Peste des petits ruminants (PPR) is an economically important disease of small ruminants such as sheep and goats. The disease is characterized by severe pyrexia, oculo-nasal discharge, pneumonia, necrosis and ulceration of the mucous membrane and inflammation of the gastro-intestinal tract leading to severe diarrhea. A SYBR Green I based real time RT-PCR targeting the N gene of PPRV has not been established for PPRV detection. Thus, the objective of present study was to develop highly sensitive N gene target SYBR Green I real time RT-PCR for specific detection and quantification of PPRV in clinical samples. A set of primers was designed to detect the nucleocapsid (N) gene of PPRV.
The assay exhibited high specificity as all the viruses which have clinical and structural similarities to PPRV including Canine distemper virus (CDV), Measles virus (MV), Bluetongue virus (BTV) and Newcastle disease virus (NDV) failed to show an amplification signal. The lower detection limit of the assay was 5.11 copies/μl (Ct value of 33.67 ± 0.5) and 0.001 TCID50/ml (Ct value of 34.7 ± 0.5) based on plasmid copy number and tissue culture infectivity titre. The assay was 3-log more sensitive than the conventional RT-PCR. The coefficient of variation (CV) values for intra- and inter-assay variability were low, ranging from 0.32% - 2.31%, and 0.71% - 5.32%, respectively. To evaluate the performance of the newly developed assay, a total of 36 clinical samples suspected of PPR were screened for the presence of PPRV in parallel with conventional RT-PCR. The real time RT-PCR assay detected PPRV in 30 (83.3%) of clinical samples compared to 16 (44.4%) by conventional RT-PCR.
The two-step SYBR Green I based real time RT-PCR assay reported here is highly sensitive, specific, reproducible and rapid for detection and quantification of PPRV nucleic acids.
SYBR Green I; Real time RT-PCR; N gene; PPRV; Detection; Quantitation
Non-tuberculous mycobacteria responsible for piscine mycobacteriosis usually produce visceral granulomas in both freshwater and marine species. In this study, the first occurrence of Mycobacterium chelonae associated with tumor-like lesions in the Russian sturgeon (Acipenser gueldenstaedtii) is reported. Fifteen sturgeons from an Italian fish farm showing skin and oral cauliflower-like masses were investigated by histopathology, bacterial culture and molecular analyses.
A total of 20 masses different in size located in the mouth and in pectoral and caudal fins (characterized by abundant calcium deposits and by mild to moderate granulomatous inflammation) were observed with a significant different degree of histological severity. All internal organs of the fish were negative for mycobacteria, Ziehl-Neelsen was positive in only one of the oral masses, whereas bacterial and PCR analyses detected the presence of M. chelonae for almost all the skin and oral masses. Based on these results, a calcinosis of dystrophic origin associated with a chronic granulomatous inflammation was considered as a primary diagnosis consequent to tissue injury in areas susceptible to trauma.
We hypothesized that the occurrence of M. chelonae in farmed sturgeons was only a secondary event related to its presence in a stressful rearing environment and subsequent to a dystrophic calcinosis occurred in previously damaged tissues.
Acipenser gueldenstaedtii; Aquaculture; Calcinosis circumscripta; Fish pathology; Mycobacteria
Enzymatic activity of Telomerase Reverse Transcriptase (TERT) is important in maintaining the telomere length and has been implicated in cancer and aging related pathology. Since cancer susceptibility as well as longevity of dogs vary between breeds, this study involved sequencing the entire TERT gene of Canis familiaris from DNA samples obtained from forty dogs, with ten dogs each of four breeds: Shih Tzu, Dachshund, Irish Wolfhound, and Newfoundland, each with different life expectancies and susceptibility to cancer.
We compared the sequences of all forty individuals amongst one another and with the published sequence of canine TERT, and analyzed relationships between members of the same or different breeds. Two separate phylogenetic trees were generated and analyzed from these individuals. Polymorphisms were found most frequently in intronic regions of the gene, although exonic polymorphisms also were observed. In many locations genotypes were observed that were either homozygous for the reference sequence or heterozygous, but the variant homozygous genotype was not observed.
We propose that these homozygous variants are likely to have adverse effects in dogs. It was also found that the polymorphisms did not segregate by breed. Because the four breeds chosen come from geographically and physiologically distinct backgrounds, it can be inferred that the polymorphic diversification of TERT preceded breed derivation.
Despite the precise pathogenesis of atopic dermatitis (AD) is unknown, an immune dysregulation that causes Th2-predominant inflammation and an intrinsic defect in skin barrier function are currently the two major hypotheses, according to the so-called outside-inside-outside model. Mast cells (MCs) are involved in AD both by releasing Th2 polarizing cytokines and generating pruritus symptoms through release of histamine and tryptase. A link between MCs and skin barrier defects was recently uncovered, with histamine being found to profoundly contribute to the skin barrier defects.
Palmitoylethanolamide and related lipid mediators are endogenous bioactive compounds, considered to play a protective homeostatic role in many tissues: evidence collected so far shows that the anti-inflammatory effect of palmitoylethanolamide depends on the down-modulation of MC degranulation.
