Serologic response and lesions in goats experimentally infected with Corynebacterium pseudotuberculosis of caprine and equine origins

Fifteen goat kids were experimentally inoculated with Corynebacterium pseudotuberculosis. Five were given a strain of caprine origin (nitrate-negative biotype) intradermally, 5 were given a strain of equine origin (nitrate-positive biotype) intradermally, and 5 were inoculated intranasally with the caprine-origin strain. Animals were monitored for 127 days. The goats given the inocula intradermally developed abscesses; those given caprine-origin strain had multiple lesions both peripherally and in visceral locations (primarily endothoracic abscesses), whereas those given the equine-origin strain had abscesses only at injection sites and draining nodes. The difference in extent of lesions could be due to biotypic bacterial differences or to the individual strains used. Intranasally inoculated goats did not develop abscesses and were essentially no different from controls. The cranial part of the respiratory tract may not be an important portal of entry for C pseudotuberculosis. Serum samples obtained monthly from all animals were subjected to the synergistic hemolysis-inhibition test, which measures antibodies to the exotoxin of C pseudotuberculosis. Animals with abscesses developed titers within 1 month of inoculation. Animals without abscesses remained seronegative. The synergistic hemolysis-inhibition test may be a reliable diagnostic assay for caseous lymphadenitis in goats.     

Molecular epidemiologic features of Corynebacterium pseudotuberculosis isolated from horses

OBJECTIVE: To characterize isolates of Corynebacterium pseudotuberculosis from horses, cattle, and sheep in Colorado, Kentucky, Utah, and California in samples collected during perceived epidemics of infection (increased numbers of cases identified) in 2002 and 2003, and determine how closely isolates were related and their possible source. SAMPLE POPULATION: 54 isolates of C pseudotuberculosis from 49 horses, 4 cattle, and 1 sheep. PROCEDURES: Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) assay, PCR assay for the gene encoding the phospholipase D (PLD) toxin, biochemical analyses, and tests for susceptibility to 17 antimicrobial drugs were performed. RESULTS: All isolates reduced nitrate to nitrite, most yielded positive results for the PLD toxin gene, and all were susceptible to antimicrobial drugs. Ten genetic types were detected by use of RAPD PCR assay; types III to X were isolated from horses, cattle, or both in 1 or more states. Types III and IX were isolated from both horses and cattle. Types VII and VIII were isolated in only 1 state, but the number of isolates in these groups was small. In contrast, all other types were isolated in 2 or more states. All isolates from Utah were type III, but the other 3 states had isolates from more than 1 type. CONCLUSIONS AND CLINICAL RELEVANCE: These data are consistent with a clonally expanding epidemic of infection in Utah and an increase in number of infections caused by multiple strains of C pseudotuberculosis not derived from a single source in the other states. The increase in number of infections could be the result of reporting bias, environmental factors facilitating infection, or host factors such as greater herd susceptibility.     

Phenotypic characterization of Brucella strains isolated from livestock in Nigeria

Isolation of brucellae from aborted fetuses, hygroma fluids, milk and vaginal swabs obtained from aborting cattle, sheep, goats, pigs, and horses in Nigeria was carried out. A total of 25 isolates, obtained mainly from cattle, sheep and horses, were biotyped. All strains belonged to one species, Brucella abortus biovar 1. The epidemiological significance of this finding is discussed. Some preliminary observations on the zoonotic and public health implications of Brucella infection in Nigerian livestock are presented. A control programme involving improved management, animal movement restrictions, public health education and mass vaccination of animals is suggested.     

