Sequence of the DNA-binding protein of a human subgroup E adenovirus (type 4): comparisons with subgroup A (type 12), subgroup B (type 7), and subgroup C (type 5)

The nucleotide sequence of the gene for the single-stranded DNA-binding protein of adenovirus type 4 (Ad4) has been determined. The gene codes for a protein of 512 amino acids. Comparison of the amino acid sequence with those previously determined for Ad5, Ad12, and Ad7 allowed identification of regions that are conserved between the four serotypes. These include stretches of 9, 9, and 12 amino acids in the carboxy-terminal domain of the protein; these sequences are similar to those identified in the single-stranded DNA-binding proteins of procaryotes as being important for interaction of the protein with single-stranded DNA. A conserved region of four amino acids in the amino-terminal domain is identical in sequence to a region of the SV40 large T antigen that has recently been implicated in the nuclear localization of the protein. Other conserved amino acids that may be important for the three-dimensional structure of the protein have also been identified. The overall homology between the DBPs of the four serotypes is 17.2% in the amino-terminal domain, 47.8% in the carboxy-terminal domain. Two-way comparisons between the DBPs of the four serotypes indicates that the DBP of Ad4 is most closely related to that of Ad7.     

Intraorbital anatomy of the koala (Phascolarctos cinereus)

This is the first documented study of the anatomical details of the contents of the normal koala orbit, excluding the bulbus oculi. Baseline data were established which are necessary for understanding and treating ocular disease in the koala (Phascolarctos cinereus). The anatomy of the orbital contents of the koala were examined and described from animals that presented dead or were euthanized for humane reasons. Dissections of the orbital cavity were performed under magnification. Polymethyl methacrylate (PMMA) casts of the nasolacrimal system and the vascular supply of the orbit were also made in order to study these systems. The superficial lymphatic drainage of the conjunctival tissues was studied by subcutaneous injection of Evan's Blue into the palpebral conjunctiva of a freshly deceased animal, and by Microfil casts of the efferent lymphatics. In general, the orbital contents of the koala are consistent with those of other carnivorous polyprotodont and herbivorous diprotodont marsupials.     

Bone marrow examination in the koala (Phascolarctos cinereus)

A technique for bone marrow aspiration from the iliac crest in the koala (Phascolarctos cinereus) is described. Bone marrow was obtained from ten healthy koalas under general anaesthesia using the combination of tiletamine HCL and zolazepam HCL. Reference ranges were wide. The mean value for M:E was 1.7 (range 0.8–2.7), the mean percentage proliferating erythroid was 11% (range 5%–16%), and the mean percentage proliferating myeloid was 25% (range 17%–35%). In addition, bone marrow aspiration was performed on 14 koalas with various haematological diseases. Results showed aspiration to be useful in the diagnosis of leukaemia, and the investigation of regenerative anaemia and dysplastic anaemia. It was of varying use in the investigation of non-regenerative anaemia.     

Spontaneous lymphoid neoplasia in the koala (Phascolarctos cinereus)

Thirteen cases of lymphoid neoplasia were examined in koalas from New South Wales and Queensland. Haematological and biochemical data were collected from 2 cases while all were subjected to detailed necropsies. Affected koalas were adults of both sexes and the disease had a variable time course. The 2 koalas examined haematologically were leukaemic. On a morphological basis, 5 cases were classified as multicentric, 7 cases as abdominal (but not necessarily with alimentary tract involvement) and one case as solitary. In most cases, the neoplastic cells appeared to be either lymphoblasts or prolymphocytes.     

Chronic peritonitis and pancreatitis in a koala (Phascolarctos cinereus)

The gross, light microscopic and ultrastructural appearance of a nodular peritonitis accompanied by pancreatic changes in a Koala (Phascolarctos cinereus) is described. Its pathogenesis in relation to pancreatitis is discussed.     

