Determination of Intestinal Viral Loads and Distribution of Bovine Viral Diarrhea Virus, Classical Swine Fever Virus, and Peste Des Petits Ruminants Virus: A Pilot Study.

The aim of this pilot study was to determine viral loads and distribution over the total length, at short distances, and in the separate layers of the intestine of virus-infected animals for future inactivation studies. Two calves, two pigs, and two goats were infected with bovine viral diarrhoea virus (BVDV), classical swine fever virus (CSFV), and peste des petits ruminants virus (PPRV), respectively. Homogenously distributed maximum BVDV viral loads were detected in the ileum of both calves, with a mean titer of 6.0 log10 TCID50-eq/g. The viral loads in colon and caecum were not distributed homogenously. In one pig, evenly distributed CSFV mean viral loads of 4.5 and 4.2 log10 TCID50-eq/g were found in the small and large intestines, respectively. Mucosa, submucosa, and muscular layer/serosa showed mean viral loads of 5.3, 3.4, and 4.0 log10 TCID50-eq/g, respectively. Homogenous distribution of PPRV was shown in the ileum of both goats, with a mean viral load of 4.6 log10 TCID50-eq/g. Mean mucosa, submucosa, and muscular layer/serosa viral loads were 3.5, 2.8, and 1.7 log10 TCID50-eq/g, respectively. This pilot study provides essential data for setting up inactivation experiments with intestines derived from experimentally infected animals, in which the level and the homogeneous distribution of intestinal viral loads are required.

Intestinal Viral Loads and Inactivation Kinetics of Livestock Viruses Relevant for Natural Casing Production: a Systematic Review and Meta-Analysis.

Animal intestines are the source of edible sausage casings, which are traded worldwide and may come from areas where notifiable infectious animal diseases are prevalent. To estimate the risks of virus contamination, knowledge about the quantity of virus and decimal reduction values of the standard preservation method by salting is of great importance. A literature search, based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, was performed in search engine CAB Abstracts to determine the viral load of 14 relevant animal viruses in natural casings or intestines. Only a very limited number of scientific publications per virus were found and viral loads in the intestines varied from high for ASFV (five publications), BVDV (3), CSFV (6), PPRV (3), RPV(2) and TGEV (3) to moderate for PEDV (2) and SVDV (3), low for HEV (2) and FMDV (5), very low for VESV (1) and negative for PrV (2) and VSV (1). PRRSV was found in intestines, however, viral titers were not published. Three viruses (BVDV, CSFV and PPRV) with high viral loads were selected to search for their inactivation kinetics. For casings, no inactivation data were found, however, thermal inactivation data of these viruses were available, but differed in quantity, quality and matrices. In conclusion, important data gaps still exist when it comes to the quantitative inactivation of viruses in sausage casings or livestock intestines.

A Pilot Study on the Contamination of Assistance Dogs' Paws and Their Users' Shoe Soles in Relation to Admittance to Hospitals and (In)Visible Disability.

(1) Background: People with disabilities may benefit from an assistance dog (AD). Despite regulations that prohibit the denial of ADs to public places, this still occurs on a regular basis. The main argument for denial of access is that dogs compromise hygiene with their presence, which could cause a health hazard. Meanwhile, people are allowed to walk into and out of public places freely. (2) Objective: As a pilot study, to investigate the number of Enterobacteriaceae and the presence of Clostridium difficile bacteria on the paws of ADs and pet dogs (PDs) as well as the shoe soles of their users and owners. With the results, an assessment can be made as to whether measures are required to reduce environmental contamination (e.g., in hospitals). (3) Methods: In total, 25 ADs, 25 PDs, and their 50 users/owners participated in the study. Each participant walked their dog for 15-30 min prior to the sampling of the front paws. Each PD owner or AD user filled out a general questionnaire about the care of their dogs, and AD users were asked to fill out an additional questionnaire on their experiences regarding the admittance of their ADs to public places (in particular, hospitals). Dutch hospitals were questioned on their protocols regarding the admittance of ADs and their visitor numbers, including the percentage of AD users, to put these numbers into perspective. (4) Results: Dog paws were more often negative for Enterobacteriaceae compared to shoe soles (72% and 42%, respectively) and also had significantly lower bacterial counts (mean of 3.54log10 and 5.03log10 colony-forming units (CFUs), respectively; p < 0.05). This was most distinct in the comparison between PDs and their owners (3.75log10 and 5.25log10 CFUs; p < 0.05); the numbers were similar between ADs and their users (3.09log10 and 4.58log10 CFUs; p = 0.2). C. difficile was found on one (4%) AD user's shoe soles. Moreover, 81% of AD users had been denied access with their current AD once or several times, the main reason being hygiene. The results of the visibly and invisibly disabled were significantly different. The number of AD users as opposed to the total number of hospital visitors was 0.03% in one hospital and is estimated to be 0.02% in the Netherlands. (5) Conclusions: The general hygiene of dogs' paws is far better than that of shoe soles, mostly demonstrated by the better general hygiene of PD paws compared with their owners' shoe soles; ADs and their users had comparable levels of general hygiene. In addition, the number of AD users amongst the total number of hospital visitors in the Netherlands is very limited. Thus, hygiene measures to reduce any contamination due to dog paws do not seem necessary.

