It is suggested that Haemophilus paragallinarum requires at least three haemagglutinins for adhesionduring infection. This paper reports the partial purification and characterization of the HA-L haemagglutininfrom H. paragallinarum strain 46-C3, a heat sensitive, trypsin sensitive haemagglutinin that hasbeen shown to be the serovar specific haemagglutinin in this organism. Using the pI and molecularmass obtained, it was shown that this protein shares similarities with other types of adhesins found in Gram-negative bacteria. The haemagglutination assay conditions were optimized at pH 7.5 at 37 °C. It was also shown that activity is enhanced by the addition of Ca2+ and Mn2+ ions.

The present study investigated the distribution of nerves in the ovary of the emu. The neuronal markers, protein gene product 9.5, neurofilament protein and neuron specific enolase demonstrated the constituents of the extrinsic and intrinsic ovarian neural systems. The extrinsic neural system was composed of ganglia in the ovarian stalk, as well as nerve bundles, which were distributed throughout the ovary. Isolated neuronal cell bodies, in the medulla and cortex, formed the intrinsic neural system. An interesting finding of the study was the presence of nerve bundles, circumscribed by lymphocytes, in the ovarian stalk. The findings of the study indicate that the distribution of nerve fibres and neuronal cell bodies in the emu ovary is similar, but not identical to that of the domestic fowl and ostrich.

The potential economic benefits of combining tactical anthelmintic treatment for gastrointestinal nematodes and nutritional supplementation with urea-molasses blocks were examined in Boer goats raised under extensive grazing conditions in the summer rainfall area of South Africa. Eight groups of nine goats were monitored over a 12-month period from 1 October 2002 to 9 October 2003. Ad libitum nutritional supplementation with urea-molasses blocks was provided when the goats were housed at night, during the summer (wet season -December 2002 to February 2003), and / or the winter (dry season -June 2003 to August 2003). All the goats were treated symptomatically for Haemonchus contortus infection when deemed necessary by clinical examination of the conjunctiva for anaemia using the FAMACHA© system. Half the groups were tactically treated for gastrointestinal nematodes in mid-summer (28 January 2003). Under the symptomatic treatment, climatic and extensive grazing conditions encountered during the trial, feed supplementation in the winter dry season had the greatest economic benefit and is therefore recommended. Tactical anthelmintic treatment afforded no additional advantage, but the nematode challenge was lo

Experimental Trichinella zimbabwensis infections were established in three baboons (Papios p.)and four vervet monkeys (Cercopithecuase thiops) and the clinical-pathological manifestations assessed. The infected animals showed clinical signs ranging from fever, diarrhoea, periorbitaol edema and muscular pain in varying degrees. One baboon became blind due to the infection. Levels of creatinine phosphokinase and lactated ehydrogenase increased to reach a peak on Day 42 post-infection(pi)for both baboons and monkeys. Blood parameters such as packed cell volume, levels of red blood cells and white blood cells did not change significantly from the normal ranges except for the levels of eosinophils which peaked above the normal ranges at Day 28 and 56 pi in baboons and at Day 56 pi in monkeys.

2-Phenoxyethanol was used as an anaesthetic to translocate 102 species of fishes representing 30 families from the Sea World aquarium on Durban's beachfront to uShaka Marine World. Most fishes responded well to a final anaesthetic concentration of 0,150 mℓ / ℓ and there were no mortalities.

