In vitro antimicrobial susceptibilities of three Porphyromonas spp and in vivo responses in the oral cavity of cats to selected antimicrobial agents

OBJECTIVES: To determine in vitro susceptibility of Porphyromonas gingivalis, P salivosa and P circumdentaria to seven antimicrobial agents by agar dilution and Epsilometer test methods and to assess the effectiveness of these antimicrobial agents in reducing the numbers of each Porphyromonas spp in the oral cavity of 16 domestic cats. DESIGN: A two-part prospective study involving in vitro antimicro-bial studies using Porphyromonas spp obtained from naturally occurring feline infections and in vivo antimicrobial response studies using client-owned cats with naturally occurring periodontal disease. PROCEDURE: Isolates (n = 25) of three feline Porphyromonas spp from the oral cavity and oral-associated disease were tested for their in vitro susceptibility to amoxycillin, amoxycillin-clavulanate, benzylpenicillin, clindamycin, doxycycline, erythromycin and metronidazole, using agar dilution and Epsilometer test methods. Digoxigenin-labelled whole chromosomal DNA probes directed against P gingivalis VPB 3492, P circumdentaria NCTC 12469T and P salivosa VPB 3313 were used to quantify organisms taken from two sample sites at the gingival margins of these cats prior to, and 5 days after, treatment with one of four commonly used antimicrobial products (amoxycillin-clavulanate, clindamycin, doxycycline or spiramycin-metronidazole). The response to treatment was assessed clinically for each cat. RESULTS: All isolates were susceptible in vitro to all seven antimicrobial agents using both methods. The numbers of P gingivalis were not reduced at the gingival sample sites by administration of amoxycillin-clavulanate for 5 days, although this treatment reduced the numbers of P salivosa and P circumdentaria to below detection levels in six of eight and two of three of sample sites, respectively; clinical improvement was not observed in cats treated with amoxycillin-clavulanate. Treatment with clindamycin, doxycycline or spiramycin-metronidazole resulted in clinical improvement and a marked reduction of all Porphyromonas isolates at the sample sites. CONCLUSION: The Epsilometer test is a simple and accurate method for determining the minimum inhibitory concentration for P gingivalis, P salivosa and P circumdentaria. All strains were susceptible in vitro to all the antimicrobial agents tested although clinical improvement of gingival disease was not noted with amoxycillin-clavulanate when given for 5 days at usual doses. This appears to be the first report of the disparity between the in vivo and in vitro susceptibility of oral bacterial strains to amoxycillin-clavulanate in the veterinary dental literature. This also appears to be the first report in which clinical and microbiological responses to commonly used antimicrobial agents for periodontal disease in cats has been documented and quantified. It was shown that treatment with clindamycin, spiramycin-metronidazole or doxycycline not only produced a substantial reduction in the number of Porphyromonas spp (in the majority of cases to below detection levels), but also resulted in substantial clinical improvement. This would indicate that these antimicrobial agents are useful adjunctive therapy to mechanical debridement in domestic cats.     

The isolation and enumeration of three feline oral Porphyromonas species from subcutaneous abscesses in cats

Samples were examined from 15 subcutaneous fight wound abscesses from 15 cats. All abscesses were closed at the time of sampling and cats had received no prior treatment. Samples were processed within 20 min and quantitative assessment made of total facultative and obligately anaerobic flora isolated. Digoxigenin labelled whole chromosomal DNA probes directed against three feline members of the genus Porphyromonas (P. gingivalis VPB 3492, P. circumdentaria NCTC 12469T and P. salivosa VPB 3313) were used to identify members of this genus and quantification of these species was made from each cat using colony lifts and southern hybridisation from nitrocellulose membranes taken from replicate plates from each abscess sample. Twelve of the 15 abscesses yielded a variety of facultative and obligately anaerobic (FOA) bacterial species and members of the genus Porphyromounas were enumerated from each of these 12 abscesses. Of the 12 abscesses in which Porphyromonas species were detected, seven contained one species only (five contained only P. gingivalis and two contained only P. salivosa) three abscesses contained two species (both P. gingivalis and P. circumdentaria) and two abscesses contained all three species of Porphyromonas. These results show that members of the genus Porphyromonas are likely to be significant contributors to the purulent disease process in subcutaneous abscesses in cats.     