Based on this background, the purpose of the present study was twofold: (a) to determine if the endogenous levels of palmitoylethanolamide and other bioactive lipid mediators are changed in the skin of AD dogs compared to healthy animals; (b) to examine if MC number is increased in the skin of AD dogs and, if so, whether it depends on MC in-situ proliferation.
The amount of lipid extract expressed as percent of biopsy tissue weight was significantly reduced in AD skin while the levels of all analyzed bioactive lipid mediators were significantly elevated, with palmitoylethanolamide showing the highest increase.
In dogs with AD, the number of MCs was significantly increased in both the subepidermal and the perifollicular compartments and their granule content was significantly decreased in the latter. Also, in situ proliferation of MCs was documented.
The levels of palmitoylethanolamide and other bioactive lipid mediators were shown to increase in AD skin compared to healthy samples, leading to the hypothesis that they may be part of the body’s innate mechanisms to maintain cellular homeostasis when faced with AD-related inflammation. In particular, the increase may be considered a temptative response to down-regulating the observed elevation in the number, functionality and proliferative state of MCs in the skin of AD dogs. Further studies are warranted to confirm the hypothesis.
Atopic dermatitis; Dog; Acylethanolamides; Palmitoylethanolamide; Mast cells
Despite great effort and investment incurred over decades to control bovine tuberculosis (bTB), it is still one of the most important zoonotic diseases in many areas of the world. Test-and-slaughter strategies, the basis of most bTB eradication programs carried out worldwide, have demonstrated its usefulness in the control of the disease. However, in certain countries, eradication has not been achieved due in part to limitations of currently available diagnostic tests. In this study, results of in-vivo and post-mortem diagnostic tests performed on 3,614 animals from 152 bTB-infected cattle herds (beef, dairy, and bullfighting) detected in 2007–2010 in the region of Castilla y León, Spain, were analyzed to identify factors associated with positive bacteriological results in cattle that were non-reactors to the single intradermal tuberculin test, to the interferon-gamma (IFN-γ) assay, or to both tests applied in parallel (Test negative/Culture + animals, T-/C+). The association of individual factors (age, productive type, and number of herd-tests performed since the disclosure of the outbreak) with the bacteriology outcome (positive/negative) was analyzed using a mixed multivariate logistic regression model.
The proportion of non-reactors with a positive post-mortem result ranged from 24.3% in the case of the SIT test to 12.9% (IFN-γ with 0.05 threshold) and 11.9% (95% CI 9.9-11.4%) using both tests in parallel. Older (>4.5 years) and bullfighting cattle were associated with increased odds of confirmed bTB infection by bacteriology, whereas dairy cattle showed a significantly lower risk. Ancillary use of IFN-γ assay reduced the proportion of T-/C + animals in high risk groups.
These results demonstrate the likelihood of positive bacteriological results in non-reactor cattle is influenced by individual epidemiological factors of tested animals. Increased surveillance on non-reactors with an increased probability of being false negative could be helpful to avoid bTB persistence, particularly in chronically infected herds. These findings may aid in the development of effective strategies for eradication of bTB in Spain.
Tuberculosis; Cattle; Diagnosis; Single tuberculin test; Interferon-gamma assay; Risk factors
Individual enteropathogen infections in healthy and clinically ill cats are well described, but prevalence and patterns of enteropathogen co-infection have only been reported on a limited basis. We studied enteropathogen co-infection in diarrhoeic UK cats using results of a real time PCR assay for 8 enteropathogenic species; feline coronavirus (Co), feline panleukopenia virus (Pa), Clostridium perfringens (Cl), Salmonella enterica (Sa), Giardia spp. (Gi), Tritrichomonas foetus (Tr), Cryptosporidium spp. (Cr), and Toxoplasma gondii (To). Age, gender, breed and history were recorded. PCR panels from 1088 diarrhoeic cats were available for analysis.
Overall enteropathogen prevalence was 56.9% (Co), 22.1% (Pa), 56.6% (Cl), 0.8% (Sa), 20.6% (Gi), 18.8% (Tr), 24.4% (Cr) and 1.0% (To). Prevalence of Co, Gi and Tr was higher in pedigree cats compared to non-pedigree cats (DSH) and prevalence decreased with increasing age for Co, Pa, Gi, Cr and Tr. Co-infection was common: ≥2 enteropathogens were detected in 62.5% of cats, and 13.3% of cats had ≥4 enteropathogens. Mean (
x¯) enteropathogen co-infection 2.01 (±1.3 SD), was significantly higher in pedigree cats (
x¯ =2.51) compared to DSH (
x¯ =1.68) and decreased with age (
x¯ =2.64 <6 months,
x¯ =1.68 for >1 yr). More cats were negative for all 8 enteropathogens tested (12.7%) than expected. When exact combinations of co-infection were examined, Tr tended to be found in combinations with Co, Cl, and Gi.
Multiple infections should be considered the most likely result of faecal testing in cats, and case management needs to take this into account. In contrast, the relatively high percentage of cats negative for all 8 enteropathogens tested could indicate an innate resistance to infection. Alternatively it could indicate a lack of exposure to these 8 enteropathogens or the presence of other enteropathogens not assessed by this assay.
