Colleen Duncan

Cryptococcosis: update and emergence of Cryptococcus gattii

Cryptococcosis is a fungal disease that occurs throughout the world. Recent reclassification of Cryptococcus species along with a change in the distribution pattern has prompted reevaluation of the organism and the diseases caused by this pathogen. This review highlights the emergence of Cryptococcus gattii as a primary pathogen in North America and summarizes our current understanding of the disease in mammals and birds.

Cryptococcus gattii in Wildlife of Vancouver Island, British Columbia, Canada

Although Cryptococcus gattii has emerged as an important pathogen of humans and domestic animals on Vancouver Island, Canada since 1999; its distribution in regional wildlife species is largely unknown. Opportunistic sampling methods were employed to obtain nasal swabs for fungal culture from wild mammal species residing within the coastal Douglas fir biogeoclimatic zone on the southeast coast of the island. Samples were collected from 91 animals representing 14 species. Cryptococcus gattii was isolated from the nasal swabs of two eastern gray squirrels (Sciurus carolinensis) trapped in Duncan, British Columbia. The relative proportion of nasal colonization in wild mammal species is consistent with findings in domestic animals, suggesting that animals may be good indicators of environmental organisms.

Comparison of Surgical Treatment Options for Cranial Cruciate Ligament Disease in Large‐Breed Dogs with Excessive Tibial Plateau Angle

OBJECTIVE To identify surgical techniques performed by veterinary surgeons for the treatment of cranial cruciate ligament disease in large-breed dogs with excessive tibial plateau angle (eTPA), compare their outcomes, identify risk factors for postoperative complications, and compare outcome and complication rate with tibial plateau leveling osteotomy (TPLO) in dogs without eTPA. STUDY DESIGN Multicenter, case-control study. ANIMALS eTPA group (TPA>or=35 degrees)=58 dogs (78 stifles); control group (TPA<or=30 degrees)=58 dogs (72 stifles). METHODS Control and eTPA group dogs were identified. Medical records, radiographic reviews, and pet-owner interviews were used to identify surgical treatments performed, their complications, and outcome. Odds ratios with 95% confidence intervals, chi(2) tests, and t-tests were calculated to discern differences between eTPA and control-group dogs. RESULTS TPLO was the most common treatment for eTPA. Owner-perceived outcome was superior for eTPA group dogs when TPLO resulted in TPA<or=14 degrees compared with those with postoperative TPA>14 degrees. Loss of tibial plateau leveling during convalescence was the most commonly observed major complication; addition of ancillary implants to TPLO significantly reduced its incidence. Postoperative complications were more common in dogs with eTPA than in dogs without eTPA; however, no difference in owner-perceived outcome was identified. CONCLUSIONS Surgical treatment of dogs with eTPA has a higher complication rate, but comparable outcome to that of dogs without eTPA. CLINICAL RELEVANCE Postoperative TPA<or=14 degrees and addition of ancillary implants are recommended when using TPLO to treat dogs with eTPA.Objective— To identify surgical techniques performed by veterinary surgeons for the treatment of cranial cruciate ligament disease in large-breed dogs with excessive tibial plateau angle (eTPA), compare their outcomes, identify risk factors for postoperative complications, and compare outcome and complication rate with tibial plateau leveling osteotomy (TPLO) in dogs without eTPA. Study Design— Multicenter, case–control study. Animals— eTPA group (TPA≥35°)=58 dogs (78 stifles); control group (TPA≤30°)=58 dogs (72 stifles). Methods— Control and eTPA group dogs were identified. Medical records, radiographic reviews, and pet-owner interviews were used to identify surgical treatments performed, their complications, and outcome. Odds ratios with 95% confidence intervals, χ2 tests, and t-tests were calculated to discern differences between eTPA and control-group dogs. Results— TPLO was the most common treatment for eTPA. Owner-perceived outcome was superior for eTPA group dogs when TPLO resulted in TPA≤14° compared with those with postoperative TPA>14°. Loss of tibial plateau leveling during convalescence was the most commonly observed major complication; addition of ancillary implants to TPLO significantly reduced its incidence. Postoperative complications were more common in dogs with eTPA than in dogs without eTPA; however, no difference in owner-perceived outcome was identified. Conclusions— Surgical treatment of dogs with eTPA has a higher complication rate, but comparable outcome to that of dogs without eTPA. Clinical Relevance— Postoperative TPA≤14° and addition of ancillary implants are recommended when using TPLO to treat dogs with eTPA.

Mycobacterium bovis BCG-mediated protection against W-Beijing strains of Mycobacterium tuberculosis is diminished concomitant with the emergence of regulatory T cells.

