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Canine leukoencephalomyelopathy (LEMP) is a juvenile-onset neurodegenerative disorder of the CNS white matter currently described in Rottweiler and Leonberger dogs. Genome-wide association study (GWAS) allowed us to map LEMP in a Leonberger cohort to dog chromosome 18. Subsequent whole genome re-sequencing of a Leonberger case enabled the identification of a single private homozygous non-synonymous missense variant located in the highly conserved metallo-beta-lactamase domain of the N-acyl phosphatidylethanolamine phospholipase D (NAPEPLD) gene, encoding an enzyme of the endocannabinoid system. We then sequenced this gene in LEMP-affected Rottweilers and identified a different frameshift variant, which is predicted to replace the C-terminal metallo-beta-lactamase domain of the wild type protein. Haplotype analysis of SNP array genotypes revealed that the frameshift variant was present in diverse haplotypes in Rottweilers, and also in Great Danes, indicating an old origin of this second NAPEPLD variant. The identification of different NAPEPLD variants in dog breeds affected by leukoencephalopathies with heterogeneous pathological features, implicates the NAPEPLD enzyme as important in myelin homeostasis, and suggests a novel candidate gene for myelination disorders in people.

Male weanling pigs (n 32) with a mean initial body weight of 7·5 kg and a mean weaning age of 28 d were used in a 31 d study to investigate the effects of feeding GM (Bt MON810) maize on growth performance, intestinal histology and organ weight and function. At weaning, the pigs were fed a non-GM starter diet during a 6 d acclimatisation period. The pigs were then blocked by weight and litter ancestry and assigned to diets containing 38·9 % GM (Bt MON810) or non-GM isogenic parent line maize for 31 d. Body weight and feed disappearance were recorded on a weekly basis (n 16/treatment), and the pigs (n 10/treatment) were killed on day 31 for the collection of organ, tissue and blood samples. GM maize-fed pigs consumed more feed than the control pigs during the 31 d study (P < 0·05) and were less efficient at converting feed to gain during days 14-30 (P < 0·01). The kidneys of the pigs fed GM maize tended to be heavier than those of control pigs (P = 0·06); however, no histopathological changes or alterations in blood biochemistry were evident. Small intestinal morphology was not different between treatments. However, duodenal villi of GM maize-fed pigs tended to have fewer goblet cells/[mu]m of villus compared with control pigs (P = 0·10). In conclusion, short-term feeding of Bt MON810 maize to weaned pigs resulted in increased feed consumption, less efficient conversion of feed to gain and a decrease in goblet cells/[mu]m of duodenal villus. There was also a tendency for an increase in kidney weight, but this was not associated with changes in histopathology or blood biochemistry. The biological significance of these findings is currently being clarified in long-term exposure studies in pigs. [ABSTRACT FROM AUTHOR]

The dynamics of Mycobacterium bovis infection in cattle can influence the proportion of infected animals that are diagnosed by ante-mortem tests in routine bovine tuberculosis (bTB) surveillance and monitoring programmes. Although the current diagnostic tests based on cell-mediated or serological responses are imperfect, they are effective in diagnosing the majority of infected animals. However, the lack of perfect sensitivity and specificity also leads to failure to diagnose all infected animals leading to persistence of infection in herds. The terms residual, subclinical, latent and anergy have been used interchangeably to denote the presence of continued undiagnosed M. bovis infection within cattle herds, which ultimately hinders the eradication of bTB and imposes substantial financial burdens on farming communities and national economies. Epidemiological data suggests the existence of M. bovis-infected, but often undetected, cattle within herds that contribute to eradication failure. This has similarities with human tuberculosis, caused by Mycobacterium tuberculosis, where latent infection is defined as the persistence of viable but quiescent bacilli for extended periods in patients without clinical symptoms but with a detectable immune response to M. tuberculosis antigens. If a similar infection state exists in cattle infected with M. bovis, the persistence of such animals in disease-managed herds is unlikely to be common given that those found to have positive immune responses to M. bovis antigens are routinely culled to minimise future risk of transmission. Apart from contributing to the burden of herd infection, such residual infection without detection may also 'seed' recipient herds following animal movements, and potentially play an important role in the overall epidemiology of bTB as the prevalence of disease decreases and the attendant altered predictive value of the diagnostic tests result in a greater proportion of infected animals remaining undetected. This review examines how the different stages of M. bovis infection in cattle may contribute to the failure to diagnose infected animals using conventional testing methodologies and the attendant risk this poses in creating prolonged or recurrent herd breakdowns., Competing Interests: Declaration of Competing Interest None of the authors of this paper has a financial or personal relationship with other people or organisations that could inappropriately influence or bias the content of the paper., (Copyright © 2024. Published by Elsevier Ltd.)

The evaluation of student and faculty opinions on curricular changes in veterinary medical education is an essential part of the quality assurance process. This study investigates if the perceived educational value of a newly introduced module (veterinary pathobiology) in the earlier stages of a veterinary curriculum changes as students progress into the later phases of their training. Data were collected by anonymous questionnaire from two sequential final year student cohorts in 2021 and 2022. These students had previously been surveyed on their views of the new integrated pathobiology module immediately after taking it in their third year. Within 2 years, student satisfaction with the module's learning objectives increased significantly. Final year students had much clearer appreciation of how this pedagogical innovation enhanced their learning and that this was particularly true of the case-based learning approach adopted. While faculty teaching these students in their final year clinical pathobiology rotation expressed the view that overall, student performance was no different when compared to previous years, this somewhat disappointing finding needed to be viewed in the context of the intervening COVID-19 pandemic where face-to-face teaching of students over many months had been severely curtailed as had student attendance of their extramural studies. This study confirms a positive effect of an integrated curricular intervention in veterinary undergraduate education and highlights the necessity of student evaluation at multiple time points as their perception of the value of such changes appears to be linked to their experience and expectations.

SARS-CoV-2 can infect wildlife, and SARS-CoV-2 variants of concern might expand into novel animal reservoirs, potentially by reverse zoonosis. White-tailed deer and mule deer of North America are the only deer species in which SARS-CoV-2 has been documented, raising the question of whether other reservoir species exist. We report cases of SARS-CoV-2 seropositivity in a fallow deer population located in Dublin, Ireland. Sampled deer were seronegative in 2020 when the Alpha variant was circulating in humans, 1 deer was seropositive for the Delta variant in 2021, and 12/21 (57%) sampled deer were seropositive for the Omicron variant in 2022, suggesting host tropism expansion as new variants emerged in humans. Omicron BA.1 was capable of infecting fallow deer lung type-2 pneumocytes and type-1-like pneumocytes or endothelial cells ex vivo. Ongoing surveillance to identify novel SARS-CoV-2 reservoirs is needed to prevent public health risks during human-animal interactions in periurban settings.

Efficient, fast, and robust scintillators for ionizing radiation detection are crucial in various fields, including medical diagnostics, defense, and particle physics. However, traditional scintillator technologies face challenges in simultaneously achieving optimal performance and high-speed operation. Herein we introduce colloidal quantum shell heterostructures as X-ray and electron scintillators, combining efficiency, speed, and durability. Quantum shells exhibit light yields up to 70,000 photons MeV -1 at room temperature, enabled by their high multiexciton radiative efficiency thanks to long Auger-Meitner lifetimes (>10 ns). Radioluminescence is fast, with lifetimes of 2.5 ns and sub-100 ps rise times. Additionally, quantum shells do not exhibit afterglow and maintain stable scintillation even under high X-ray doses (>10 9  Gy). Furthermore, we showcase quantum shells for X-ray imaging achieving a spatial resolution as high as 28 line pairs per millimeter. Overall, efficient, fast, and durable scintillation make quantum shells appealing in applications ranging from ultrafast radiation detection to high-resolution imaging., (© 2024. UChicago Argonne, LLC, Operator of Argonne National Laboratory.)