Glycophorins are the major sialoglycoproteins in red blood cell membranes, possessing various physiological and pathological roles. We examined membrane glycoproteins in canine red cells and cloned cDNAs for two major glycophorins, glycophorins A (GPA) and C (GPC) from bone marrow cells. Periodic acid-Schiff staining and immunoblotting analyses showed that canine red cell membranes contained several glycoproteins immunoreactive to an anti-bovine GPC antibody, whereas the most abundant sialoglycoproteins, the candidates for GPA, did not react with an anti-human GPA antibody. The amino acid sequences of the extracellular domains of GPA and GPC had no significant homology to those from other mammalian species, including humans, and had O-linked and/or N-linked glycosylation sites. On the other hand, the C-terminal cytoplasmic domain and/or the transmembrane helices of GPA and GPC were conserved among species, indicating some functional significance of those regions in red cell membranes that include dimerization of GPA in the membrane-spanning region, and association of GPC with membrane skeletal proteins through binding with protein 4.1 and p55 in the cytoplasmic domain. These findings provide insights for clinical studies to evaluate the involvement of GPA and GPC in the pathogenesis of red cell diseases.

The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Qsup(245)TVTKKsup(250) on SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV.

In the present study, we employed flow cytometry to evaluate the level of parasitemia of Babesia gibsoni infecting canine erythrocytes in vivo and in vitro by using fluorescent nucleic acid staining. Peripheral blood samples from a B. gibsoni-infected dog and cultured B. gibsoni parasitizing in canine erythrocytes were stained with a membrane-permeable fluorescent nucleic acid stain, SYTO16. In this study, we utilized normal canine erythrocytes (LK erythrocytes) and canine erythrocytes containing high concentrations of potassium, reduced glutathione, and some free amino acids (HK erythrocytes) as host cells for culture. In vivo parasitized cells were discriminated completely from unparasitized cells and a correlation (r=0.998) between the percentage of SYTO16-positive cells and parasitemia in vivo was observed. On the other hand, in vitro erythrocytes could not be divided clearly into parasitized and unparasitized cells. However, when LK erythrocytes were used as host cells, the percentage of SYTO16-positive cells was the almost same as, and was well correlated (r=0.932) with the level of parasitemia. When HK erythrocytes were used as host cells, the percentage of SYTO16-positive cells was almost half of, but was correlated (r=0.982) with the level of parasitemia. Therefore, we attempted to observe the changes in the percentage of parasitized cells after treatment with antiprotozoal drug or mitochondria inhibitors by using flow cytometry. The changes in the percentage of SYTO16-positive cells corresponded well with the changes of the level of parasitemia when the parasites in HK erythrocytes were cultured with each compound. The present results suggest that flow cytometric detection using SYTO16 is a rapid and reliable method for monitoring parasitemia both in vivo and in vitro.

Bovine nasal and oral discharges were used as samples for bovine viral diarrhea virus (BVDV) gene detection. Viral genes in serum (S), nasal discharge (N) and oral discharge (O) were quantified with real-time polymerase chain reaction using SYBR Green by the relative quantification method, and findings were compared among samples. Although the quantity of the BVDV gene in S was greater than those in N and O, all samples were available to identify persistently infected (PI) cattle with BVDV by reverse transcription polymerase chain reaction (RT-PCR). The swab samples were able to be stored for a few days at 4degC with a little decrease of amplification signal in RT-PCR. Oral swab sampling was easier than nasal swab sampling, and was also less uncomfortable for the cattle than other sampling methods without pain or unnecessary retention. This sampling method can be performed without any special technique and equipment. Therefore, the oral swab sampling method is useful for screening to detect BVDV PI cattle by RT-PCR.

Although mammals produce either sperm or eggs depending on their sex, newborn MRL/MpJ male mice contain oocytes within their testes. In our previous study, the testicular oocyte appears as early as day 0 afterbirth and has morphological characteristics as an oocyte such as zona pellucida and follicular epithelial cells. Based on the observation of F1 between MRL/MpJ and C57BL/6, one of the genes causing the appearance of testicular oocyte exists on the Y chromosome. In the present study, we found testicular oocytes within newborn AKR mice. We have also analyzed the Sry genes from several inbred mouse strains and identified a shortened glutamine repeat near the C-terminal region that is unique to MRL and AKR. These results suggest that polymorphism of glutamine repeat within SRY correlates with the appearance of testicular oocyte and this phenotype is derived from AKR, one of the original strains of MRL mice.