Detection of retrovirus sequences in budgerigars with tumours

Renal tumours are a common neoplastic disease of budgerigars. Although a retro-virus has been implicated as the aetiological agent, there is no definitive proof for this hypothesis. Sixteen birds suspected to have renal tumours were examined in an attempt to elucidate the possible role of retroviruses. Thirteen birds had renal tumours and the majority of these birds showed abdominal enlargement and paresis. Renal masses were detected by radiography in nine birds. Post-mortem examination confirmed the presence of abdominal tumours which were mostly confined to the kidneys. All of the renal tumours were carcinomas. ELISA tests to detect the presence of p27 of avian leukosis virus and virus isolation attempts were negative. DNA from eight tumours was examined by dot-blot hybridization for the presence of sequences hybridizing with a full length clone of the RAV-2 strain of the avian leukosis virus. A positive reaction was detected with DNA from 6/8 tumours. Southern blot hybridization demonstrated the presence of a 7.2 kb fragment following restriction with BamHI and a 4.6 kb fragment in an additional tumour following digestion with EcoRI that were recognized by the RAV-2 probe. These results suggest the presence of a retrovirus in tumours of budgerigars.     

Bone marrow examination in the koala (Phascolarctos cinereus)

A technique for bone marrow aspiration from the iliac crest in the koala (Phascolarctos cinereus) is described. Bone marrow was obtained from ten healthy koalas under general anaesthesia using the combination of tiletamine HCL and zolazepam HCL. Reference ranges were wide. The mean value for M:E was 1.7 (range 0.8–2.7), the mean percentage proliferating erythroid was 11% (range 5%–16%), and the mean percentage proliferating myeloid was 25% (range 17%–35%). In addition, bone marrow aspiration was performed on 14 koalas with various haematological diseases. Results showed aspiration to be useful in the diagnosis of leukaemia, and the investigation of regenerative anaemia and dysplastic anaemia. It was of varying use in the investigation of non-regenerative anaemia.     

Detection of porcine endogenous retrovirus (PERV) using highly specific antisera against Gag and Env

Porcine endogenous retroviruses (PERV) are considered an obstacle to the safe use of cells, tissues, and organs from pigs in the course of xenotransplantation. Thus, the detection of viral proteins and of a potential PERV infection is of major interest. Recently, we have published the generation of a highly specific antiserum directed against the nucleocapsid (p10) of PERV (Xenotransplantation 7 (2000), 221). Here we present new peptide-antisera specific to the capsid protein (p30) and the surface molecule of PERV class B (SU, gp70(B)) as well as the transmembrane moiety of the envelope protein (TM, p15E) of PERV which showed functionality in several immunological assays, such as immunoblots, immunofluorescence, and immunogold staining. Thus, these antisera can be used as tools for the identification of viral proteins in basic research as well as clinical trials.     

The stomach gland patch of the koala (Phascolarctos cinereus)

A large gland patch is found in the submucosa of the lesser curvature of the koala stomach near the entrance of the esophagus. It measures approximately 3 cm in diameter and is comprised of numerous invaginations continuous with the surrounding mucosa. The glandular invaginations drain into the lumen of the stomach by 25 to 30 large orifices. The mucosa of the gland patch invaginations differs from the mucosal lining of the stomach chiefly in thickness. The elongate gastric glands that comprise the invaginations exhibit an increased number of parietal and chief cells. Ultrastructural examination of the gland patch illustrates that the parietal, chief, and mucous neck cells are similar in structure to those of other mammalian species.     

