Outer membrane protein antigens of Moraxella bovis

Outer membranes were isolated from bovine isolates and type strains of Moraxella bovis, M phenylpyruvica, M lacunata, and M ovis by sodium N lauroyl sarcosinate extraction and differential centrifugation. Analysis of outer membranes from these organisms by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis revealed that all M bovis isolates shared a common polypeptide pattern that was readily distinguishable from other Moraxella spp. Nine major outer membrane protein bands were identified by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis analysis of M bovis. Immunoblotting of protein antigens of M bovis revealed several outer membrane proteins that seemed to be common antigens of all M bovis isolates.     

Bacteriological investigation of infectious keratoconjunctivitis in Norwegian sheep

Contagious keratoconjunctivitis is a rather common disease in Norwegian sheep. Since the knowledge of its aetiology is limited, the present study was performed to determine the microorganisms involved. Local veterinarians throughout the country collected conjunctival swabs from both sick (n = 43) and healthy (n = 42) sheep on 15 farms with outbreaks of ovine keratoconjunctivitis, and further from healthy sheep (n = 50) on 17 farms not showing any signs of conjunctival disease. All samples were cultivated for bacteria and mycoplasma. Listeria monocytogenes was isolated from 3 cases (1%) in one single herd. Staphylococcus aureus (5%), Corynebacterium spp. (2%) and Escherichia coli (4%) were isolated only in herds with keratoconjunctivitis, but from both sick and healthy animals. Moraxella (Branhamella) ovis was isolated from 28% of sampled animals in affected herds and from 10% of sampled animals in healthy herds. The corresponding numbers for Moraxella spp. were 9%/12%, for Pseudomonas spp. 7%/8%, for Staphylococcus spp. 22//22%, for Bacillus spp. 12%/14%, for Micrococcus spp. 6%/2% and for Streptococcus/Enterococcus spp. 2%/2%. Mycoplasma conjunctivae was isolated from 16 animals with keratoconjunctivitis (37%) and from 3 animals without clinical signs (7%) in farms with keratoconjunctivitis. In farms without clinical signs of keratoconjunctivitis, M. conjunctivae was isolated in 4 animals (8%). To our knowledge, this is the first time M. conjunctivae has been isolated in Norway. Other predisposing agents found were Moraxella (Branhamella) ovis and Listeria monocytogenes. The etiological importance of different microorganisms in ovine keratoconjunctivitis seems to vary; some are probably only present as secondary invaders. Other possible causes of ovine keratoconjunctivitis in Norway, such as Chlamydia psittaci, remain to be investigated.     

Identification and characterization of complete RTX operons in Moraxella bovoculi and Moraxella ovis

To determine if Moraxella bovoculi (M. bovoculi), a recently characterized coccoid Moraxella that was isolated from the eyes of calves affected with infectious bovine keratoconjunctivitis (IBK), and Moraxella ovis (M. ovis), originally isolated from sheep with conjunctivitis, possessed genes encoding RTX proteins, genomic DNA was amplified with oligonucleotide primers targeting RTX operon genes of Moraxella bovis (M. bovis). Complete classical RTX operons composed of RTXCABD genes closely linked to a putative secretion accessory protein encoding gene (tolC) were identified in M. bovoculi and M. ovis and were designated mbvCABDtolC and movCABDtolC, respectively. These genes were closely related to M. bovis mbxCABDtolC. Polyclonal rabbit antiserum against the carboxy terminus of M. bovoculi MbvA neutralized hemolytic activity of both M. bovoculi and M. ovis; this antiserum did not neutralize the hemolytic activity of M. bovis. M. bovoculi and M. ovis possess genes that encode proteins related to pathogenic factors of M. bovis.     

Isolation of Mycoplasma bovoculi from cases of infectious bovine keratoconjunctivitis.

