Influence of fibrolytic enzymes on the hydrolysis and fermentation of pure cellulose and xylan by mixed ruminal microorganisms in vitro

A series of in vitro studies was conducted to determine the effects of adding a commercial enzyme product on the hydrolysis and fermentation of cellulose, xylan, and a mixture (1:1 wt/wt) of both. The enzyme product (Liquicell 2500, Specialty Enzymes and Biochemicals, Fresno, CA) was derived from Trichoderma reesei and contained mainly xylanase and cellulase activities. Addition of enzyme (0.5, 2.55 and 5.1 microL/g of DM) in the absence of ruminal fluid increased (P < 0.001) the release of reducing sugars from xylan and the mixture after 20 h of incubation at 20 degrees C. Incubations with ruminal fluid showed that enzyme (0.5 and 2.55 microL/g of DM) increased (P < 0.05) the initial (up to 6 h) xylanase, endoglucanase, and beta-D-glucosidase activities in the liquid fraction by an average of 85%. Xylanase and endoglucanase activities in the solid fraction also were increased (P < 0.05) by enzyme addition, indicating an increase in fibrolytic activity due to ruminal microbes. Gas production over 96 h of incubation was determined using a gas pressure measurement technique. Incremental levels of enzyme increased (P < 0.05) the rate of gas production of all substrates, suggesting that fermentation of cellulose and xylan was enzyme-limited. However, adding the enzyme at levels higher than 2.55 microL/g of DM failed to further increase the rate of gas production, indicating that the maximal level of stimulation was already achieved at lower enzyme concentrations. It was concluded that enzymes enhanced the fermentation of cellulose and xylan by a combination of pre- and postincubation effects (i.e., an increase in the release of reducing sugars during the pretreatment phase and an increase in the hydrolytic activity of the liquid and solid fractions of the ruminal fluid), which was reflected in a higher rate of fermentation.     

Radial carpal bone fractures in 15 dogs

Between 1977 and 1992, 15 adult dogs were presented to the Universities of Bristol, Glasgow, Liverpool and London with fractures of the radial carpal bone which had occurred without a known episode of violent trauma. Eleven of the dogs were boxers and the other breeds affected were a Pharaoh hound, rottweiler, springer spaniel and a greyhound. The condition was bilateral in three cases. Eleven dogs were treated surgically by cast immobilisation (four cases), fragment removal (one case), lag screw fixation (three cases), pan-carpal arthrodesis (one case) and lag screw reduction followed by pan-carpal arthrodesis (two cases). The remaining four dogs were treated conservatively with rest and tactical use of non-steroidal anti-inflammatory drugs. Follow-up periods ranged from two months to 11 years. Varying degrees of lameness, which tended to be exacerbated by exercise, persisted in all cases. The radial carpal bone appears to have at least three separate centres of ossification which eventually fuse; their planes of fusion correspond approximately to the two main fracture lines. These areas of fusion could be weak points within the radial carpal bone.     

Use of contoured bar transhock external fixators in 17 cats

Seventeen cats with injuries involving the distal tibia, talocrural joints, tarsus and metatarsus were treated with contoured connecting bar transhock external fixators as either primary or secondary fixation. The assemblies stabilised all the injuries successfully. Fracture union was achieved in 11 out of 12 cases. Excellent results were achieved in eight cats and the outcome was satisfactory in a further five. Those cats with injuries involving talocrural joint luxation with displacement and significant periarticular soft tissue disruption had poorer long-term results. Complications occurred in seven cases.     