Feline; Enteropathogen; Co-infection
Forty dogs presented for brachycephalic airway obstructive syndrome with laryngeal collapse not over 1st degree (saccule eversion) underwent glottis endoscopic and radiographic skull measurements before surgery. Fifteen Pugs, fifteen French and ten English Bulldogs were included. The goals were prospectively to compare three common brachycephalic breeds for anatomical differences regarding glottis and skull measurements, and to assess if any correlation between glottis and skull measurements was present. Linear measurements were used to obtain glottis and skull indices. Correlations between glottis and skull indices and glottic measurements were evaluated. Finally, glottis indices were compared among the three breeds.
No correlation was found for glottis and skull indices. The glottic index differed among the three breeds (smaller in Pugs and higher in English Bulldogs), ultimately representing a morphologic indicator of the different larynx shape in the three breeds (more rounded in English Bulldogs, more elliptical in Pugs and in-between in French Bulldogs).
The lack of correlation between skull/glottic indices does not support skull morphology as predictor of glottic morphology. As Pugs had the lowest glottic index, it may be speculated that Pugs’ original narrow glottic width may predispose to further progressive respiratory deterioration more easily than in the other two breeds.
Canine brachycephalic airway obstructive syndrome; Skull measurements; Glottic measurements; Glottic index; Laryngeal collapse
There is a substantial paucity of studies concerning musculoskeletal injuries in harness Standardbred racehorses. Specifically, little is known about the epidemiology of exercise-related musculoskeletal injuries. Most studies on this subject involve Thoroughbred racehorses, whose biomechanics and racing speed differ from Standardbred, making comparisons difficult. Here, a population of Standardbred racehorses trained at the same racecourse was studied over four years and a classification system for exercise-related musculoskeletal injuries was designed. The incidence rates of musculoskeletal injuries causing horses’ withdrawal from training for 15 days or longer were investigated. A mixed-effects Poisson regression model was used to estimate musculoskeletal injury rates and to describe significance of selected risk factors for exercise-related injuries in this population.
A total of 356 trotter racehorses from 10 different stables contributed 8961 months at risk of musculoskeletal injuries. Four-hundred-and-twenty-nine injuries were reported and classified into 16 categories, based on their aetiology and anatomical localisation. The overall exercise-related injury rate was 4.79 per 100 horse months. When considering risk factors one by one in separate univariable analyses, we obtained the following results: rates did not differ significantly between genders and classes of age, whereas one driver seemed to cause fewer injuries than the others. Racing speed and racing intensity, as well as recent medical history, seemed to be significant risk factors (p < 0.001), while being shod or unshod during racing was not. On the other hand, when pooling several risk factors in a multivariable approach, only racing intensity turned out to be significant (p < 0.001), since racing speed and the racing intensity were partially confounded, being strongly correlated to one another.
Characterizing epidemiology of exercise-related musculoskeletal injuries in trotter racehorses provides baseline incidence rate values. Incidence rates of stress fracture are lower in Standardbreds compared to Thoroughbreds, whereas the opposite is true for tendon and suspensory ligament injuries. In addition to identification of risk factors for musculoskeletal injuries among Standardbred racehorses, results suggest that racing intensity seems to be a protective predictor of risk and recent medical history could be used to identify horses at risk of injury.
Standardbred racehorses; Epidemiology; Trotter; Musculoskeletal injuries; Exercise-related injuries
Wider adoption of reporting guidelines by veterinary journals could improve the quality of published veterinary research. The aims of this study were to assess the knowledge and views of veterinary Editors-in-Chief on reporting guidelines, identify the policies of their journals, and determine their information needs. Editors-in-Chief of 185 journals on the contact list for the International Association of Veterinary Editors (IAVE) were surveyed in April 2012 using an online questionnaire which contained both closed and open questions.
The response rate was 36.8% (68/185). Thirty-six of 68 editors (52.9%) stated they knew what a reporting guideline was before receiving the questionnaire. Editors said they had found out about reporting guidelines primarily through articles in other journals, via the Internet and through their own journal. Twenty of 57 respondents (35.1%) said their journal referred to reporting guidelines in its instructions to authors. CONSORT, REFLECT, and ARRIVE were the most frequently cited. Forty-four of 68 respondents (68.2%) believed that reporting guidelines should be adopted by all refereed veterinary journals. Qualitative analysis of the open questions revealed that lack of knowledge, fear, resistance to change, and difficulty in implementation were perceived as barriers to the adoption of reporting guidelines by journals. Editors suggested that reporting guidelines be promoted through communication and education of the veterinary community, with roles for the IAVE and universities. Many respondents believed a consensus policy on guideline implementation was needed for veterinary journals.
Further communication and education about reporting guidelines for editors, authors and reviewers has the potential to increase their adoption by veterinary journals in the future.
Veterinary journals; Veterinary research; Reporting guidelines; Reporting quality; Editors; Editorial policies; Views; Barriers
Within a feasibility study the use of antibiotics in pigs and cattle was determined in 24 veterinary practices in Lower Saxony and on 66 farms in North Rhine-Westphalia in Germany. Focus was laid on the comparison of the Used Daily Doses (UDD) (dose per animal and day prescribed by the veterinarians) with the Defined Animal Daily Doses (ADD) (dose per animal and day calculated by means of recommended dosages and estimated live weights).