Five outbreaks of infectious bovine keratoconjunctivitis were examined for bacteria and mycoplasmas. Mycoplasma bovoculi was demonstrated in four of the five outbreaks. Other mycoplasmatales were represented by Ureaplasma in one sample. Moraxella bovis and Neisseria ovis were found in all the outbreaks, the former being present in the vast majority of the animals. Transmission experiments with Mycoplasma bovoculi and Moraxella bovis in combination were carried out on four young, colostrumdeprived calves. Mycoplasma bovoculi appeared to have an enhancing effect on the pathogenicity of Moraxella bovis.     

Differentiation of Moraxella bovoculi sp. nov. from other coccoid moraxellae by the use of polymerase chain reaction and restriction endonuclease analysis of amplified DNA.

Moraxella ovis was historically the only coccoid Moraxella identified in cultures of ocular fluid from cattle with infectious bovine keratoconjunctivitis (IBK) and could be morphologically and biochemically differentiated from Moraxella bovis. Moraxella bovoculi sp. nov. is a recently characterized Moraxella isolated from ulcerated eyes of calves with IBK in northern California in 2002. Like Moraxella ovis, M. bovoculi sp. nov. is a gram-negative coccus/diplococcus. All 18 original isolates of M. bovoculi sp. nov. possessed phenylalanine deaminase (PADase) activity and could therefore be differentiated from M. ovis and M. bovis. During the characterization of 44 additional isolates of hemolytic gram-negative cocci that were cultured from ulcerated eyes of IBK-affected calves, 2 PADase-negative isolates were identified that could not be differentiated biochemically from M. ovis; however, the DNA sequence of the 16S-23S intergenic spacer region (ISR) of the isolates matched the 16S-23S ISR DNA sequence of M. bovoculi sp. nov. To facilitate the identification of PADase-negative moraxellae, a polymerase chain reaction (PCR) coupled with restriction enzyme digestion analysis of amplified DNA was developed. Amplification of the 16S-23S ISR followed by AfaI digestion of amplified DNA could differentiate M. bovoculi sp. nov. from M. ovis and other moraxellae. The DNA sequence analysis of the amplified 16S-23S ISR from the 42 PADase-positive isolates of hemolytic gram-negative cocci indicated that all were M. bovoculi sp. nov. and all possessed an AfaI site. A PCR coupled with restriction analysis of amplified DNA can aid in identifying M. bovoculi sp. nov.     

Etiologic significance of bacterial isolates from rams with palpable epididymitis

Identified and partly identified bacterial isolates were obtained from 48 rams of various breeds that had unilateral or bilateral epididymitis. Most of the animals were approximately 1 year of age; a few were older. Brucella ovis, Actinobacillus spp, Corynebacterium spp, Haemophilus spp, Acinetobacter spp, Escherichia coli, Moraxella spp, Staphylococcus spp, Pasteurella spp, Streptococcus spp, and Chlamydia psittaci were isolated. A vaccine strain of B ovis, isolated species of bacteria, and mixtures of isolates of tissue homogenates containing all isolates except B ovis and C psittaci were inoculated via the mucous membranes of the eyes, nares, and prepuce. Palpable epididymitis was not produced by the inoculations. The vaccine strain of B ovis induced complement-fixation reaction in 11 of 20 rams.     

Molecular studies on Babesia,Theileria and Hepatozoon in southern Europe. Part I. Epizootiological aspects

