Distemper virus as a cause of central nervous disease and death in badgers (Meles meles) in Denmark

During the summer of 2002 a distemper-like disease was observed in the free-ranging badger population in Denmark. It was characterised by grand seizures, abnormal behaviour and death; the badgers all had severe chronic pneumonia and some had non-suppurative encephalomyelitis. In this study, eight of the affected badgers were examined by gross pathological, histological, immunohistological, bacteriological, parasitological and virological methods, and were diagnosed with distemper; canine distemper virus was identified.     

Streptococci from the bovine udder in dairy herds in Khartoum Province, Sudan

Five hundred and seventy-nine milk samples were collected from dairy cows on seven farms in Khartoum North area and one farm in Omdurman and examined by bacteriological cultures for the presence of streptococci. One hundred and ninety-three (33.33%) isolates were recovered and identified on the basis of bacteriological characteristics and biochemical reactions as: S. pyogenes, S. agalactiae, S. dysgalactiae, S. faecalis, S. faecium, S. bovis, S. equi, S. lactis and S. uberis. Fifty-seven isolates representing the preliminary identification were tested by the latex-agglutination test to determine the serological groups. It was found that 39 strains belonged to group B, 3 strains to group C. Four strains gave a weak reaction with group D sera and were identified by biochemical tests as S. uberis. Two isolates could not be identified by the available sera. The isolation of S. uberis, S. bovis, S. equi, S. lactis, S. faecalis, S. faecium and S. pyogenes from cows in the Sudan was reported for the first time.     

Amplification of 16S ribosomal RNA gene sequences for the identification of streptococci of Lancefield group B.

The 16S r RNA gene of 49 streptococci of serological group B isolated from various origins was amplified by polymerase chain reaction (PCR) and subsequently digested with the restriction enzymes Rsa I and Msp I. The restriction profiles of all group B streptococci appeared to be identical indicating no intraspecies sequence variations of this gene. A fragment of the gene of two group B-streptococcal reference strains, including the hypervariable V2 region, could be amplified by PCR and sequenced. The sequence appeared to be identical and allowed the design of species-specific oligonucleotide primers. The primer pair used produced an amplicon with a size of 1250 bp and correctly identified all 49 group B-streptococci investigated but none of the control strains of various species and serogroups. This primer could be used in a multiplex PCR and allowed a rapid identification of bacteria of this species.     

The effect of a high dose bovine herpes virus 1 marker vaccine in pregnant heifers: virological, bacteriological, immunological, and pathological findings

To determine a possible relationship between the compulsory vaccination against bovine herpesvirus 1 (BHV1) and cattle wasting disease, the effects of BHV1 vaccination on heifers were investigated. Twenty heifers in the third trimester of pregnancy were randomly allotted to a vaccine and a control group. The vaccine group was vaccinated twice with a 50-fold dose of BHV1 vaccine and the control group was inoculated with the diluent. The experiment was performed double blind. After vaccination, the cows were examined daily and condition scores were determined weekly. Blood, milk, and faeces samples were collected weekly for virological, bacteriological, and immunological investigation. The heifers were euthanized either 9 or 13 weeks after the first inoculation and pathological, virological, and bacteriological examination was performed. No differences were detected between the vaccine group and the control group. No concurrent infections were detected and there were no indications of immunosuppression after vaccination. No relationship between the BHV1 vaccination and wasting disease in cattle was detected.     

Abortion in goats by Caprine alphaherpesvirus 1 in Spain

An abortion outbreak occurred in a goat herd of Murciano‐Granadina breed in Almeria Region in Spain where 80 pregnant females aborted. All bacteriological and parasitological examinations resulted negative, whereas virological investigations and real‐time PCR assay showed the presence of Caprine alphaherpesvirus 1 DNA in the pathological specimens from aborted foetuses. Nucleotide sequence analysis revealed that the DNA was highly close related to the Swiss strain E‐CH (99.7%) and a little less extent to the Italian BA.1 strain (99.4%). Histopathological examination revealed multifocal, well‐circumscribed, 50‐ to 200‐μm‐diameter foci of coagulative necrosis in the liver, lungs and kidneys of three foetuses. In the periphery of the necrosis, there were frequently epithelial cells with the chromatin emarginated by large, round, amphophilic intranuclear viral inclusion bodies. The source of the infection in the herd could not clearly find out even some hypothesis were formulated. This seems to be the first report of an abortion outbreak due to Caprine alphaherpesvirus 1 in a goat herd in Spain.     

