Corynebacterium equi in cattle and pigs

Summary The cervical lymph nodes of pigs, the retropharyngeal and submandibular lymph nodes of cattle and faecal samples from both animal species were examined for the presence of Corynebacterium equi. The organism was recovered from 19 (35 per cent) of 54 porcine cervical lymph nodes and from 0 of 54 bovine retropharyngeal and submandibular lymph nodes. Fifteen (50 per cent) of 30 bovine faecal and 11 (35 per cent) of 31 porcine faecal samples yielded C. equi.     

Systemic Listeria monocytogenes infection and concurrent pleural mesothelioma in a cat

Herein we describe a rare case of systemic Listeria monocytogenes infection with concurrent pleural mesothelioma in a stray cat that was found dead and submitted for autopsy. Gross pathology changes consisted of thoracic clear yellow fluid admixed with suspended fibrin strands; clear-to-tan, variably sized, <3 mm diameter pulmonary nodules; and enlargement of the submandibular, retropharyngeal, and prescapular lymph nodes. Histologic changes consisted of extensive areas of suppurative inflammation and necrosis with mineralization that partially effaced the pulmonary parenchyma and lymph nodes. Random, distinct necrotic foci were present throughout the hepatic parenchyma. Extending from the pleura, within perinecrotic alveolar spaces, and infiltrating the submandibular, retropharyngeal, and prescapular lymph nodes were dense sheets of neoplastic epithelioid cells with moderate pleomorphism and occasional karyomegaly and multinucleation. Neoplastic cells exhibited immunolabeling for pancytokeratin AE1/AE3 and vimentin, consistent with pleural mesothelioma. Aerobic bacterial culture of lung yielded heavy growth of L. monocytogenes. Immunohistochemistry (IHC) for L. monocytogenes revealed clusters of bacteria in the lung, lymph node, and liver. Pathologic changes were consistent with systemic listeriosis, confirmed by bacterial culture and IHC, and concurrent pleural mesothelioma.     

Sequelae and complications of Streptococcus equi subspecies equi infections in the horse

Streptococcus equi ssp. equi infection in the horse, or strangles, commonly results in abscessation of the submandibular, submaxillary or retropharyngeal lymph nodes. Although this classical presentation of strangles is associated with a low mortality rate, complications and sequlae may worsen the prognosis and increase mortality rates. This article reviews sequelae and complictions of S. equi ssp. equi infection in the horse, including guttural pouch empyema, bastard strangles and immune mediated diseases such as purpura haemorrhagica, myopathies and myocarditis.     

Frequency of latent equine herpesvirus type-1 infection among a sample of horses in the central North Island of New Zealand

ABSTRACT

Aim: To estimate the frequency of infection with equine herpesvirus type-1 (EHV-1) among horses from the central North Island of New Zealand, including the frequency of detection of the D₇₅₂ genotype.

Methods: Samples of retropharyngeal lymph nodes (RLN) and submandibular lymph nodes (SLN) were dissected from the heads of 63 horses that were humanely killed for various unrelated reasons between March and November 2015. DNA extracted from these tissues was subjected to enrichment for EHV-1 sequences by hybridisation with biotin-labelled EHV-1 specific probe, followed by recovery of EHV-1 sequences on streptavidin-coated magnetic beads. Enriched samples were tested for the presence of EHV-1 using nested quantitative real-time PCR. The EHV-1 amplicons were sequenced to determine the genotype of the virus.

Results: The median age of the horses was 6 (min 2, max 30) years, and 47/63 (75%) were Thoroughbreds. EHV-1 DNA was detected in RLN samples from 6/63 (10%) horses, and three of these horses were also positive for EHV-1 DNA in SLN. The remaining horses were negative for EHV-1 DNA in both RLN and SLN samples. The N₇₅₂ genotype was detected in all positive samples and the D₇₅₂ genotype was not detected in any of the samples.