The energetic stability of positron-dianion systems [A-; e+; A-] is studied via many-body theory, where A- includes H-, F-, Cl-, and the molecular anions (CN)- and (NCO)-. Specifically, the energy of the system as a function of ionic separation is determined by solving the Dyson equation for the positron in the field of the two anions using a positron-anion self-energy as constructed in Hofierka et al. [Nature 606, 688 (2022)] that accounts for correlations, including polarization, screening, and virtual-positronium formation. Calculations are performed for a positron interacting with H22-, F22-, and Cl22- and are found to be in good agreement with previous theory. In particular, we confirm the presence of two minima in the potential energy of the [H-; e+; H-] system with respect to ionic separation: a positronically bonded [H-; e+; H-] local minimum at ionic separations r ∼ 3.4 Å and a global minimum at smaller ionic separations r ≲ 1.6 Å that gives overall instability of the system with respect to dissociation into a H2 molecule and a positronium negative ion, Ps-. The first predictions are made for positronic bonding in dianions consisting of molecular anionic fragments, specifically for (CN)22- and (NCO)22-. In all cases, we find that the molecules formed by the creation of a positronic bond are stable relative to dissociation into A- and e+A- (positron bound to a single anion), with bond energies on the order of 1 eV and bond lengths on the order of several ångstroms., (© 2024 Author(s). Published under an exclusive license by AIP Publishing.)

Colloidal quantum shells are spherical semiconductor quantum wells, which have shown strong promise as optical materials, particularly in classes of experiments requiring multiple excitons. The two-photon properties of CdS/CdSe/CdS quantum shell samples are studied here to demonstrate large non-linear absorption cross-sections while retaining advantageous multiexciton physics conferred by the geometrical structure. The quantum shells have large two-phonon cross sections (0.4-7.9 × 10 6 GM), which highlights their potential use in upconversion imaging in which large per particle two-photon absorption is critical. Time-resolved measurements confirmed that the quantum shells have long biexciton lifetime (>10 ns in the largest core samples reported here) and large gain bandwidth (>300 meV). The combination of these attributes with large two-photon cross sections makes the CdS/CdSe/CdS quantum shells excellent gain media for two-photon excitation. With a broad gain bandwidth and long gain lifetime, quantum shell solids support multimodal amplified spontaneous emission from excitons, biexcitons, and higher excited states. Thresholds for amplified spontaneous emission and lasing, which are as low as 1 mJ cm -2 , are comparable to, or lower than, the thresholds reported for other colloidal materials.

Background: True congenital pancreatic cysts are a rare pathological process reported within feline and human literature. To date there has been no documented case of a true congenital cyst affecting a canine patient. The objective of this case report is to document the clinical findings, diagnostic investigations, surgical treatment, histopathological diagnosis and long-term outcome of a dog with a true pancreatic cyst., Case Presentation: A 5-month-old crossbreed dog was presented with a six-week history of abdominal pain, apparent bilateral pelvic limb weakness, reluctance to walk and intermittent vomiting and diarrhoea. An abdominal ultrasound examination performed by the dog's primary care veterinarian identified a large intra-abdominal structure of unclear origin. A computed tomographic examination identified a large ovoid structure measuring 156 mm in length, 95 mm in height and 89 mm in width and apparently originating from the left limb of the pancreas. An exploratory coeliotomy was performed and a partial pancreatectomy was performed to allow complete removal of the cystic structure. Histopathological analysis of sections of the wall of the large fluid-filled cyst identified a thick fibromuscular wall lined by a well regimented hyperplastic tall columnar epithelium with basally located round to ovoid nuclei featuring fine chromatin stippling and abundant apically located and surface mucin, concurrent with a true congenital pancreatic cyst. A long-term follow-up of twenty-nine months identified no clinical signs of recurrence., Conclusion: A partial pancreatectomy and en bloc excision of a true pancreatic cyst provided an excellent long-term outcome in a dog., (© 2022. The Author(s).)

The School of Veterinary Medicine, University College Dublin, Ireland, restructured the teaching of general pathology, parasitology, and microbiology in third year in 2018 as part of the development of an outcome-based curriculum. A new integrated teaching module was created, called Veterinary Pathobiology, which encompassed the three paraclinical subjects, worth 20 ECTS credits. Subject integration was driven and supported by case-based learning (CBL) activities, and practical classes, which were aimed at facilitating the understanding of basic disease processes, infectious agents, and the application of diagnostic tests. The disciplines maintained their identities within lectures which were aligned by content. The restructuring led to a reduction of contact hours by 20% and of assessment time by 40%. The examinations included integrated questions with an emphasis on the material students had covered in their CBL. Despite positive outcomes, which included equivalent examination scores and positive written feedback by students on teaching and learning, understanding, assessment, relevance, CBL, group work, and generic skills, the average scores for overall student satisfaction dropped dramatically in the second academic year of implementation. This followed the introduction of new regulations by the University relating to student progression, which was capped at "carrying" 10 ECTS credits, thus preventing students that failed the new module from progressing. Other criticisms of the new module by students included too little communication on the changes implemented in its first iteration and a workload perceived to be too heavy. Further restructuring is therefore necessary. This study highlights the process/pitfalls of integration/curricular innovation.

Background: Intraosseous epidermoid cyst (IEC) is a rare, non-neoplastic, pathology in animals and humans that most commonly affects the distal phalanx. In dogs, it is important to differentiate this lesion from malignant digital tumours causing bone lysis. In previous reports, IEC has been described to affect only a single digit at the time of diagnosis which is usually based on histopathology. This is the first case report to describe immunohistochemically confirmed IECs affecting simultaneously multiple digits., Case Presentation: A 4-and-a-half-year-old female spayed Great Dane was presented with a 2-month history of progressive swelling of the distal phalanx (PIII) of digits IV and V of the right pelvic limb. Eleven weeks prior to presentation, the dog had a low-grade cutaneous mast cell tumour completely excised from the craniolateral base of its left pinna. A history of trauma to 1 of the nails of the same pes 4 years prior to referral was also reported. Examination of the right pelvic limb identified firm non-painful swelling of PIII of digits IV and V, with concurrent deformation of the nails. Radiographs of the right pes obtained by the primary veterinarian identified an expansile lesion of PIII of digits IV and V. Computed tomography identified large expansile lesions of PIII of digits IV and V, with associated cortical thinning and soft tissue swelling. Neoplasia was considered the most likely radiographic diagnosis. Histopathology of Jamshidi bone biopsies was consistent with intraosseous epidermoid cyst, which was confirmed with immunohistochemistry. Amputation of PIII of digits IV and V at the level of mid-PII was performed as definitive treatment. No recurrence of the lesion occurred during the 10-month follow-up period., Conclusions: Intraosseous epidermoid cysts should be included in the differential diagnosis for expansile lesions affecting the canine digit. It is important to differentiate them from other digital lesions, with bone involvement, such as malignant digital tumours, which often require more extensive surgery for definitive treatment. The case herein highlights that this lesion can affect simultaneously multiple digits. Definitive diagnosis can be achieved by identification of keratin-producing epithelial cells on histopathology and confirmed by pancytokeratin labelling.