The significance of the incidence of coccidia of the Cryptosporidium species in the etiopathogenesis of neonatal diarrhea syndrome and enteritis in calves on the basis of simultaneous histopathologic and parasitologic studies

In 1985, a histopathological examination was conducted in the ileum and a parasitological examination in the ileum and rectum of 345 emergency slaughtered calves. The pathogenic role of cryptosporidia was compared in two groups of calves: group A contained 184 calves (53.3%) with scours and enteritis, group B contained 161 calves (46.7%) with other types of disease. In both groups the calves were divided into four age categories: 7 to 10 days (7.8%), 11 to 14 days (29.3%), 15 to 21 days (53.0%), 22 days and older (9.9%). During the parallel examination by both methods, cryptosporidia were detected in 124 cases, i.e. at the total invasion extensity of 35.93%; in group A the cryptosporidia were found in 76 cases (22.02%) and in the group B in 48 cases (13.91%). As indicated by the results, cryptosporidiosis is a disease of polyfactorial origin and cryptosporidia must be taken into account as one of the enteropathogenic factors in the etiology of scours. It is confirmed by the positive findings of cryptosporidia in the calves of group B (29.81%) and the negative finding in group A (31.3%) that no clear relationship was demonstrated to exist between the positive findings of these protozoans and the clinical symptoms of the diseases and enteritis. The pathogenesis of cryptosporidiosis includes significant regressive changes in the microvillous layer and enterocytes, which were a common finding at heavy invasions. The total positivity of the findings of cryptosporidia in the ileum was significantly higher (by 14.8%) during the histopathological examination, as compared with the parasitological examination. Out of the total positive findings, 39 cases (31.5%) were demonstrated histopathologically. During the parasitological examination, the positivity of the findings of cryptosporidia was higher by 4.3% in the rectum than in the ileum. When there was conducted a parallel parasitological examination of the ileum and rectum, the total positivity was higher by 7.5% than at the histopathological examination in the ileum. The highest invasion extensity was found at the age of 7 to 10 days in both groups of calves. The highest invasion intensity was found at the age of 11 to 21 days in both groups. The age until the 21st day when the parasitosis is intensively spread is considered as the most dangerous age of the calves from the epizootological point of view. Cryptosporidia occurred all the year round, no characteristic seasonal patterns were observed. The total average invasion extensity of the 24 farms from where the calves came was 42.3%, its range being from 0 to 85.7%.     

Distribution of the hyaluronate lyase encoding gene hylB and the insertion element IS1548 in streptococci of serological group B isolated from animals and humans

The present study was performed to investigate streptococci of serological group B obtained from various sources and group B streptococcal reference strains for serotype, hyaluronate lyase enzyme activity, the occurrence of the hylB gene and the insertion sequence IS1548. All group B streptococci were identified by cultural, biochemical, and serological properties and by polymerase chain reaction amplification of species-specific parts of the 16S-23S rDNA intergenic spacer region, the 16S rRNA gene and the CAMP-factor (cfb) gene. Of the 73 group B streptococci investigated, 59 strains displayed hyaluronate lyase enzyme activity. All hyaluronate-lyase-positive strains and three phenotypically hyaluronate-lyase-negative strains had a hylB gene with an amplicon size of 3.3kb. Eleven of the 14 phenotypically hyaluronate-lyase-negative strains generated a hylB gene PCR product with a size of 4.6kb, and 10 of these strains displayed a IS1548 amplicon with a size of 0.98kb. The hyaluronate-lyase-negative isolates were mainly observed among group B streptococci of serotype III/Rib. All strains harbouring IS1548 had an additional copy of IS1548 located downstream of the C5a peptidase (scpB) gene.     

Primary culture of chicken bursal plical epithelium

Plical epithelial cells were obtained by trypsin-EDTA treatment of chicken bursa of Fabricius and cultured in the presence of type IV collagen. The culture became confluent six to seven days after seeding. The grown cells showed a positive reaction for cytokeratin by immunostaining and had ultrastructural characteristics of the epithelial cells in vivo. The cell culture will be useful for parasitological and virological studies.     

Influence of sampling time on bacteriological diagnosis of goat intramammary infection

Intramammary infection in seven commercial goat herds was studied using premilking and postmilking samples for purposes as bacteriological diagnosis. Using a positive result on both premilking and postmilking samples as the definitive diagnosis, we compared the efficacy of single samples collected either premilking or postmilking. With this aim, 2268 bacteriological culture results were compared. The kappa values (0.60) showed moderate agreement between the two samples. Specificity and positive predictive value were higher for postmilking samples than for premilking samples. Specificity of postmilking samples was 99.4% for coagulase-negative staphylococci, 99.9% for Gram-negative bacilli, 100% for streptococci, 99.9% for corynebacteria and 100% for mixed cultures. Premilking samples specificity was 96.6, 99.5, 99.7, 99.8 and 99.8%, respectively. False positive diagnoses were more frequent for premilking samples. The results suggest that postmilking samples should be used to diagnosis of goat intramammary infection.     