Identification of a novel B variant allele at the ABO locus in Chinese Han individuals with B subgroup

We studied the molecular genetic background of the B subgroup in the Chinese Han population and identified a novel allele at the ABO locus. Ten control samples from randomly selected blood donors of normal B phenotype and 6 samples from individuals diagnosed as B subgroup by serological tests were genotyped by PCR-SSP and direct DNA sequencing at exons 6 and 7 of the ABO gene. Exons 6 and 7 and the intervening intron 6 of B alleles from the 6 B subgroup samples were analyzed by cloning and haplotype-sequencing. A novel B variant allele was identified in 2 individuals who were serologically-determined as members of the B(x) and B(w) subgroups, respectively. The novel B allele differs from allele B101 by a single 695T>C missense mutation in exon 7. The family of the individual with B(x) subgroup was studied; among 8 family members tested, 4 had the novel B variant allele. No mutation at exon 6 or 7 of the ABO gene was detected in the 10 control samples or in the other 4 B subgroup samples. Mutation at position 695 where T is replaced by C results in an amino acid change from Leu to Pro, which is predicted to diminish B transferase activity. This indicates that alteration of the amino acid at position 232 is critical to the activity of glycosyltransferases.     

Petting zoos as sources of Shiga toxin-producing Escherichia coli (STEC) infections

Despite their general low incidence, Shiga toxin-producing Escherichia (E.) coli (STEC) infections are considered an important public health issue due to the severity of illness that can develop, particularly in young children. We report on two Austrian petting zoos, one in Tyrol (2015) and one in Vorarlberg (2016), which were identified as highly likely infection sources of STEC infections. The petting zoo related cases involved a case of hemolytic uremic syndrome (HUS) due to STEC O157:HNM in 2015 and an outbreak of STEC O157:H7 infections affecting five young children and two adults in 2016. The HUS case accounted for 2.8% of the 36 STEC O157:HNM/H7 infections notified in Austria in 2015 (5,9% of 17 HUS cases). The seven cases described for 2016 accounted for 4.0% of the 177 human STEC infections documented for Austria in 2016, and for 19.4% of the 36 STEC O157:HNM/H7 infections notified that year. The evaluation of the STEC infections described here clearly underlines the potential of sequence-based typing methods to offer suitable resolutions for public health applications. Furthermore, we give a state-of-the-art mini-review on the risks of petting zoos concerning exposure to the zoonotic hazard STEC and on proper measures of risk-prevention.     

Prevalence of antibody to group B (atypical) rotavirus in humans and animals.

Enzyme-linked immunosorbent assays were developed for the detection of group B rotavirus antigen and antibody. The specificities of both assays were evaluated for antigens and serum specific for rotavirus groups A to D. Serum collected in the United Kingdom from different animal species exhibited the following high prevalence of group B rotavirus-specific antibody: pigs, 97%; cattle, 71%; sheep, 91%; and goats, 91%. In human serum, a lower prevalence of group B-specific antibody was detected; serum from blood donors showed 10% prevalence, and serum from veterinarians showed 4% prevalence.     

The zona pellucida of the koala (Phascolarctos cinereus): its morphogenesis and thickness

In this study the ultrastructural organization of the koala oocyte and the thickness of the surrounding extracellular coat, the zona pellucida, has been determined to ascertain whether there is coevolution of the morphology of the female gamete with that of the highly divergent male gamete that is found in this marsupial species. Ovaries from several adult koalas were obtained and prepared for transmission electron microscopy. Oocytes in large tertiary follicles were somewhat smaller than those of most other marsupials, although their ultrastructural organization appeared similar and included many yolk vesicles. The zona pellucida surrounding the oocytes in tertiary follicles was approximately 8 microm thick and thus is of similar thickness to that of some eutherian mammals but at least twice as thick as that of most marsupial species so far studied. The results indicate that the koala oocyte is unusually small for a marsupial species whereas the zona pellucida is, by contrast, much thicker. How this relates to sperm-egg interaction at the time of fertilization has yet to be determined.     