The association of two recombinant proteinases of a feline strain of Porphyromonas gingivalis with periodontal disease in cats

Serum from 40 domestic cats with various grades of periodontal disease was used to probe two recombinant functional proteinases from feline strain VPB 3457 of Porphyromonas gingivalis expressed in E. coli. One recombinant proteinase (VPB 2856) was constructed using polymerase chain reaction and had 91% DNA identity with the prtC collagenase gene of the human type strain of P. gingivalis, while the other proteinase (VPB 2814) was isolated from a size selected genomic library and had an amino-terminal sequence with no significant identity with deposited sequences. Thirteen of 40 cats showed a serum antibody response to VPB 2856 using Western immunoblot detection. All the 13 cats had an overall periodontal grade of 3 or greater and greater than 1.68x10(5) cfu P. gingivalis at the canine and premolar periodontium sample sites. Fourteen of 40 cats showed a serum antibody response to VPB 2814. Thirteen of these 14 cats had an overall periodontal grade of 3 or greater. Regression analysis of overall periodontal grade against the serum antibody response showed significant positive relationships for both VPB 2856 (r2 = 0.351; p<0.001) and VPB 2814 (r2 = 0.247; p<0.001). Regression analysis of the total colony forming units of feline strain P. gingivalis against the grade of serum antibody response showed a positive relationship for both VPB 2856 (r2 = 0.662; p<0.001) and VPB 2814 (r2 = 0.531; p<0.001). These data provide strong evidence that the recombinant proteinases of feline P. gingivalis expressed in E. coli clones VPB 2856 and VPB 2814 are associated with periodontal disease in cats.     

Serum antibody responses of cats to soluble whole cell antigens of feline Porphyromonas gingivalis

The whole cell soluble antigens of two strains (VPB 3457 and VPB 3492) of feline Porphyromonas gingivalis were analysed by Western blotting using serum taken from 40 domestic cats with various grades of periodontal disease. Five strongly immunogenic protein bands (70, 34, 27, 24 and 19kDa) from VPB 3457 and seven from VPB 3492 (58, 44, 34, 27, 25, 24 and 21kDa) were selected for further study. A significant positive correlation was found between the serum antibody response to the 70, 34, 27, 24 and 19kDa bands of VPB 3457 and the 58, 44, 25, 24 and 21kDa bands of VPB 3492 and the overall periodontal grade. A significant positive correlation was also found between the serum antibody response to the 24kDa band of VPB 3457 and the total colony forming units of P. gingivalis. N-terminal sequencing of the 44kDa band of VPB 3492 showed 75% identity with the translated amino acids from the hag A (haemagglutinin) gene of a human strain of P. gingivalis and N-terminal amino acid sequence of the 27kDa band of VPB 3457 showed 88% identity with the amino acid sequences translated from DNA of purported genes coding for variously named proteinases of human strains of P. gingivalis.     

Ecology of genus Porphyromonas in canine periodontal disease.

Asaccharolytic pigmented Porphyromonas species, including P. endodontalis, P. gingivalis, P. circumdentaria and unclassified species, were isolated from the plaque of adult dogs, but not from any oral sites of puppies and adolescent dogs. With age-dependency, the proportion of Porphyromonas species in the flora of plaque increased. Isolation of the genus Porphyromonas was clearly associated with the progress of periodontol disease. We suggested that Porphyromonas is the exogenous organism and obligate pathogen for canine periodontal diseases.     

Serum antibody responses of cats to soluble whole cell antigens and isolated fimbriae of feline Porphyromonas salivosa (macacae) and associations with periodontal disease