Salt inactivation of classical swine fever virus and African swine fever virus in porcine intestines confirms the existing in vitro casings model.

Natural casings, to be used as sausage containers, are being traded worldwide and may be contaminated with contagious viruses. Standard processing of such natural casings is by salt treatment with a duration of 30 days before shipment. Since information is lacking about the efficacy of these virus inactivation procedures, an in vitro 3D collagen matrix model, mimicking natural casings, was developed previously to determine the efficacy of salt to inactivate specific viruses. To validate this model, a comparison in vivo experiment was performed using intestines of pigs experimentally infected with African swine fever virus (ASFV) and classical swine fever virus (CSFV). Decimal reduction (D) values, were determined at 4 °C, 12 °C, 20 °C and 25 °C. The standard salt processing procedure showed an efficient inactivation of ASFV and CSFV over time in a temperature dependent way. Dintestine values of both viruses, treated with the standard salt treatment, were in line with the Dcollagen values. It was concluded that these results underline the suitability of the 3D collagen matrix model to determine virus inactivation and to replace animal experiments. Furthermore, an increase in storage time for standard salt processed casings derived from CSFV endemic regions is highly recommended for an efficient inactivation of CSFV.

Validation of a High-Throughput Sausage Casing Model for the Assessment of Bacterial Inactivation Affected by Salt Concentration, pH, and Temperature.

Previous studies have shown the efficacy of high concentrations of salt as the main preservative against vegetative bacteria present on natural sausage casings. These studies were limited in the number of variables and the interactions between these variables that were assessed. To remedy this situation, a MicroCasing high-throughput model was developed and validated to study the inactivation kinetics of various combinations of parameters (salt concentration, pH, and temperature) on eight bacterial isolates of Salmonella enterica, Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes over a prolonged period. A Weibullian power model was the best fit to show the trends in sensitivity of each bacterial isolate to salt, pH, and temperature over time. The inactivation kinetics generated with this novel approach could serve as a predictive model for the required salting period for casings. The actual bacterial contamination of the product can vary with the respective production step during processing from animal intestine into sausage casings (initial level, ∼105 CFU/g; level after salting, <102 CFU/g). Subsequent selection and grading of these casings will require complete removal of all salt, and upon completion of this production step, the casings will be resalted. By determining the actual contamination level before the salting process, the minimum storage period in salt can be calculated and potentially optimized by adjusting the pH and temperature. As a result, a standard holding period of at least 30 days may no longer be necessary to produce salted natural casings in accordance with validated quality and food safety criteria.

The Translation and Validation of the Dutch Monash Dog⁻Owner Relationship Scale (MDORS).

The Monash Dog-Owner Relationship Scale (MDORS) is a questionnaire that is used to evaluate the perceived relationship between humans and their dog. This questionnaire was originally only formulated and validated in English, which limits its use among non-English speaking individuals. Although a translation could be made, the translation of questionnaires without additional validation often impairs the reliability of that questionnaire. Therefore, the aim of this study was to validate a translation of the MDORS that is suitable for use among native Dutch speakers. To achieve this, a Dutch translation of the MDORS was made and checked for spelling/grammar mistakes, readability, feasibility, and clarity. A test-retest comparison was subsequently performed on the translation together with a calculation of Cronbach's alpha score and principal component analysis (PCA). Through the PCA, we found that the three-factor model of the original MDORS was also largely present in the Dutch translation. However, deviations were also found, as several questions did not achieve high PCA scores in their original factor. Therefore, we propose that these questions are excluded from the Dutch MDORS.

Erratum: The study of service dogs for veterans with Post-Traumatic Stress Disorder: a scoping literature review.

[This corrects the article DOI: 10.1080/20008198.2018.1503523.].

The study of service dogs for veterans with Post-Traumatic Stress Disorder: a scoping literature review.