Clinical and anatomical features of lymphosarcoma in 118 cats

Objective To determine patients' characteristics and anatomical distribution of lesions in cats with lymphosarcoma. Design Prospective multi-institutional study of naturally occurring feline lymphosarcoma. Methods Veterinarians in Sydney were provided with free diagnostic laboratory services for suspect cases of feline lym-phosarcoma. Lymphosarcoma was diagnosed based on physical findings, radiographic and/or ultrasonographic images and results of cytological or histopathological examination. When owners were not interested in pursuing an antemortem diagnosis, suspect cases were collected for necropsy. Patients' characteristics and physical findings were recorded. A modified scheme for anatomical classification of lesions was devised including a ‘mixed’ category for cases which involved two or more anatomical forms. Results One hundred and eighteen cases were accrued over an 18 month period. The median age was 120 months and range 5 to 212 months. Age distribution was bimodal, with a small peak for cats less than 24 months, and a normal distribution centred on 97 to 120 months. Eighty cats were domestic crossbreds, 22 were Siamese or Oriental cats (including crosses), 6 were Burmese, 5 were purebred longhairs and the remaining 5 were one of a number of purebred shorthaired breeds. In comparison to 1017 consecutive cases admitted to our hospital for conditions other than lymphosarcoma, Siamese/Oriental cats were over-represented amongst lymphosarcoma cases (P = 0.0006). Male cats were also over-represented, accounting for 72 of 118 cases (P = 0.05). Abdominal lymphosarcoma was the most common anatomical form (43 cats), followed by mixed (39), nodal (20), mediastinal (9) and atypical (involving non-lymphoid organs, 7) forms. When analysed for specific organ involvement, 29 (25%) had mediastinal involvement, 71 (60%) had abdominal involvement including 60 (51%) with involvement of the intestinal tract and/or mesenteric lymph nodes and 36 (31%) with bilateral renal involvement, and 47 (40%) had peripheral lymph node involvement. No case of primary lymphoid leukaemia was identified. A noticeable subgroup of cats younger than 24 months had involvement of the anterior mediastinum with or without concurrent enlargement of cervical or axillary lymph nodes; Siamese/Oriental cats were over-represented in this subgroup. Among cases with nodal involvement, lymph nodes of the head and neck were frequently involved, mandibular nodes most commonly, followed by superficial cervical nodes. In seven cases a solitary node was affected. Conclusions Compared with similar surveys overseas, our cats were older and male cats were over-represented. There was a notable subgroup of young cats with mediastinal involvement. Siamese/Oriental cats were over-represented in this subgroup as well as in the larger population of cats with lymphosarcoma. Compared with overseas surveys, renal involvement, mixed cases and atypical cases (including nasal lymphosarcoma) were more common. A new subcategory of nodal lymphosarcoma, with involvement restricted to node(s) of head and neck, was identified.     

Prevalence of megabacteria in budgerigar colonies

Objective To measure the prevalence of megabacteria in budgerigar-breeding colonies and to evaluate possible methods to reduce the prevalence.
Design A monitoring study over several years.
Sample population Two budgerigar ( Melopsittacus undulatus ) colonies with over 300 birds each.
Procedure The prevalence of megabacteria in the faeces in two budgerigar breeding colonies, colony 1 and 2, was determined by faecal examination of each bird. Following an initial survey (1990), most of the birds that were scored 2+ or more were culled and a management practice was implemented to discriminate against positive birds. Consecutive yearly surveys (1991, 1992) were conducted on the young birds bred in these colonies. The prevalence of megabacteria in colony 2 was also evaluated in 1994 and 1996 after all the birds were treated with amphotericin B administered in drinking water.
Results The prevalence of megabacteria in the two colonies was significantly (P < 0.001) different. Overall the prevalence of megabacteria adjusted for colony differences was significantly higher (P < 0.025) in males compared to females. Age was not an influencing factor. After the initial survey, the prevalence in the offspring did not significantly (P > 0.05) decrease in the following two annual breeding seasons but by inference it did significantly decrease after amphotericin B treatment.
Conclusion The practice of culling most birds with more megabacteria in faeces and discriminating against positive birds when selecting birds for breeding or culling birds on show quality does not decrease megabacteria prevalence in the offspring. However, a reduction in prevalence does occur with administration of amphotericin B. Birds may have amphotericin B-resistant organisms and these birds need to be identified and culled.     

