[Show abstract][Hide abstract] ABSTRACT: This study was conducted to determine the occurrence of zinc and copper resistances in methicillin-resistant Staphylococcus aureus (MRSA) from swine and veal calves in a global strain collection.The test population consisted of 476 porcine MRSA isolates from ten European countries, 18 porcine MRSA isolates from Canada and seven MRSA from China, 92 MRSA and 60 methicillin-susceptible S. aureus (MSSA) isolates from veal calves in the Netherlands and 88 porcine MSSA isolates from four European countries. Most porcine MRSA (n = 454) and all bovine MRSA belonged to clonal complex (CC) 398 whereas 37 of the pig MRSA from Europe and the seven Chinese isolates belonged to other CCs and 3 isolates were not classified into a CC.All isolates were tested for susceptibility to zinc chloride and copper sulphate using agar dilution and tested by PCR for the czrC gene encoding zinc resistance.Phenotypic zinc resistance (MIC > 2 mM) was observed in 74% (n = 324) and 42% (n = 39) of European MRSA CC398 from pigs and veal calves, respectively, and in 44% of the Canadian isolates (n = 8), but not among the Chinese isolates. Almost all (99%) zinc-resistant MRSA carried czrC. Of the 37 European non-CC398 MRSA, 62% were resistant to zinc, but only 46% of them carried czrC. The MICs of the MSSA isolates to zinc chloride ranged from 1 to 4 mM and none carried czrC. The MICs of copper sulphate were associated neither with methicillin resistance nor with the detection of czrC.This study showed that zinc resistance and the czrC gene are widespread among CC398 MRSA isolates. This suggests that the use of zinc in feed might have contributed to the emergence of MRSA.
[Show abstract][Hide abstract] ABSTRACT: Two groups of one-day-old Peking ducklings (Groups I and II, 12 birds/group) were inoculated orally with Brachyspira pilosicoli and two groups with B. alvinipulli (Groups III and IV, 12 birds/group). T-2 toxin was added to the feed of Groups II and IV in a dose of 1 mg/kg of feed. Groups V and VI served as uninfected control groups (ducks of Group VI received T-2 toxin). The body weight gain of the ducks was measured and clinical signs were monitored continuously. The birds were sacrificed and necropsied on days 7, 14, 21, and 28 post infection (PI). The liver, spleen, kidney, thymus, bursa of Fabricius, ileum, caecum and colon were examined histologically. Culturing of Brachyspira spp. and immunohistochemistry were performed from the sampled parts of the intestines as well. No gross pathological or histological lesions that could be associated with B. pilosicoli or B. alvinipulli were detectable in the intestinal mucous membrane including the colonised intestinal glands. Mortality did not occur during the experimental period. Decrease in body weight gain was significant in the T-2-toxin-treated groups, and it was slight (not significant) in the Brachyspira-infected groups. Crust on the beaks, necrosis, crusting and ulceration in the mucous membrane of the oral cavity and on the skin of the feet, atrophy of the thymus and bursa of Fabricius due to the effect of T-2 toxin, accompanied by lymphocyte depletion, were observed. These lesions were most prominent on days 14 and 21 PI but were seen on day 28 PI as well. Immunohistochemical detection and reisolation of B. pilosicoli and B. alvinipulli were successful on days 7, 14, 21 and 28 days from different segments of the intestine of certain birds, but no significant difference was observed in the colonisation rate between the T-2-toxin-treated and the untreated groups.
[Show abstract][Hide abstract] ABSTRACT: Campylobacter spp. are the most common cause of bacterial enteritis in Hungary, and the aim of this study was to identify the distribution, genotypes, and antimicrobial susceptibility of Campylobacter species in the most important food-producing animals at the time of slaughter during 2008 and 2009. Of 1,110 samples, 266 were identified as Campylobacter coli (23.9%) and 143 as C. jejuni (12.9%) by real-time PCR. Resistance to enrofloxacin-ciprofloxacin and nalidixic acid was significant, especially in C. jejuni (73.3%) and C. coli (77.2%) from broilers. Higher erythromycin (P = 0.043) and tetracycline (P = 1.865e-14) resistance rates were found among C. coli isolates (9.7 and 74.1%, respectively) than among C. jejuni isolates (3.1 and 36.6%, respectively). A total of 47 fla short variable region sequences were identified among 73 selected C. coli and C. jejuni isolates, with 35 fla types detected only once. At the nucleotide level, fla types A66 and A21 were the most common. Using the pulsed-field gel electrophoresis method, 66% of strains exhibited unique profiles after Sma I digestion. Forty-two isolates assigned to 18 Sma I clusters were further typed by Kpn I, and of these, 24 were assigned to 10 Kpn I clusters. For isolates in five Kpn I clusters, epidemiological links were observed. Stable C. jejuni and C. coli clones were detected, indicating that further studies involving broiler and human isolates need to be conducted to elucidate the importance of these stable clones in human infections.
[Show abstract][Hide abstract] ABSTRACT: During 2008 and 2009, within the framework of the Hungarian monitoring program of antibiotic resistance of zoonotic agents from food-producing animals, a significant number (43 strains) of Campylobacter lanienae were detected for the first time in Hungary. The isolates were genotyped using partial 16S rRNA gene sequencing and pulsed-field gel electrophoresis using three different restriction enzymes. The antimicrobial resistance of the isolates was determined by microtiter broth dilution. C. lanienae isolation was successful only from swine but not from other animal species. According to phylogenetic analysis, clustering of the isolates shows the same extensive genetic diversity as other Campylobacter species. Sequence analysis of the partial 16S rRNA gene showed that additional variations exist in variable regions Vc2 and Vc6. SmaI restriction enzyme proved to be the most efficient for pulsed-field gel electrophoresis analysis of C. lanienae. A significant tetracycline resistance (60.9%) and the presence of erythromycin-, enrofloxacin-, and multiresistant C. lanienae strains were found. Although the pathogenic potential of C. lanienae in humans is currently unknown, this study demonstrates that C. lanieanae is common in pigs in the country, provides further details on the genotypic and phenotypic properties of C. lanienae, and offers a genotyping method for use in source tracing.