The whole cell soluble antigens of two strains (NCTC 11632 and VPB 3313) of feline Porphyromonas salivosa (macacae) were analyzed by Western blotting using serum taken from 40 domestic cats with various grades of periodontal disease. Nine strongly immunogenic protein bands (66, 52, 42, 29, 27, 23, 22, 21 and 19kDa) were selected from both strains for further study. Both strains showed a significant association between overall periodontal grade and serum responses to the 66 and 21kDa bands with significant responses across both strains to all other bands except the 52kDa band. Similarly, both strains showed a significant association between the total colony forming units and serum responses to the 66 and 42kDa bands with significant responses across both strains to all other bands except the 19kDa band. When sera from 25 of these cats were tested by Western blotting against the isolated fimbriae of VPB 3313, there was a significant association between the grade of response of cats to the 42kDa fimbrial preparation and (1) the total reactivity of the mouth (the sum of the responses to all individual whole cell antigens), (2) the total colony forming units of P. salivosa (macacae) at the premolar site, and (3) to their responsiveness to the 42kDa band in the soluble whole cell antigen preparations. These findings suggest that P. salivosa (macacae) is a strong immunogen in the mouths of cats and those cats with more severe periodontal disease have a greater serum antibody reactivity to various soluble whole cell antigens, specifically including the fimbriae of this organism, than those with less severe periodontal disease. Overall, the findings suggest that this organism may be a contributor to periodontal disease in cats.     

Pigmented-anaerobic bacteria associated with canine periodontitis

The etiology of human periodontal disease has been the focus of considerable research, yet relatively little is known about the causative agents of companion animal periodontitis. In humans, Porphyromonas gingivalis, a black-pigmented anaerobic bacteria (BPAB), has been implicated as the primary periopathogen. It has been demonstrated that BPAB are also found in companion animal periodontal pockets. While some animal BPAB have been individually identified, a study to identify the most frequently isolated subgingival BPAB has not been completed using genetic tools. The objective of this work was to identify the types and relative frequencies of pigmented anaerobic bacteria found in the periodontal pockets of dogs. Porphyromonas salivosa, Porphyromonas denticanis (a novel species) and Porphyromonas gulae were found to be the most frequently isolated BPAB associated with canine periodontitis.     

Cloning and expression of the superoxide dismutase gene of the feline strain of Porphyromonas gingivalis: immunological recognition of the protein by cats with periodontal disease

Recent evidence suggests that feline members of the genus Porphyromonas are of consequence in periodontal disease in cats. Several possible virulence factors from feline strains of Porphyromonas gingivalis have been described that have similarities to those of human P. gingivalis. Both human and feline strains of P. gingivalis produce superoxide dismutase (SOD) which has been proposed as modulator of the inflammatory response during infection. The objective of this study was to clone the superoxide dismutase gene of feline P. gingivalis, to compare the characteristics of its product with that of the native enzyme and to determine its immunoreactivity in cats with periodontal disease. The sod gene of the feline strain Veterinary Pathology and Bacteriology (VPB) 3457 of P. gingivalis was amplified by PCR and cloned in frame with the alpha-peptide of the LacZ gene of E. coli in plasmid pUC19. This construct expressed SOD activity in E. coli with characteristics similar to those of the native SOD enzyme of P. gingivalis human strain 381 and the parent feline strain VPB 3457. The recombinant SOD had an apparent molecular weight of 54,700+/-1300 (S.E.M.) and was inactivated by 5mM hydrogen peroxide but not by 2mM KCN. There was a significant association (P=0.005) between the immunoreactivity of cats to P. gingivalis VPB 3457 soluble whole cell proteins on immunoblots and their responsiveness to the SOD protein. This suggests that cats showing a marked serum responsiveness to P. gingivalis itself, react to the SOD enzyme and further supports the role of feline P. gingivalis in periodontal disease.     

Evaluation of cross-protection by immunization with an experimental trivalent companion animal periodontitis vaccine in the mouse periodontitis model

Companion animal periodontal disease is one of the most prevalent diseases seen by veterinarians. The goal of this study was to evaluate the vaccine performance of a trivalent canine periodontitis vaccine in the mouse oral challenge model of periodontitis. Mice vaccinated subcutaneously with an inactivated, whole-cell vaccine preparation of Porphyromonas denticanis, Porphyromonas gulae, and Porphyromonas salivosa displayed significantly reduced alveolar bone loss in response to heterologous and cross-species challenges as compared to sham vaccinated animals. Based on the results of these studies, a periodontitis vaccine may be a useful tool in preventing the initiation and progression of periodontitis caused by the most commonly isolated pigmenting anaerobic bacteria in animals.     