Molecular epizootiology of piroplasmids (Babesia spp., Theileria spp.) and Hepatozoon canis was studied in mammals from southern Europe (mainly from Spain, but also from Portugal and France). Partial amplification and sequencing of the 18s rRNA gene was used for molecular diagnosis. In some particular cases (B. ovis and B. bovis) the complete 18s rRNA gene was sequenced. Blood samples were taken from domestic animals showing clinical symptoms: 10 dogs, 10 horses, 10 cows, 9 sheep and 1 goat. In addition, DNA samples were isolated from blood of 12 healthy dogs and from spleen of 10 wild red foxes (Vulpes vulpes). The results of the survey were the following: Piroplasmid infections: Approximately from 50 to 70% of wild or domestic mammals (symptomatic) were infected.Piroplasmids detected in ruminants were:COW: B. bovis, T. annulata and Theileria sp. (type C). Sheep and goat: B. ovis. Piroplasmids present in canids were: Babesia canis vogeli, Babesia canis canis, Theileria annae and B. equi. The only piroplasmid found in asymptomatic dogs was B. equi. Piroplasmids found in horse were: B. equi and B. canis canis.H. canis infections in canids: H. canis was absent of domestic dog samples, whereas all foxes studied were infected by this protozoa.Genetic analysis showed that most of piroplasmid and Hepatozoon isolates from southern Europe matched unambigously with previously described species, as demonstrated by the high level sequence identity between them, usually between 99 and 100%. Minor differences, usually detected in hypervariable regions of 18s rRNA gene are probably due to strain variations or rare genetic polymorphisms. A possible exception was B. bovis, which shows a relatively lower degree of homology (94%) with regard to other B. bovis isolates from several countries. The same is true for B. ovis, that showed a 94% identity with regard to Babesia sp. from South African cow and a 92% with rapport to B. bovis from Portugal.     

Internal parasites in calves and heifers in a central rearing barn

In the specialised rearing house for young cattle of all age groups the occurrence of endoparasites was followed. In the age group of 21 to 90 days, 56% of calves aged about 30 days eliminated oocysts of coccidia. On the whole, 49.3% of calves were infested by coccidia, 3% by enterohelminths. 90.3% of calves aged from three to six months were parasitised by coccidia, 7.4% by enterohelminths. Young cattle, aged from 6 to 12 months, were parasitised in 94.6% by coccidia and in 14.1% by enterohelminths. 86.6% of pregnant heifers at the age of 14 to 19 months had coccidia, 7.7% enterohelminths. The total farm capacity was about 950 animals. Throughout the year the number of examined animals amounted to 1,195, out of this number 958 animals, i. e. 80.2%, were infested by coccidia, 55.6% of animals by the species Eimeria bovis, 47.0% by E. zuernii, 43.3% by E. auburnensis, 39.4% by E. ellipsoidalis, 25.2% by E. cylindrica, 13.6% by E. subspherica, 3.1% by E. bukidnonensis, 1.7% by Isospora spp., 0.4% by E. brasiliensis, 0.08% by E. pellita. Enterohelminths were observed only in 96 animals, i. e. in 8.03%. Trichocephalus was found in 3.9% of animals, Cooperia in 2.4%, Ostertagia in 0.6%, Chabertia in 0.4%, Nematodirus in 0.3%, Capillaria in 0.2%, Oesophagostomum in 0.2%, Bunostomum in 0.1% and Trichostrongylus in 0.1%. The highest elimination of oocysts of coccidia was observed in August, January and February, of eggs of enterohelminths from August to October. In all age groups the most frequent was the occurrence of pathogenic species of coccidia.     

Biological reactivity of Moraxella bovis lipopolysaccharide

Lipopolysaccharide (LPS) was isolated from Moraxella bovis 118F and ATCC 10900, M ovis ATCC 33078, and M phenylpyruvica ATCC 23333 by hot phenol-water extraction. In silver-stained sodium dodecyl sulfate polyacrylamide electrophoresis gels, M bovis 118F LPS had a smooth profile, whereas the other Moraxella preparations appeared to be rough. The LPS preparations induced pyrogenicity and dermal Shwartzman reactions in rabbits, and induced production of tumor necrosis factor and interleukin-1 in vitro. Induction of tumor necrosis factor appeared to be among the most potent biological activities of M bovis LPS.     