ABSTRACT Despite issues relating to variable efficacy in the past, the Mycobacterium bovis BCG vaccine remains the basis for new-generation recombinant vaccines currently in clinical trials. To date, vaccines have been tested mostly against laboratory strains and not against the newly emerging clinical strains. In this study, we evaluated the ability of BCG Pasteur to protect mice from aerosol infections with two highly virulent W-Beijing clinical strains, HN878 and SA161. In a conventional 30-day protection assay, BCG was highly protective against both strains, but by day 60 of the assay, this protection was diminished. Histological examination of the lungs of vaccinated animals showed reduced lung consolidation and smaller and more-organized granulomas in the vaccinated mice after 30 days, but in both cases, these tissues demonstrated worsening pathology over time. Effector T cell responses were increased in the vaccinated mice infected with HN878, but these diminished in number after day 30 of the infections concomitant with increased CD4+ Foxp3+ T cells in the lungs, draining lymph nodes, and the spleen. Given the concomitant decrease in effector immunity and continued expansion of regulatory Foxp3+ cells observed here, it is reasonable to hypothesize that downregulation of effector immunity by these cells may be a serious impediment to the efficacy of BCG-based vaccines.

Cutaneous neoplastic lesions of equids in the central United States and Canada: 3,351 biopsy specimens from 3,272 equids (2000–2010)

OBJECTIVE To identify epidemiological trends in cutaneous neoplasms affecting equids in central North America and compare them with previously reported trends. DESIGN Retrospective case series. SAMPLE 3,351 cutaneous biopsy specimens from 3,272 equids with a neoplastic diagnosis. PROCEDURES Diagnostic reports from 2 diagnostic laboratories (Colorado State University and Prairie Diagnostic Services Inc) were reviewed for frequency of specific lesions and epidemiological trends. Variables included in analyses (if known) were age, sex, breed, geographic location, date of diagnosis, location of neoplasm on the body, and presence or absence of ulceration. RESULTS Sarcoid, squamous cell carcinoma (SCC), and melanoma were the 3 most common tumors diagnosed. Tumors associated with UV radiation (SCC, SCC in situ, hemangioma, hemangiosarcoma) were 2.3 (95% confidence interval [CI], 1.8 to 3.0) times as common in biopsy specimens received by Colorado State University than in specimens received by Prairie Diagnostic Services Inc. Appaloosa horses and American Paint horses, respectively, were 7.2 (95% CI, 5.6 to 9.2) and 4.4 (95% CI, 3.6 to 5.4) times as likely as other breeds to have tumors associated with UV radiation. Thoroughbreds were predisposed to cutaneous lymphoma, whereas Arabians were more likely to have melanomas. Draft and pony breeds were 3.1 (95% CI, 1.9 to 5.1) times as likely as other breeds to have benign soft tissue tumors. Morgans and pony breeds more commonly had basal cell tumors. Tumors in the perianal region were significantly more likely to be SCC or melanoma while tumors on the limbs were more likely to be giant cell tumor of soft parts. CONCLUSIONS AND CLINICAL RELEVANCE Signalment, anatomic location of the mass, and geographic location of the horse can be used to help equine practitioners formulate differential diagnoses for cutaneous masses. Further research is necessary to identify the biological basis for the development of many equine cutaneous neoplasms.

Prevalence of selected zoonotic and vector-borne agents in dogs and cats in Costa Rica.

To estimate the prevalence of enteric parasites and selected vector-borne agents of dogs and cats in San Isidro de El General, Costa Rica, fecal and serum samples were collected from animals voluntarily undergoing sterilization. Each fecal sample was examined for parasites by microscopic examination after fecal flotation and for Giardia and Cryptosporidium using an immunofluorescence assay (IFA). Giardia and Cryptosporidium IFA positive samples were genotyped after PCR amplification of specific DNA if possible. The seroprevalence rates for the vector-borne agents (Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis, and Anaplasma phagocytophilum) were estimated based on results from a commercially available ELISA. Enteric parasites were detected in samples from 75% of the dogs; Ancylostoma caninum, Trichuris vulpis, Giardia, and Toxocara canis were detected. Of the cats, 67.5% harbored Giardia spp., Cryptosporidium spp., Ancylostoma tubaeforme, or Toxocara cati. Both Cryptosporidium spp. isolates that could be sequenced were Cryptosporidium parvum (one dog isolate and one cat isolate). Of the Giardia spp. isolates that were successfully sequenced, the 2 cat isolates were assemblage A and the 2 dog isolates were assemblage D. D. immitis antigen and E. canis antibodies were identified in 2.3% and 3.5% of the serum samples, respectively. The prevalence of enteric zoonotic parasites in San Isidro de El General in Costa Rica is high in companion animals and this information should be used to mitigate public health risks.