Comparative phospholipid analogue distributions of Porphyromonas gingivalis isolated from cats in Australia and the USA

DNA-DNA homology measurements and phospholipid (PL) analogue profiling have shown heterogeneity of Porphyromonas gingivalis. The aim of this study was to determine whether there were differences between cat strains of P. gingivalis from Australia and USA with respect to PL analogue distribution. Lipids were extracted with chloroform-methanol and examined by fast atom bombardment-mass spectrometry (FAB-MS) in negative-ion mode, using published methods. For PL analogues, the major anions included those with mass-to-charge (m/z)=634, 648, 662, 705, 932, 946 and 960, respectively, corresponding to expected presence of PE (28:0), PE (29:0), PE (30:0), PG (32:1), and three unknown homologues of a glycero-phospholipid with a single nitrogen. Analyses were compared to calculate a matrix of Pearson coefficients of linear correlation from which a dendrogram was produced of strains clustered by single linkage. One cluster was comprised solely of Australian cat-to-cat bite isolates and a second cluster included exclusively USA cat- and dog-to-human bite isolates except for one Australian cat-to-cat bite isolate (VPB 5089). The US cluster included three outliers, one of which was the Australian cat isolate VPB 5089. The human type strain (ATCC 33277) was quite remote from all dog and cat strains. It was shown that P. gingivalis human and non-human animal isolates have distinct PL analogue profiles from each other. Furthermore, the cat strains from the USA and those from Australia showed quantitative differences in polar lipid profiles that correlated largely with country of isolation.     

Role of ovarian progesterone and potential role of prostaglandin F2alpha and prostaglandin E2 in modulating the uterine response to infectious bacteria in postpartum ewes

In sheep and cattle, the postpartum uterus is resistant to bacterial challenge until after corpora lutea develop. A 2 x 2 factorial arrangement of treatments was used to determine whether prostaglandins may mediate the effects of progesterone in transforming the postpartum uterus from resistant to susceptible. On d 14 postpartum, ewes (n = 6/group) were ovariectomized or sham ovariectomized, and the vena cava was catheterized for daily collection of uteroovarian-enriched blood. From d 15 to 20, ewes received twice daily intramuscular injections of progesterone in sesame oil or plain sesame oil. On d 20, each uterus received 75 x 10(7) cfu of Arcanobacterium pyogenes and 35 x 10(7) cfu of Escherichia coli. Uteri were collected on d 25 and examined for signs of infection. For each blood sample, unstimulated and mitogen-stimulated lymphocyte proliferation was measured as [3H]thymidine incorporation, smears were prepared for differential white blood cell (WBC) counts, and progesterone, prostaglandin F2alpha, (PGF2alpha), and prostaglandin E2 (PGE2) were quantified. All 12 progesterone-treated, but only two of the 12 oil-treated, ewes developed uterine infections (P < 0.001). Progesterone treatment increased (P < 0.001; 3.1 vs 1.5 ng/mL) and ovariectomy decreased (P < 0.001; 3.7 vs 0.9 ng/mL) vena caval progesterone. Progesterone treatment reduced (P < 0.01) PGF2alpha, (303.9 vs 801.3 pg/mL), and PGF2alpha was greater (P < 0.05) before than after inoculation (626.4 vs 478.8 pg/mL). The PGE2 concentration was greater in progesterone-treated, ovary-intact ewes than in ewes in the other groups (ovariectomy x progesterone treatment; P < 0.01). Ovariectomy increased (P < 0.005; 4.4 vs 2.9 pmol) and progesterone treatment decreased (P < 0.05; 3.2 vs 4.1 pmol) concanavalin A-stimulated lymphocyte proliferation. Ovariectomy increased lipopolysaccharides-stimulated proliferation (P < 0.05; 2.4 vs 1.9 pmol). For neutrophils per 100 WBC, the ovariectomy x progesterone and progesterone x period interactions were significant (P < 0.01). The ovariectomy x progesterone interaction was significant (P < 0.01) for lymphocytes per 100 WBC. Ovariectomy decreased monocytes (P < 0.001; 10 vs 13) and increased eosinophils (P < 0.001; 10 vs 5) per 100 WBC. Progesterone makes the postpartum uterus in ewes susceptible to infection, but ovariectomy allows ewes to remain resistant; uterine prostaglandins may mediate this change. This model creates opportunities to determine the mechanisms responsible for the shift from resistance to susceptible.     