Genome variants of human adenovirus 40 (subgroup F)

Twenty-six strains of faecal adenovirus from three continents (two from Europe, 16 from South Africa, and eight from Canada) were typed as Ad40 by restriction enzyme analysis of the DNA using Sma I. The DNA of the strains was further compared by digestion with six other enzymes, namely Bam HI, Eco RI, Bgl I, Xho I, Sst I, and Hind III. At least two different restriction profiles were found to exist for each enzyme. Nineteen of the 26 strains studied had the same combination of profiles for all the enzymes, within the limits of resolution of the technique. This genome type was detected from all three continents and included the Finnish reference strain Hovi-X. The other seven strains were genome variants, representing five other genome types. Three genome types were detected only in South African specimens, whereas one genome type was detected in single specimens from both South Africa and Canada. Strain Dugan from Holland, indistinguishable from other Ad40 strains in neutralization tests using polyclonal antibody preparations [de Jong et al, 1983], deviated from the common Hovi-X-like genome on digestion of the DNA with any of the enzymes except Sma I.     

Histologic lesions in cynomolgus monkeys (Macaca fascicularis) naturally infected with simian retrovirus type D: comparison of seropositive, virus-positive, and uninfected animals

Simian retrovirus (SRV) type D is a common cause of simian acquired immunodeficiency syndrome (SAIDS), a usually fatal immunosuppressive disease of macaques. Associated gross and histologic lesions have been well described for the rhesus macaque (Macaca mulatta) in experimental and natural infections. However, morphologic changes induced by this virus at the gross and light-microscopic level have not been documented in the cynomolgus macaque (Macaca fascicularis). In 1996, sporadic cases of anemia, weight loss, and diarrhea were noted in a colony of cynomolgus macaques in our research facility. Out of 28 animals, 24 tested positive for SRV by serology or virus isolation. Animals could mainly be classified into 1 of 2 categories: 1) positive for virus isolation but negative for SRV antibody and 2) negative for virus isolation but antibody positive. During the process of eliminating the virus from the colony, a complete postmortem examination was performed on the 24 infected animals that had to be culled. Twelve SRV-negative animals were available as controls. Minimal to mild follicular lymphoid infiltrates were seen in various organ systems in 75% of the negative animals, compared with moderate to marked infiltrates in 83% of infected animals. Lymphoid infiltrates were more common in the brain, bone marrow, and salivary gland of viremic animals and were rare to nonexistent in seropositive or negative animals. Lymphoid hyperplasia was present in 38% of the infected animals, whereas lymphoid depletion was seen in 47% of the infected animals. Overall, lesions were of greater severity in viremic animals than in virus-negative or seropositive animals. Overall, infected animals had lower, statistically significant hematocrit and lymphocyte values. Viremic animals had significantly lower hematocrit, white blood cell, lymphocyte, and neutrophil values than did controls. Only 1 out of 24 infected animals had clinical signs that were consistent with the definition of SAIDS, and none had evidence of opportunistic infections. Lesions were similar to those already reported in other species of macaques, but the absence of severe illness that was consistent with SAIDS in most viremic animals suggests that there may be a different manifestation of disease in the cynomolgus.     

Detection of a novel border disease virus subgroup in Tunisian sheep

Summary. Nine pestiviruses isolated from different batches of a contaminated Tunisian sheep pox vaccine and one Tunisian field ovine isolate of border disease virus (BDV) were studied at the antigenic and molecular levels. Seroneutralization tests were carried out on three vaccine isolates, the Tunisian field isolate and representative reference strains of the different pestivirus groups. The antigenic study showed that the Tunisian isolates were closer to the two BDV reference strains than to the Alfort-187 and the NADL reference strains. Comparison of the nucleotide sequences of the 5-non coding regions of all the Tunisian isolates to those of other pestiviruses have shown that these isolates were distinct from the established pestivirus species. The entire N^pro–E2 coding sequences of four Tunisian isolates were determined and compared to other pestiviruses. Segregation of these pestiviruses based on the N^pro–E2 region was identical to that obtained with the 5UTR sequences. The phylogenetic tree obtained with these sequences showed that the Tunisian isolates formed a separate branch between the BDV and CSFV groups, and consequently a possible new species within the pestivirus genus. However, as indicated by the antigenic study and the host origin of the isolates, the Tunisian isolates were assigned to a novel subgroup within the BDV species.