T cell-mediated protection against Mycobacterium tuberculosis (Mtb) is dependent upon the ability to localize within the site of pulmonary infection and directly interact with infected cells. In turn, vaccine strategies to improve rapid T cell targeting of Mtb-infected cells after pulmonary exposure are being actively pursued. Given parenterally, the subunit vaccine H56:CAF01 elicits polyfunctional CD4 T cells that localize to the lung parenchyma and confer durable protection. Here, we find that airway mucosal boosting of parenteral H56:CAF01 immunization greatly enhances the population of long-lived lung-resident T cells (Trm) and increases early vaccine T cell responses to pulmonary Mtb challenge in multiple mouse models. However, mucosal boosting does not alter the Th1/17 vaccine signature typical of H56:CAF01 and does not further improve durable control of pulmonary infection following aerosol Mtb-challenge. Additional mucosal boosting with H56:CAF01 further enhances the Trm response without further improving protection, while blocking the recruitment of non-Trm with FTY720-treatment failed to exposed Trm-mediated protection in mucosally boosting animals. These results demonstrate the limitations of maximizing lung-localized Trm in vaccine control of pulmonary Mtb infection, especially within an immunization protocol that is already optimized for the induction of mucosal-homing Th17 cells.

The development of a vaccine against genital chlamydia in women is advancing, and the evaluation of in situ immune responses following vaccination and challenge infections is crucial for development of a safe and protective vaccine. This study employs the sexually mature minipig model to characterize the genital in situ immune response to Chlamydia trachomatis infection in pigs previously immunized intramuscularly with UV-inactivated C. trachomatis serovar D (UV-SvD) adjuvanted/formulated with CAF01 adjuvant compared to a CAF01-alone control group. Pigs immunized with UV-SvD were significantly protected against vaginal challenge with C. trachomatis on day 3 post inoculation and showed significantly higher cervical infiltrations of approximately equal numbers of CD4 + and CD8 + T-cells, and IgG + and IgA + plasma cells compared to adjuvant-alone immunized controls. These immunological signatures correspond to findings in mice and are similar to those described in female chlamydia patients. This proves important potential for the pig model in elucidating immunological in situ signatures in future translational research in chlamydia vaccinology.

Background: Bovine respiratory disease (BRD) remains among the leading causes of death of cattle internationally. The objective of this study was to identify risk factors associated with exposure to BRD pathogens during the peri-weaning period (day (d)-14 to d 14 relative to weaning at 0) in dairy bull calves using serological responses to these pathogens as surrogate markers of exposure. Clinically normal Holstein-Friesian and Jersey breed bull calves (n = 72) were group housed in 4 pens using a factorial design with calves of different breeds and planes of nutrition in each pen. Intrinsic, management and clinical data were collected during the pre-weaning (d - 56 to d - 14) period. Calves were gradually weaned over 14 days (d - 14 to d 0). Serological analysis for antibodies against key BRD pathogens (BRSV, BPI3V, BHV-1, BHV-4, BCoV, BVDV and H. somni) was undertaken at d - 14 and d 14. Linear regression models (for BVDV, BPI3V, BHV-1, BHV-4, BCoV and H. somni) and a single mixed effect random variable model (for BRSV) were used to identify risk factors for changes in antibody levels to these pathogens., Results: BRSV was the only pathogen which demonstrated clustering by pen. Jersey calves experienced significantly lower changes in BVDV S/P than Holstein-Friesian calves. Animals with a high maximum respiratory score (≥8) recorded significant increases in H. somni S/P during the peri-weaning period when compared to those with respiratory scores of ≤3. Haptoglobin levels of between 1.32 and 1.60 mg/ml at d - 14 were significantly associated with decreases in BHV-1 S/N during the peri-weaning period. Higher BVDV S/P ratios at d - 14 were significantly correlated with increased changes in serological responses to BHV-4 over the peri-weaning period., Conclusions: Haptoglobin may have potential as a predictor of exposure to BHV-1. BRSV would appear to play a more significant role at the 'group' rather than 'individual animal' level. The significant associations between the pre-weaning levels of antibodies to certain BRD pathogens and changes in the levels of antibodies to the various pathogens during the peri-weaning period may reflect a cohort of possibly genetically linked 'better responders' among the study population.

Chronic inhalation of crystalline silica and silicates may lead to severe lung disease in humans, termed silicosis. The disease is an occupational health concern in miners and related professions worldwide. Silicosis is also a strong risk factor for tuberculosis in humans. Due to its subterranean lifestyle, the European badger (Meles meles) is continuously exposed to environmental dust, while this species is also susceptible to tuberculosis, caused by Mycobacterium bovis. To date, a thorough investigation of mineral dust retention and its possible implication as a risk factor for mycobacterial infection in badgers has not been performed. The aims of this retrospective histological study were (1) to describe the systemic tissue distribution of silica-laden macrophages (SLMs) in badgers; (2) to compare the amount of SLMs in tissues of badgers of differing M. bovis infection status, pulmonary SLM burden and age; and (3) to assess whether inflammation was associated with SLMs. We assessed lung, lymph nodes, liver and spleen of 60 wild-caught badgers of known M. bovis infection status for the presence of SLMs using polarizing light microscopy. SLMs were consistently present within the lungs and were widely distributed throughout the lymphatic system. No inflammatory reaction to SLMs, as occurs in human silicosis, was observed in any tissue. Distribution and amount of SLMs were similar between M. bovis positive and negative badgers, and we were not able to show an association between the amount of SLMs and M. bovis infection status. The amount of SLMs within intra- and extrathoracic lymph nodes was positively associated with the amount of pulmonary SLMs, and with age. This is the first report of substantial and systemic tissue retention of mineral dust particles in a mammalian species lacking associated chronic inflammation (i.e. silicosis). We further highlight different pathogenetic mechanisms underlying silicosis and benign SLM accumulations following siliceous dust inhalation.

Mycobacterium tuberculosis (Mtb), the etiologic agent of tuberculosis (TB), causes 1.8M deaths annually. The current vaccine, BCG, has failed to eradicate TB leaving 25% of the world's population with latent Mtb infection (LTBI), and 5-10% of these people will reactivate and develop active TB. An efficient therapeutic vaccine targeting LTBI could have an enormous impact on global TB incidence, and could be an important aid in fighting multidrug resistance, which is increasing globally. Here we show in a mouse model using the H56 (Ag85B-ESAT-6-Rv2660) TB vaccine candidate that post-exposure, but not preventive, vaccine protection requires low vaccine antigen doses for optimal protection. Loss of protection from high dose post-exposure vaccination was not associated with a loss of overall vaccine response magnitude, but rather with greater differentiation and lower functional avidity of vaccine-specific CD4 T cells. High vaccine antigen dose also led to a decreased ability of vaccine-specific CD4 T cells to home into the Mtb-infected lung parenchyma, a recently discovered important feature of T cell protection in mice. These results underscore the importance of T cell quality rather than magnitude in TB-vaccine protection, and the significant role that antigen dosing plays in vaccine-mediated protection.