Mitotic index is predictive for survival for canine cutaneous mast cell tumors

Mitotic index (MI) is an indirect measure of cell proliferation that has been demonstrated to be a strong predictor of outcome for several human and canine cancers. The purpose of this study was to evaluate the utility of MI as a predictor of biologic behavior and survival in dogs with cutaneous mast cell tumors (MCTs). Medical records from 148 dogs with histologically confirmed MCTs were reviewed. Information regarding tumor grade, local recurrence, metastatic disease, date of death/last follow-up, and outcome was obtained. The region of the tumor with the highest overall mitotic activity was chosen for evaluation, and the MI value was defined as the number of mitotic figures/10 high-power fields (400x, 2.7 mm(2)). A Cox proportional hazards regression model was used to compare MI with survival data. A Mann-Whitney test was used to compare MI on the basis of the development of local recurrence and metastatic disease. The MI correlated directly with tumor grade (P < .0001). The median survival time for dogs with an MI < or =5 was significantly longer (70 months) than for those with an MI >5 (2 months), regardless of grade (P < .001). For grade II tumors with an MI < or =5, the median survival time (MST) was 70 months, compared with 5 months for those with an MI >5 (P < .001). For grade III tumors with an MI < or =5, the MST was not reached, compared with <2 months for those with an MI >5 (P < .001). In conclusion, MI is a strong predictor of overall survival for dogs with cutaneous MCTs and should be included as a prognostic indicator when determining therapeutic options.     

Radiographic evaluation of destructive periodontal disease in blue mink in relation to age and blood morphology

In this study, blood samples and jaws were collected from 2 genotypes of blue mink (n = 289) in order to examine phenotypic expression of specific characteristics of Chediak-Higashi Syndrome (C-HS). Blood samples were subjected to differential counts to assess the proportion of abnormal polymorphonuclear leukocytes characteristic for CH-S (C-HS-leukocytes). Abnormal leukocytes with characteristic signs of C-HS were found in blood smears from all mink included in this study. Four teeth in one half of the mandible (P3, P4, M1, M2) were subjected to quantitative radiographic evaluation of alveolar bone loss and tooth loss. There was a high prevalence of destructive periodontal disease among blue mink included in this study. Mild to moderate periodontal disease (defined by less than 50% alveolar bone loss related to 1 or more teeth) affected 73.7% of young mink (age = 7 mo) and 67.9% of older animals (age > or = 19 mo). Severe periodontal disease (defined by more than 50% bone loss related to one or more teeth) was not detected in mink aged 7 mo, but affected 15.3% of mink aged 19 mo and 39.6% of mink aged 31 mo. The positive relationship between age and periodontal disease was statistically significant (P < 0.01). The prevalence of tooth loss was found to be high among blue mink aged > 19 mo (21.6%) and was also significantly related to age (P < 0.01). A significant positive interaction between alveolar bone loss and tooth loss (P < 0.01), implies that the highly prevalent tooth loss in the mink was related to and possibly caused by destructive periodontal disease. There was no significant difference in the prevalence of periodontal disease between the 2 genotypes and age was found to be the only statistical predictor of poor production results (P < 0.01) in blue mink.     

Effects of live cultures of Lactobacillus acidophilus (strains NP45 and NP51) and Propionibacterium freudenreichii on performance, carcass, and intestinal characteristics, and Escherichia coli strain O157 shedding of finishing beef steers