For piglets and calves most of the UDD (50% and 46% of nUDD, respectively) were above the ADD (i.e. UDD/ADD-ratio above 1.25). Regarding sows, fattening pigs, dairy and beef cattle, most of the UDDs (49% to 65% of nUDD) were lower than the respective ADD (i.e. UDD/ADD-ratio below 0.8). In pigs, the UDDs of beta-lactams, fluoroquinolones and cephalosporins, and in cattle, those of macrolides and beta-lactams were often below the ADDs. Tetracyclines were frequently used above the recommended dose.
Enteric diseases were more often treated below the recommended dose than respiratory diseases, possibly due to overestimation of the live weight (diarrhea in young animals, respiratory diseases in elder animals) and consequently overestimation of the recommended dose.
Comparisons between UDD and ADD can be used to observe differences between antimicrobials and trends in the usage of antibiotics. But individual treatment comparisons of UDD and ADD must be interpreted carefully, because they may be due to lower live weights than estimated. Correlating such data with data on the occurrence of resistant bacteria in future may help to improve resistance prevention and control.
VetCAb; Livestock; Tetracyclines; Fluoroquinolones; Macrolides; Cephalosporins
Bovine herpesvirus type 1 (BoHV-1) is the causative agent of respiratory and genital tract infections; causing a high economic loss in all continents. Use of marker vaccines in IBR eradication programs is widely accepted since it allows for protection of the animals against the disease while adding the possibility of differentiating vaccinated from infected animals.
The aim of the present study was the development and evaluation of safety and efficacy of a glycoprotein E-deleted (gE-) BoHV-1 marker vaccine strain (BoHV-1ΔgEβgal) generated by homologous recombination, replacing the viral gE gene with the β-galactosidase (βgal) gene.
In vitro growth kinetics of the BoHV-1ΔgEβgal virus was similar to BoHV-1 LA. The immune response triggered by the new recombinant strain in cattle was characterized both as live attenuated vaccine (LAV) and as an inactivated vaccine. BoHV-1ΔgEβgal was highly immunogenic in both formulations, inducing specific humoral and cellular immune responses. Antibody titers found in animals vaccinated with the inactivated vaccine based on BoHV-1ΔgEβgal was similar to the titers found for the control vaccine (BoHV-1 LA). In the same way, titers of inactivated vaccine groups were significantly higher than any of the LAV immunized groups, independently of the inoculation route (p < 0.001). Levels of IFN-γ were significantly higher (p < 0.001) in those animals that received the LAV compared to those that received the inactivated vaccine. BoHV-1ΔgEβgal exhibited an evident attenuation when administered as a LAV; no virus was detected in nasal secretions of vaccinated or sentinel animals during the post-vaccination period. BoHV-1ΔgEβgal, when used in either formulation, elicited an efficient immune response that protected animals against challenge with virulent wild-type BoHV-1. Also, the deletion of the gE gene served as an immunological marker to differentiate vaccinated animals from infected animals. All animals vaccinated with the BoHV-1ΔgE βgal strain were protected against disease after challenge and shed significantly less virus than control calves, regardless of the route and formulation they were inoculated.
Based on its attenuation, immunogenicity and protective effect after challenge, BoHV-1ΔgEβgal virus is an efficient and safe vaccine candidate when used either as inactivated or as live attenuated forms.
BoHV-1ΔgEβgal; BoHV-1 marker vaccine; Inactivated/live attenuated vaccine; Bovines
One of the most common diseases in intact bitches is pyometra– a potentially life-threatening disease associated with a variety of clinical and laboratory findings. The aims of the present study were to describe complications of the disease and to investigate clinically useful indicators associated with peritonitis and/or prolonged postoperative hospitalization.
A retrospective study was performed using records from 356 bitches diagnosed with pyometra during the years 2006–2007 at the University Animal Hospital, Swedish University of Agricultural Sciences, Uppsala, Sweden. Of the 356 bitches, 315 were surgically treated by ovariohysterectomy, 9 were medically treated and 32 were euthanized without treatment. In the surgically treated bitches, univariable associations between clinical and laboratory data, risk for prolonged hospitalization (≥ 3 days) and/or signs of peritonitis, were analyzed by Chi-square and Fisher’s exact test. Logistic regression models were used to assess multivariable associations.
The most common complication observed in surgically treated bitches was peritonitis (40 bitches), followed by urinary tract infection (19 bitches), wound infection (8 bitches), uveitis (6 bitches), and cardiac arrhythmia (5 bitches). Leucopenia and fever/hypothermia were associated with increased risk for peritonitis (18-fold and three-fold, respectively). Moderate to severe depression of the general condition, pale mucous membranes and leucopenia were associated with increased risk (seven-fold, three-fold, and over three-point-five-fold, respectively) for prolonged postoperative hospitalization.
Several clinically useful indicators were identified. Leucopenia was the most important marker, associated with 18-fold increased risk for peritonitis and an over three-point-five increased risk for prolonged hospitalization. Fever/hypothermia, depression and pale mucous membranes were associated with increased risk for peritonitis and/or prolonged hospitalization. The results of the present study may be valuable for identifying peritonitis and predicting increased morbidity in surgically treated bitches with pyometra.
