Anaplasma infection in free-ranging Iberian red deer in the region of Castilla-La Mancha, Spain

Organisms in the genus Anaplasma are obligate intracellular pathogens that multiply in both vertebrate and invertebrate hosts. The type species, Anaplasma marginale, causes bovine anaplasmosis and infects erythrocytes of the vertebrate host and undergoes a complex developmental cycle in ticks which serve as biological vectors. Infected cattle, wild ruminants and ticks can all serve as reservoirs of A. marginale. In this study, hunter killed Iberian red deer (Cervus elaphus hispanicus) from the region of Castilla-La Mancha in southwestern Spain were tested for Anaplasma infection. We found that 10% of the deer examined were seropositive for Anaplasma. Three A. marginale strains were subsequently obtained from salivary glands of Hyalomma marginatum that were removed from these deer, and the sequence of the major surface protein (msp)4 gene was determined for each strain and used for phylogenetic studies. Maximum parsimony analyses of msp4 sequences from H. marginatum ticks in comparison with New World cattle and bison isolates reported previously, suggested different origins for these Spanish A. marginale strains. The results of this study demonstrated that Iberian red deer are naturally infected with Anaplasma, and may therefore serve as a wildlife reservoir of the pathogen. Although the link between deer infection and the strains of A. marginale identified in ticks was not established, H. marginatum and Rhipicephalus bursa were identified as potential biological vectors for A. marginale in this region and may effect transmission of A. marginale between deer and cattle populations.     

Real-time polymerase chain reaction testing for the detection of Mycobacterium genavense and Mycobacterium avium complex species in avian samples

Diagnosis of avian mycobacteriosis, caused by Mycobacterium genavense or species belonging to the Mycobacterium avium complex (MAC), is problematic. Polymerase chain reaction (PCR) offers rapid and sensitive detection of minute quantities of DNA, and conventional protocols have been used for evaluating avian specimens. The recent development of real-time PCR offers several advantages over conventional PCR. In attempts to improve diagnosing avian mycobacteriosis, a real-time TaqMan PCR assay was developed targeting the 65-kD heat shock protein gene of M. genavense and MAC spp. Nineteen reference isolates, 16 clinical isolates, and 32 avian tissue samples were used to evaluate the assay. When sufficient amplicons were produced, the species of mycobacteria was determined by standard sequencing of TaqMan PCR products and compared with results from commercial mycobacteriology laboratories and/or standard sequencing of conventional PCR products. The TaqMan PCR detected DNA from reference isolates of M. genavense, MAC spp., and Mycobacterium tuberculosis complex spp. Of the clinical isolates, the TaqMan PCR detected DNA from 10 of 12 Mycobacterium avium avium isolates and two of three Mycobacterium avium intracellulare isolates. For the tissue samples, the TaqMan PCR amplified DNA in six of nine samples that were identified by sequencing of conventional PCR products and/or by commercial mycobacteriology laboratories as being MAC spp. positive and three of four samples that were positive for M. genavense. There was some disagreement between speciation results from the TaqMan PCR and those from commercial mycobacteriology laboratories or conventional PCR or both. This disagreement was suspected to be because of relatively small numbers of base pairs in the TaqMan PCR products. The TaqMan PCR may provide a useful tool for evaluating clinical samples for DNA from mycobacteria species that most commonly infect birds; however, further refinement is needed in order to improve sensitivity and provide more accurate speciation.     

The effect of hoof trimming on radiographic measurements of the front feet of normal Warmblood horses

The understanding of the normal position of the third phalanx (P3) and the distal sesamoid bone in relation to the size and shape of the hoof capsule in sound horses is helpful in the diagnosis of equine foot lameness. Some measurements on radiographs used to define the position of the pedal bone within the hoof capsule are significantly influenced by hoof trimming and the height of the withers. In this study, the front hooves of 40 Warmblood horses were radiographed twice, eight weeks apart, both before and after their hooves were trimmed by an experienced farrier. Using the software programme Metron PX, 22 parameters on the lateromedial view and 16 parameters on the dorsopalmar view were measured and the effect of hoof trimming and height of the withers were calculated, respectively. Some of the hoof parameters showed mild positive correlation with the height of the withers. In 70% of the horses the left hoof capsule and P3 were significantly larger than the right. Hoof trimming had a remarkable influence on hoof conformation, especially for parameters in the toe region. Of all the measurements that describe the position of the third phalanx (P3) in relation to the hoof capsule, the distances between the distal tip of P3 to the solar surface of the foot, P3 to the tip of the toe and P3 to the point of break-over showed the greatest differences before and after trimming. The database of the present study can be used by farriers and veterinarians as a guideline for routine and corrective shoeing of Warmblood horses.     