Conclusions: EHV-1 continues to circulate among horses in New Zealand. The frequency of latent EHV-1 infection among sampled horses may have been underestimated due to the sensitivity limit of the assay or because of the limited anatomical sites sampled in the study. Lack of detection of the D₇₅₂ genotype suggests that infection with this genotype is not common in horses in New Zealand.

Clinical Relevance: If live animals are tested for EHV-1 using SLN biopsy it should be kept in mind that negative results do not rule out the presence of latent EHV-1 infection at other sites inaccessible for testing. The RLN appear to be the preferred sample for detection of EHV-1 DNA in horses following recent euthanasia.     

AN ABATTOIR SURVEY OF TUBERCULOSIS IN FERAL BUFFALOES

Tuberculosis lesions were found in 193 (1.7%) of 11,322 buffaloes examined during routine post-mortem inspection at 2 export abattoirs. The prevalence of tuberculosis in buffaloes supplied from 17 separate farms ranged from 0.3% to 8.22%, with the highest levels occurring on the coastal plains. Lesions were confined to one major body region in 50 of 72 randomly chosen cases of tuberculosis and to 2 or more regions in 22 cases. Thoracic lesions occurred in 65 of the 72 cases, abdominal lesions in 19, head lesions in 18 and carcase lesions in 9. In the thoracic cavity, lesions occurred most frequently in mediastinal and bronchial lymph nodes. In the head region the retropharyngeal lymph node was most frequently involved, in the abdominal cavity, the liver, and in the carcase, the deep inguinal lymph node. Tuberculosis lesions in buffaloes had a lardaceous consistency and were paler in colour and less calcified than those normally exhibited by cattle. Mycobacteria were isolated from 30 of 31 lesion samples submitted for bacteriological examination. Of the isolates, 25 were identified as Mycobacterium bovis, 3 as M. avium-intracellulare-scrofulaceum complex, one as M. fortuitum and one as M. flavescens. The M. bovis isolates from buffaloes showed minor cultural differences to those normally characteristic of bovine isolates.     

Farm and slaughter survey of bovine tuberculosis in captive deer in Switzerland

In 1998, a survey was conducted by postal questionnaire to gather basic knowledge about the management, health and productivity of captive deer in Switzerland. In addition, lymph nodes were collected from slaughtered deer from 124 of the 262 holdings surveyed, and tested for Mycobacterium bovis and Mycobacterium tuberculosis. The total farmed deer population was 8389 animals kept on 485 holdings; 87 per cent were fallow deer, 8 per cent red deer, 4 per cent sika deer, and there were small numbers of other species. The median herd sizes were 12 for fallow deer and eight for red deer. Few owners had handling facilities or crushes. In none of the lymph nodes examined were lesions typical of bovine tuberculosis observed, and neither M bovis nor M tuberculosis was cultivated from any of the samples.     

Parallel faecal and organ Mycobacterium avium subsp. paratuberculosis culture of different productivity types of cattle