Bitch; Uterine inflammation; Surgical treatment; Hospitalization; Peritonitis; Risk; Outcome; Dogs
The importance of the wild boar as a reservoir of Lawsonia intracellularis was assessed by investigating the seroprevalence of this pathogen among wild boars in the Republic of Korea. The extent of exposure to L. intracellularis among wild boars (Sus scrofa coreanus) was monitored by a country-wide serological survey using an immunoperoxidase monolayer assay.
In this study, antibodies to L. intracellularis were observed in 165 of 716 clinically healthy wild boars tested. The overall apparent prevalence calculated directly from the sample and the true prevalence calculated based on the accuracy of the test method were 23.0% (95% confidence interval: 20.0-26.3%) and 25.6% (95% confidence interval: 23.9-27.2%), respectively. Serologically positive animals were found in all the tested provinces.
Our results confirm that L. intracellularis is present in the wild boar population worldwide, even in Far East Asia. Despite the high seroprevalence shown in wild boars, further studies are warranted to evaluate their potential as a reservoir species because seroprevalence does not prove ongoing infection nor shedding of the bacteria in amounts sufficient to infect other animals. It should also be determined whether the wild boar, like the domestic pig, is a natural host of L. intracellularis.
Lawsonia intracellularis; Serology; Wild boar
Tooth root problems and periodontal diseases are common in South American camelids (SAC). The objective was to evaluate and optimize the imaging technique for dental radiography in SAC and to describe the radiographic and computed tomographic (CT) anatomy of normal teeth at different ages. In this study, the heads of 20 healthy SAC slaughtered for meat production or euthanized for reasons not related to dental problems included 7 female and 10 male llamas and 3 male alpacas. Using a standardized protocol, radiographs and CT scans of the 20 specimen were performed.
The most useful radiographic projections for mandibular and maxillary cheek teeth evaluation turned out to be lateral30°ventral - laterodorsal and lateral30°dorsal - lateroventral with slight separation of the dental arcades respectively. Digital radiographic and CT appearance of the mandibular and maxillary teeth were described from the beginning of mineralization till maturity. In addition the normal range of the CT radio density of different cheek teeth and different dental tissues were measured. Hounsfield units of different dental tissues of SAC turned out to be similar to equids.
Deviation, shortening and partial destruction of the distal tooth root of mandibular 09′s and 10′s and of maxillary 09′s was observed and the existence of a common pulp chamber in younger teeth was revealed.
The present study provides information about the dental imaging morphology in clinically healthy SAC. This basic information provides fundamental knowledge for evaluating images and planning treatments in clinically affected animals.
Llama; Alpaca; Teeth; Radiography; CT
Pain due to spontaneous intervertebral disc (IVD) disease is common in dogs. In chondrodystrophic (CD) dogs, IVD disease typically develops in the cervical or thoracolumbar spine at about 3–7 years of age, whereas in non-chondrodystrophic (NCD) dogs, it usually develops in the caudal cervical or lumbosacral spine at about 6–8 years of age. IVD degeneration is characterized by changes in the biochemical composition and mechanical integrity of the IVD. In the degenerated IVD, the content of glycosaminoglycan (GAG, a proteoglycan side chain) decreases and that of denatured collagen increases. Dehydration leads to tearing of the annulus fibrosus (AF) and/or disc herniation, which is clinically characterized by pain and/or neurological signs. Current treatments (physiotherapy, anti-inflammatory/analgesic medication, surgery) for IVD disease may resolve neurological deficits and reduce pain (although in many cases insufficient), but do not lead to repair of the degenerated disc. For this reason, there is interest in new regenerative therapies that can repair the degenerated disc matrix, resulting in restoration of the biomechanical function of the IVD. CD dogs are considered a suitable animal model for human IVD degeneration because of their spontaneous IVD degeneration, and therefore studies investigating cell-, growth factor-, and/or gene therapy-based regenerative therapies with this model provide information relevant to both human and canine patients. The aim of this article is to review potential regenerative treatment strategies for canine IVD degeneration, with specific emphasis on cell-based strategies.
Intervertebral disc; Degeneration; Spine; Dog; Regenerative medicine; Mesenchymal stem cell; Notochordal cell
Toll-like receptor (TLR) agonists reportedly have potent antiviral and antitumor activities and may be a new kind of adjuvant for enhancing immune efficacy. Resiquimod (R848) is an imidazoquinoline compound with potent antiviral activity and functions through the TLR7/TLR8 MyD88-dependent signaling pathway. Polyinosinic-polycytidylic acid [poly(I:C)] is a synthetic analog of double-stranded RNA that induces the production of pro-inflammatory cytokines by the activation of NF-κB through TLR3. This study investigated the potential of R848 and poly(I:C) as an adjuvant 146S foot-and-mouth disease virus (FMDV) vaccine formulated with aluminum hydroxide (Al(OH)3).
Antibody titers to FMDV and CD8+ T cells were markedly enhanced in mice immunized to 146S FMDV + Al(OH)3 + R848 + poly(I:C) compared with mice immunized to FMDV + ISA206. IFN-γ secretion substantially increased compared with IL-4 secretion by splenic T cells stimulated with FMDV antigens in vitro, suggesting that R848, poly(I:C), and with Al(OH)3 together biased the immune response toward a Th1-type direction.