Visceral caseous lymphadenitis in thin ewe syndrome: isolation of Corynebacterium, Staphylococcus, and Moraxella spp from internal abscesses in emaciated ewes

The relationship between the visceral form of caseous lymphadenitis and a chronic debilitating condition of mature sheep designated as the thin ewe syndrome was investigated. Internal abscesses were found during necropsy in 81% of animals with thin ewe syndrome and Corynebacterium pseudotuberculosis (C ovis) was recovered from 86% of the animals with internal abscesses. Other pyogenic bacteria, including C pyogenes, C equi, Staphylococcus epidermis, S aureus, and Pseudomonas aeruginosa were often recovered in association with C pseudotuberculosis. Moraxella sp was recovered in 41% of the animals with internal abscesses. In some abscesses, Moraxella sp was the dominant microorganism isolated and in others, they were outnumbered only by C pseudotuberculosis. Species isolated included M bovis, M osloensis, and M nonliquefaciens. The potential importance of Moraxella sp to the cause and pathogenesis of the thin ewe syndrome is not known. The results of the present study indicate that visceral caseous lymphadenitis is either an important contributing factor to the development of thin ewe syndrome or that the presence of thin ewe syndrome may predispose affected sheep to the development of visceral caseous lymphadenitis. A skin test reagent prepared by sonicating C pseudotuberculosis was of limited value in detecting animals with visceral caseous lymphadenitis. Only 56% of the animals with abscesses caused by C pseudotuberculosis gave positive delayed-type hypersensitivity skin test responses.     

A field trial of autogenous Moraxella bovis bacterin administered through either subcutaneous or subconjunctival injection on the development of keratoconjunctivitis in a beef herd

The primary purpose of these experiments was to evaluate an autogenous Moraxella bovis bacterin administered through 2 separate routes of inoculation. An autogenous bacterin was manufactured by using M. bovis recovered from the herd. The bacterin was administered by subcutaneous injection or subconjunctival injection. In each experiment, unvaccinated animals served as controls. Random selection methods were used to place calves into a vaccination or control group. There was no statistical difference in development of infectious keratoconjunctivitis between the vaccinated and unvaccinated calves. There was a statistically significant difference between the sexes; heifers had a higher rate of keratoconjunctivitis.     

新疆南疆部分地方品种羊无浆体的分子检测与鉴定

为了解新疆南疆部分地方品种羊的无浆体感染情况和分子特征,用PCR法检测新疆南疆5种地方品种绵羊共100份血液DNA样本,发现无浆体总感染率为67.0％(67/100).以多浪羊感染率最高,为100％(20/20),和田羊感染率最低,为44.0％(11/25);散养和圈养羊的无浆体感染率分别为74.0％(37/50)和6...     

Mycoplasma bovoculi infection increases ocular colonization by Moraxella ovis in calves

To determine whether infection with Mycoplasma bovoculi increases ocular colonization of cattle eyes with Moraxella bovis and other bacteria, colonization of ocular gram-negative bacteria were measured in eyes of cattle infected with Mycoplasma bovoculi. Strains of Moraxella ovis were chosen because these are among the most commonly isolated species of gram-negative bacteria from cattle eyes. Five strains of M ovis were characterized biochemically and by pilus structure, permitting the recognition of 2 biotypes. All strains were tested in a mouse corneal pathogenicity model. One strain of each biotype was selected for testing in calves. All 5 strains were apathogenic for mice, and the 2 strains tested in cattle did not induce keratitis. Infection of calves with Mycoplasma bovoculi increased the amount and persistence of colonization with the strains of M ovis.     

Epidemiology of infectious keratoconjunctivitis in cattle in south-east Ethiopia

The aim of the study was to determine the prevalence of infectious keratoconjunctivitis on cross-breed dairy farms and to evaluate factors that influence its distribution. A total of 11 dairy farms and one cattle cross-breeding farm in different parts of Arsi region, south-east Ethiopia were clinically and bacteriologically investigated from February 1989 to December 1990. Out of 5221 local zebu and cross-breed dairy animals, 110 were found to have different degrees of infectious bovine keratoconjunctivitis. Of these, 94 (85.5%) were unilateral and 16 (14.5%) were bilateral. The prevalence of the disease was significantly (P < 0.001) influenced by age and breed; high exotic blood level and younger age groups were mainly affected. However, severity of the disease was not significantly associated with age (P > 0.750). Moraxella bovis was isolated from 88 (80%) of the 110 bacteriologically positive cases. Of these isolates, 61 (69%) were haemolytic and 27 (31%) were non-haemolytic strains. Infection with M. bovis was unilateral. Concurrent infections with M. bovis included Actinomyces pyogenes 23 (21%), Staphylococcus aureus 17 (15%), Pasteurella haemolytica nine (8%), Escherichia coli eight (7%) and Proteus spp. four (3.6%).     

