the potential vector,Culex pipiens pallens

Two western equine encephalomyelitis virus (WEEV) strains have been isolated in China. Our previous studies have verified that the mosquito Culex pipiens pallens Coquillett (Diptera: Culicidae) infected with WEEV was capable of transmitting this arbovirus, but it was not clear how the sequential multiplication and spread of virus occurred within the mosquito. In this study, we observed the distribution of WEEV antigen in orally‐infected Cx. p. pallens by immunohistochemistry in order to better understand the initial infection, dissemination, and transmission of WEEV in the potential vector. Orally‐infected WEEV dissemination varied within the different tissues of Cx. p. pallens, with virus antigen consistently observed in the salivary glands, foregut, midgut epithelial cells, Malpighian tubules, hindgut, and ovarian follicles of some individuals after various days of extrinsic incubation. We suggest that Cx. p. pallens, the potential vector of WEEV, has the ability to harbor the virus through the alimentary system, and the midgut epithelial cell may be the initial site of WEEV replication after ingestion of a viremic blood meal.     

A comparison of the occurrence of common dental abnormalities in stabled and free-grazing horses

The purpose of this study was to gain evidence on the prevalence of dental abnormalities in stable-kept horses in comparison with free-living horses. It is expected that free-living horses that graze for as much as 16 h/day will have fewer dental abnormalities than stable-kept horses. In this study, the latter group was fed a diet that was based on a relatively high-energy, cereal-based feed. This was thought to be a representative of common practice in domesticated, stable-kept horses. Compound diets such as this have previously been shown to increase the frequency of chewing cycles and decrease mediolateral excursion. The occurrence of 10 named dental abnormalities present in the dentition of 60 Thoroughbred-type horses was recorded. Half of the population was at grass all year round in New South Wales, Australia. The remainder were stabled for 24 h/day in Gloucestershire, UK. All horses were between 5 and 15 years of age. The same, experienced, equine dental technician examined all horses using a full-mouth speculum and produced a routine dental chart. Stable-kept horses had a significantly higher total occurrence of abnormalities (P < 0.001) than free-living horses. The stable-kept group had a significantly higher prevalence of exaggerated transverse ridging across the occlusal surface of the cheek teeth, focal or ramped overgrowths of the cheek teeth and periodontal disease (P < 0.01 in all cases). All horses in both groups had some occurrence of sharp edges of the buccal and lingual edges of the cheek teeth. The results are in agreement with anecdotal evidence that a fibre-based, grazed diet results in fewer dental abnormalities. However, sharp edges may occur even with what is perceived as a 'natural' diet.     

Ddi1, a eukaryotic protein with the retroviral protease fold

Retroviral aspartyl proteases are homodimeric, whereas eukaryotic aspartyl proteases tend to be large, monomeric enzymes with 2-fold internal symmetry. It has been proposed that contemporary monomeric aspartyl proteases evolved by gene duplication and fusion from a primordial homodimeric enzyme. Recent sequence analyses have suggested that such "fossil" dimeric aspartyl proteases are still encoded in the eukaryotic genome. We present evidence for retention of a dimeric aspartyl protease in eukaryotes. The X-ray crystal structure of a domain of the Saccharomyces cerevisiae protein Ddi1 shows that it is a dimer with a fold similar to that of the retroviral proteases. Furthermore, the double Asp-Thr-Gly-Ala amino acid sequence motif at the active site of HIV protease is found with identical geometry in the Ddi1 structure. However, the putative substrate binding groove is wider in Ddi1 than in the retroviral proteases, suggesting that Ddi1 accommodates bulkier substrates. Ddi1 belongs to a family of proteins known as the ubiquitin receptors, which have in common the ability to bind ubiquitinated substrates and the proteasome. Ubiquitin receptors contain an amino-terminal ubiquitin-like (UBL) domain and a carboxy-terminal ubiquitin-associated (UBA) domain, but Ddi1 is the only representative in which the UBL and UBA domains flank an aspartyl protease-like domain. The remarkable structural similarity between the central domain of Ddi1 and the retroviral proteases, in the global fold and in active-site detail, suggests that Ddi1 functions proteolytically during regulated protein turnover in the cell.     

Melatonin Decreases Nitric oxide Production, Inducible Nitric oxide Synthase Expression and Lipid Peroxidation Induced by Venezuelan Encephalitis Equine Virus in Neuroblastoma Cell Cultures

Increased expression of inducible nitric oxide synthase has been shown in murine Venezuelan equine encephalitis (VEE) virus infection. In this experimental model, melatonin (MTL) treatment has shown to be beneficial. The aim of this study was to determine the effect of VEE virus on the nitric oxide (NO) production and lipid peroxidation in neuroblastoma cell cultures, and to investigate the role of MTL during cell-virus interaction. Neuroblastoma cells were co-cultured with VEE virus and treated with MTL at doses ranging from 0 to 1.8 mM, for 6, 12, 24 and 48 h. NO and lipid peroxidation were measured in culture supernatants and in the cellular content by nitrite concentration and thiobarbituric acid assay, respectively. Expression of inducible nitric oxide synthase (iNOS) was determined by indirect immunofluorescence. Increased production of NO and lipid peroxidation products were found in supernatants and cellular contents of VEE virus treated cultures. Both NO and lipid peroxidation were decreased by MTL treatment in a time dependent manner. Increased iNOS expression was observed in VEE virus infected cultures that was reduced by MTL treatment. These results could be related to the beneficial role of MTL in the VEE experimental disease and address the possible therapeutic potential of the hormone in human VEE virus infection.