Toxoplasmosis in Pallas' cats (Otocolobus felis manul) at the Denver Zoological Gardens.

In May 1996 the Denver Zoological Gardens obtained two male and two female Pallas' cats (Otocolobus felis manul) that were wild-caught in the Ukraine. These animals were part of a group of 16 wild-caught adults (eight male and eight female) imported to the United States and Canada between 1995 and 1996. The Denver Zoological Gardens cats were quarantined at the zoo hospital for approximately I mo. During the quarantine period they were immobilized for physical examination, and sera were obtained from them to evaluate for exposure to Toxoplasma gondii. All cats were positive for T. gondii antibodies by latex agglutination (titers from 1:512 to 1:1,024). After being paired for breeding, one pair produced two litters, and another pair produced four litters, a total of 17 kittens between 1997 and 2001. Four kittens and two young adults died from a disseminated granulomatous and necrotizing inflammation consistent with toxoplasmosis. Toxoplasma gondii infection was confirmed in all six deceased cats by polymerase chain reaction performed on formalin-fixed tissues. An additional five kittens disappeared and were not available for necropsy. The fatality rate from toxoplasmosis was 35.3% (6/17) for cats that were available for necropsy and could have been as high as 64.7% (11/17) if it were assumed that the disappeared kittens were also affected. The Pallas' kitten survival rate at the Denver Zoological Gardens was 35.3%. This article describes the clinical and pathologic features of toxoplasmosis in a group of Pallas' cats at the Denver Zoological Gardens.     

Retracted: Mycobacterium bovis BCG Danish Strain 1331 isolated from a periarticular lesion in a domestic cat

A 7-year-old male neutered domestic shorthair outdoor cat was referred for chronic left forelimb lameness, which had been treated with intra-articular injections of triamcinolone acetonide. A soft tissue swelling around the elbow joint, extending from the distal humerus to the proximal ulna, was surgically explored and biopsy samples obtained. Mycobacterium bovis was cultured from samples from the soft tissue and bone. The mycobacteria from the media were killed and the DNA extracted and tested on a multiplex real-time PCR for the absence of specific genes and the presence of mycobacterial genus markers. The PCR revealed bacillus Calmette-Guérin Danish Strain 1331; this was also isolated from the prescapular lymph node, muscle and bone, obtained at post mortem examination. Badgers had been vaccinated with the bacillus Calmette-Guérin vaccine SSI (Statens Serum Institute) in the area where the cat lived, in the spring and autumn of the previous year. To the authors' knowledge, this is the first report of infection with M. bovis bacillus Calmette-Guérin Danish Strain 1331 in a domestic cat, potentially associated with annual vaccination of badgers in the proximity of the cat's home.     

Variation in E(rns) viral glycoprotein associated with failure of immunohistochemistry and commercial antigen capture ELISA to detect a field strain of bovine viral diarrhea virus

Bovine viral diarrhea virus (BVDV) affects cattle populations causing clinical signs that range from subclinical immunosuppression to severe reproductive and respiratory problems. Detection and removal of persistently infected (PI) calves is the single most important factor for control and eradication of BVDV. Current testing strategies to detect PI calves rely heavily on immunohistochemistry (IHC) and a commercially available antigen capture ELISA (ACE) assay. These viral assays depend on 1 or 2 monoclonal antibodies which target the E(rns) glycoprotein of BVDV. The sensitivity and specificity of these two tests have been reported previously. The purpose of this research was to characterize a strain of BVDV (AU501) that was undetectable using IHC and ACE based on a single monoclonal antibody, but was consistently detected in samples from a Holstein steer using virus isolation and PCR testing. Sequencing of this AU501 viral isolate revealed a unique mutation in the portion of the genome coding for the E(rns) glycoprotein. This unique field strain of BVDV demonstrates the risk of relying on a single monoclonal antibody for detection of BVDV. Multiple testing strategies, including polyclonal or pooled monoclonal antibodies that detect more than one viral glycoprotein may be necessary to detect all PI calves and facilitate eradication of BVDV.     