Differential Bos taurus cattle response to Babesia bovis infection

Bovine babesiosis is a tick-borne disease caused by Babesia spp. haemoprotozoans. The disease is of great importance at tick enzootic unstable areas and hampers cattle production in several developing countries. The available immunisation alternatives are pre-immunition and attenuated vaccines. Despite being efficient and protective, they are unsafe as they use cattle blood cells as inoculum and may potentially spread other diseases. Another alternative to help in babesiosis control would be the identification of genetically resistant cattle to Babesia bovis infection. The objective of this work was to phenotype cattle based on primary response against B. bovis infection. Two-hundred and forty half-sib Hereford and Aberdeen Angus heifers (120 animals from each breed), 12-18-month-old na?ve cattle, originated from a tick-free area in Southern Brazil, were used in the experiment. Animals were monitored following an inoculation with 1x10(7)B. bovis parasitised erythrocytes. Results showed three different phenotypes: 1-'susceptible', animals with babesiosis clinical signs that received treatment to avoid death; 2-'intermediate', animals with clinical signs: parasitaemia, >or=21.5% reduction in packed cell volume (PCV) and increase in body temperature when compared to their pre-challenge physiological parameters, no specific treatment was needed as animals self recovered from the disease, and 3-'resistant', animals without clinical signs that showed B. bovis presence in blood smears, <21.5% PCV reductions, with little or no increase in body temperature and no need for babesiosis treatment. The frequencies of each phenotype were: 45.4, 26.7, and 27.9%, respectively, demonstrating the existence of phenotypic variation for B. bovis in Bos taurus cattle.     

Bacterial Identity and Characteristics in Healthy and Unhealthy Respiratory Tracts of Sheep and Calves

Barbour, E.K., Nabbut, N.H., Hamadeh, S.K. and Al-Nakhli, H.M., 1997. Bacterial identity and characteristics in healthy and unhealthy respiratory tracts of sheep and calves. Veterinary Research Communications, 21 (6), 421-430The aim of this study was to compare different bacteriological aspects of the respiratory systems of healthy (H) versus unhealthy (UH) animals with respiratory signs. The prevalence of different bacterial species was determined in the upper and lower respiratory tract of H and UH Najdi sheep, Somali sheep and Holstein calves. The characteristics of Pasteurella spp. isolates, and the biotype of Pasteurella haemolytica were identified in H and UH animals. Eighteen out of 28 (64.3%) of the identified bacterial species in the upper respiratory tract were more prevalent in the nasal cavities of UH Najdi and Somali sheep and Holstein calves with respiratory signs than in apparently healthy animals; four of the most prevalent bacteria in the upper respiratory system of UH sheep were Moraxella spp., Pseudomonas pseudomallei, Erysipelothrix spp., and Pasteurella multocida, while three of the most prevalent bacteria in UH calves were Pasturella haemolytica, Actinomyces spp., and Pseudomonas aeruginosa. The prevalence of six different bacterial species was greater in the lungs of UH animals, namely Actinomyces pyogenes, Erysipelothrix spp., P. haemolytica, Pasteurella ureae, Staphylococcus aureus, and Staphylococcus epidermidis, which could be risk factors in the complexity of the prevalent respiratory diseases of the animals surveyed.Of the biochemical, cytological and colonial characteristics studied in the identified P. haemolytica and P. multocida, two characters were significantly different (p < 0.05) in organisms isolated from UH as compared to those from H animals. These were the higher loss of haemolytic power by the strains of P. haemolytica and the decreased fermentation of trehalose by all the strains of P. multocida recovered from healthy animals.The only biotype of P. haemolytica isolated from H animals was biotype A, while both biotypes A (88.0% of the isolates) and T (12.0% of the isolates) were recovered from UH animals.     

