Polymorphism of ompH gene of Pasteurella multocida serotype A strains isolated in Iran

One of the most frequent causes of respiratory infection and death in sheep and goats is Pasteurella multocida. In humans, it has been associated with diseases of the respiratory tracts, arthritis, osteomyelitis and meningitis. Outer membrane protein H (OmpH) has a role in immunogenicity and pathogenicity of P. multocida. The aim of this study was to characterize the genetic diversity of ompH gene of a panel of P. multocida serotype A strains isolated in sheep. Forty P. multocida serotype A strains isolated in previous study were selected and analyzed by restriction fragment length polymorphism (RFLP) of a species-specific PCR assay. RFLP amplified fragment produced five different cleavage patterns. On the basis of combinations resulting from ompH gene digestion, the 40 P. multocida isolates were classified in six RFLP type. It seems that isolates with variants genetic profile represent different pathogenecity. New vaccine formulation should consider multivariants of P. multocida in order to confer a wider protection.

Mycobacterial infections in European wild boar (Sus scrofa) in the CzechRepublic during the years 2002 to 2005

A total of 842 wild boar of differing ages, originating from 29 (37.7%) of the 77 districts in the Czech Republic, were examined during the hunting seasons from 2002 to 2005. Of them, 274 (32.5%) of the animals were wild specimens and 568 (67.5%) from game parks. Out of 786 animals, the following were included in the study: 668 piglets, 61 juveniles, 32 adult males and 25 adult females. A total of 2 704 samples from various tissues and faeces were examined: 309 separately collected faecal samples from 309 (36.7%) animals, 2 332 samples from various tissues and 63 faecal samples from 533 (63.3%) animals. Mycobacteria were isolated from 75 (8.9%) animals from 11 of the districts. Neither a causative agent of bovine tuberculosis, nor any other members of Mycobacterium tuberculosis complex were isolated from any of the animals. From one (0.1%) animal, M. avium subsp. paratuberculosis of IS900 RFLP type A-C10 was isolated from intestinal lymph nodes, which was also isolated within the same district during other studies of cattle and free living ruminants. The causative agent of avian tuberculosis, M. a. avium (IS901+ and IS1245+), was isolated from 7 (0.8%) animals; among them tuberculous lesions were detected in intestinal lymph nodes, with gross tuberculous lesions visible on two animals. The causative agent of avian mycobacteriosis M. a. hominissuis (IS901– and IS1245+) was detected in lymph nodes without gross lesions in one (0.1%) animal. From 45 (5.5%) animals without lesions, atypical mycobacteria of the following nine species were isolated from pulmonary lymph nodes, small and large intestine, intestinal mucosa and faeces: M. fortuitum, M. chelonae, M. scrofulaceum, M. triviale, M. terrae, M. phlei, M. abscessus, M. flavescens, and M. smegmatis. Due to a high density of wild boar and their large migration radius, they can be viewed as a potential source for mycobacterial infections as well as other infectious agents.

Human-to-human and human-to-dog Mycobacterium tuberculosis transmission studied by IS6110 RFLP analysis: a case report

This study reports on the transmission of Mycobacterium tuberculosis of the same IS6110 RFLP type between two acquaintances with open pulmonary tuberculosis and a five-year-old Doberman bitch. No clinical signs, gross lesions at necropsy or histopathological lesions were observed in the infected lungs and gastrointestinal tract of the dog, although M. tuberculosis was directly detected by IS6110 PCR and culture examinations in the respiratory and gastrointestinal tracts. IS6110 PCR positivity in the faeces and blood of the dog poses a risk of M. tuberculosis transmission between the dog and humans.

Genotyping of Mycobacterium avium subsp. avium isolates from domestic animals in Slovenia by IS901 RFLP