Cochlosoma infections in finches

Objective To survey finches in pet shops for Cochlosoma infection and evaluate the efficacy of antiprotozoal therapy with metronidazole or ronidazole.
Design A survey of pet shop finches and drug efficacy trials.
Procedure Finches in pet shops were randomly selected and their faeces examined microscopically for motile Cochlosoma sp trophozoites. Drug trials were carried out on 60 adult finches with naturally occurring infections. Body weight was measured and the faeces of each bird was examined for trophozoites at the beginning and 7 days after the end of treatment. In some birds, additional daily faecal examinations were done until three consecutive negative results were obtained. Metronidazole was administered at various dose rates by crop gavage or in drinking water to eight groups of five to ten finches each. Ronidazole was given in water for 7 days to ten finches. In addition, six finches whose faeces tested positive were necropsied and their tissues collected for histological examination.
Results Motile flagellates in the faeces were identified as C anatis -like protozoa. Red-headed parrot-finches, Bengalese and Lady Gould finches were found to be most commonly infected. Cochlosoma sp was also found in the blue-faced parrot-finch, zebra finch, painted finch, nutmeg mannikin and double-barred finch. Metronidazole and ronidazole were found to be effective against Cochlosoma sp. Histological findings on infected adult finches were normal, except for the presence of numerous flagellates between the colorectal villi and cloacal mucosal folds.
Conclusions Cochlosoma anatis -like organisms can infect several species of finches and in adult finches are confined to the colorectum and cloaca. Infection in adult finches was mostly subclinical and could be treated effectively with metronidazole or ronidazole.     

Growth and enzyme production by Bacillus subtilis and Flavobacterium pectinovorum in Picea sitchensis

Summary When Sitka spruce (Picea sitchensis) is stored in water, an increase takes place in the permeability of the wood. This increase is due to bacterial degradation of the tori and bordered pit membranes and partial degradation of the crossfield pits. It has been demonstrated that two bacteria—Bacillus subtilis and Flavobacterium pectinovorum—isolated from this water stored wood, grow and produce amylase, xylanase and pectinase in sapwood blocks suspended in sterile pond water. Since these organisms have also been shown to increase the permeability of sapwood blocks, it has been concluded that they are primarily responsible for the permeability changes that occur when spruce poles are stored in lake water.When the two organisms are incubated together in sapwood blocks, growth of F. pectinovorum is synergistically affected, while growth of B. subtilis is antagonised. However, no antagonism of enzyme production was evident. The levels of biological activity in the sapwood in these laboratory experiments were much higher than the values observed in the natural lake system. The concentration effect obtained in the laboratory system, in contrast to the dilution effect occurring in the lake environment, is undoubtedly responsible for this variance. Results indicate that the rate at which permeability changes occur could possibly be accelerated by replacing the natural lake system with an enclosed tank system.     

Slow viscous flow and the organization of the cell wall in conifer tracheids

Summary In view of the known orientation of microfibrils about pits in conifer tracheids a model is presented which is consistent with the assumption that this pattern derives from the slow viscous flow of a fluid about a cylindrical obstacle.At present at the Botany Department, La Trobe University, Bundoora, 3083, Victoria, AustraliaThe authors wish to thank Dr. W. B. Thompson and Professor A. B. Wardrop for their constructive criticism.     

Incubation of ovine scrapie with environmental matrix results in biological and biochemical changes of PrPSc over time

Ovine scrapie can be transmitted via environmental reservoirs. A pool of ovine scrapie isolates were incubated on soil for one day or thirteen months and eluted prion was used to challenge tg338 mice transgenic for ovine PrP. After one-day incubation on soil, two PrP^Sc phenotypes were present: G₃₃₈ or Apl₃₃₈ii. Thirteen months later some divergent PrP^Sc phenotypes were seen: a mixture of Apl₃₃₈ii with either G₃₃₈ or P_338, and a completely novel PrP^Sc deposition, designated Cag₃₃₈. The data show that prolonged ageing of scrapie prions within an environmental matrix may result in changes in the dominant PrP^Sc biological/biochemical properties.