These results indicated that the R848 and poly(I:C) together with Al(OH)3 enhanced humoral and cellular immune responses to immunization with 146S FMDV antigens. Thus, this new vaccine formulation can be used for FMDV prevention.
R848; Poly(I:C); Aluminum hydroxide; FMDV
Molecular techniques that detect canine lymphoma cells by their clonal antigen receptor gene rearrangement play an increasing role for diagnosis as well as for monitoring minimal residual disease during and after cytostatic therapy. However, the methods currently available are time-consuming and/or cost-intensive thus impeding the use in clinical routine. The aim of the present study was to develop and evaluate a real-time polymerase chain reaction (PCR) with subsequent melting curve analysis (MCA) for the detection of clonally rearranged antigen receptor genes in dogs with B and T cell lymphoma on non formalin-fixed and paraffin-embedded lymph node samples.
In lymph node aspirates from 30 dogs with multicentric B cell lymphoma, real-time PCR with MCA detected clonal rearrangement in 100% and conventional PCR with polyacrylamide gel electrophoresis (PAGE) in 93% of samples. Both methods correctly identified clonality in 80% of lymph node aspirates of 10 dogs with T cell lymphoma. None of the two PCR systems detected clonal rearrangement in samples from 9 dogs with lymph node hyperplasia. Using a dilutional series with regular lymphoid desoxyribonucleic acid (DNA), detection limits of lymphoma DNA were as low as 0.8% and 6.25% for B and T cell clonal rearrangement with real-time PCR and MCA and at 3.13% and 12.5% with the conventional system. Median absolute detection limits of lymphoma DNA were shown to be at 0.1 ng and 1 ng for the B and T cell immunophenotype with the real-time PCR system and at 10 ng each with conventional PCR and PAGE.
Real-time PCR with MCA is a convenient and reliable method with a good analytical sensitivity. Thus, the method may assist the detection of clonal antigen receptor gene rearrangement in canine lymphoma patients in a clinical setting also in the presence of small amounts of neoplastic cells.
Lymphoma; Dog; Real-time polymerase chain reaction; Melting curve analysis
Thrombosis has been associated to some diseases like hyperadrenocorticism (HAC). Several drugs can alter the balance, such as the corticosteroid prednisone, used mainly for its anti-inflammatory and immunosuppressive effects. It is known that hypercortisolism can stimulate thrombi formation by increasing coagulation factors and decreasing fibrinolysis. However it is not known how prednisone administration affects hemostasis in dogs and if it is dose dependent. The aim of this study, therefore, was to demonstrate the effects of prednisone administration on dogs’ hemostatic profile.
Significant decrease of antithrombin levels was observed in both groups (anti-inflammatory and immunosuppressive doses) after 15 days of treatment. An increase of platelet aggregation was observed in dogs receiving immunosuppressive doses of prednisone (Group II).
From the results obtained in our study, it is not possible to infer that hypercortisolism can increase the thromboembolic risk, despite the decreased anticoagulant factors (antithrombin levels).
Platelets; Hemostasis; Prednisone; Thromboembolism
Bovine tuberculosis (bTB) is a chronic debilitating disease and is a cause of morbidity and mortality in livestock, wildlife and humans. This study estimated the prevalence and risk factors associated with bovine tuberculosis transmission in indigenous cattle at the human-animal interface in the Serengeti ecosystem of Tanzania.
A total of 1,103 indigenous cattle from 32 herds were investigated for the presence of bTB using the Single Intradermal Comparative Tuberculin Test. Epidemiological data on herd structure, management and grazing system were also collected.
The apparent individual animal prevalence of tuberculin reactors was 2.4% (95% confidence interval (CI), 1.7 – 3.5%), whereas the true prevalence was 0.6% CI, 0.6 – 0.7% as indicated by a reaction to avian tuberculin purified protein derivatives (PPD) which is more than 4 mm greater than the reaction to avian tuberculin PPD. The results showed that 10.6% (117/1,103) showed non-specific reactions (atypical mycobacterium). The herd prevalence of 50% (16/32) was found. Tuberculin skin test results were found to be significantly associated with age, location, size of the household and animal tested. Of 108 respondents, 70 (64.8%) individuals had not heard about bovine tuberculosis at all. Thirty five percent (38/108) of respondents at least were aware of bTB. About 60% (23/38) of respondents who were aware of bTB had some knowledge on how bTB is spread. Eighty one percent (87/108) of respondents were not aware of the presence of bTB in wildlife. There is regular contact between cattle and wild animals due to sharing of grazing land and water sources, with 99% (107/108) of households grazing cattle in communal pastures.
The study has demonstrated a high reported interaction of livestock with wildlife and poor knowledge of most cattle owners concerning bTB and its transmission pathways among people, livestock and wildlife. Although the overall proportion of animals with bTB is relatively low, herd prevalence is 50% and prevalence within herds varied considerably. Thus there is a possibility of cross transmission of bTB at wildlife-livestock interface areas that necessitates use of genetic strain typing methods to characterize them accurately.