Background: Ovine pulmonary adenocarcinoma (OPA), caused by Jaagsiekte sheep retrovirus (JSRV), is characterised by the development of invariably fatal lung tumours primarily in adult sheep. High infection rates and disease prevalence can develop during initial infection of flocks, leading to on-farm economic losses and animal welfare issues in sheep with advanced disease. The disease has been reported in Ireland and is notifiable, but the presence of JSRV has never been confirmed using molecular methods in this country. Additionally, due to the difficulties in ante-mortem diagnosis (especially of latently-infected animals, or those in the very early stages of disease), accurate information regarding national prevalence and distribution is unavailable. This study aimed to confirm the presence of JSRV in Ireland and to obtain estimates regarding prevalence and distribution by means of an abattoir survey utilising gross examination, histopathology, JSRV-specific reverse transcriptase polymerase chain reaction (RT-PCR) and SU protein specific immunohistochemistry (IHC) to examine the lungs of adult sheep., Results: Lungs from 1911 adult sheep were examined macroscopically in the abattoir and 369 were removed for further testing due to the presence of gross lesions of any kind. All 369 were subject to histopathology and RT-PCR, and 46 to IHC. Thirty-one lungs (31/1911, 1.6%) were positive for JSRV by RT-PCR and/or IHC but only ten cases of OPA were confirmed (10/1911, 0.5%) Four lung tumours not associated with JSRV were also identified. JSRV-positive sheep tended to cluster within the same flocks, and JSRV-positive sheep were identified in the counties of Donegal, Kerry, Kilkenny, Offaly, Tipperary, Waterford and Wicklow., Conclusions: The presence of JSRV has been confirmed in the Republic of Ireland for the first time using molecular methods (PCR) and IHC. In addition, an estimate of OPA prevalence in sheep at slaughter and information regarding distribution of JSRV infection has been obtained. The prevalence estimate appears similar to that of the United Kingdom (UK). Results also indicate that the virus has a diverse geographical distribution throughout Ireland. These data highlights the need for further research to establish national control and monitoring strategies., Competing Interests: No ethical approval was necessary as no animal was euthanased or slaughtered for the purpose of this study. Permission was granted by the supervising DAFM veterinarians of Kildare Chilling Co. for removal of tissue and collection of relevant traceability information.Not applicable.The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

There is evidence from epidemiology studies of a negative association between infection with helminth parasites and the development of allergy and asthma. Here, we demonstrate that the excretory/secretory products of the helminth Fasciola hepatica (FHES) protected mice against ovalbumin (OVA)-induced allergic asthma when administered at time of allergen sensitization. FHES reduced the accumulation of mucus, eosinophils and lymphocytes into the airways of allergen-challenged mice. Furthermore, FHES treatment suppressed Th2 responses in the airways. Interestingly, systemic administration of FHES at allergen challenge had no effect on airway inflammation, demonstrating that alum-induced Th2 response is set following initial allergen sensitization. Our findings highlight the immunomodulatory potential of molecules secreted by F. hepatica., (© 2017 John Wiley & Sons Ltd.)

The bovine paranasal sinuses are a group of complex cavernous air-filled spaces, lined by respiratory epithelium, the exact function of which is unclear. While lesions affecting these sinuses are occasionally reported in cattle, their microbial flora has not been defined. Furthermore, given that the various bacterial and viral pathogens causing bovine respiratory disease (BRD) persist within herds, we speculated that the paranasal sinuses may serve as a refuge for such infectious agents. The paranasal sinuses of clinically normal cattle (n = 99) and of cattle submitted for post-mortem examination (PME: n = 34) were examined by microbial culture, PCR and serology to include bacterial and viral pathogens typically associated with BRD: Mycoplasma bovis, Histophilus somni, Mannheimia haemolytica and Pasteurella multocida, bovine respiratory syncytial virus (BRSV) and bovine parainfluenza-3 virus (BPIV-3). Overall, the paranasal sinuses were either predominantly sterile or did not contain detectable microbes (83.5%: 94.9% of clinically normal and 50.0% of cattle submitted for PME). Bacteria, including BRD causing pathogens, were identified in relatively small numbers of cattle (<10%). While serology indicated widespread exposure of both clinically normal and cattle submitted for PME to BPIV-3 and BRSV (seroprevalences of 91.6% and 84.7%, respectively), PCR identified BPIV-3 in only one animal. To further explore these findings we investigated the potential role of the antimicrobial molecule nitric oxide (NO) within paranasal sinus epithelium using immunohistochemistry. Expression of the enzyme responsible for NO synthesis, inducible nitric oxide synthase (iNOS), was detected to varying degrees in 76.5% of a sub-sample of animals suggesting production of this compound plays a similar protective role in the bovine sinus as it does in humans.

We examined the pathogens, morphologic patterns, and risk factors associated with bovine respiratory disease (BRD) in 136 recently weaned cattle ("weanlings"), 6-12 mo of age, that were submitted for postmortem examination to regional veterinary laboratories in Ireland. A standardized sampling protocol included routine microbiologic investigations as well as polymerase chain reaction and immunohistochemistry. Lungs with histologic lesions were categorized into 1 of 5 morphologic patterns of pneumonia. Fibrinosuppurative bronchopneumonia (49%) and interstitial pneumonia (48%) were the morphologic patterns recorded most frequently. The various morphologic patterns of pulmonary lesions suggest the involvement of variable combinations of initiating and compounding infectious agents that hindered any simple classification of the etiopathogenesis of the pneumonias. Dual infections were detected in 58% of lungs, with Mannheimia haemolytica and Histophilus somni most frequently recorded in concert. M. haemolytica (43%) was the most frequently detected respiratory pathogen; H. somni was also shown to be frequently implicated in pneumonia in this age group of cattle. Bovine parainfluenza virus 3 (BPIV-3) and Bovine respiratory syncytial virus (16% each) were the viral agents detected most frequently. Potential respiratory pathogens (particularly Pasteurella multocida, BPIV-3, and H. somni) were frequently detected (64%) in lungs that had neither gross nor histologic pulmonary lesions, raising questions regarding their role in the pathogenesis of BRD. The breadth of respiratory pathogens detected in bovine lungs by various detection methods highlights the diagnostic value of parallel analyses in respiratory disease postmortem investigation.

Bovine respiratory disease (BRD) control poses significant challenges to the cattle industry worldwide. The sometimes complex interactions of factors associated with the animal, the pathogen and the environment complicate the implementation of effective control measures. Blanket vaccination or mass medication provides inconsistent control and the effective tackling of BRD will require innovative, evidence-based and targeted interventions which, if employed sensibly, offer useful alternatives for addressing this disease. This review appraises the role of the specific interventions employed in BRD control to assess how our understanding of their role and efficacy has evolved in recent years., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Bovine respiratory disease (BRD) is one of the most commonly diagnosed causes of morbidity and mortality in cattle and interactions of factors associated with the animal, the pathogen and the environment are central to its pathogenesis. Emerging knowledge of a role for pathogens traditionally assumed to be minor players in the pathogenesis of BRD reflects an increasingly complex situation that will necessitate regular reappraisal of BRD pathogenesis and control. This review appraises the role of selected key pathogens implicated in BRD pathogenesis to assess how our understanding of their role has evolved in recent years., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: An unusual presentation of skin disease was identified in two related neonatal Pedigree Limousin calves presented to University Veterinary Hospital, University College Dublin, following detailed post mortem examination a diagnosis of dermatosparaxis was made. Dermatosparaxis in animals or Ehlers Danlos Syndrome, which is the analogous condition seen in humans, is a connective tissue disorder characterised by extreme skin fragility. To the authors' knowledge this is the first report of such a diagnosis in the Limousin breed and the features of this lethal phenotype were severe in comparison to previous reports of the condition., Case Presentation: Two calves, which were full siblings, a pedigree Limousin bull (Calf A) and pedigree Limousin heifer (Calf B) were examined clinically after presenting collapsed since birth, both had grossly abnormal skin with multiple skin fissures visible and both calves were subsequently euthanised. Both calves underwent gross post mortem examination, after which histological samples were reviewed and electron microscopical examination of selected skin samples was carried out. Histological features of dysplastic dermal collagen were identified. The diagnosis of dermatosparaxis in the Limousin breed was confirmed. Genetic testing was conducted to determine if the current cases had the same mutation as has previously been described in Belgian Blue cattle. Some common parentage was traced but genetic testing did not show a similar mutation to that previously described in cattle. The specific genetic cause in this case is unknown., Conclusions: This is the first report of dermatosparaxis in the Limousin and the presentation of the dermatosparaxis phenotype has some noteworthy features thus further genetic testing is required to pinpoint the causative mutation or other genetic defect. Given the popularity of the breed and the lethal nature of the phenotype in this case it is important to raise awareness of the condition.