Diversity among isolates of Actinobacillus equuli and related organisms as revealed by ribotyping

Objective The objective of this work was to examine the diversity within Australian isolates of Actinobacillus equuli and related organisms by the genotypic method of ribotyping.
Design Ribotyping, performed using the enzyme Hae III, was used to examine the diversity in 12 field isolates of A equuli (five being capable of fermenting L-arabinose), one field isolate of Pasteurella caballi and two unclassifiable field isolates. Isolates were obtained from Australian horses, except for three isolates of A equuli (one L-arabinose positive and two L-arabinose negative) which were obtained from horses and a pig in Africa. In addition, the type strains for A equuli and P caballi and a reference strain for Bisgaard Taxon 9 were included in the study.
Results The ribotype patterns were analysed by computerised cluster analysis, yielding five clusters (A to E). All five of the L-arabinose positive A equuli were assigned to cluster A, with all the other seven A equuli isolates (all L-arabinose negative) and the type strain being assigned to cluster B. One of the two unclassified isolates formed cluster C along with the reference strain for Bisgaard Taxon 9. The remaining unclassified isolate formed cluster D. Cluster E consisted of the field isolate and reference strain of P caballi .
Conclusion The results of this study indicate that A equuli is a diverse species, with L-arabinose positive isolates of A equuli being quite distinct from typical L-arabinose negative isolates. Ribotyping appears to be a useful tool in confirming the identity of A equuli -like organisms from horses.     

The in vitro effect of six antimicrobials against Mycoplasma putrefaciens, Mycoplasma mycoides subsp. mycoides LC and Mycoplasma capricolum subsp. capricolum isolated from sheep and goats in jordan

Respiratory disease in sheep and goats is a major problem in Jordan and is often associated with Mycoplasma species. Without effective vaccines, control is mainly by chemotherapy, but the uncontrolled use of antimicrobials has led to concerns about the potential development of antimicrobial resistance. The in vitro effect of chloramphenicol, florfenicol, enrofloxacin, tylosin, erythromycin and oxytetracycline was determined against 32 isolates of Mycoplasma species-M. mycoides subsp. mycoides LC (6), M. capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions of Jordan. The antimicrobial susceptibility showed some Mycoplasma species-specific differences, with M. capricolum subsp. capricolum being more susceptible to tylosin and erythromycin. Chloramphenicol and florfenicol were the least effective for all three Mycoplasma species. No trends or significant differences in antimicrobial susceptibilities were observed between sheep and goat isolates, between milk or nasal swab isolates, or between isolates from different regions of Jordan. Some isolates of M. capricolum subsp. capricolum and M. putrefaciens showed higher MIC levels with oxytetracycline, as did two isolates of M. mycoides subsp. mycoides LC with tylosin, possibly indicating signs of development of antimicrobial resistance.     

Prevalence of Brucellosis in Horse North-East of Iran

Brucella preferentially infects cattle, swine, sheep, and goats. However, some epidemiological surveys have been carried out to investigate nonruminants, such as horses. Horse brucellosis has been found in clinical cases, but there are few epidemiologic patterns. Between May 2008 and April 2009, a total of 120 horses were screened for brucella infections in Mashhad, Iran, by the rose bengal test and the tube agglutination test. Sera from three horses were found positive by rose bengal test and tube agglutination test, and therefore the prevalence rate was 2.5%. In horses, the highest individual seroprevalence was in an animal kept close under the intensive system, with other animals such as cattle, sheep, and goats. The zoonotic aspects of brucellosis from the horse must, therefore, be considered because the disease is important from a public health standpoint. The present study documents the first serological evidence of Brucella spp. infection in horses in Iran.     

Serological survey of adenovirus antibodies in domestic animals in Nigeria

Serum samples collected from 1,197 goats, 586 sheep, 254, cattle, 55 dogs and 44 horses were examined for antibodies to adenovirus by the agar-gel precipitation test. Results show that 17.7% of the goats, 18.4% of the sheep, 4.3% of the cattle, and 4.5% of the horses had precipitating antibodies. None of the dog sera examined was positive. The results seem to indicate a moderate level of previous exposure to adenovirus infection especially among goats and sheep in Nigeria.     