In Exp. 1, 240 beef steers (initial BW = 332.8 kg) were used to determine the effects of Lactobacillus acidophilus (LA) plus Propionibacterium freudenreichii (PF) on performance, carcass, and intestinal characteristics; serum IgA concentrations; and the prevalence of Escherichia coli O157 (EC). Cattle were fed a steam-flaked corn-based, 92% concentrate diet, and the four direct-fed microbial (DFM) treatments (12 pens/treatment) included in a randomized complete block design were as follows: 1) control, lactose carrier only (CON); 2) 1 x 10(9) cfu of LA NP51 plus 1 x 10(6) cfu of LA NP45 plus 1 x 10(9) cfu of PF NP24 per animal daily (LA45-51H); 3) 1 x 10(9) cfu of LA NP51 plus 1 x 10(9) cfu of PF NP24 per animal daily (LA51); and 4) 1 x 10(6) cfu of LA NP51 plus 1 x 10(6) cfu of LA NP45 plus 1 x 10(9) cfu of PF NP24 per animal daily (LA45-51L). No differences (P > 0.10) were detected for pen-based performance data. The average lamina propria thickness for LA51 and LA45-51H steers was less (P = 0.02) than the average for CON and LA45-51L steers. Moreover, LA51 and LA45-51H steers had a lower (P = 0.06) prevalence of EC shedding than CON and LA45-51L steers. In Exp. 2, 660 steers fed 91% concentrate, steam-flaked corn-based diets were used to determine the effects of the following DFM treatments (10 pens/treatment) on performance, carcass, and intestinal characteristics: 1) control, lactose carrier only (CON); 2) 5 x 10(6) cfu of LA NP51 plus 5 x 10(6) cfu of LA NP45 plus 1 x 10(9) cfu of PF NP24 per animal daily (LA45-51L); and 3) 1 x 10(9) cfu of LA NP51 plus 5 x 10(6) cfu of LA NP45 plus 1 x 10(9) cfu of PF NP24 per animal daily (LA45-51H). Steers were from two weight groups (WG). One group (SDOF; BW at arrival = 351.5 kg) had grazed before arrival, and the other group (LDOF; BW at arrival = 314 kg) had been in a grower yard. A split plot was used with WG as the whole-plot factor and DFM in the split plot. There was an interaction of WG and DFM for ADG (P = 0.05) and for carcass-adjusted ADG (P = 0.08). The simple-effect ADG and carcass-adjusted ADG means for DFM treatments differed (P < or = 0.01) between WG classifications. Within SDOF, ADG for CON and LA45-51L did not differ (P = 0.70), but both were less (P < or = 0.08) than for LA45-51H. Overall, these data indicate that live cultures of LA plus PF did not greatly affect feedlot performance and carcass characteristics. Some of the DFM used decreased fecal EC shedding, which might be related to the results for ileal lamina propria thickness.     

Molecular and antigenic similarities of the fimbrial major components between Porphyromonas gulae and P. gingivalis

Porphyromonas gulae is black-pigmented anaerobic bacteria associated with canine periodontitis. There is little information available about the specific identify and relative occurrence of pigmented anaerobes in companion animals. Our aim was to clarify the factor involved in the adherence and colonization of the organism in the oral cavity. Fimbrial protein was purified from P. gulae ATCC 51700. The molecular mass of this protein was approximately 41kDa as estimated by SDS-PAGE. An antibody against 41-kDa fimbrial protein from P. gingivalis ATCC 33277 reacted with fimbrillin of P. gulae ATCC 51700. Immunogold electron microscopy revealed that the anti-41kDa fimbrial serum bound to fimbria on the cell surface of P. gulae ATCC 51700. Thus, fimbrial protein of P. gulae ATCC 51700 had the same size and antigenicity as 41-kDa fimbriae of P. gingivalis ATCC 33277. The nucleotide sequence of the fimA gene from P. gulae ATCC 51700 showed 94% homology with that of P. gingivalis ATCC 33277. Moreover, the deduced amino acid sequences have 96.8% identity. P. gulae has adherent ability to gingival epithelial cells. The properties of P. gulae fimbriae are similar to those of P. gingivalis fimbriae. We suggest that the surface structure of P. gulae may play a role in the colonization of this organism in periodontal pockets in companion animals.     