Bovine respiratory disease (BRD) is a multifactorial disease and the primary cause of both bovine morbidity and mortality in Ireland. The risk factors associated with a primary necropsy diagnosis of BRD among cattle in the traditional (non-feedlot) husbandry systems prevalent in Ireland have not been investigated previously. The aim of this case-control study was to investigate those risk factors among cattle of all ages over an 8 year period. A total of 3,090 BRD cases and 5,236 controls were matched by submitting veterinary practitioner. Univariable and multivariable analyses were performed to examine the association of selected animallevel, herd-level and environmental risk factors with case or control status using a conditional logistical regression model. Male cattle aged more than 31 days were significantly more likely to record a primary necropsy diagnosis of BRD than female cattle. Older cattle of both sexes were at increased odds of a BRD necropsy diagnosis than younger calves with the exception of female cattle aged greater than 165 days. The risk of a primary necropsy diagnosis of BRD increased with increasing herd size and decreased with increasing time in days since the last animal movement into the submitting herd. There were significantly reduced odds of a primary necropsy diagnosis of BRD in the summer (June to August) when compared with the autumn (September to November). These findings identify significant risk factors for a necropsy diagnosis of BRD under non-feedlot-type husbandry conditions., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Chlamydia is one of the most common sexually transmitted diseases in humans worldwide, causing chronic lesions in the reproductive tract. Due to its often asymptomatic course, there is limited knowledge about the initial changes in the genital tract following infection. This study employs a novel sexually mature minipig model to investigate the initial histopathological changes following vaginal infection with Chlamydia trachomatis serovar D., Results: A vaginal inoculation resulted in an infection primarily affecting the lower genital tract. The histopathological changes were characterized by a subepithelial inflammation consisting of neutrophils and mononuclear cells, followed by an increase in the number of plasma cells within the sub-epithelial stroma of the vagina. Detection of Chlamydia was associated with expression of cyclooxygenase-2 and interleukin-8 by superficial epithelial cells. The infection was self-limiting, with a duration of 7 days., Conclusion: Neutrophils, plasma cells and IL-8 have been linked with Chlamydia genital infection of unknown duration in human patients. In this study, we observe a similar pattern of local immune response/inflammation following experimental inoculation suggesting this porcine model shows promise as a model for translational chlamydia research.

Bovine besnoitiosis, caused by the apicomplexan protozoan parasite Besnoitia besnoiti, was diagnosed in an Irish dairy herd. This is the first diagnosis of besnoitiosis in Ireland or the UK and the most northerly European outbreak yet described. The diagnosis occurred following a farm investigation in June 2015 into an unusual dermatological problem that had been ongoing since 2010. On an annual basis, 1-2 per cent of cows in the herd exhibited clinical signs, including skin thickening, alopecia, weight loss and poor performance. Others displayed pyrexia, limb oedema, respiratory distress and reduced milk yield. Histopathological examination of skin revealed granulomatous and eosinophilic dermatitis, with characteristic intradermal protozoal cysts, consistent with cutaneous besnoitiosis. Follow-up serological testing and clinical examination of cattle (n=228) on the farm found that 68 per cent (144/212) were seropositive for B. besnoiti In addition, 51 per cent (117/228) had characteristic scleral conjunctival cysts and 68 per cent (134/198) had vulval cysts. Postmortem examination of a severely affected animal revealed typical gross and histopathological lesions of B. besnoiti infection. These results confirmed endemic infection with B. besnoiti The identification of this exotic disease highlights the importance of veterinary surveillance at both local and national level, particularly in relation to emerging diseases., (British Veterinary Association.)

Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne's disease (paratuberculosis), a chronic wasting disease in cattle with important welfare, economic and potential public health implications. Current tests are unable to recognise all stages of the disease, which makes it difficult to diagnose and control. This review explores emerging diagnostic techniques that could complement and enhance the diagnosis of MAP infection, including bacteriophage analysis, new MAP-specific antigens, host protein expression in response to infection, transcriptomic studies, analysis of microRNAs and investigation of the gastrointestinal microbiome. It emphasises the inherent challenges of diagnosing bovine Johne's disease and investigates novel areas which may have the potential both to advance our understanding of the immunopathology of MAP infection and to augment current diagnostic tests., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: During the FMD outbreak in Ireland and the UK in 2001, there was significant uncertainty amongstveterinary practitioners and government veterinary inspectors surrounding the clinical diagnosis of FMD insheep. This situation was complicated by reports of idiopathic oral ulcers that closely resembled FMD ongross appearance which at that time were referred to as ovine mouth and gum obscure disease., Methods: A field and abattoir study was carried out to determine the frequency, appearance and significance of oraland digital lesions in sheep in Ireland. A total of 3, 263 sheep were examined in 22 flocks, including 1, 969lambs and 1, 294 adults. A further 2,403 animals were examined by abattoir inspections. Animals bearing lesions of interest were identified, samples of the lesions were taken and subsequently examined by bacteriology, electron microscopy, serology, immunohistochemistry and histopathology., Results: Forty four oral and 20 digital lesions were identified and characterised. Oral lesions were recorded mostfrequently in lambs, where the most common cause was orf virus infection. The majority of the oral lesions recorded in the adults was idiopathic and consistent with a diagnosis of idiopathic oral ulceration. A variety of digital lesions was observed, consistent with scald, foot-rot and contagious ovine digital dermatitis (CODD). All of the animals with lesions were seronegative to FMD virus (FMDV)., Conclusions: There was no difficulty in differentiating these lesions from those caused by FMDV on the basis of flockhistory and careful clinical examination.

Bordetella pertussis causes whooping cough, an infectious disease of the respiratory tract that is re-emerging despite high vaccine coverage. Here we examined the role of Toll-like receptor (TLR) adapter protein Mal in the control of B. pertussis infection in the lungs. We found that B. pertussis bacterial load in the lungs of Mal-defective (Mal(-/-)) mice exceeded that of wild-type (WT) mice by up to 100-fold and bacteria disseminated to the liver in Mal(-/-) mice and 50% of these mice died from the infection. Macrophages from Mal(-/-) mice were defective in an early burst of pro-inflammatory cytokine production and in their ability to kill or constrain intracellular growth of B. pertussis. Importantly, the B. pertussis bacterial load in the lungs inversely correlated with the number of alveolar macrophages. Despite the maintenance and expansion of other cell populations, alveolar macrophages were completely depleted from the lungs of infected Mal(-/-) mice, but not from infected WT mice. Our findings define for the first time a role for a microbial pattern-recognition pathway in the survival of alveolar macrophages and uncover a mechanism of macrophage-mediated immunity to B. pertussis in which Mal controls intracellular survival and dissemination of bacteria from the lungs.