Fluorangiographic study of the ocular fundus in normal horses

Purpose:  To describe the protocol and appearance of fluorescein angiography (FA) in normal horses.
Animals:  A total of 25 healthy horses aged between 5 and 15 years.
Materials and Methods:  The horses were sedated with 15 µg/kg detomidine and 50 µg/kg butorphanol and dilated with topical tropicamide 1%. All angiograms were recorded after intravenous bolus injection of 10 mg/kg of fluorescein sodium solution.
Results:  Two successive angiographic phases could be discerned: the choriopapillary phase, starting at 46.95 ± 9.48 s, and the retinal vascular phase, starting at 47.79 ± 10.38 s. The retinal vascular phase was divided in three parts: filling phase , maximum fluorescence point , and fading phase . During the filling phase, the dye progressed into the retinal vessels, obtaining maximum fluorescence at 59.79 ± 10.39 s, termed the maximum fluorescence point. The fading phase started immediately following the maximum fluorescence point. During this phase, vascular fluorescence decreased to complete reduction at 74.76 ± 9.81 s. Also, areas of delayed choroidal filling, the presence of short retinal vessels in the ventral region of the optic disc, and a particular filling of the optic disc were also observed.
Conclusions:  The normal angiographic sequence was described in horses. FA may be a useful method for studying the integrity of the blood–retinal barriers in horses.     

Using BCG,MPT-51 and Ag85 as antigens in an indirect ELISA for the diagnosis of bovine tuberculosis

This study evaluated the Mycobacterium tuberculosis protein antigen MPT-51, the trimeric antigen 85 (Ag85) complex, and Bacillus Calmette-Guérin (BCG) in an indirect ELISA to diagnose bovine tuberculosis (TB) from serum samples. Serum was collected from 208 intra-dermal tuberculin test (ITT)-positive and 54 ITT-negative animals from a region where bovine TB is endemic. Using the Ag85 and BCG antigens, the indirect ELISA was able to discriminate ITT-positive from ITT-negative animals. This level of discrimination was not achieved when using the MPT-51 antigen. The highest sensitivity (Se) and specificity (Sp) of the test was found when BCG was used (Se, 82%; Sp, 91%). Further work in different epidemiological settings and with larger numbers of animals will be required to validate these findings.     

Use of the distomedial-proximolateral oblique radiographic view of the elbow joint for examination of the medial coronoid process in dogs

OBJECTIVE: To describe and evaluate a new radiographic view of the elbow joint in dogs that would potentially enhance observation of the medial coronoid process (MCP). SAMPLE POPULATION: 20 cadaver limbs from 10 dogs and clinical examination of 100 elbow joints of 53 dogs. PROCEDURE: Twenty elbow joints from 10 cadavers were imaged by use of mediolateral, flexed mediolateral, craniocaudal, craniolateral-caudomedial oblique (Cr15L-CdMO), and distomedial-proximolateral oblique (Di35M-PrLO) radiographic views before and after placement of 3 lead pellets placed on the cranial, medial, and craniodistal aspect of the MCP. Three examiners independently reviewed these radiographs. One hundred elbow joints of 53 dogs with forelimb lameness and signs of pain elicited on palpation of the elbow joint were examined. These joints were radiographed and treated by use of arthroscopy. Three examiners independently graded the radiographs. RESULTS: The MCP was identified on all Di35M-PrLO views made during the anatomic study. The Di35M-PrLO view had the largest area under the receiving operating characteristic (ROC) curve for detection of abnormalities of the MCP. Fractured and nonfractured MCP could only be significantly differentiated on Di35M-PrLO and mediolateral views. The Di35M-PrLO view had a higher agreement between examiners than other radiographic views for detection of fractures of the MCP. CONCLUSION AND CLINICAL RELEVANCE: The Di35M-PrLO view enhances the identification of anomalies and fragmentation of the MCP in dogs, compared with other radiographic views. The Di35M-PrLO view may be of benefit for early screening of dogs potentially affected with elbow dysplasia.     

Clostridium septicum myositis in a western lowland gorilla (Gorilla gorilla gorilla).