Ticks on livestock in Barbados

Cattle, sheep, goats, and horses in most areas of Barbados were examined for ticks. About 68% of the cattle, mostly Holstein dairy cows, and 23% of the sheep, mostly purebred or crossbred black belly sheep, were infested with the southern cattle tick, Boophilus microplus (Canestrini), a vector of bovine babesiosis and anaplasmosis. About 13% of the horses were infested with the tropical horse tick, Anocentor nitens (Neumann), a vector of equine babesiosis. Cattle owners used a variety of acaricides as dusts, sprays, or washes on their cattle for the control of ticks.     

A recently introduced Dichelobacter nodosus strain caused an outbreak of footrot in Norway

Background

In 2008, an outbreak of ovine footrot occurred in Norway. Dichelobacter nodosus isolates collected between 2008 and 2011 have been characterised. Isolates defined as virulent by the gelatin gel test (GG-test) were only found in sheep in Rogaland County, where the severe cases of footrot were registered. The majority (96%) of the virulent isolates belonged to serogroup A. It is suspected that they represent a newly introduced strain, and the aim of the present study was to investigate whether they are genetically similar. Sixty-one virulent isolates from sheep and 116 benign isolates from sheep, cattle and goats were included. Four GG-test virulent isolates from Danish sheep were also included. All isolates were genotyped by pulsed-field gel electrophoresis (PFGE) and by PCR for pgr variant determination.

Results

The Norwegian virulent isolates were assigned to 8 pulsotypes (PTs), while the benign isolates were assigned to 66 PTs. Thirty-seven (68.5%) of the 54, virulent, serogroup A isolates belonged to the same PT, and included isolates from 2008 through 2011. Isolates belonging to this PT were defined as the outbreak strain. The remaining virulent serogroup A isolates belonged to 4 PTs differing by ≤3 bands from the outbreak strain. Two virulent, Danish, serogroup A isolates differed by 2 bands from the Norwegian outbreak strain. All but 3 (95%) of the virulent isolates had the pgrA variant while 85% of the benign isolates had the pgrB variant.

Conclusion

This study provides evidence that the footrot outbreak in Norway in 2008 most likely was caused by new introduction and local spread of one virulent D. nodosus strain.     

Fluorescence spectra and measurement of phylloerythrin (phytoporphyrin) in plasma from clinically healthy sheep, goats, cattle and horses

AIM: To measure the background concentration of phylloerythrin in plasma from clinically healthy sheep, goats, cattle and horses on pasture. METHODS: Blood samples were taken from 34 sheep of the Dala breed, 20 female Norwegian dairy goats, 35 Norwegian Red cows and 34 horses of different breeds. All animals were grazing green pasture when blood samples were taken. Blood samples were collected from each of four clinically healthy newborn lambs, goats, calves and foals, and pooled into one sample per species. Plasma samples were analysed for phylloerythrin by fluorescence spectroscopy, using a Perkin-Elmer LS-50B luminescence spectrometer equipped with a red-sensitive photomultiplier. The fluorescence spectra of phylloerythrin in plasma from the adult ruminants were compared with those in plasma from the neonatal ruminants, to which a known concentration of phylloerythrin had been added. RESULTS: Plasma obtained from the adult ruminants had spectral characteristics similar to those of phylloerythrin, namely weak emission peaks at 650 and 711 nm, when excited at 425 nm. Emission spectra obtained from plasma from the neonatal ruminants showed no fluorescence at these wavelengths. On average, 0.012 (SD 0.004), 0.06 (SD 0.04), and 0.05 (SD 0.03) micromol/l phylloerythrin were present in plasma samples from the sheep, goats, and cattle, respectively. The fluorescence spectra of plasma from the newborn foals were similar to spectra of plasma from adult horses, with weak emission at 669 nm. CONCLUSION: Small concentrations of phylloerythrin were detected in plasma from clinically healthy sheep, goats and cattle, but none could be detected in plasma from clinically healthy horses.     

Fluorescence spectra and measurement of phylloerythrin (phytoporphyrin) in plasma from clinically healthy sheep,goats, cattle and horses

AIM: To measure the background concentration of phylloerythrin in plasma from clinically healthy sheep, goats, cattle and horses on pasture.