Faecal (at least 3 months before slaughtering) and organ examinations were carried out in 611 animals (497 dairy, 69 dual-purpose and 44 beef cattle) originating from eight paratuberculosis infected cattle herds. The diagnosis in cattle was established by routine intestinal culture (ileum and the adjacent lymph nodes) after slaughter. In selected 132 animals, post-mortem intensive culture was performed on tissue samples collected from the gastrointestinal tract (duodenum, jejunum, ileum, ileocecal valve, caecum, rectum) and the corresponding lymph nodes, submandibular, retropharyngeal, tracheobronchial, liver and supramammary lymph nodes, kidney, liver and spleen. In 251 (41.1%) of all 611 animals, Mycobacterium avium subspecies paratuberculosis could be isolated from the faeces; in 164 (65.7%) out of 251 shedding animals the infection was detected in the ileum and adjacent lymph nodes. The detection of M. paratuberculosis by routine intestinal culture of faecal culture positive animals varied from 46.0% in animals shedding 1 CFU (colony forming unit), to 94.7% in massive shedders. On the contrary, M. paratuberculosis was detected by routine intestinal culture in 92 (25.5%) of the 360 faecal culture negative animals. Shedding animals had significantly higher (P<0.01) number of organisms in their organs than non-shedding animals. During the intensive tissue cultivation from selected 132 animals, 72 (54.5%) of them were positive. For the negative animals, no significant difference was found between the detection rate in organs examined after slaughter with routine and intensive method. However, in the subgroup of tissue culture positive animals a highly significant difference (P<0.01) was found by intensive examination (83.0%) compared with the routine examination (60.4%). Out of 72 tissue culture positive animals 73.6% of them harboured M. paratuberculosis in the gastrointestinal tract, 16.7% in the gastrointestinal tract and the parenchymatous organs, tracheobronchial and mandibular lymph nodes. The rest of the 9.7% of the infection was detected in the lymph nodes of head and lungs. Our study concerning the distribution of M. paratuberculosis by intensive examinations revealed a minimum effect of breed and production type on localisation of the agent. Thus, the results suggest that in case of an active infection, M. paratuberculosis can be localised in different organs of animals irrespective of their breed or production type.     

Radiographic lymphangiographyin the dog using iodized oil

The objective of this study was to develop a clinically applicable technique to visualize the medial retropharyngeal, superficial cervical, axillary, superficial inguinal and medial iliac lymph nodes on radiographs. Direct and indirect lymphangiographic methods using iodized oil were repeated for a minimum of five times at eight different locations to enhance the various lymph nodes, using 16 healthy research dogs. Direct lymphangiography, although more invasive than indirect lymphangiography, resulted in uniform contrast uptake by an increased number of nodes and increased enhancement of the lymphatic vasculature, and is recommended for imaging the medial iliac and superficial cervical lymph nodes. Side effects were more frequent after indirect lymphangiography (10/20 injection sites) than after direct lymphangiography (3/16 injection sites). The small size of afferent lymphatic vessels did not allow use of direct lymphangiography for the medial retropharyngeal, axillary and superficial inguinal lymph nodes; however, indirect techniques allowed adequate visualization of these nodes.     

Pulmonary lesions and Mycobacterium bovis excretion from the respiratory tract of tuberculin reacting cattle

Although it is generally recognised that tuberculous lesions are present in lymph nodes associated with the respiratory tract in approximately 90 per cent of reactors with confirmed infection, lung lesions are found in only 1 to 2 per cent of such cases during abattoir examination. When lung lesions are not detected, it has been claimed that such cattle are non-excretors and thus unimportant in the epidemiology of the disease. In this study the lungs of 55 reactor cattle were sliced into sections approximately 0.5 cm thick. Tuberculous lesions were evident in over 70 per cent of lungs from reactors with concurrent lesions in lymph nodes of the respiratory system. Further, M bovis was isolated from single samples of nasal and, or, tracheal mucus taken at slaughter in 19 per cent of confirmed cases. Several of these reactors had a clear tuberculin test less than six months previously indicating recent infection. This study confirms the continued importance of the infected bovine in the epidemiology and current eradication of bovine tuberculosis. It is suggested that all tuberculous cattle with lesions in respiratory lymph nodes, rather than being regarded as non-excretors, should be considered as possible excretors and thus important sources of infection for other cattle both within and between herds.     

Early pathogenesis of equine Streptococcus equi infection (strangles)