The therapeutic application of human-animal interaction has gained interest recently. One form this interest takes is the use of service dogs as complementary treatment for veterans with Post-Traumatic Stress Disorder (PTSD). Many reports on the positive effect of PTSD Service Dogs (PSDs) on veterans exist, though most are indirect, anecdotal, or based on self-perceived welfare by veterans. They therefore only give a partial insight into PSD effect. To gain a more complete understanding of whether PSDs can be considered an effective complementary treatment for PTSD, a scoping literature review was performed on available studies of PSDs. The key search words were 'dog', 'canine', 'veteran', and 'PTSD'. This yielded 126 articles, of which 19 matched the inclusion criteria (six empirical studies). Recurrent themes in included articles were identified for discussion of methodology and/or results. It was found that results from most included studies were either applicable to human-animal interaction in general or other types of service animals. They therefore did not represent PSDs specifically. Studies which did discuss PSDs specifically only studied welfare experience in veterans, but used different methodologies. This lead us to conclude there is currently no undisputed empirical evidence that PSDs are an effective complementary treatment for veterans with PTSD other than reports on positive welfare experience. Additionally, the lack of development standardization and knowledge regarding welfare of PSDs creates risks for both human and animal welfare. It is therefore recommended that a study on the effect of PSDs be expanded to include evaluation methods besides self-perceived welfare of assisted humans. Future studies could include evaluations regarding human stress response and functioning, ideally conducted according to validated scientific methodologies using objective measurement techniques to identify the added value and mechanisms of using PSDs to assist treatment of PTSD in humans.

La aplicación terapéutica de la interacción humano-animal ha ganado interés en los últimos años. Una forma que toma este interés es el uso de perros de servicio como tratamiento complementario para veteranos con Trastorno de Estrés Postraumático (TEPT). Existen muchos reportes del efecto positivo de los Perros de Servicio en TEPT (PSDs, en su sigla en inglés) en los veteranos, aunque la mayoría son indirectos, anecdóticos o basados en la autopercepción de bienestar de los veteranos. Por lo tanto, sólo entregan una visión parcial sobre el efecto de los PSD. Para obtener una comprensión más completa sobre si los PSDs pueden ser considerados un tratamiento complementario efectivo para el TEPT, se realizó una revisión exploratoria de la literatura de los estudios disponibles de PSDs. Las palabras clave de búsqueda fueron 'perro', 'canino' 'veterano' y 'TEPT', lo que arrojó 126 artículos, de los cuales 19 cumplieron los criterios de inclusión (6 estudios empíricos). Los temas recurrentes en los artículos incluidos fueron identificados para discusión de la metodología y/o resultados. Se encontró que los resultados de la mayoría de los estudios incluídos eran aplicables a la interacción humano-animal en general o en otro tipo de animales de servicio. Por lo tanto, no representaban a los PSDs específicamente. Los estudios que discutían acerca de PSDs en forma específica solo estudiaron la experiencia de bienestar en los veteranos, aunque usaron diferentes metodologías entre ellos. Esto lleva a concluir que actualmente no hay evidencia empírica indiscutible de que los PSDs sean un tratamiento complementario efectivo para los veteranos con TEPT más allá de los reportes de una experiencia positiva de bienestar. Adicionalmente, la falta de estandarización del desarrollo y conocimiento acerca del bienestar de los PSDs genera riesgos para el bienestar de ambos, humano y animal. Por lo tanto es recomendable que el estudio del efecto de los PSDs sea ampliado para incluir métodos de evaluación mas allá del bienestar auto-percibido de los humanos asistidos. Estudios futuros podrían incluir evaluaciones en relación a la respuesta al estrés y funcionamiento humanos, idealmente conducidos de acuerdo a metodologías científicas validadas usando técnicas de medición objetivas para identificar el valor agregado, y mecanismos, del uso de PSDs para asistir el tratamiento del TEPT en humanos.

Inactivation of foot-and-mouth disease virus in various bovine tissues used for the production of natural sausage casings.

Bovine intestines, bladders and oesophagus are used for the production of natural casings ("beef casings") as edible sausage containers. Derived from cattle experimentally infected with FMDV (initial dosage 10(4) TCID(50)/mL, strain A Iran 97), these beef casings were treated with sodium chloride (NaCl) or phosphate supplemented salt (P-salt). In addition, different in-vitro experiments using beef casings were done on a small scale with other FMDV strains (A Turkey 06, C-Oberbayern and O(1) Manisa) as "proof of principle". Based on the combined results of the in-vivo and in-vitro experiments, it can be concluded that the storage period of 30 days at 20 °C in NaCl is sufficiently effective to inactivate a possible contamination with FMDV in beef casings and that the usage of P-salt does not clearly enhance the inactivation of FMDV infectivity. Storage of salted beef casings at about 20 °C for 30 days is already part of the Standard Operating Procedures (included in HACCP) of the international casing industry and can therefore be considered as a protective measure for the international trade in natural casings.