Persistent Bovine Viral Diarrhea virus Infection in Wild Cervids of Colorado

Bovine viral diarrhea virus (BVDV) is a significant viral pathogen of domestic cattle. Worldwide, there is evidence of BVDV exposure and infection in wild ungulates; however, the frequency and significance of such events are unknown. To determine the prevalence and distribution of Colorado deer, elk, and moose persistently infected (PI) with BVDV, a cross-sectional study was conducted using full-thickness ear tissue samples collected from animals presented to the Colorado Division of Wildlife for chronic wasting disease surveillance in the 2005–2006 hunting season. Tissue from 5,597 harvested animals (2,934 mule deer, 2,516 elk, 141 white-tailed deer, and 6 moose) was paraffin-embedded and stained for BVDV using immunohistochemistry. A single adult male mule deer had BVDV antigen in the skin; staining distribution was consistent with that seen in PI cattle. Skin and lymph node were also positive for viral RNA by polymerase chain reaction, and the virus was determined to be a type 1. The prevalence of BVDV PI cervids in Colorado is very low. However, the identification of a naturally infected adult PI animal in the wild suggests that the virus infects free-ranging populations. The source of the BVDV is unknown and is assumed to be spillover from cattle or maintenance within wildlife populations. Consideration of a potential wild animal reservoir is important in the design and implementation of BVDV management practices in cattle.

A mouse model of tuberculosis reinfection

Recent clinical observations shows that individuals treated with chemotherapy for tuberculosis who live in endemic areas are four times more likely to develop secondary disease, often as not caused by exogenous reinfection. In a mouse model described here, we show that mice infected with the virulent W-Beijing Mycobacterium tuberculosis strain HN878, then given chemotherapy to clear the infection, were resistant to re-challenge with the same organism thereafter. This resistance, which was mediated by rapid expression of CD4 T cells expressing markers consistent with both central and effector memory immunity, was only transient however. After 20-30 days of the reinfection the numbers of these cells steadily declined, the bacterial load in the lungs surged up, and the lung tissues became increasingly consolidated. No evidence was found for a regulatory T cell response in these mice, but many T cells harvested from the lungs showed evidence of increased PD-1 expression, indicating exhaustion. These data indicate that the memory T cell response to reinfection may not be as stable and long lived as previously thought, a finding with obvious implications for vaccine development.

Histopathologic and immunohistochemical findings in two white-tailed deer fawns persistently infected with Bovine viral diarrhea virus

Bovine viral diarrhea virus (BVDV) is an important pathogen of domestic cattle. Serologic, experimental, and individual case studies explored the presence and pathogenesis of the virus in wild ungulates; however, there remain large gaps in knowledge regarding BVDV infection in nonbovine species. Live twins were born from a white-tailed deer (Odocoileus virginianus) doe infected with noncytopathic BVDV during its first trimester of pregnancy. The twins died at 1 day of age from trauma unrelated to the infection, and tissues were collected for histologic and immunohistochemical examination. The most significant histologic abnormality was diffuse depletion of B-lymphocytes in both fawns. The BVDV antigen was distributed widely throughout many tissues and cell types, most notably epithelium and vascular endothelium, consistent with that reported in cattle. In contrast to cattle, lymphocytes exhibited only very rare positive staining.

Coxiella burnetii in Northern Fur Seal (Callorhinus ursinus) Placentas from St. Paul Island, Alaska

The decline in the number of northern fur seal (NFS; Callorhinus ursinus) pups on St. Paul Island, Alaska, has led to multidisciplinary research, including investigation into issues of reproductive health and success. Given the recent identification of Coxiella burnetii in the placenta of two other marine mammal species, NFS placentas were collected from Reef rookery on St. Paul Island, Alaska, during the 2010 pupping season, examined histologically, and tested for C. burnetii using polymerase chain reaction (PCR). Of 146 placentas examined, gram-negative intratrophoblastic bacteria that were positive for C. burnetii on immunohistochemistry were observed in 5 (3%) placentas. Placental infection was usually devoid of associated inflammation or significant ancillary pathology. One hundred nine (75%) of the placentas were positive for C. burnetii on PCR. C. burnetii is globally distributed and persists for long periods in the environment, providing ample opportunity for exposure of many species. The significance of this finding for the declining fur seal population, potential human exposure and infection, and impact on other sympatric marine mammal or terrestrial species is unclear; further investigation into the epidemiology of Coxiella in the marine ecosystem is warranted.