Background: There is a need to improve vaccination against respiratory pathogens in calves by stimulation of local immunity at the site of pathogen entry at an early stage in life. Ideally such a vaccine preparation would not be inhibited by the maternally derived antibodies. Additionally, localized immune response at the site of infection is also crucial to control infection at the site of entry of virus. The present study investigated the response to an intranasal bovine parainfluenza 3 virus (BPI3V) antigen preparation encapsulated in PLGA (poly dl-lactic-co-glycolide) nanoparticles in the presence of pre-existing anti-BPI3V antibodies in young calves and comparing it to a commercially available BPI3V respiratory vaccine., Results: There was a significant (P < 0.05) increase in BPI3V-specific IgA in the nasal mucus of the BPI3V nanoparticle vaccine group alone. Following administration of the nanoparticle vaccine an early immune response was induced that continued to grow until the end of study and was not observed in the other treatment groups. Virus specific serum IgG response to both the nanoparticle vaccine and commercial live attenuated vaccine showed a significant (P < 0.05) rise over the period of study. However, the cell mediated immune response observed didn't show any significant rise in any of the treatment groups., Conclusion: Calves administered the intranasal nanoparticle vaccine induced significantly greater mucosal IgA responses, compared to the other treatment groups. This suggests an enhanced, sustained mucosal-based immunological response to the BPI3V nanoparticle vaccine in the face of pre-existing antibodies to BPI3V, which are encouraging and potentially useful characteristics of a candidate vaccine. However, ability of nanoparticle vaccine in eliciting cell mediated immune response needs further investigation. More sustained local mucosal immunity induced by nanoparticle vaccine has obvious potential if it translates into enhanced protective immunity in the face of virus outbreak.

Protein energy malnutrition (PEM) increases susceptibility to infectious diseases, including tuberculosis (TB), but it is not clear how PEM influences vaccine-promoted immunity to TB. We demonstrate that PEM during low-level steady-state TB infection in a mouse model results in rapid relapse of Mycobacterium tuberculosis, as well as increased pathology, in both Mycobacterium bovis BCG-vaccinated and unvaccinated animals. PEM did not change the overall numbers of CD4 T cells in BCG-vaccinated animals but resulted in an almost complete loss of antigen-specific cytokine production. Furthermore, there was a change in cytokine expression characterized by a gradual loss of multifunctional antigen-specific CD4 T cells and an increased proportion of effector cells expressing gamma interferon and tumor necrosis factor alpha (IFN-γ(+) TNF-α(+) and IFN-γ(+) cells). PEM during M. tuberculosis infection completely blocked the protection afforded by the H56-CAF01 subunit vaccine, and this was associated with a very substantial loss of the interleukin-2-positive memory CD4 T cells promoted by this vaccine. Similarly, PEM during the vaccination phase markedly reduced the H56-CAF01 vaccine response, influencing all cytokine-producing CD4 T cell subsets, with the exception of CD4 T cells positive for TNF-α only. Importantly, this impairment was reversible and resupplementation of protein during infection rescued both the vaccine-promoted T cell response and the protective effect of the vaccine against M. tuberculosis infection., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Tuberculosis (TB) does not always develop in people or cattle exposed to the disease and some exposed individuals may not exhibit evidence of infection. Such variability in susceptibility may be mediated through host innate immunity, non-specific inflammatory responses that may successfully eliminate infection or at least reduce the infectious load, thus modulating and easing the burden on the subsequent acquired immune response. Assessing evidence from research in man, cattle and laboratory animal models, this review appraises the role of innate immunity in TB including the role of particular leucocytes (i.e. macrophages, neutrophils, γδ-T lymphocytes and natural killer cells), endogenous host defence compounds (i.e. cathelicidin, human neutrophil peptide, lipocalin and natural resistance-associated membrane protein-1) and, in particular, vitamin D. Innate responses may be particularly important in neonatal animals and people where adaptive responses have not yet established and their success in preventing the establishment of infection may be predicated on dose and/or route of infection as well as on characteristics of the infecting isolate. Innate defences could potentially be exploited in novel vaccination and immunotherapeutic approaches to disease control, modulating their effectiveness through the use of defined mycobacterial peptides as adjuvants or therapeutics. Such novel immunomodulatory compounds may be particularly relevant in countering emerging multi- and extremely drug-resistant strains of Mycobacterium tuberculosis (Mtb)., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Vaccine adjuvants are typically designed to stimulate both systemic and mucosal immune responses. Polymeric nanoparticles have been used as adjuvants in the development of vaccines against a number of viral pathogens and tested in laboratory animals. The objective of the study was to assess if synthetic bovine parainfluenza virus type-3 (BPI3V) peptide motifs and solubilised BPI3V proteins encapsulated in poly (dl-lactic-co-glycolide) (PLGA) nanoparticles (NPs) induce specific humoral immune responses in a mouse model following intranasal administration. BPI3V-specific and peptide specific IgG ELISAs were used to measure serum IgG levels to BPI3V. Intranasal delivery of PLGA nanoparticles encapsulating BPI3V proteins elicited an early, gradually increasing BPI3V-specific IgG response that persisted over the subsequent 6 weeks, suggesting slow, persistent release of antigen. PLGA-BPI3V particles administered intranasally induced a stronger IgG antibody response at an earlier time point compared with solubilised BPI3V antigen alone. Such an approach could be deployed in the development of new generation vaccines., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In 2006 bluetongue virus serotype 8 (BTV 8) was identified for the first time in the Netherlands causing a major epidemic in sheep and cattle that quickly spread to neighbouring Belgium, Germany and beyond to France and the UK. This resulted in severe animal health and welfare problems as well as substantial economic losses to the agrifood industries of these countries. Given that the early diagnosis of BTV infection 'in-the-field' is extremely useful to its subsequent management and control, this study was established to design a novel, sensitive and rapid nucleic acid diagnostic test for the serotype-specific detection of BTV 8, which could be used without the use of advanced laboratory support and equipment. Primers for the detection of BTV 8 were based on genome segment 2 of the virus, the VP2 gene. The assay was assessed using a full panel of BTV reference strains and clinical samples. Positive amplification was observed using a fluorescent detection reagent. The sensitivity of the RT-LAMP assay was 102 copies of RNA. The assay did not amplify the closely related orbivirus EHDV. This novel RT-LAMP offers a sensitive, specific and rapid method of detecting BTV 8. The approach is inexpensive and easy to use and could potentially be used in a 'pen-side' setting 'in the field' or by smaller less well-equipped laboratories in developing countries., (Copyright © 2014 Elsevier B.V. All rights reserved.)