Association of sexual experience with isolation of various bacteria in cases of ovine epididymitis

Testicles from rams in flocks experiencing ram epididymitis in Idaho and eastern Oregon were cultured. Twenty-six breeding rams from 6 flocks were cultured and only Brucella ovis was isolated. Virgin rams (65) harbored numerous species of small fastidious gram-negative rods, including Actinobacillus actinomycetemcomitans, Haemophilus spp, Moraxella spp, and Pasteurella spp. Thus, there appeared to be 2 separate disease entities, dependent on sexual experience of the animal.     

Molecular and antigenic characterization of a Mycoplasma bovis strain causing an outbreak of infectious keratoconjunctivitis.

An unusually high incidence of infectious keratoconjunctivitis followed by pneumonia and arthritis was observed in beef calves of a managed herd. No Moraxella spp. or bacteria other than Mycoplasma spp. were obtained from conjunctival and nasal swabs. A strategy was designed for characterization of bovine mycoplasmas at species and strain level on the basis of a combination of molecular tools and the immunoblotting method. The strategy made it possible to rapidly assign the bacterium responsible for this outbreak to one of the phylogenetic clusters of bovine mycoplasmas delineated in this study and then to identify it as Mycoplasma bovis. The strain, designated Sar 1, showed a 100% 16S rDNA sequence identity with two European strains (120/81 and MC3386) isolated in Germany and Ireland, respectively, and hosts a vsp gene analog to the vspA, vsp422-4, and vsp422-8 genes of the M. bovis reference strain PG45T and of the field strain 422. The use of a cross-reactive rabbit serum developed against the Mycoplasma agalactiae immunodominant antigen P48 confirmed the molecular findings. The immunological response of calves against M. bovis was also investigated. This is the first report on the occurrence of M. bovis on the Island of Sardinia (Italy).     

Molecular and phenotypic analysis of Moraxella spp. associated with infectious bovine keratoconjunctivitis in Uruguay

Infectious bovine keratoconjunctivitis (IBK) is a common ocular disease of cattle, which is generally thought to be caused by Moraxella bovis. However, a recently characterized Moraxella, M. bovoculi, has been isolated from animals with IBK. The aim of this study was to identify and characterize strains of Moraxella spp. obtained from IBK cases in different geographic locations within Uruguay. Ribosomal gene sequencing indicated that there were two groups of isolates that showed homology with either M. bovis or M. bovoculi. Phylogenetic analysis confirmed the presence of two species as the isolates grouped in different branches of the dendrogram. Conventional biochemical characterization did not distinguish between the species; only 9/25 isolates which had genetic homology with M. bovoculi showed any differences in biochemistry.     

Molecular characterization of Cryptosporidium spp. in grazing beef cattle in Japan

Cattle are major hosts of Cryptosporidium spp. Cryptosporidiosis in neonatal calves is associated with retarded growth, weight loss and calf mortality, and zoonotic infections in humans. In many areas, cow-calf glazing system is an important beef cattle rearing method with distinct advantages in terms of cost and the labor required. However, few epidemiologic studies of Cryptosporidium spp. have been conducted in this system, especially using molecular diagnostic tools. To understand the transmission of Cryptosporidium spp. in a grazing system, we followed cryptosporidiosis on a grazing farm in Osaki City, Miyagi Prefecture, in northwest Japan for one year. Fecal samples were collected from Japanese Black and Japanese Shorthorn cattle and examined by PCR-RFLP and sequence analyses. Of 113 fecal samples collected in October 2010, 23 (20%) were positive for Cryptosporidium, including 15 samples (13%) having C. bovis, 6 (5%) having C. ryanae, and 2 (2%) having mixed infections of both species. Additionally, C. bovis or C. ryanae was detected on all other sampling dates involving smaller numbers of animals. The infection rate of C. bovis was significantly different among age groups, and calve-to-calve infection might be the major route of cryptosporidiosis transmission in beef cattle. Interestingly, one animal had C. bovis infection or re-infection for one year. Our results suggest that C. bovis and C. ryanae are distributed in Japan, but might have low level of detection in grazing beef cattle.