A 10-yr-old male gorilla (Gorilla gorilla gorilla) with a history of conspecific bite wounds was evaluated for acute onset of depression, anorexia, and right hemiparesis. The animal was immobilized for diagnostic examination and treatment for suspected toxic shock from a necrotizing, emphysematous wound infection, but was euthanized due to complications during recovery. Gross and histopathologic examination revealed acute necrotizing myositis, fasciitis, cellulitis, and emphysema in the affected wound area, with large numbers of large Gram-positive rods among necrotic muscle fibers. Severe pulmonary edema with airways containing fibrin, acute hemorrhage in multiple body sites, thrombosis in blood vessels in the skeletal muscle, liver, and lung, and lymph node hyperplasia with lymphoid necrosis and hemorrhage. Immunohistochemical fluorescent antibody staining of muscle from the wound site was positive for     

Effect of environmental factors and changes in the body condition score on the onset of the breeding season in mares

Several methods have been proposed to advance the onset of the breeding season in horses. Most of them are based on the exposure to an artificial lighting period combined with hormonal treatments. Mares exposed to an artificial photoperiod are most often housed indoors where the ambient temperature is often higher than the outside temperature. Mares held in barns are also exposed to different daylight intensities than horses kept outside, depending on the architecture. In the current study, we evaluated the impact of ambient temperature, daylight intensity and changes in body condition score (BCS) on the timing of first ovulation after winter anestrus in mares exposed to an artificial photoperiod. Mares (n = 211) were housed in barns with different ambient temperature and daylight exposure but with the same artificial photoperiod exposure (except for a natural photoperiod control group). Artificial photoperiod as well as an increase in BCS over the winter significantly advanced the first spring ovulation. The BCS at the start and end of the anestrus period did not have an effect on the interval to first ovulation and neither did the modest increase in ambient temperature in the barn. However, a higher light intensity during the daytime significantly advanced the first spring ovulation. The results of this study suggest that exposure to more sunlight advances the onset of the breeding season. This effect is likely mediated through the biological effect of short wavelength blue light and its impact on melatonin suppression and biological rhythms. We suggest that greater/direct exposure to the blue light component of daylight improves the response to the artificial photoperiod. The results of the present study can further assist to optimize the conditions that lead to an efficient spring transition of breeding mares.     

Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?

Sperm from fertile donkeys have been successfully frozen in absence of permeable cryoprotectants. The aim of this study was to determine whether this cryopreservation method is suitable for subfertile donkeys in comparison to conventional sperm freezing with glycerol. Ejaculates were collected from four Andalusian Donkeys: three fertile and one subfertile. Semen was frozen with an extender containing glycerol (GLY), or adding instead sucrose 0.25 molar and 1% bovine serum albumin (SUC) as non‐permeable cryoprotectants. After thawing, samples were assessed for total (TM, %) and progressive (PM, %) sperm motility by CASA, plasma membrane integrity (PMI, %) by epifluorescence microscopy and DNA integrity (DFI, %) by SCSA. Results (mean ± SD) were compared between extenders in fertile and subfertile donkeys using the Student's t test. No differences between GLY and SUC treatments were found in the fertile group for the sperm parameters assessed. In subfertile donkey ejaculates, GLY resulted in significantly higher values than SUC for TM (25.5 ± 3.1 vs. 19.6 ± 1.9) and PM (13.3 ± 5.1 vs. 4.0 ± 1.2), respectively. In conclusion, considering all the sperm parameters assessed, sperm freezing in absence of permeable cryoprotectants may not be still an option for cryopreservation of subfertile donkey sperm.     

Demographic, clinical, and radiographic features of bronchiectasis in dogs: 316 cases (1988-2000)

OBJECTIVE: To determine demographic, clinical, and radiographic features of bronchiectasis in dogs. DESIGN: Retrospective study. ANIMALS: 289 dogs identified through the Veterinary Medical Database (VMDB) and 27 dogs examined at the North Carolina State University Veterinary Teaching Hospital. PROCEDURE: Demographic characteristics of dogs identified through the VMDB were compared with characteristics of the entire population of dogs entered in the VMDB. Medical records of dogs examined at the teaching hospital were reviewed; the diagnosis was confirmed through review of thoracic radiographs. RESULTS: Analysis of data from the VMDB indicated that American Cocker Spaniels, West Highland White Terriers, Miniature Poodles, Siberian Huskies, English Springer Spaniels, and dogs > 10 years old had an increased risk of bronchiectasis. Among dogs examined at the teaching hospital, coughing was the most common clinical sign. There was evidence for excessive airway mucus but not hemorrhage. A variety of bacterial organisms were isolated from tracheal wash and bronchoalveolar lavage samples. On thoracic radiographs, cylindrical bronchiectasis, generalized disease, and right cranial lung lobe involvement were most common. Seven of 14 dogs for which follow-up radiographs were available did not have any progression of radiographic lesions. Median duration of clinical signs prior to diagnosis of bronchiectasis was 9 months (range, 1 day to 10 years). Median survival time was 16 months (range, 2 days to 72 months). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that despite substantial clinical abnormalities, dogs with bronchiectasis may survive for years. Certain purebred dogs and older dogs may have an increased risk of developing bronchiectasis.