METHODS: Blood samples were taken from 34 sheep of the Dala breed, 20 female Norwegian dairy goats, 35 Norwegian Red cows and 34 horses of different breeds. All animals were grazing green pasture when blood samples were taken. Blood samples were collected from each of four clinically healthy newborn lambs, goats, calves and foals, and pooled into one sample per species. Plasma samples were analysed for phylloerythrin by fluorescence spectroscopy, using a Perkin-Elmer LS-50B luminescence spectrometer equipped with a red-sensitive photo- multiplier. The fluorescence spectra of phylloerythrin in plasma from the adult ruminants were compared with those in plasma from the neonatal ruminants, to which a known concentration of phylloerythrin had been added.

RESULTS: Plasma obtained from the adult ruminants had spectral characteristics similar to those of phylloerythrin, namely weak emission peaks at 650 and 711 nm, when excited at 425 nm. Emission spectra obtained from plasma from the neonatal ruminants showed no fluorescence at these wavelengths. On average, 0.012 (SD 0.004), 0.06 (SD 0.04), and 0.05 (SD 0.03) μmol/l phylloerythrin were present in plasma samples from the sheep, goats, and cattle, respectively. The fluorescence spectra of plasma from the newborn foals were similar to spectra of plasma from adult horses, with weak emission at 669 nm.

CONCLUSION: Small concentrations of phylloerythrin were detected in plasma from clinically healthy sheep, goats and cattle, but none could be detected in plasma from clinically healthy horses.     

Isolation of tibrogargan virus,a new Australian rhabdovirus,from Culicoides brevitarsis

CSIRO 132 virus, which is new to science in Australia, and probably the world, has been isolated from Culicoides brevitarsis. Electron micrographs show that it resembles a rhabdovirus. Antibodies to the new virus have been detected in water buffaloes and cattle, but not in 58 human beings, 14 camels, 21 dogs, 67 goats, 15 horses, 43 pigs, 154 sheep, 98 wallabies or 38 possums. The distribution of antibodies in cattle lies within the distribution range of C. brevitarsis. It has not so far been associated with disease. The name Tibrogargan is proposed for the new virus.     

Evaluation and application of ribotyping for epidemiological studies of Actinobacillus pleuropneumoniae in Denmark

The aim of the present study was to evaluate ribotyping as an epidemiological tool for Actinobacillus pleuropneumoniae and apply the method in studies of A. pleuropneumoniae infections in Danish pig herds. The evaluation of ribotyping was based on the 13 international reference strains and 106 epidemiologically unrelated Danish field strains representing the nine serotypes of biotype 1 (1, 2, 5A/B, 6, 7, 8, 10, 12, and K2:O7) and one serotype 14 of biotype 2. Enzymes CfoI and HindIII were chosen for generation of ribotype patterns. Ribotyping of the reference strains resulted in 10 CfoI types and 11 HindIII types. Ribotyping of the Danish strains resulted in 17 different CfoI ribotypes and 24 different HindIII ribotypes. Combining HindIII- and CfoI-ribotyping divided the Danish strains into 26 different types. The stability, reproducibility and typability of ribotype patterns were good, and the discriminatory power was between 0.85–0.89. The relatively low discriminatory power was caused by four predominant types, containing 61% of the isolates. The typing system was applied in studies of routes of infection of specific pathogen-free (SPF) pig herds and included 112 strains of A. pleuropneumoniae. Airborne transmission from neighboring conventional pig farms was investigated in 12 cases of infected SPF herds. Transmission via vehicles transporting pigs between SPF herds was investigated in nine cases while transmission by trading of pigs between SPF herds was investigated in two cases. Serotype 2 was isolated from all SPF herds included in this study, except one, emphasizing the high prevalence of this serotype in Denmark. By ribotyping, airborne transmission was indicated in five of 12 cases, transmission via pig transporting vehicle was indicated in six of nine cases, and transmission via trading was indicated in one of two cases. In many cases findings of predominant ribotypes made interpretations of suspected routes of transmission difficult. The relationship of strains based on ribotypes was calculated using Dices coefficient and clustered by UPGMA. HindIII ribotypes of serotype 2 strains were closely related, though only showing 43% similarity to HindIII ribotypes of remaining serotypes.     