ABSTRACT: Apart from birds, <I>Mycobacterium avium</I> subsp. <I>avium (MAA) </I> is often isolated from granulomatous lesions in pigs and occasionally from cattle and other animals. The objectives of this study were the detection of IS<I>901</I> restriction fragment length polymorphism (RFLP) types of <I>MAA</I> isolates from different species of domestic animals between the years 1998 and 2004 and the comparison of the detected RFLP types with previously described RFLP types collected in the database of the OIE Reference Laboratory for Avian Tuberculosis (Brno, Czech Republic). Furthermore, the RFLP types of the isolates obtained from <I>MAA</I> outbreaks on one of the largest pig farms in Slovenia were also investigated. A total of 62 isolates (56 from pigs, five from poultry and one from cattle) were identified with IS<I>901</I> PCR and IS<I>901</I> RFLP typed using restriction endonucleases <I>Pvu</I>II and <I>Pst</I>I. Seven <I>Pvu</I>II RFLP and 11 <I>Pst</I>I RFLP types resulted in 12 combined <I>Pvu</I>II <I>Pst</I>I types; none of these matched the combined RFLP types described in previous studies. Our contributions to the database were two new <I>Pvu</I>II and eight new <I>Pst</I>I RFLP types. Identical RFLP types were found among isolates of animals originating from individual farms. Finding of identical RFLP types within a farm is not surprising because the animals were epidemiologically related and infected with one strain. A unique RFLP type F-A17 was detected in isolates from different pig herds and also in isolates from poultry. Detection of identical RFLP types on different farms may reflect one <I>MAA</I> source. The other combined <I>Pvu</I>II <I>Pst</I>I RFLP types were identified only once which indicates considerable variety of <I>MAA</I> RFLP types in Slovenia.

Distribution and transmission of Mycobacterium avium subspecies paratuberculosis in farmed red deer (Cervus elaphus) studied by faecal culture, serology and IS900 RFLP examinations

The objectives of this study were the determination of <I>Mycobacterium avium</I> subsp. <I>paratuberculosis</I> (MAP) distribution in organs of farmed red deer (<I>Cervus elaphus</I>) and the investigation of its vertical and horizontal spread among animals, using serology, cultivation and the standardized IS<I>900</I> RFLP method. During the three year of study, the production of antibodies for <I>MAP</I> increased from 0 in the first year to 7.7% (positive) and 0 to 88.5% (dubious) in the third year of the study. The first performed global culture examination of faecal samples from 28 animals was negative for <I>MAP</I>. In the three subsequent examinations of animals, the following positivity was found: 5.9%, 34.6%, and 36.8%, respectively. In the last year of the study, clinical signs such as diarrhoea were observed in four animals. The animals with clinical symptoms and those that were found to be infected with <I>MAP</I> by serology or faecal culture were euthanized. <I>MAP</I> was isolated from the intestinal tract and pulmonary lymph (tracheobronchial or mediastinal lymph nodes) nodes of all studied animals. Apart of this <I>MAP</I> was also isolated from reproductive organs, such as the mammary gland, milk, uterus, amniotic fluid and testicles. Application of the IS<I>900</I> RFLP method revealed that the prevailing <I>MAP</I> isolates were of RFLP type B-C1; this profile was found in all types of tissue samples as well as in faeces, milk and amniotic fluid. In five animals a mixed infection of two profiles B-C1 and B-C5 or B-C1 and B-C16 was detected.

A mixed infection of Mycobacterium avium subsp. paratuberculosis and M. a. hominissuis in one red deer (Cervus elaphus) studied by IS900 BstEII and IS1245 PvuII RFLP analyses: a case report

A mixed infection with <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> (<i>MAP</i>) and <i>Mycobacterium avium</i> subsp. <i>hominissuis</i> (<i>MAH</i>) in one naturally infected red deer stag from a game park is described. The animal was euthanized because of symptoms of poor condition, weight loss and chronic diarrhoea. In spite of that, pathological lesions were observed only in the mesenteric lymph nodes, which were five to ten times enlarged with confluent caseous granulomas of 1 to 10 mm in size. Mycobacteria were isolated from all studied samples: a mixed infection of <i>MAP</i> and <i>MAH</i> was confirmed by multiplex PCR for the detection of IS <i>900</i>, IS<i>901</i>1, IS<i>1245</i> and <i>dnaJ</i>. MAP</i> of the identical IS<i>900</i> <i>BstE</i>II RFLP type C1 was isolated from all tissue samples and faeces. <i>MAH</i> isolates were detected in six examined tissue samples, including three mesenteric lymph nodes with caseous granulomas. Only minor differences in the band numbers and position of four different IS<i>1245</i> <i>Pvu</i>II RFLP patterns of <i>MAH</i> isolates were found. It follows from these results that red deer may potentially be infected with <i>MAH</i>, when a <i>MAP</i> infection is under way.