Risk factors; Bovine tuberculosis; Mycobacterium bovis; Human-animal interface; Serengeti ecosystem; Wildlife
Escherichia coli O157 (E. coli O157) has been isolated from bison retail meat, a fact that is important given that bison meat has been implicated in an E. coli O157-multistate outbreak. In addition, E. coli O157 has also been isolated from bison feces at slaughter and on farms. Cattle are well documented as E. coli O157 reservoirs, and the primary site of E. coli O157 persistence in such reservoirs is the rectoanal junction (RAJ), located at the distal end of the bovine gastrointestinal tract. Since bison and cattle share many genetic similarities manifested as common lineage, susceptibility to infection and the nature of immune responses to infectious agents, we decided to evaluate whether the RAJ of these animals were comparable both in terms of cellular architecture and as sites for adherence of E. coli O157. Specifically, we compared the histo-morphologies of the RAJ and evaluated the E. coli O157 adherence characteristics to the RAJ squamous epithelial (RSE) cells, from these two species.
We found that the RAJ of both bison and cattle demonstrated similar distribution of epithelial cell markers villin, vimentin, cytokeratin, E-cadherin and N-cadherin. Interestingly, N-cadherin predominated in the stratified squamous epithelium reflecting its proliferative nature. E. coli O157 strains 86–24 SmR and EDL 933 adhered to RSE cells from both animals with similar diffuse and aggregative patterns, respectively.
Our observations further support the fact that bison are likely ‘wildlife’ reservoirs for E. coli O157, harboring these bacteria in their gastrointestinal tract. Our results also extend the utility of the RSE-cell assay, previously developed to elucidate E. coli O157-cattle RAJ interactions, to studies in bison, which are warranted to determine whether these observations in vitro correlate with those occurring in vivo at the RAJ within the bison gastrointestinal tract.
O157:H7; Bovine; Bison; Tissue; Epithelia; Markers; Adherence
Presently, few data exist on the level and duration of anti-protective antigen (PA) IgG in vaccinated livestock. Various adaptation of enzyme-linked immunosorbent assays (ELISAs) have been developed in studies to assess immune response following vaccination, albeit mostly in laboratory rodent models. The quantitative anti-anthrax IgG ELISA in this study describes a method of enumerating the concentration of anti-PA specific IgG present in sera of immunized goats, with the aid of an affinity-purified caprine polyclonal anti-anthrax PA-83 IgG standard. This was compared with the anthrax toxin neutralization assay (TNA) which measures a functional subset of toxin neutralizing anti-PA IgG.
The measured concentrations obtained in the standard curve correlated with the known concentration at each dilution. Percentage recovery of the standard concentrations ranged from 89 to 98% (lower and upper asymptote respectively). Mean correlation coefficient (r2) of the standard curve was 0.998. Evaluation of the intra-assay coefficient of variation showed ranges of 0.23-16.90% and 0.40-12.46% for days 28 and 140 sera samples respectively, following vaccination. The mean inter-assay coefficient of variation for triplicate samples repeated on 5 different days was 18.53 and 12.17% for days 28 and 140 sera samples respectively. Spearman’s rank correlation of log-transformed IgG concentrations and TNA titres showed strong positive correlation (rs = 0.942; p = 0.01).
This study provides evidence that an indirect ELISA can be used for the quantification of anti-anthrax PA IgG in goats with the added advantage of using single dilutions to save time and resources. The use of such related immunoassays can serve as potential adjuncts to potency tests for Sterne and other vaccine types under development in ruminant species. This is the first report on the correlation of polyclonal anti-anthrax PA83 antibody with the TNA in goats.
Protective antigen; Indirect ELISA; Toxin neutralization assay; Anthrax; Immunoglobulin; Sterne vaccine; Goats
African swine fever (ASF) is a contagious viral disease which can cause up to 100% mortality among domestic pigs leading to serious socio-economic impact on people’s livelihoods. ASF is endemic in Uganda and there is paucity of information on the epidemiology of the disease. The major aim of this study was to determine the seroprevalence and prevalence of African swine fever virus (ASFV) in apparently healthy slaughter pigs at Wambizi slaughterhouse in Kampala city, Uganda. We also estimated the presence of ASFV antibodies and circulating viral antigens in pigs from selected districts of Uganda during targeted surveillance. We analysed 540 and 181 blood samples collected from slaughter pigs and pigs from targeted surveillance districts respectively.
The prevalence of ASFV in slaughter pigs was 52.96% (95% CI, 48.75-57.14) and 11.5% (95% CI, 9.06-14.45) by ELISA and PCR respectively. In surveillance districts, the proportion of ASFV positive pigs was 53.59% (95% CI, 46.33-60.71) and 0.55% (95% CI, 0.1-3.06) by ELISA and PCR respectively.
The study has found out a high seroprevalence of ASFV antibodies in apparently healthy slaughter pigs and also a high proportion of ASFV antibody seropositive pigs in surveyed districts in Uganda indicating exposure to ASFV. However, there was a lower prevalence of ASFV infection implying that there could be low virulent strains of ASFV circulating in domestic pigs in Uganda which requires further investigation.