Serotypes of Pasteurella haemolytica and Pasteurella trehalosi isolated from farm animals in Hungary.

The biochemical and serological characteristics of 486 P. haemolytica and 31 P. trehalosi strains (517 in total) isolated from different lesions of cattle, sheep, goats, pigs and poultry were examined. A total of 476 P. haemolytica strains (97.9%) showed the characteristics typical of the former biotype A of P. haemolytica, while 10 isolates (2.1%), all from poultry, could not be biotyped. A total of 481 strains (93.0%) could be assigned to one of the 17 serotypes of P. haemolytica-P. trehalosi and 36 strains (7.0%) could not. The majority (83.6%) of the cattle isolates were serotypes A1 and A2. Among strains isolated from sheep all serotypes of P. haemolytica could be identified with the exception of A14, but serotypes A1, A2, A6, A8 and A5 were the most frequent. The overwhelming majority (94%) of the caprine isolates were A2, other serotypes occurred only sporadically. The pig isolates, which could be isolated only very rarely, represented different serotypes, while none of the 10 strains isolated from poultry could be biotyped or serotyped.     

Antimicrobial susceptibility patterns and sensitivity to tulathromycin in goat respiratory bacterial isolates

Bacterial pneumonia is a common and often life-threatening respiratory problem in both meat and dairy goats. Options for approved antibiotic therapy in goats to combat these bacterial infections are severely limited and frequently drugs must be used in an extra-label manner. Tulathromycin, a triamilide macrolide antimicrobial drug shown to be effective against swine and cattle respiratory bacterial agents, has been identified as a potentially useful drug in caprines. The present study was conducted to determine the susceptibility of recognized bacterial respiratory pathogens to commonly prescribed antimicrobials, with a particular emphasis on the efficacy of tulathromycin against these agents. Minimum inhibitory concentration (MIC) testing using microbroth dilution was performed on a collection of 45 Mannheimia haemolytica, 11 Pasteurella multocida, and 11 Bibersteinia trehalosi isolates from the lungs of goats with clinical pneumonia. To further characterize efficacy of tulathromycin against these pathogens, minimum bactericidal concentration (MBC) testing and kinetic killing assays were conducted. Most isolates were susceptible to the antimicrobials tested; however, increased resistance as demonstrated by higher MIC values was seen in all species to penicillin, in P. multocida to sulfadimethoxine, and in B. trehalosi to the tetracyclines. All isolates were susceptible to tulathromycin, which demonstrated a high killing efficiency in both bactericidal assays. Results of this study indicate that most goat pneumonic bacterial pathogens remain susceptible to commonly prescribed antibiotics, although some evidence of resistance was seen to certain drugs; and that tulathromycin is highly effective against goat respiratory pathogens which could make it a valuable medication in this species.     

The majority of atypical cpb2 genes in Clostridium perfringens isolates of different domestic animal origin are expressed

This study examined the prevalence and expression of the "consensus" and the "atypical"cpb2 genes in Clostridium perfringens isolates from cattle, chickens, dogs, goats, horses, pigs and sheep using polymerase chain reaction (PCR), sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western blotting. Almost all porcine isolates (12/14) carried and expressed the consensus form of cpb2 but, when present in 108 non-porcine isolates, the gene was usually the atypical form (40 atypical versus 9 consensus). Western blotting showed expression in 30 of 40 (75%) atypical cpb2-positive isolates, considerably more frequently than reported previously. CPB2 was expressed by almost all (20/21) the consensus cpb2-positive isolates, regardless of source.     