Reasons for performing study: Little is known about entry and subsequent multiplication of Streptococcus equi following exposure of a susceptible horse. This information would have value in design of intranasal vaccines and understanding of shedding and protective immune responses. Objectives: To determine entry points and sites of subsequent replication and dispersion of S. equi at different times after intranasal infection or commingling exposure. Methods: Previously unexposed horses and ponies were subjected to euthanasia 1, 3, 20 or 48 h following intranasal inoculation with biotin labelled or unlabelled S. equi CF32. Some ponies were inoculated with suspensions of equal numbers of CF32 and its mutants lacking capsule, S. equi M‐like protein or streptolysin S. Others were infected by commingling exposure and subjected to euthanasia after onset of fever. Tonsils and lymph nodes were cultured for S. equi and tissues sectioned for histopathological examination and fluorescent microscopy. Results: Tonsillar tissues of both the oro‐ and nasopharynx served as portals of entry. Entry was unexpectedly rapid but involved few bacteria. Small numbers of organisms were detected in tonsillar crypts, in adjacent subepithelial follicular tissue and draining lymph nodes 3 h after inoculation. By 48 h, clumps of S. equi were visible in the lamina propria. At onset of fever, tonsillar tissues and one or more mandibular and retropharyngeal lymph nodes were heavily infiltrated by neutrophils and long chains of extracellular S. equi. Mutant S. equi lacking virulence factors were not seen in draining lymph nodes. Conclusions: Although very small numbers of S. equi entered the lingual and nasopharyngeal tonsils, carriage to regional lymph nodes occurred within hours of inoculation. This observation, together with visual evidence of intracellular and extracellular multiplication of S. equi in tonsillar lymphoid tissue and lymph nodes over the following days, indicates involvement of potent antiphagocytic activity and failure of innate immune defences. Relevance: Future research should logically address the tonsillar immune mechanisms involved including identification of effector cell(s) and antigens.     

Detection of Mycoplasma mycoides subspecies mycoides in tissues from an outbreak of contagious bovine pleuropneumonia by culture, immunohistochemistry and polymerase chain reaction

Postmortem observations of 37 cattle from an outbreak of contagious bovine pleuropneumonia (CBPP) in north Italy in 1993 were made at the abattoir, where samples of lung and tracheobronchial lymph node tissues were taken for culture and identification of Mycoplasma mycoides subspecies mycoides (MmmSC), immunohistochemistry with the peroxidase anti-peroxidase (PAP) system, and molecular detection by the polymerase chain reaction (PCR) amplification of specific DNA from MmmSC. Nasal swabs were also taken for testing by PCR Lung pathology typical of CBPP was observed in 38 per cent of the animals, and MmmSC was isolated from 19 per cent DNA of MmmSC was detected by PCR in 64 per cent of lung samples and 35 per cent of the nasal swabs. Staining of lung tissue and lymph node tissue by PAP was positive in 27 per cent and 30 per cent of cases, respectively, and was a useful back-up test. These results suggest that PCR amplification from lung tissue may be used as a rapid and accurate confirmatory test for cases with pathology resembling CBPP.     

Comparative scanning electron microscope analysis of the enamel of permanent human,bovine and porcine teeth

BackgroundBovine and porcine teeth are often used in in vitro experiments as substitutes of human teeth.ObjectivesThe aim of the present study was to perform a comparative analysis of enamel morphology of permanent human, bovine and porcine teeth under the scanning electron microscope.MethodsAs many as 10 human, 10 bovine, and 10 porcine teeth were studied. All the teeth were sectioned and the halves were randomly divided into 2 groups according to the examined tissue (vestibular enamel at the mid-height of the dental crown and in the cervical area). Human and bovine enamel was etched for 15 sec and porcine enamel for 30 sec. The scanning electron microscope analysis was performed. The length and width of enamel prisms were determined with the “Met-Ilo” 1.1 computer program.ResultsAll enamel samples revealed the same etching pattern—Silverstone''s type 2. Bovine enamel showed a similar porosity and the amount of interprismatic enamel compared to human enamel while the amount and width of interprismatic enamel bands in porcine enamel were evidently greater. The shape of the porcine prisms was visually similar to human prisms, although dimensions were significantly different. However, bovine prisms differed in form and appeared to be distinctly elongated.ConclusionsReported findings indicate that the results of experimental studies carried out on bovine and porcine enamel should not be compared with the results obtained on human enamel.     