African swine fever virus; ELISA; PCR; Prevalence; Seropositive; Slaughter pigs; Surveillance
Epilepsy is one of the most common chronic neurological disorders in dogs characterized by recurrent seizures. The endocannabinoid (EC) system plays a central role in suppressing pathologic neuronal excitability and in controlling the spread of activity in an epileptic network. Endocannabinoids are released on demand and their dysregulation has been described in several pathological conditions. Recurrent seizures may lead to an adverse reorganization of the EC system and impairment of its protective effect. In the current study, we tested the hypothesis that cerebrospinal fluid (CSF) concentrations of the endocannabinoids anandamide (AEA) and 2-arachidonoyl glycerol (2AG) are altered in epileptic dogs. Concentrations of AEA and total AG (sum of 2AG and 1AG) were measured in 40 dogs with idiopathic epilepsy and in 16 unaffected, healthy control dogs using liquid chromatography combined with tandem mass spectrometry.
AEA and total AG were measured at 4.94 (3.18 – 9.17) pM and 1.43 (0.90 – 1.92) nM in epileptic dogs and at 3.19 (2.04 – 4.28) pM and 1.76 (1.08 – 2.69) nM in the control group, respectively (median, 25 – 75% percentiles in brackets). The AEA difference between epileptic and healthy dogs was statistically significant (p < 0.05). Values correlated with seizure severity and duration of seizure activity. Dogs with cluster seizures and/or status epilepticus and with seizure activity for more than six months displayed the highest EC concentrations.
In conclusion, we present the first endocannabinoid measurements in canine CSF and confirm the hypothesis that the EC system is altered in canine idiopathic epilepsy.
Endocannabinoids; Anandamide; 2-arachidonyl glycerol; Epilepsy; Cerebrospinal fluid; Canine
The bacteriophage therapy is an effective antimicrobial approach with potentially important applications in medicine and biotechnology which can be seen as an additional string in the bow. Emerging drug resistant bacteria in aquaculture industry due to unrestricted use of antibiotics warrants more sustainable and environmental friendly strategies for controlling fish infections.
The isolated bacteria from fish lesions was characterised based on isolation on selective and differential medium like Pseudomonas agar, gram staining, biochemical tests and 16SrRNA sequencing. The metallo-beta-lactamase (MBL) producing bacterial isolate was evaluated using Imipenem - Ethylenediaminetetraacetic acid (EDTA) disk method. The specific bacteriophage was isolated and concentrated using coal bed developed in our lab at CSIR-NEERI. The isolated and enriched bacteriophage was characterised by nucleotide sequencing and electron microscopy. The phage therapy was applied for treating ulcerative lesion in fish.
The pathogenic bacterium responsible for causing ulcerative lesions in catfish species (Clarias gariepinus) was identified as Pseudomonas aeruginosa. One out of twenty P. aeruginosa isolate showing multi drug resistance (MDR) was incidentally found to be MBL producing as determined by Imipenem-EDTA disk method. The phage therapy effectively cured the ulcerative lesions of the infected fish in 8–10 days of treatment, with a sevenfold reduction of the lesion with untreated infection control.
Bacteriophage therapy can have potential applications soon as an alternative or as a complement to antibiotic treatment in the aquaculture. We present bacteriophage therapy as a treatment method for controlling MDR P. aeruginosa infection in C. gariepinus. To the best of our knowledge this is a first report of application of phage therapy against MBL producing P. aeruginosa isolated from aquatic ecosystem.
P. aeruginosa; Multi drug resistance; Metallo-β-lactamase; Bacteriophage therapy; Catfish
Propofol is a widely used injectable anesthetic agent for induction and short-term maintenance in dogs. A multi-dose formulation of propofol (MDP) has been developed which includes 2% benzyl alcohol as a preservative. In order to document the use of the product under clinical conditions, MDP was tested in a prospective clinical trial conducted at six sites within the United States. One hundred thirty-eight healthy, client-owned dogs were assigned to one of six treatment groups based on premedicants (none, acepromazine/buprenorphine, midazolam/buprenorphine, medetomidine/buprenorphine) and maintenance agents (MDP, inhaled anesthetic). Anesthesia was induced by the intravenous administration of MDP given to effect. Physiological indices including heart rate, respiratory rate and blood pressure were monitored prior to and during anesthesia induction, maintenance and recovery. Adverse events, defined for severity by pre-established limits of these physiological values, as well as side effects, defined as any observation outside the normal range, were noted.
The mean intubation dose was 7.6 ± 2.1 mg/kg for MDP alone and 4.7 ± 1.3, 4.0 ± 1.0 mg/kg and 3.2 ± 1.4 mg/kg when buprenorphine was used in combination with midazolam, acepromazine and medetomidine, respectively. Of the 32 adverse events, apnea (12 incidents), bradycardia (9 incidents) and hypotension (7 incidents) were most frequently recorded. Emesis, cyanosis and second degree heart block were each noted once and successfully resolved. The cause of a single death 2 days post-anesthesia was assessed as a surgical complication.
MDP was found to be acceptable for use in healthy dogs for induction and short term maintenance of anesthesia when used alone and in combination with premedicants and inhaled anesthetics.
Dog; Anesthesia; Clinical trial; Propofol