Minimum inhibitory concentrations of 20 antimicrobial agents against Staphylococcus aureus isolated from bovine intramammary infections in Japan

Minimum inhibitory concentrations (MICs) of 20 antimicrobial agents were determined against 51 isolates of Staphylococcus aureus from bovine intramammary infections. Fourteen (27.4%) isolates were resistant to benzylpenicillin, but none of the isolates was resistant to cloxacillin, nafcillin, or cephems. Among aminoglycosides, gentamicin was the most active, with an MIC50 of 0.2 microg/ml, followed by kanamycin, with an MIC50 of 0.78 microg/ml. Five isolates (9.8%) were resistant to dihydrostreptomycin, three isolates (5.9%) to kanamycin and two isolates (3.9%) to gentamicin. Resistance to erythromycin was observed in two isolates (3.9%). Tylosin was less active than erythromycin, with MIC50s of 1.56 microg/ml versus 0.39 microg/ml, but none of the isolates was resistant to this antibiotic. Oxytetracycline MICs were situated in the range of 0.39-1.56 microg/ml for 48 susceptible isolates. Although 19 (37.3%) isolates were resistant to one or more antimicrobial agents, a single resistance pattern was most frequent: benzylpenicillin (12 isolates), dihydrostreptomycin (two isolates) and kanamycin (one isolate). There were no isolates resistant to antimicrobial agents such as methicillin, lincomycin, clindamycin, chloramphenicol, florfenicol and virginiamycin, which have not been approved for use in cattle husbandry in Japan.     

New techniques to measure blood cholinesterase activity in domesticated animals.

A macromethod and a semimicromethod were developed to measure erythrocyte acetylcholinesterase activity in cattle, sheep, goats, horses, dogs, and swine, and to measure plasma cholinesterase activity in horses, dogs, and swine. Comparison of the 2 methods with erythrocytes of sheep, cattle, goats, and horses indicated both methods gave similar results. They can be done in a shorter time and are more sensitive than Michel's method. Normal deltapH values per minutes, with standard deviations for blood cholinesterase activity of animals of different ages, sexes, breeds, and species, were: 0.76 +/- 0.12/30; 0.65 +/- 0.10/15; 0.69 +/- 0.19/45; 0.78 +/- 0.11/45; 0.63 +/- 0.11/45; and 0.71 +/- 0.06/25 for sheep, cattle, goats, horses, dogs, and swine erythrocyte acetylcholinesterase, respectively; and 0.66 +/- 0.18/20; 0.67 +/- 0.20/30, and 0.46 +/- 0.05/60 for horses, dogs, and swine plasma cholinesterase, respectively. It was shown that either the chloride or the iodide salt of acetylcholine can be used as the enzyme substrate. tin blood samples stored at 5 C for 24 hours, there was no significant change of the enzymatic activity.     

Molecular and biological characterization of vascular endothelial growth factor of parapoxviruses isolated from wild Japanese serows (Capricornis crispus)

Contagious dermatitis in domestic and wild ruminants caused by parapoxvirus (PPV) occurs worldwide. Although PPV infections appear in cattle, sheep and goats, the papular, nodular, pustular and ulcerated skin lesions of wild Japanese serows (Capricornis crispus) are significantly more severe than those of other animals. To determine the factors involved in the severity of these skin lesions, we compared the molecular characteristics of 4 PPV isolates from Japanese serows and 2 isolates from sheep in Japan, and also investigated the biological properties of primary endothelial cells from different host species. All of the 6 Japanese isolates harbored a vascular endothelial growth factor (VEGF) gene, which is known as one of the virulence factors of PPVs. Three different amino acid sequences of viral VEGF were identified and the sequence was identical among the 4 isolates from the Japanese serows. The temporal expression pattern of viral VEGF mRNA was almost the same among 3 isolates encoding the 3 different VEGFs in infected cells from different host species. Recombinant forms of the 3 different VEGFs showed the ability to induce vascular permeability and endothelial cell proliferation. Primary endothelial cells from Japanese serows were most responsive to recombinant viral VEGF compared to cells from cattle, sheep, and goats. These results suggest that not only the biological activity of viral VEGF but also the high responsiveness to viral VEGF of endothelial cells might be involved in the severe proliferative skin lesions induced by PPV infection in Japanese serows.