Virus inactivation by salt (NaCl) and phosphate supplemented salt in a 3D collagen matrix model for natural sausage casings.

Due to possible presence and spread of contagious animal viruses via natural sausage casings the international trade in these food products is subject to veterinary and public health requirements. In order to manage these restrictions we determined the effect of casing preservation on four highly contagious viruses for livestock: foot-and-mouth-disease virus (FMDV), classical swine fever virus (CSFV), swine vesicular disease virus (SVDV) and African swine fever virus (ASFV). We used an in vitro 3D collagen matrix model in which cells, infected with the four different viruses were embedded in a bovine collagen type I gel matrix and treated with either saturated salt (NaCl) or phosphate supplemented saturated salt at four different temperatures (4, 12, 20 and 25 °C) during a period of 30 days. The results showed that all viruses were faster inactivated at higher temperatures, but that stability of the various viruses at 4 °C differed. Inactivation of FMDV in the 3D collagen matrix model showed a clear temperature and treatment effect on the reduction of FMDV titres. At 4 and 12 °C phosphate supplemented salt showed a very strong FMDV inactivation during the first hour of incubation. Salt (NaCl) only had a minor effect on FMDV inactivation. Phosphate supplemented salt treatment increased the effect temperature had on inactivation of CSFV. In contrast, the salt (NaCl) treatment only increased CSFV inactivation at the higher temperatures (20 °C and 25 °C). Also SVDV inactivation was increased by phosphate supplemented salt, but salt (NaCl) treatment only resulted in a significant decrease of SVDV titre at a few time points. The ASFV results showed that both salt (NaCl) and phosphate supplemented salt were capable to inactivate ASFV within 48 h. In contrast to the other viruses (FMDV, CSFV and SVDV), ASFV was the most stable virus even at higher temperatures. The results obtained in this in vitro model underline the efficacy of a combined treatment using phosphate supplemented salt and storage at 20 °C or higher for a period of 30 days. This treatment may therefore be useful in reducing the animal health risks posed by spread of contagious animal viruses by international trade of natural sausage casings.

Quantitative histological analysis of bovine small intestines before and after processing into natural sausage casings.

A histological study was undertaken to determine the efficiency in the removal of the mucosa and Peyer's patches by standard processing of bovine intestines into natural sausage casings. The second objective was to calculate the quantity of lymphoid and nervous tissue per consumable sausage. For the histological analysis, intestinal samples were collected from 80 beef cattle during the slaughter process. Fresh and cleaned intestines were compared in analyzing the thickness of the intestinal wall, weight reduction during cleaning, removal of the mucosal layer, and the presence of lymphoid and neural tissue after cleaning. The obtained data indicate a weight reduction of about 50% during standard cleaning procedures, as 90% of the mucosa and 48% of the lymphoid tissue are removed. Based on the quantitative histological image analysis, it was calculated that 1 m of cleaned casings, weighing on average 64 g, contains about 2.8 g of mucosa, 0.3 g of lymphoid tissue, and 0.1 g of neural tissue. Assuming, in a worst-case scenario, that the sausage casing is ingested when consuming 200 g of sausage at one meal, this consumption includes 0.09 g of lymphoid tissue and 0.02 g of neural tissue as part of the sausage casing. These data can be included in a risk assessment on the potentialexposure of consumers to bovine spongiform encephalopathy infectivity after eating sausages in beef casings.

Modulation of the cytokine responses in equine macrophages following TACE-inhibition.

The detrimental effects of the pro-inflammatory cytokine TNF-alpha during equine acute abdominal disease are well known. Its pivotal role in many human diseases has led to various in-depth studies regarding its release mechanism, in particular by TNF-alpha converting enzyme (TACE). In this study we investigated the inhibitory effect of a TACE-inhibitor on cytokine release (TNF-alpha, IL-1alpha and IL-6) in three different cell models, including U937 cells, a recently established equine macrophage cell line, known as eCAS, and primary equine PBMC. The aim was to show the similarity of TNF-alpha release through the TACE mechanism in human and equine cells after stimulation with LPS. Results indicate that, by using a TACE-inhibitor, TNF-alpha, IL-1alpha and IL-6 release can be reduced in both equine cell models and achieved comparable results in the human U937 cells. These results suggest that equine TNF-alpha, like its human homologue, is released from its membrane-bound position by TACE.