Heterogeneity of fibronectin reactivity among streptococci as revealed by binding of fibronectin fragments

Streptococci belonging to serological groups A, B, C, G, L and U were studied for their interaction with 125I-labelled fibronectin and its fragments. Fibronectin purified from humans plasma by affinity chromatography on gelatin-agarose and heparin-agarose was cleaved by thrombin into a 29,000 Dalton and a 210,000 Dalton fragments. Terminal analysis of purified fragments indicated that 29,000 fragment was from amino-terminal and 210,000 fragment from carboxyl-terminal domain of fibronectin. Binding of fibronectin was observed in all streptococci except those of group B. Streptococci of groups A, G, L, U and S. equisimilis reacted only with 29,000 Dalton fragment whereas S. dysgalactiae, S. zooepidermicus and S. equi reacted only with 210,000 Dalton fragment. The streptococcal binding sites for these two fragments were distinct from each other. Fibrinogen blocked the binding of 210,000 Dalton fragment but not of 29,000 Dalton fragment. Trypsinization of streptococci did not affect their binding sites for 210,000 Dalton fragment but destroyed those for 29,000 Dalton fragment. The results indicate that the streptococci of group A and G as well as S. equisimilis which are mainly pathogenic in humans bind amino-terminal fragment of fibronectin. This may facilitate the adherence of these pathogens. On the other hand, the streptococci isolated from animal infections had different binding sites recognized only by carboxyl-terminal part of fibronectin.     

Immune responses of immunocompetent and immunocompromised mice experimentally infected with Bartonella henselae

The aim of this study is to understand host immune responses in immunocompetent and immunocompromised mice against Bartonella henselae infection. BALB/c and nude (BALB/c nu/nu) mice were inoculated intraperitoneally with 10(8) colony forming units of B. henselae (Houston-1 strain). Blood, brain, liver, spleen, kidney and bone marrow samples were collected 0, 3, 7, 14, 21 and 28 days after infection and submitted to bacteriological, serological and genetical examinations. B. henselae was isolated only from the liver 3 days after infection. DNA of the inoculums was detected by polymerase chain reaction from blood, liver, and spleen samples collected from BALB/c and blood from nude mice 3 and 7 days after infection. No bacterial DNA was detected from both BALB/c and nude mice thereafter during 4 weeks observation periods. These results indicate that the T-cell may not participate in the effective elimination of the organisms from mice. In addition, western blot analysis revealed that the antigens of 27.3- and 31.5-kDa reacted with IgM antibodies from the blood of BALB/c and nude mice after 3 days of infection, suggesting that these antigens were recognized by thymus-independent mechanism. Furthermore the antigens were detected from the culture-supernatants of B. henselae, indicating that these antigens were secreted from the organisms.     

Relationship between group B streptococcal serotypes and cell surface hydrophobicity.

Cell surface hydrophobicities of streptococci of serological group B were determined by the adherence of the bacteria to hexadecane droplets. A significant adherence to hexadecane was observed with the group B streptococcal type reference strains Ib, V, Ic, R and X, but not with those of serotype Ia, II, III and IV. Cultivation of the bacteria in microcapsule-inducing media reduced the hexadecane adherence properties. The adherence to hexadecane was not related to fibrinogen binding properties of the cultures. Screening a large number of group B streptococci isolated from humans and bovines revealed that those with polysaccharide type antigen alone were generally hydrophilic, those with protein antigen alone or with protein antigen in combination with polysaccharide antigen were mostly hydrophobic. Cultivation of the bacteria under microaerobic conditions or after a single mouse passage enhanced microcapsule production and correspondingly reduced the hexadecane adherence values. Treatment of the bacteria by guanidinium chloride or by neuraminidase enhanced the hexadecane adherence. The hydrophobic component on group B streptococcal surface appeared to be partly inactivated by heat or proteolytic treatment of the bacteria.     

Lancefield group B and C streptococci in East African camels (Camelus dromedarius)

Seventeen Lancefield group C streptococci (13 Streptococcus equi zooepidemicus and four Streptococcus dysgalactiae equisimilis) and 185 Lancefield group B streptococci (Streptococcus agalactiae) were isolated from camels (Camelus dromedarius) in Kenya and Somalia; 59 of the isolates were from healthy nasopharynx, vaginal and rectal mucosa and from non-abscessed lymph nodes, and the other 143 isolates were from clinical infections of the respiratory tract, tick bite lesions, abscessed lymph nodes, abscesses and other purulent skin lesions, periarthritis and arthritis, puerperal infection and gingivitis. The role of Lancefield group B and C streptococci as commensals and common opportunistic pathogens in East African camels is described.     