Infectivity and persistence of an outbreak strain of Salmonella enterica serotype Typhimurium DT160 for house sparrows (Passer domesticus) in New Zealand

AIM: To examine the infective dose, incubation period and disease progression of an isolate of Salmonella enterica serotype Typhimurium definitive type 160 (DT160) originating from a naturally-infected house sparrow (Passer domesticus) during an outbreak of the disease in New Zealand. METHODS: Thirty-six house sparrows captured from the wild and free of Salmonella spp were divided into six groups of six birds, housed individually, and inoculated orally with phosphate buffered saline (PBS) or 10(1), 10(2), 10(3), 10(5), 2 x 10(8) colony forming units (cfu) of the outbreak strain of S. Typhimurium DT160. The birds were observed for 10 days for clinical signs and/or mortality, and faecal samples were collected to determine excretion of S. Typhimurium. The birds were euthanised 11 days post-inoculation (p.i.) and a wide range of tissue samples were collected for histopathological examination, and culture and typing of Salmonella spp. Macro-restriction profiling by pulsed-field gel electrophoresis (PFGE) using XbaI was performed for the epidemiological typing of S. Typhimurium DT160 isolates. RESULTS: Mortality in house sparrows inoculated with S. Typhimurium DT160 was dose-dependent, and 2/6 birds inoculated with 10(5) cfu and all six birds inoculated with 2 x 10(8) cfu died during the study. Infected sparrows displayed few clinical signs, apart from diarrhoea and/or polyuria, fluffed plumage, and sitting on the floor of the cage. Faecal excretion of DT160 occurred briefly in two birds inoculated with 10(2) cfu and four birds inoculated with 10(3) cfu, on most days in five birds inoculated with 10(5) cfu, and continuously in six birds inoculated with 2 x 10(8) cfu. DT160 was isolated from the livers of three birds which received 10(3) cfu, five birds dosed with 10(5) cfu, and all six birds given 2 x 10(8) cfu. Following necropsy, histopathological lesions similar to those seen in the natural disease were observed in the liver or spleen of three birds which received 10(3) cfu, and all birds dosed with > or =10(5) cfu. CONCLUSION: The results indicate that an isolate of S. Typhimurium DT60 originating from house sparrows in New Zealand is pathogenic to these birds and that the response is dose dependent. The persistence and excretion of the pathogen may last for at least 10 days. This confirms that sparrows infected with DT160 could be a source of infection to humans and other in-contact animals.     

Human Foetal Sacral Length Measurement for the Assessment of Foetal Growth and Development by Ultrasonography and Dissection

This study investigated the value of prenatal ultrasonographic measurements of sacrum as a predictor of in utero development and gestational age (GA), and to make a comparison between the data obtained by ultrasonography and dissection. A prospective cross-sectional study of ultrasonography was conducted in 186 pregnant women with uneventful single pregnancies. GA ranged from 14 to 40 weeks, and the relationships of sacrum length (SL) with GA, femur length (FL), biparietal diameter (BPD), head circumference (HC) and abdominal circumference (AC) were evaluated. In addition, the sacral bones of 101 spontaneously aborted foetuses aged from 13 to 39 weeks were dissected and measured. The relationship of the results with GA was investigated. There was a linear relationship between the ultrasonographic measurements of the SL and the GA (R2 = 0.93, P < 0.001), the FL (R2 = 0.93, P < 0.001), the BPD (R2 = 0.93, P < 0.001), the HC (R2 = 0.92, P < 0.001) and the AC (R2 = 0.90, P < 0.001). The rate of increase of SL was significantly higher before 28 weeks of gestation than in later pregnancy, with a correlation coefficient between SL and GA of 0.96 and 0.88, respectively. The SL measurements obtained by dissection were similar to those of ultrasonography. There was a linear relationship between the SL and the GA (R2 = 0.96, P < 0.001) and the correlation coefficients between SL and GA were 0.97 before 28 weeks of gestation, and 0.94 in later weeks. In conclusion, this study demonstrates a high correlation between the SL, GA and other standard measurements of foetal growth, and suggests that SL measurement is a valuable predictor of GA in the foetuses with normal growth.     

Development of a multivariate yield grade equation to predict compositional traits in mature cow carcasses.