The Corynebacterium pyogenes infection in cattle. 1. Incidence of the pathogen]

Reported are the incidence of Corynebacterium pyogenes together with different pathological changes as well as the existence of latent Corynebacterium pyogenes infections and their widespread occurrence. Corynebacterium pyogenes was established from 609 in 2,130 samples of pathological processes, accounting for 28.6%. The pathogen was cultivated from various processes, including enlarged tail lymph nodes (61.1%), tail phlegmons (56.3%), abscesses (49.1%), epiphysiolyses (45.2%), liver abscesses (31.8%), panaritia at beginning of fattening (20.5%), aborted foetuses (14.9%), foetal membranes in cases of incarcerated placenta (12.0%), and panaritia on end of fattening (3.4%). The same pathogenic microorganism was recorded from nine per cent of apparently intact heifer udders, before pasturing. Corynebacterium pyogenes was cultivated also from nasal mucous membrane (8.4%) and retropharyngeal lymph nodes (37.2%). The highest detection rate was 71.6%, obtained from the tonsils.     

Indications of a relationship between buying-in policy and infectious diseases on dairy farms in Wales

During the period February to May 2008, bulk milk samples were collected from 57 dairy farms throughout Wales in the framework of a voluntary somatic cell count project. Bulk milk samples were tested for antibodies to bovine viral diarrhoea virus (BVDV), bovine herpesvirus type 1 (BHV-1) and Leptospira Hardjo, and samples were also tested for the presence of BVDV antigen by PCR. A questionnaire was used to determine whether the herd was open or closed, what the vaccination status was, and to obtain general farm information such as the herd size and average milk yield. Vaccination against BVD, infectious bovine rhinotracheitis and leptospirosis was practised on 37, 12 and 35 per cent of the farms, respectively. The presence of bulk milk antibodies on farms that did not use vaccination was 75 per cent for BVDV, 54 per cent for BHV-2 and 76 per cent for L Hardjo. Open herds had 10 times the odds (95 per cent confidence interval [CI] 1.7 to 59.4)of having bulk milk antibodies for BVDV and 16.7 times the odds (95 per cent CI 2.0 to 49.7) of having bulk milk antibodies to BHV-1 compared with closed herds. A farm with bulk milk antibodies to one disease had significantly higher odds of having bulk milk antibodies to a second disease (P<0.05).     

First case of a dog infected with Aspergillus (Phialosimplex) caninus in Australasia

ABSTRACT

Case history: A 2-year-old Rottweiler dog from Perth (WA, Australia) was referred for assessment of a chronic productive cough and weight loss.

Clinical findings: Severely enlarged bilateral superficial cervical lymph nodes and severely enlarged abdominal organs were present. The body condition score was poor and there was moderate muscle wasting. Thoracic and abdominal computed tomography images revealed severe diffuse enlargement of thoracic and abdominal lymph nodes, hepatomegaly and diffuse splenomegaly. A diffuse bronchial pattern with severe multifocal saccular bronchiectasis was identified in the lungs.

Diagnostic findings: Fungal organisms were seen within macrophages on cytological preparations and on histopathological sections of biopsies of the superficial cervical lymph node. Macrophages contained intracytoplasmic, non-filamentous round-to-ovoid organisms, which varied in size from 5–30?µm in diameter with variable morphology. Budding was not observed, and no hyphae were present. Fungal culture of lymph node tissue resulted in growth of Aspergillus (Phialosimplex) caninus which was confirmed by amplification and sequencing of a segment of the 16S-23S rRNA internal transcribed spacer. Concurrent bacterial bronchitis was diagnosed on culture of broncho-alveolar fluid.

Diagnosis: Disseminated aspergillosis caused by Aspergillus caninus.

Clinical relevance: This is believed to be the first report of infection caused by A. caninus in a dog in Australasia. The dog was treated with itraconazole for 7 months and was still alive 7 months after the start of treatment.     