Use of a polymerase chain reaction assay to detect bovine herpesvirus type 2 DNA in skin lesions from cattle suspected to have pseudo-lumpy skin disease

Beef cattle from a herd in north Alabama were examined because of an outbreak of nonfatal skin disease characterized by discrete circumscribed areas of inflammation that developed on the skin from the neck to the hips. Areas of inflammation, which tended to be superficial, underwent necrosis and scabbed over. The scabs eventually dropped off leaving discrete, round, whitish, hairless lesions that were 1.2 to 2.5 cm diameter. Because clinical signs were consistent with those expected with pseudo-lumpy skin disease (PLSD) caused by bovine herpesvirus type 2 (BHV-2), samples from 16 representative animals were submitted for BHV-2 testing. All 16 animals were seropositive for BHV-2, but the virus could not be isolated from skin biopsy specimens or buffy coat samples. Results of a polymerase chain reaction assay incorporating primers designed to amplify 2 DNA sequences from BHV-2 were positive for 3 of the 10 cattle, suggesting that skin lesions in these cattle were a result of PLSD. Our findings suggest that PLSD may be more common and widespread in the United States than suggested by the frequency with which BHV-2 has been isolated from cattle with PLSD-like skin lesions.     

Induction of protective immunity to Dictyocaulus viviparus in calves while under treatment with endectocides

Three groups of five parasite-naive calves were used. The treatments were: (a) Group 1 calves were weighed on Day 0 and injected with doramectin at 200 microg/kg. From Day 1 to 19 they were dosed orally with 2000 infective larvae of Dictyocaulus viviparus. On Day 28 they were again injected with doramectin, and infected with D. viviparus larvae from Days 33 to 41. They were then left untreated until Day 81 when they were infected with 20 infective larvae of D. viviparus per kg body weight. They were killed on Day 110 and lungworms were counted; (b) Group 2 calves were immunised with oral lungworm vaccine on Days 0 and 28, and infected and slaughtered as Group 1 on Days 81 and 110, respectively; (c) Group 3 calves acted as infection controls. Blood samples were taken at Days 0, 21, 49, 77 and 110 for antibody tests to D. viviparus. At autopsy there were no significant differences between the number of lungworms from Groups 1 and 2 (Means 17.4 and 31.3, respectively); Group 1 had significantly less value than Group 3 (Mean 228) (p < 0.05). Increased antibody titres to the larval sheath of the infective larvae were observed from Groups 1 and 2, showing that the larvae in Group 1 had penetrated the intestine before being killed by the circulating anthelmintic. This experiment shows that if calves are exposed to infective larvae while under systemic endectocide cover, an immune reaction is stimulated.     

Isolation of antigenic proteins from erythrocytes parasitised with Babesia divergens and a comparison of their immunising potential

Soluble proteins present in the supernatant prepared from a sonicated lysate of concentrated erythrocytes infected with Babesia divergens were separated by isoelectric focussing into acidic and non acidic fractions. The acidic fraction was further separated by passage through Concanavalin A Sepharose (Con A) into two fractions, the first (Fraction 2) consisting of proteins plus one glycoprotein which did not bind to Con A, the second of glycoproteins (Fraction 1) which were eluted after binding to Con A. One group of six cows was treated with three injections of Fraction 2 with incomplete Freunds adjuvant (IFA); two other groups received the same number of injections of Fraction 1, one with IFA, the other with glucan as adjuvant. These cows plus an untreated control group were infected with 10(5) erythrocytes infected with a heterologous isolate of B. divergens. Only the group treated with Fraction 2 (Group 1) resisted lethal multiplication of the parasite. There were significant differences in magnitude of parasitaemia, reduction of packed cell volume and antibody titres between Group 1 and the other groups.     

Effects of Nylon and Polyglactin 910 Suture Material on Perilimbal Corneal Wound Healing in the Dog

The effects of fine suture materials on corneal wound healing in the dog were studied. In 20 dogs, standardized perforating perilimbal clear corneal wounds were made and closed with either monofilament Polyglactin 910 or nylon. Five dogs each were euthanized 8,12,16, and 21 days postoperatively. Results of gross, biomicroscopic, and histologic examinations at 8,12,16, and 21 days showed nylon and Polyglactin 910 to cause similar inflammatory responses. Epithelialization and suppuration around the suture tracts were observed more frequently when Polyglactin 910 was used. Both materials were associated with a foreign body (granulomatous) response. There was no loss of wound integrity with either material. Strengths of the incised corneal tissues with and without the sutures intact were determined. At day 16 postsurgery, there was a statistically significant difference in tissue strengths between cases in which sutures were and were not intact. This difference was not apparent at 21 days postsurgery, which suggested that the suture dependent phase ends between the 16th and 21st postsurgical days. In addition, at day 16 there was no statistical difference between the tissue strength of wounds sutured with nylon or Polyglactin 910. Corneal suture materials studied should remain in place for at least 16 days, and absorbable material is only appropriate if it retains tensile strength for 16 days.