The wildlife hosts of Mycobacterium avium subsp. paratuberculosis in the Czech Republic during the years 2002–2007

The objective of this study was to determine the wildlife hosts of Mycobacterium avium subsp. paratuberculosis (MAP) in the Czech Republic. A total of 8 796 wildlife animals were examined by culture of faecal or tissue samples during the years 2002–2007. MAP was isolated from 12 (0.5%) out of 2 296 red deer (Cervus elaphus), two (0.2%) out of 835 roe deer (Capreolus capreolus), 78 (5.7%) out of 1 381 fallow deer (Dama dama), 28 (3.2%)out of 866 mouflons (Ovis musimon), four (2.5%) out of 162 chamois (Rupicapra rupicapra) and from one (0.1%) out of 805 wild boar (Sus scrofa). MAP was not cultured from 82 badgers (Meles meles), 55 martens (Martes foina), one pine marten (Martes martes), 25 brown hares (Lepus europaeus), five rabbits (Oryctolagus cuniculus), nine European polecats (Mustela putorius), two steppe polecats (Mustela eversmannii), two American minks (Mustela vison), four raccoon dogs (Nyctereutes procyonoides) and four Eurasian otters (Lutra lutra). MAP was isolated from three (2.0%) out of 149 small terrestrial mammals: one (5.9%) out of 17 brown rats (Rattus norvegicus), one (1.7%) out of 59 common voles (Microtus arvalis) and one (2.6%) out of 39 lesser white-toothed shrews (Crocidura suaveolens). Culture examinations of 34 house mice (Mus musculus) and 2 113 pigeons (Columba livia f. domestica) were negative. All 123 in vitro growing MAP isolates from wild ruminants were of IS900 RFLP type B-C1. One mouflon infected with a MAP strain which did not grow on the tested media was after IS1311-PRA-PCR assessed as being infected with a “sheep” strain. The RFLP type of the MAP isolate from the wild boar was of the RFLP type A-C10. Although the detection of MAP in wildlife in the Czech Republic was not very high, their role as a potential risk factor for cattle should be considered.

Genetic variability of Mycobacterium avium subsp. avium of pig isolates

The genetic diversity of 132 pig isolates of <i>Mycobacterium avium</i> subsp. <i>avium</i> (MAA) from the Czech Republic, the Slovak Republic and Slovenia was examined by IS<i>901</I> restriction fragment length polymorphism (RFLP) analysis with restriction endonuclease <i>PvuII</i>. A total of 18 RFLP types were detected. The occurrence frequency of respective RFLP types varied between respective pig farms, with the exception of one RFLP type F found in 21 (34.4%) of 61 farms and in 10 (55.6%) of 18 farms in the Czech Republic and Slovak Republic, respectively. Two different RFLP types were detected in 5 (33.3%) of the 15 studied farms, from which more than one isolate were examined. These results show the low variability of the <i>MAA</i> isolates among the pig farms and the possibility of various sources of infection for pigs from infected farms.

Temporal trends in circulating Bordetella pertussis strains in Australia

Australia experienced a resurgence of pertussis in the 1990s despite improved vaccine coverage. Although much of the increase was attributable to increased detection of cases in older persons with waning immunity by serology, vaccine changes or alterations in circulating Bordetella pertussis strains may also have contributed. We determined the frequency of variants of B. pertussis pertactin (prn), and pertussis toxin subunit 1 (ptxS1) genes, restriction fragment length polymorphism (RFLP) types and fimbrial serotypes prevalent in Australia prior to, and during the 1990s. Ampoules of the whole-cell vaccine in use prior to 1999 and 84 B. pertussis isolates stored between 1967 and 1998 by laboratories around Australia were analysed. One pertactin allele, Prn3, not detected before 1985, was found in 24 out of 57 (42%) isolates between 1989 and 1998 (P<0·0001). PtxS1A was found in all isolates. IS1002 type 29, found in 17 out of 31 (55%) isolates tested, was the predominant RFLP type. The only difference in fimbrial serotype distribution between the time-periods was an increase in serotype 3 (P=0·054). The whole-cell vaccine contained only the alleles prn1 and ptxS1A. Antigenic shift in B. pertussis may have contributed to the re-emergence of pertussis in Australia. Monitoring these trends will be important as acellular vaccines are introduced and changes are made to pertussis vaccine schedules.