Distribution of angiostrongylosis in Cornwall

One hundred and ninety-seven faecal samples were examined for Angiostrongylus vasorum larvae, from dogs throughout Cornwall, which were presented with one or more clinical signs consistent with infection, were known to eat slugs or snails or were related to an infected dog. Twenty samples (10-1 per cent) contained nematode larvae, of these A vasorum larvae were identified in eight (4-1 per cent) and larvae of Filaroides species in eight (4-1 per cent). Seven of the eight dogs positive for A vasorum were found to have been infected within an area six miles in diameter incorporating Redruth. Statistical analysis suggested that this aggregation of cases was unlikely to be a random event (P < 0–01). No correlation between published data on the distribution of the intermediate host, slugs and snails and the cases of A vasorum infection reported in this paper could be found. It is concluded that A vasorum infection is enzootic within a small area around Redruth in the mid-west of Cornwall.     

Critical points in meat production lines regarding the introduction of listeria monocytogenes

Summary

In order to elucidate critical points concerning Listeria monocytogenes during bovine and porcine slaughter, cutting and processing, 843 samples were obtained from carcasses, primal cuts, products at retail and from environmental surfaces. Only 2–7% of the carcasses and 0–10% of the environmental samples in the ‘clean’ part of the pork slaughterline were found to be positive for L. monocytogenes. The incidence of L. monocytogenes was increased after chilling and cutting. In the cutting room 11–36% of the primal cuts and 71–100% of the environmental samples were found positive for L. monocytogenes. Our findings indicate that contamination of pork meat with L. monocytogenes orginates from the processing environment of the chilling or cutting room. The incidence of L. monocytogenes in the bovine cutting and meat processing line (0–60%) was lower than in the porcine cutting and meat processing line (11–100%).     

Direct Detection of Mycobacterium bovis in Bovine Lymph Nodes by PCR

Thirty‐five lymph node samples were taken from animals with macroscopic lesions consistent with Mycobacterium bovis infection. The animals were identified by postmortem examination in an abattoir in the northwestern region of state of Paraná, Brazil. Twenty‐two of the animals had previously been found to be tuberculin skin test positive. Tissue samples were decontaminated by Petroff’s method and processed for acid‐fast bacilli staining, culture in Stonebrink and Lowenstein‐Jensen media and DNA extraction. Lymph node DNA samples were amplified by PCR in the absence and presence (inhibitor controls) of DNA extracted from M. bovis culture. Mycobacterium bovis was identified in 14 (42.4%) lymph node samples by both PCR and by culture. The frequency of PCR‐positive results (54.5%) was similar to that of culture‐positive results (51.5%, P > 0.05). The percentage of PCR‐positive lymph nodes increased from 39.4% (13/33) to 54.5% (18/33) when samples that were initially PCR‐negative were reanalysed using 2.5 μl DNA (two samples) and 1 : 2 diluted DNA (three samples). PCR sensitivity was affected by inhibitors and by the amount of DNA in the clinical samples. Our results indicate that direct detection of M. bovis in lymph nodes by PCR may be a fast and useful tool for bovine tuberculosis epidemic management in the region.     

Quantitation and localization of immunoglobulin E containing cells in bovine lymphoid tissues

Although recent studies have begun to describe and quantify IgE responses in bovine serum and secretions, little is known about the distribution and quantity of IgE containing cells in cattle. In the present study, cells with cytoplasmic IgE were quantitated in bovine lymphoid tissues, using immunoperoxidase staining and evaluation by an image analysing computer (Quantimet). Frozen sections from retropharyngeal, bronchial and mesenteric lymph nodes, tonsil and spleen were stained from 11 calves, some of which had been exposed to antigen by aerosol or injection. Although individual variability was considerable, bronchial and mesenteric lymph nodes generally contained the greatest percentage of IgE containing cells, while retropharyngeal lymph node, tonsil, and spleen had less. Parenteral immunization with ovalbumin appeared to increase the splenic percentage, while aerosol exposure to ovalbumin was associated with a greater percentage of IgE containing cells in bronchial lymph nodes. Comparison of the present results with those reported for other species shows some similar trends in IgE localization.