A Preliminary Study of 13C-Phenylalanine and 13C-Dipeptide Breath Tests in Horses

This study aimed to establish a standard dose and sample collection time for ¹³C phenylalanine and ¹³C-Dipeptide breath test in horses. To evaluate dose-dependent effects, healthy horses received 2.5 mg/kg, 5 mg/kg, and 10 mg/kg ¹³C phenylalanine dissolved in 1 ml/kg distilled water and 1.25 mg/kg, 2.5 mg/kg, and 5 mg/kg ¹³C dipeptide dissolved in 2 ml/ kg distilled water. Tmax was observed during the sample collection time. For ¹³C phenylalanine, the standard deviation of Cmax at 5 mg/kg was lower than that of 10 mg/kg. For ¹³C dipeptide, the standard deviation of Tmax was the lowest at 5 mg/kg. This study revealed that an optimal dose for breath tests with ¹³C phenylalanine and ¹³C dipeptide may be 5 mg/kg in horses.     

Detection of Corynebacterium bovis infection in athymic nude mice from a research animal facility in Korea

Corynebacterium (C.) bovis infection in nude mice causes hyperkeratosis and weight loss and has been reported worldwide but not in Korea. In 2011, nude mice from an animal facility in Korea were found to have white flakes on their dorsal skin. Histopathological testing revealed that the mice had hyperkeratosis and Gram-positive bacteria were found in the skin. We identified isolated bacteria from the skin lesions as C. bovis using PCR and 16S rRNA sequencing. To the best of our knowledge, this is the first report of C. bovis infection in nude mice from Korea.     

Reemergence of Dourine in Italy: Clinical Cases in Some Positive Horses

In 2011, Trypanosoma equiperdum reemerged in Italy, almost 10 years after its last appearance. A total of eight infected horses have been observed to date. Six horses were affected by natural outbreaks of the disease, whereas two were infected experimentally. The aim of this study was to offer a recent perspective on clinical cases of dourine in Europe. Investigation of the clinical aspects confirmed the three stages reported in the literature: stage 1 (genital lesions), stage 2 (cutaneous signs), and stage 3 (nervous signs). The most common signs in the horses under study were notable weight loss, edematous skin eruptions and oedemas of the abdomen, mammary glands and hind legs. Three animals presented neurological signs (lip ptosis of lower lip and ataxia). Infections were paucisymptomatic or asymptomatic in some animals. Hyperthermia was not reported in infected animals and considerable anemia was observed. High antibody titers did not always correspond to clinical signs. Positive polymerase chain reaction test results of blood or tissue (skin, eye swab) often correspond to an advanced stage of the disease. Dourine is a variable disease; owing to its low prevalence and chronic manifestation, it can be difficult to make a quick diagnosis when facing a Dourine-positive horse.     

Control of Mycobacterium bovisinfection in two sika deer herds in Ireland

In a number of countries, tuberculosis (due to infection with Mycobacterium bovis) is a significant health problem of captive deer. This paper describes outbreaks of bovine tuberculosis in sika deer (Cervus nippon) on two farms in Ireland and the methods used to control the disease. On Farm A, infection was first detected during 1993. The infection was eradicated using a programme of test and removal, in association with segregation of young animals. A second outbreak (also due to infection with M. bovis, but a different RFLP profile) was detected in 2002. In the latter outbreak, infection was particularly prevalent in two groups of young deer. M. bovis with the same RFLP profile was also isolated in a badger found dead on the farm. Control was achieved by test and removal in association with herd management changes. In Herd B, infection was first detected in 1995, and subsequently eradicated using test and removal alone. In Herd A, re-infection remains an ongoing risk. Control rather than eradication of infection may more realistic in the short-to medium-term.     

Extent of Mycobacterium bovis infection in dairy cattle herds subject to partial culling as determined by an interferon-gamma assay

The interferon-gamma (IFN-γ) assay is employed as a complementary diagnostic test for bovine tuberculosis (BTB) in many countries. To simplify this assay, we established a 96-well plate format using the ESAT-6 and CFP-10 antigens and then employed it to determine the extent of Mycobacterium (M.) bovis infection in dairy herds with a history of BTB outbreaks in a country where only selective culling is practiced. The sensitivity and specificity of this IFN-γ assay were 85.9% and 100%, respectively, based on comparison with the conventional single intradermal tuberculin test (SIDT). The IFN-γ assay was also positive in 30.4% and 36.8% of SIDT-negative animals from herds with recent and remote BTB outbreaks, respectively. Of 14 SIDT-negative, IFN-γ positive cattle, five (35.7%) were culture positive and an additional six were positive based on a polymerase chain reaction-based test for M. bovis. Therefore, the IFN-γ assay has the potential to serve as a specific and sensitive test for M. bovis infection in dairy cattle. Further, the results indicated that a substantial portion of SIDT-negative animals in herds with previous BTB outbreaks were actually infected with M. bovis. Accordingly, the present selective-culling strategy may require modifications to include this more sensitive assay.     