Data from 68 cow carcasses were used to develop a new yield grading system. First principal component (FPC) values for compositional attributes (LNFT = separable lean weight/[lean+fat weight] x 100, LNBN = separable lean weight/[lean+bone+connective tissue weights] x 100, and BTPR = [defatted lean from the round, loin, rib, and chuck]/side weight x 100) were determined. The first component explained 83.5% of the standardized variance and load values were .63, -.52, and .58, respectively. The resulting FPC values ranged from -1.93 to 1.89. The linear regression of LNFT, LNBN, and BTPR (dependent variables) on FPC (independent variable) explained a significant amount of variation (P less than .001) in each case and resulted in R2-values of .98, .67, and .85, respectively. A best-fit yield grade equation, developed to predict FPC, included adjusted fat thickness (ADF), percentage of kidney, pelvic, and heart fat (KPH), and overall muscling grade (OM). The equation, FPC = 2.04 - (.67 x ADF) - (.21 x KPH) - (.0016 x OM), explained a significant amount (P less than .001) of variation in FPC with R2 = .94 and residual standard deviation = .25. Simple correlations for ADF, KPH, and OM with FPC were -.87, -.71, and -.80, respectively. Cow carcasses were assigned to one of three grades based on FPC values that corresponded with predetermined levels of LNFT, LNBN, and BTPR. These grades generally had smaller CV than existing grades. When used in conjunction with quality grades, proposed grades could be more useful to the cow meat industry.     

Experimental infection with BCG as a model of tuberculosis in the brushtail possum (Trichosurus vulpecula)

AIMS: To study the development and progression of lesions produced following experimental inoculations of possums with Bacille Calmette-Guérin (BCG) Pasteur Strain 1173P2 and to compare these with lesions that occurred following natural Mycobacterium bovis infection. METHODS: Possums were inoculated with 5 x 106 colony forming units (cfu) of BCG via the intra-dermal (I/D) route into the dorsum of the neck (n=38) or the left brachium (n=7),orally (n=10), via the endobronchial (E/B) route (n=12), or intravenously (I/V) (n=10, half of which received 5 x 106 cfu and half of which received 5 x 107 cfu of BCG). The possums were humanely killed between 1-4 weeks post inoculation (p.i.), and the nature and distribution of lesions examined grossly and histopathologically. RESULTS: The distribution of lesions following I/D inoculation via either route was similar to that of the natural disease, but there were few lesions in the lung. Endobronchial inoculation resulted in pulmonary disease but produced few lesions outside the respiratory tract. Lesions produced by I/V inoculation were similar in distribution to those seen in terminally ill tuberculous possums. No lesions were produced following oral inoculation. Regression of lesions commenced after 3 weeks p.i. CONCLUSIONS: Although the phenomenon of lesion resolution restricts the use of BCG to the study of early lesion development, it avoids the overwhelming disease induced using M. bovis and thus allows the early phases of the development and progression of tuberculosis in this species to be observed. Intradermal inoculation produced evidence that infection through the skin is associated with lesions in superficial lymph nodes, whereas pulmonary disease was associated with E/B inoculation. The results are consistent with the hypothesis that both percutaneous and respiratory routes are important in natural infection of possums with M. bovis.     

Bacteriological quality of raw cow's milk from four dairy farms and a milk collection centre in and around Addis Ababa

Bacteriological quality of raw cow's milk taken at different sampling points from four dairy farms and a milk collection centre in and around Addis Ababa was evaluated. Milk samples were aseptically collected from udder, bucket, storage container before and after cooling and upon arrival at the processing plant. A high increase in the mean total aerobic plate count was observed in milk samples taken from the bucket (1.1 x 10(5) cfu/ml), storage container before cooling (4 x 10(6) cfu/ml) and upon arrival at the processing plant (1.9 x 10(8) cfu/ml). The mean coliform counts ranged from 1.3 x 10(4) cfu/ml (storage container before cooling) to 7.1 x 10(4) cfu/ml (upon arrival at the processing plant). The hygienic quality of raw milk from the collection centre was poor with a mean total bacterial count of 1.3 x 10(7) cfu/ml. Milk sampled from the udder contained mainly staphylococci and micrococci as udder-specific bacteria, while samples taken at later stages were additionally contaminated with bacteria of environmental origin (especially Enterobacteriaceae). Lack of knowledge about clean milk production, use of unclean milking equipment and lack of potable water for cleaning purposes were some of the factors which contributed to the poor hygienic quality of raw milk in the study farms.