CD4 T cells are crucial to the control of Mycobacterium tuberculosis infection and are a key component of current vaccine strategies. Conversely, immune-mediated pathology drives disease, and recent evidence suggests that adaptive and innate responses are evolutionarily beneficial to M. tuberculosis. We compare the functionality of CD4 T cell responses mounted against dominant and cryptic epitopes of the M. tuberculosis 6-kDa early secreted Ag (ESAT-6) before and postinfection. Protective T cells against cryptic epitopes not targeted during natural infection were induced by vaccinating mice with a truncated ESAT-6 protein, lacking the dominant epitope. The ability to generate T cells that recognize multiple cryptic epitopes was MHC-haplotype dependent, including increased potential via heterologous MHC class II dimers. Before infection, cryptic epitope-specific T cells displayed enhanced proliferative capacity and delayed cytokine kinetics. After aerosol M. tuberculosis challenge, vaccine-elicited CD4 T cells expanded and recruited to the lung. In chronic infection, dominant epitope-specific T cells developed a terminal differentiated KLRG1(+)/PD-1(lo) surface phenotype that was significantly reduced in the cryptic epitope-specific T cell populations. Dominant epitope-specific T cells in vaccinated animals developed into IFN-γ- and IFN-γ,TNF-α-coproducing effector cells, characteristic of the endogenous response. In contrast, cryptic epitope-specific CD4 T cells maintained significantly greater IFN-γ(+)TNF-α(+)IL-2(+) and TNF-α(+)IL-2(+) memory-associated polyfunctionality and enhanced proliferative capacity. Vaccine-associated IL-17A production by cryptic CD4 T cells was also enhanced, but without increased neutrophilia/pathology. Direct comparison of dominant/cryptic epitope-specific CD4 T cells within covaccinated mice confirmed the superior ability of protective cryptic epitope-specific T cells to resist M. tuberculosis infection-driven T cell differentiation.

Forty eight individual pigs (8.7±0.26 kg) weaned at 28±1 d of age were used in a 22-d study to evaluate the effect of oral administration of a Bacillus pumilus spore suspension on growth performance and health indicators. Treatments (n = 16) were: (1) non-medicated diet; (2) medicated diet with apramycin (200 mg/kg) and pharmacological levels of zinc oxide (2,500 mg zinc/kg) and (3) B. pumilus diet (non-medicated diet + 10(10) spores/day B. pumilus). Final body weight and average daily gain tended to be lower (P = 0.07) and feed conversion ratio was worsened (P<0.05) for the medicated treatment compared to the B. pumilus treatment. Ileal E. coli counts were lower for the B. pumilus and medicated treatments compared to the non-medicated treatment (P<0.05), perhaps as a result of increased ileal propionic acid concentrations (P<0.001). However, the medicated treatment reduced fecal (P<0.001) and cecal (P<0.05) Lactobacillus counts and tended to reduce the total cecal short chain fatty acid (SCFA) concentration (P = 0.10). Liver weights were lighter and concentrations of liver enzymes higher (P<0.05) in pigs on the medicated treatment compared to those on the non-medicated or B. pumilus treatments. Pigs on the B. pumilus treatment had lower overall lymphocyte and higher granulocyte percentages (P<0.001) and higher numbers of jejunal goblet cells (P<0.01) than pigs on either of the other two treatments or the non-medicated treatment, respectively. However, histopathological examination of the small intestine, kidneys and liver revealed no abnormalities. Overall, the B. pumilus treatment decreased ileal E. coli counts in a manner similar to the medicated treatment but without the adverse effects on growth performance, Lactobacillus counts, cecal SCFA concentration and possible liver toxicity experienced with the medicated treatment.

The ESX systems from Mycobacterium tuberculosis are responsible for the secretion of highly immunogenic proteins of key importance for bacterial survival and growth. The two prototypic proteins, ESAT-6 (EsxA from ESX-1) and TB10.4 (EsxH from ESX-3) share a lot of characteristics regarding genome organization, size, antigenic properties, and vaccine potential but the two molecules clearly have very different roles in bacterial physiology. To further investigate the role of ESAT-6 and TB10.4 as preventive and post-exposure tuberculosis vaccines, we evaluated four different fusion-protein vaccines; H1, H4, H56 and H28, that differ only in these two components. We found that all of these vaccines give rise to protection in a conventional prophylactic vaccination model. In contrast, only the ESAT-6-containing vaccines resulted in significant protection against reactivation, when administered post-exposure. This difference in post-exposure activity did not correlate with a difference in gene expression during infection or a differential magnitude or quality of the vaccine-specific CD4 T cells induced by ESAT-6 versus TB10.4-containing vaccines. The post-exposure effect of the ESAT-6 based vaccines was found to be influenced by the infectious load at the time-point of vaccination and was abolished in chronically infected animals with high bacterial loads at the onset of vaccination. Our data demonstrate that there are specific requirements for the immune system to target an already established tuberculosis infection and that ESAT-6 has a unique potential in post-exposure vaccination strategies.

Avian tuberculosis rarely affects ratites compared to other bird species and is typically caused by Mycobacterium avium species. This study describes the pathological and microbiological findings in three adult ostriches with mycobacteriosis, in one of which Mycobacterium bovis was isolated from the lesions. Post mortem examinations on ostriches from two different zoological collections in Ireland revealed multifocal caseous granulomas affecting the spleen and liver in all cases, with additional involvement of intestines in two cases. In one case, granulomas were present within the pharynx, at the thoracic inlet and multifocally on the pleural surface. Acid-fast bacilli were observed in all lesions. Mycobacterium sp. of the M. avium complex was isolated from the intestinal lesions in the two cases with intestinal involvement, and M. bovis sp. oligotype SB0140 was cultured from the liver of the third ostrich. This represents the first reported case of M. bovis infection in an ostrich. Avian tuberculosis due to M. bovis is rare and to date has been reported in only parrots and experimentally inoculated birds. Mycobacterium bovis needs to be considered as a possible cause of tuberculosis in ostriches because the lesions are similar to those observed with M. avium complex infection.

The role of selenium (Se) supplementation in cancer prevention is controversial; effects often depend on the nutritional status of the subject and on the chemical form in which Se is provided. We used a combination of in vitro and in vivo models to study two unique therapeutic windows for intervention in the process of cutaneous melanomagenisis, and to examine the utility of two different chemical forms of Se for prevention and treatment of melanoma. We studied the effects of Se in vitro on UV-induced oxidative stress in melanocytes, and on apoptosis and cell cycle progression in melanoma cells. In vivo, we used the HGF transgenic mouse model of UV-induced melanoma to demonstrate that topical treatment with l-selenomethionine results in a significant delay in the time required for UV-induced melanoma development, but also increases the rate of growth of those tumors once they appear. In a second mouse model, we found that oral administration of high dose methylseleninic acid significantly decreases the size of human melanoma xenografts. Our findings suggest that modestly elevation of selenium levels in the skin might risk acceleration of growth of incipient tumors. Additionally, certain Se compounds administered at very high doses could have utility for the treatment of fully-malignant tumors or prevention of recurrence.