A Nonlinear Model for Estimating Hoof Surface Area in Unshod Meat-type Horses

The aim of this study was to develop a simple, inexpensive, and noninvasive technique for estimating the hoof surface area (HSA) of unshod horses under field conditions. It was hypothesized that HSA in horses can be extrapolated from simple linear hoof measurements. Two linear measurements—lateromedial width and dorsoplantar width—were obtained from all four feet of 57 unshod meat-type horses. Different algorithms for determining HSA were developed. The most simple equation was HSA = (1.229 × LM)^1.071 × (DP)^0.772 (where LM = lateromedial width and DP = dorsoplantar width), representing a good nonlinear model with a standard deviation of error for the estimate of 5.150 and a coefficient of multiple determination of 0.9384. This formula may be helpful as a repeatable, nonintrusive, and easily performed in vivo estimation of the hoof surface of unshod horses, at least in the studied breed.     

An epidemiological evaluation of Mycobacterium bovis infections in wild game animals of the Spanish Mediterranean ecosystem

Recreational hunting of indigenous wild artiodactyls has been one of the most lucrative and rapidly growing industries in Western Spain over the last five years. In the absence of careful ecological management, one consequence of the commercial exploitation of this natural resource has been the appearance of outbreaks of infectious disease; most notably bovine tuberculosis. From the outset of the study in 1997, we have observed a steady increase in prevalence of Mycobacterium bovis (M. bovis) in both species reaching 1.74 (+/-0.17) in deer in 2002 and 2.32 (+/-0.24) in wild boar. The latter species seems to be most severely affected with pulmonary lesions appearing more chronic than those observed in deer. In this study, we describe the epidemiology of M. bovis in European wild boar (Sus scrofa) and Iberian red deer (Cervus elaphus hispanicus) in Extremadura (W. Spain); a region where there are large areas of natural habitat for these species.     

The efficacy of eprinomectin extended-release injection against Hypoderma spp. (Diptera: Oestridae) in cattle

The efficacy of eprinomectin in an extended-release injection (ERI) formulation was determined in cattle harboring naturally acquired infestations of first- or second- and third-stage larvae of Hypoderma spp. in three studies conducted according to the same protocol in the USA (two studies) and Germany (one study). Thirty cattle sourced from herds with a history of Hypoderma infestation were included in each study. Cattle were formed into replicates of three animals each on the basis of pre-treatment anti-Hypoderma antibody titers. Within replicates each animal was randomly allocated to one of the following treatments: ERI vehicle (control) at 1 mL/50 kg bodyweight, administered once on Day 0; Eprinomectin 5% ERI at 1 mL/50 kg bodyweight (1.0 mg eprinomectin/kg), administered once on Day 0 (when larvae were expected to be first instars); or Eprinomectin 5% ERI at 1 mL/50 kg bodyweight (1.0 mg eprinomectin/kg), administered once when larvae were second or third instars (study dependent, Day 73, 119, or 140). Treatments were administered by subcutaneous injection in front of the shoulder. In all studies, emerging and/or expressed Hypoderma larvae were recovered, speciated, and counted and viability was determined. Eprinomectin LAI treatment was 100% (p < 0.05) efficacious against first- and second- or third-stage larvae of Hypoderma bovis (two studies) and Hypoderma lineatum (one study). All animals accepted the treatment well. No adverse reaction to treatments was observed in any animal in any study.     

Outbreak of Lice in Horses: Epidemiology,Diagnosis, and Treatment

The aim of this study is to report an outbreak of lice (Bovicola equi) in horses in southern Brazil, focusing on epidemiology, diagnosis, and treatment. Twenty-three horses in a stud farm presented with intense itching, crusted lesions, focal alopecia, and scaly skin. The outbreak occurred during the winter and probably due to the absence of sanitary barriers on the property. The diagnosis was carried out based on clinical signs associated with macroscopic and microscopic visualization of lice identified as B. equi. The treatment was performed with a single dose of Fipronil (Topline® Merial Brasil, Campinas, São Paulo, Brazil) spray (0.5%), eliminating immature and adult stages of lice. The clinical signs caused by the lice in horses disappeared few days after treatment.     

Chronic pneumonia in calves after experimental infection with Mycoplasma bovis strain 1067: Characterization of lung pathology,persistence of variable surface protein antigens and local immune response

Background

Mycoplasma bovis is associated with pneumonia in calves characterized by the development of chronic caseonecrotic lesions with the agent persisting within the lesion. The purposes of this study were to characterize the morphology of lung lesions, examine the presence of M. bovis variable surface protein (Vsp) antigens and study the local immune responses in calves after infection with M. bovis strain 1067.

Methods

Lung tissue samples from eight calves euthanased three weeks after experimental infection with M. bovis were examined by bacteriology and pathology. Lung lesions were evaluated by immunohistochemical (IHC) staining for wide spectrum cytokeratin and for M. bovis Vsp antigens and pMB67 antigen. IHC identification and quantitative evaluation of CD4⁺ and CD8⁺ T lymphocytes and immunoglobulin (IgG1, IgG2, IgM, IgA)-containing plasma cells was performed. Additionally, expression of major histocompatibility complex class II (MHC class II) was studied by IHC.