Isolation of Mycoplasma arthritidis from the joint fluid of boars

During an outbreak of a disease in a swine insemination centre in Bavaria, Fed. Rep. of Germany, characterized by conjunctivitis, severe polyarthritis and infertility mycoplasmas have been isolated from the joint fluids of the three boars investigated. Two of the isolate could be typed as Mycoplasma (M.) arthritidis which causes arthritis in rats, mice and rabbits, the third as M. collis, a probably apathogenic rodent mycoplasma species. One of the two isolated M. arthritidis strains (strain D 263) was injected intravenously in rats and mice, which developed mild to severe polyarthritis or even died, depending on the numbers of organisms inoculated. Since the bacteriological and virological investigations of the joints of boars did not yield a causative agent, it is to suppose that M. arthritidis played the substantial role in the production of the disease of the boars. It is very likely that the boars caught the mycoplasmas from rodents infected with the isolated species.     

Infectious agents associated with rabbit pneumonia: isolation of amyxomatous myxoma virus strains

Sixty-six rabbits, with no history of vaccination against myxomatosis and which had died of pulmonary lesions, were submitted for virological and serological tests for Myxoma virus (MV) infection and for bacteriological examinations. At post mortem, the diagnoses based on observed lesions were as follows: acute haemorrhagic pneumonia (38%); acute suppurative bronchopneumonia (35%); and fibrinohaemorrhagic bronchopneumonia with fibrinous pleuritis (27%). MV was isolated from 10% of the rabbits, mainly from those with acute haemorrhagic pneumonia. Serological evidence of MV infection was demonstrated in 44% of rabbits. Pathogenic bacteria species isolated from lungs were Pasteurella (spp. and multocida), Escherichia coli, Bordetella bronchiseptica and Pseudomonas aeruginosa, respectively, from 41, 11, 7 and 6% of samples. No relationship could be established between the presence of specific antibodies to MV and the observed pulmonary lesions or the results of the bacteriological examinations. A significant trend was established between the severity of the lesions and the results of the bacteriological examinations.     

Control of ovine gastrointestinal helminthiasis by the use of 'clean' grazing and strategic dosing in the field

Despite the widespread adoption of clean grazing systems in lowland sheep flocks, detailed parasitological investigations had not previously been carried out on such flocks. A trial was therefore conducted on two commercial flocks: a traditional permanent pasture flock (A) and one operating a system of clean grazing (B), and on an East of Scotland College flock (C) which had operated a clean grazing system for eight years. Ewe and lamb worm egg output, pasture larval levels and lamb liveweight gains were monitored and tracer lambs were grazed during July and August on each farm. Under clean grazing conditions on farm C all parasitological parameters were lower than on both commercial farms. However, in the commercial flocks comparable contamination was evident from midsummer onwards and tracer lambs grazed during August on farm B had significantly greater worm burdens than on the other two farms. The differences observed between the flocks were thought to be due to greater residual contamination by overwintered larvae in both commercial flocks while the higher worm burdens in August on farm B probably resulted partly from incomplete control of the periparturient rise in ewe faecal egg output and partly to autoinfection of the lamb crop. It was concluded that farm C grazing was the cleanest. Considerable contamination was present on farm A while farm B occupied an intermediate position which resulted in considerable worm burdens in lambs grazing during the latter part of the season.     

Molecular characterisation of a field strain of bubaline herpesvirus isolated from buffaloes (Bubalus bubalis) after pharmacological reactivation

Two healthy buffaloes (Bubalus bubalis) in a herd which had not been vaccinated against infectious bovine rhinotracheitis (IBR), were selected for their seropositivity for anti-bovine herpesvirus type 1 (BoHV-1) glycoprotein E antibodies, and injected intramuscularly daily with dexamethasone for five consecutive days (day 1 to day 5) to reactivate any latent herpesvirus. Blood samples and nasal and vaginal swabs were collected daily from day 5 to day 15 from each buffalo for virological examination. All the vaginal swabs and blood samples were negative, but 13 of the 22 nasal swabs were positive; a cytopathic effect was observed in primary cultures of bovine fetal lung cells, and the viral isolates were identified as a herpesvirus by PCR. The viral strains were characterised by the sequence analysis of the genes coding for glycoproteins D and B, and the gene sequences were then used for phylogenetic analysis. The isolates from both buffaloes appeared identical at the level of the two genes, and were more closely related to bovine herpesvirus type 5 than to BoHV-1.     

Diarrhoea in adult horses: a survey of clinical cases and an assessment of some prognostic indices

Samples of faeces and blood were obtained from 66 adult horses with diarrhoea. The results of routine bacteriological, parasitological, haematological and biochemical tests were correlated with the outcome of the cases. Twenty-two (33 per cent) of the horses died or were destroyed as a consequence of the diarrhoea. A diagnosis was reached in only 23 cases (35 per cent), and in nine of them only at post mortem examination. Salmonella typhimurium was isolated from five cases. Statistical analysis revealed significant differences between the horses which survived and those which died in their packed cell volumes, white blood cell counts, neutrophil counts, serum albumin concentrations and alkaline phosphatase activities.