This study assessed the effect of feeding genetically modified maize expressing a truncated form of the Cry1Ab protein from Bacillus thuringiensis (Bt MON810 maize) to sows during gestation and lactation and their offspring from weaning to 115 d postweaning on offspring growth and health. After weaning at approximately 28 d of age (d 0), individually penned, mixed sex pigs (approximately 8 kg BW) from sows fed isogenic or Bt maize diets were blocked by sow treatment, sex, and BW and randomly assigned to Bt or isogenic maize diets as follows: i) isogenic maize-fed sow/isogenic maize-fed offspring (iso/iso); ii) isogenic maize-fed sow/Bt maize-fed offspring (iso/Bt); iii) Bt maize-fed sow/isogenic maize-fed offspring (Bt/iso); and iv) Bt maize-fed sow/Bt maize-fed offspring (Bt/Bt). Growth performance was recorded at intervals to harvest at approximately 105 kg BW (n=15/treatment) and blood samples were taken for biochemical analysis on d 0, 30, 70, 100, and 115 postweaning (n=10/treatment). Pigs were harvested on d 115 postweaning (n=10/treatment), and carcass weight, backfat depth, and organ weights (heart, kidney, spleen, and liver) were recorded. Kidney, liver, lymph nodes, and small intestine were collected for histological analysis. Offspring from Bt maize-fed sows were heavier than offspring from isogenic maize-fed sows on d 30 (P<0.05), 100 (P<0.05), and 115 postweaning (P<0.05) and had greater overall ADG (P<0.05). Overall ADFI was greater for offspring from sows fed Bt maize (P<0.05) and for Bt maize-fed pigs (P<0.05). Offspring from Bt maize-fed sows had greater carcass (P<0.05) and lighter spleen (P<0.05) weights. Dressing percentage was greater for Bt maize-fed pigs than isogenic maize-fed pigs (P<0.05), and livers were lighter for pigs in the Bt/Bt group than pigs in the iso/Bt or Bt/iso group (P<0.05). Offspring from Bt maize-fed sows also had greater duodenal crypt depths (P<0.05) and lower villus height/crypt depth ratios (P<0.05). No pathology was observed in the organs, and serum biochemistry values generally remained within normal limits and no overall differences were observed, with the exception of overall γ glutamyltransferase, which was less for pigs on the Bt/Bt treatment than pigs on the iso/Bt and Bt/iso treatments. These results indicate that transgenerational consumption of Bt maize diets is not detrimental to pig growth and health.

A total of 72 male weaned pigs were used in a 110-day study to investigate the effect of feeding genetically modified (GM) Bt MON810 maize on selected growth and health indicators. It was hypothesised that in pigs fed Bt maize, growth and health are not impacted compared with pigs fed isogenic maize-based diets. Following a 12-day basal period, pigs (10.7 ± 1.9 kg body weight (BW); ∼40 days old) were blocked by weight and ancestry and randomly assigned to treatments: (1) non-GM maize diet for 110 days (non-GM), (2) GM maize diet for 110 days (GM), (3) non-GM maize diet for 30 days followed by GM maize diet up to day 110 (non-GM/GM) and (4) GM maize diet for 30 days followed by non-GM maize diet up to day 110 (GM/non-GM). BW and daily feed intake were recorded on days 0, 30, 60 and 110 (n = 15). Body composition was determined by dual energy X-ray absorptiometry (n = 10) on day 80. Following slaughter on day 110, organs and intestines were weighed and sampled for histological analysis and urine was collected for biochemical analysis (n = 10). Serum biochemistry analysis was performed on days 0, 30, 60, 100 and 110. Growth performance and serum biochemistry were analysed as repeated measures with time and treatment as main factors. The slice option of SAS was used to determine treatment differences at individual time points. There was no effect of feeding GM maize on overall growth, body composition, organ and intestinal weight and histology or serum biochemistry on days 60 and 100 and on urine biochemistry on day 110. A treatment × time interaction was observed for serum urea (SU; P < 0.05), creatinine (SC; P < 0.05) and aspartate aminotransferase (AST; P < 0.05). On day 30, SU was lower for the non-GM/GM treatment compared with the non-GM, GM and GM/non-GM treatments (P < 0.05). On day 110, SC was higher for the non-GM/GM and GM/non-GM treatments compared with non-GM and GM treatments (P < 0.05). Overall, serum total protein was lower for the GM/non-GM treatment compared with the non-GM/GM treatment (P < 0.05). The magnitude of change observed in some serum biochemical parameters did not indicate organ dysfunction and the changes were not accompanied by histological lesions. Long-term feeding of GM maize to pigs did not adversely affect growth or the selected health indicators investigated.

Background: Historically, quantitative clinical diagnostic assays (QCDAs) have not been accepted for use in pharmacokinetic or bioequivalence studies because they do not fully comply with the US Food and Drug Administration (FDA) Guidance for Industry: Bioanalytical Method Validation (e.g., full calibration curve not generated with each analytical run). Samples from a bioequivalence study were analysed for insulin and C-peptide concentrations with QCDAs and guidance-conforming radioimmunoassays (RIAs) and the results compared across and within assays., Methods: Serum samples (n=1913) from study MKC-TI-142 were analysed first using the Roche E170 electrochemiluminescence immunoassay (ECLIA) for insulin concentration and the Immulite 2000 chemiluminescence immunoassay (CLIA) for C-peptide, and then using the corresponding Millipore RIAs., Results: The insulin assays were highly correlated: r2=0.92 excluding samples requiring dilution and R2=0.88 including all samples. There was increasing negative bias of the ECLIA compared with the RIA with increasing insulin, especially with samples that required dilution for the RIA. The ECLIA had significantly fewer below-quantifiable-limit samples, a larger dynamic analysis range without dilution, and tighter agreement within incurred sample reanalysis (ISR) as compared with the RIA. The C-peptide assays showed good agreement but the CLIA method produced ISR results that were in closer agreement with the original values., Conclusions: The science indicates that the QCDAs are appropriate for the quantification of serum insulin (ECLIA) and C-peptide (CLIA) concentrations in human pharmacokinetic and bioequivalence studies even though the calibration curve is not generated in each analytical run.

Purpose: To determine distribution and deposition of Technosphere® Insulin (TI) inhalation powder and the rate of clearance of fumaryl diketopiperazine (FDKP; major component of Technosphere particles) and insulin from the lungs., Methods: Deposition and distribution of (99m)pertechnetate adsorbed onto TI immediately after administration using the MedTone® inhaler was quantified by gamma-scintigraphy. Clearance from the lungs was studied in a second experiment by serial bronchoalveolar lavage (BAL) after administration of TI inhalation powder and assay of the recovered fluid for FDKP and insulin., Results: Following inhalation, ~60% of radioactivity (adsorbed on TI) emitted from the inhaler was delivered to the lungs; the remainder of the emitted dose was swallowed. Clearance from the lung epithelial lining fluid (ELF) of FDKP and insulin have a half-life of ~1 hour., Conclusion: TI inhalation powder administered via the MedTone inhaler was uniformly distributed throughout the lungs; ~40% of the initial cartridge load reached the lungs. Insulin and FDKP are quickly cleared from the lungs, mainly by absorption into the systemic circulation. The terminal clearance half-life from the lung ELF, estimated from sequential BAL fluid measurements for both components, was ~1 hour. Since there is an overnight washout period, the potential for accumulation on chronic administration is minimal.

All tuberculosis vaccines currently in clinical trials are designed as prophylactic vaccines based on early expressed antigens. We have developed a multistage vaccination strategy in which the early antigens Ag85B and 6-kDa early secretory antigenic target (ESAT-6) are combined with the latency-associated protein Rv2660c (H56 vaccine). In CB6F1 mice we show that Rv2660c is stably expressed in late stages of infection despite an overall reduced transcription. The H56 vaccine promotes a T cell response against all protein components that is characterized by a high proportion of polyfunctional CD4(+) T cells. In three different pre-exposure mouse models, H56 confers protective immunity characterized by a more efficient containment of late-stage infection than the Ag85B-ESAT6 vaccine (H1) and BCG. In two mouse models of latent tuberculosis, we show that H56 vaccination after exposure is able to control reactivation and significantly lower the bacterial load compared to adjuvant control mice.