Results

Suppurative pneumonic lesions were found in all calves. In two calves with caseonecrotic pneumonia, necrotic foci were surrounded by epithelial cells resembling bronchial or bronchiolar epithelium. In all calves, M. bovis Vsp antigens were constantly present in the cytoplasm of macrophages and were also present extracellularly at the periphery of necrotic foci. There was a considerable increase in numbers of IgG1- and IgG2-positive plasma cells among which IgG1-containing plasma cells clearly predominated. Statistical evaluation of the numbers of CD4⁺ and CD8⁺ T cells, however, did not reveal statistically significant differences between inoculated and control calves. In M. bovis infected calves, hyperplasia of bronchus-associated lymphoid tissue (BALT) was characterized by strong MHC class II expression of lymphoid cells, but only few of the macrophages demarcating the caseonecrotic foci were positive for MHC class II.

Conclusions

The results from this study show that infection of calves with M. bovis results in various lung lesions including caseonecrotic pneumonia originating from bronchioli and bronchi. There is long-term persistence of M. bovis as demonstrated by bacteriology and immunohistochemistry for M. bovis antigens, i.e. Vsp antigens and pMB67. The persistence of the pathogen and its ability to evade the specific immune response may in part result from local downregulation of antigen presenting mechanisms and an ineffective humoral immune response with prevalence of IgG1 antibodies that, compared to IgG2 antibodies, are poor opsonins.     

Retrospective Study on Seroprevalence of Borrelia burgdorferi Antibodies in Horses in Minnesota

To determine the seroprevalence of Borrelia burgdorferi antibodies in horses in Minnesota, the database of the Veterinary Diagnostic Laboratory, University of Minnesota, was searched over a 10-year period (May 2001 to May 2010). A total of 1,260 equine serum samples submitted by 112 veterinary clinics were tested using an indirect fluorescent antibody test. Samples with titers of ≥1:320 were considered positive. The average rate of seroprevalence was 58.7%, indicating high exposure of horses to B burgdorferi in Minnesota. Our results indicate that borreliosis should be considered as a differential in cases of horses with undiagnosed musculoskeletal or neurologic disease.     

Calcium ions are involved in egress of Babesia bovis merozoites from bovine erythrocytes

Bovine babesiosis is a livestock disease known to cause economic losses in endemic areas. The apicomplexan parasite Babesia bovis is able to invade and destroy the host’s erythrocytes leading to the serious pathologies of the disease, such as anemia and hemoglobinuria. Understanding the egress mechanisms of this parasite is therefore a key step to develop new therapeutic strategies. In this study, the possible involvement of Ca²⁺ in the egress of B. bovis merozoites from infected erythrocytes was investigated. Egress was artificially induced in vitro using calcium ionophore A23187 and thapsigargin to increase Ca²⁺ concentration in the cytosol of the parasite cells. The increased intracellular Ca²⁺ concentration following these treatments was confirmed using live cell Ca²⁺ imaging with confocal laser scanning microscopy. Based on our findings, we suggest a Ca²⁺ signalling pathway in the egress of B. bovis merozoites.     

Classification of worldwide bovine tuberculosis risk factors in cattle: a stratified approach

The worldwide status of bovine tuberculosis (bTB) as a zoonosis remains of great concern. This article reviews the main risk factors for bTB in cattle based on a three-level classification: animal, herd and region/country level. A distinction is also made, whenever possible, between situations in developed and developing countries as the difference of context might have consequences in terms of risk of bTB. Recommendations are suggested to animal health professionals and scientists directly involved in the control and prevention of bTB in cattle. The determination of Millenium Development Goals for bTB is proposed to improve the control/eradication of the disease worldwide.     

Epidemiological Aspects in the Leptospira spp. and Toxoplasma gondii Infection in Horses from Botucatu,São Paulo,Brazil

Leptospirosis and toxoplasmosis are zoonoses with high importance because of the economic and public health impact. This study was aimed to determine the seroprevalence of leptospirosis and toxoplamosis in 714 serum samples of horses from different farms from Botucatu, São Paulo, Brazil. The samples were researched for Toxoplasma gondii antibodies by indirect immunofluorescence antibody test (IFAT) and for Leptospira spp. antibodies by microscopic agglutination test. Of 714 serum samples, 128 (17.9%; 95% CI: 15.3%-20.9%) were positive for one or more serovars of Leptospira spp., with icterohaemorrhagiae, canicola, and castellonis as the most prevalent serovars, whereas 42 (5.9%; 95% CI: 4.4%-7.9%) were positive for T gondii, of which 33 samples (78.57%; 95% CI: 64.0%-88.2%) presented a titer of 16, 7 (16.7%; 95% CI: 8.4%-30.7%) a titer of 64, and 1 (2.38%; 95% CI: 0.6%-12.3%) a titer of 256. No significant difference was found among the results obtained and the associated variables such as age and sex.