[Show abstract][Hide abstract] ABSTRACT: The aetiology of increased mortality observed in two breeder duck flocks (Flock A consisting of 3500 laying ducks and Flock B comprising 4300 laying ducks) during the first egg-laying season was studied. In Flocks A and B, 773 ducks and 715 ducks (18.4% and 16.6%) died within a 24-week and a 20-week period, respectively. Death was preceded by clinical signs including movement difficulties, lack of appetite and depression lasting for 1 to 2 days. Diarrhoea was not observed. On gross pathological examination, the ducks were found to have haemorrhagic to fibrinonecrotic typhlocolitis, renal degeneration accompanied by fibrosis and mineralization, hepatic and splenic amyloidosis, and swelling of some of the metatarsal and phalangeal joints. Histopathological and immunohistochemical examination consistently demonstrated spirochaetes in the mucous membrane of the affected large intestine. On the basis of their cultural and biochemical properties and polymerase chain reaction sequencing analysis, four out of seven spirochaete strains isolated from the ducks (Flock A) by culture on special media under anaerobic conditions were identified as Brachyspira hyodysenteriae, and five out of eight strains (Flock B) were identified as Brachyspira pilosicoli. This is the first report on the isolation of B. hyodysenteriae and B. pilosicoli from laying ducks affected by fibrinonecrotic typhlocolitis.
[Show abstract][Hide abstract] ABSTRACT: The authors present examination on intestinal sprirochaetosis (brachyspirosis) in Hungarian laying hen flocks. They detected profuse diarrhoea in meat hybrid parent flocks ("A", "B" and "C") of different age (50, 87 and 47 weeks) and number (30000, 35000 and 19000). It occurred occasionally in 10-50% of the animals. The faeces of involved birds contained undigested feed and watery phase or it was brownish-yellow and smelly. The litter was moistened and the environment, the feathers and the eggs were contaminated by liquid faeces. Decrease in egg production and hatchability reached 15-20% after reoccurrence of diarrhoea. Eggshells became thin, got fragile and the colour became light. The hens weakened, moved with difficulty, sat frequently. The sole thickened, got chapped and frequently inflamed due to secondary infections. Daily mortality rate has not changed in flocks "A" and "B", while slightly increased in flock "C" (from 0.3-0.4% per week to 0.7-0.8%). On the basis of biochemical and sequence analysis, they determined: in flock "A" 2 Brachyspira spp., 1 B. intermedia strains, in flock "B" 2 B. hyodysenteriae and 1 B. innocens strains, in flock "C" 3 B. hyodysenteriae strains. Following treatment with tiamulin and doxycyclin for 5-7 days the number of birds suffering of diarrhoea decreased, but after finishing treatment it reoccurred. Following change in feeding diarrhoea stopped and has not reoccurred. In the feedingstuffs; given to flock "C" at the time of diarrhoea 0.25-0.5 ppm deoxynivalenol (DON) and 0.15-0.5 ppm F2 (zearalenon) contamination was detected. In the feedingstuffs; given to flock "A" and "B" no decrease in quality was detected by microbiological methods. From Brachyspira species pathogenic to birds, according to scientific literature (B. pilosicoli, B. intermedia, B. alvinipulli, B. hyodysenteriae) the first three Brachyspira species have been detected abroad from hens suffering of diarrhoea, while pathogenicity of B. hyodysenteriae to hens was demonstrated only by experimental infections. Recently (2008) Dutch authors demonstrated B. hyodysenteriae also from natural infections in hens, our present examination confirm it.
Magyar Allatorvosok Lapja 01/2009; 131(6):323-330. · 0.19 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In this report we examined the glycopeptide susceptibility of enterococci, isolated in 2005, from slaughtered animals, within the confines of Hungarian Antibiotic Resistance Monitoring System. We determined the presence of the van genes as well as their genetic relatedness in enterococci from poultry. Enterococcus sp. strains (n=175) were collected from intestinal samples of slaughtered poultry in 2005. The origin of the samples was registered at county level. After screening the strains with 30 mg vancomycin disc 19 (86%) intermediate resistant and 4 (3%) fully resistant strains were found. The distribution of minimum inhibitory concentration (MIC)-values among 23 enterococcus strains which were intermediate or resistant to vancomycin were 0.25 mg/L (4.4%), 2 mg/L (8.6%), 4 mg/L (8.6%), 8 mg/L (61%), 16 mg/L (8.6%) and 256 mg/L (8.6%). The MICs of teicoplanin were 0.25 mg/L (4.3%), 1 (8.6%), 4 mg/L (78.3%), 16 mg/L (4.3%) and 256 mg/L (4.3%). The two most vancomycin-resistant strains were vanA carriers (1 E. faecalis and 1 E. faeciuum). The farms that produced these strains can be reservoirs of VRE and the affected farms should change the technology of disinfection and breeding in order to prevent the emergence of high numbers of human VRE isolates in Hungary.
[Show abstract][Hide abstract] ABSTRACT: The presence of the vanA gene was determined in enterococci from healthy poultry, originating from the Hungarian resistance monitoring system between 2001 and 2004. Enterococci (n = 562) were collected from intestinal samples of slaughtered broiler chickens. The presence of van genes was detected by polymerase chain reaction (PCR). The vancomycin-resistant enterococcus (VRE) strains carried only the vanA gene. Genus- and species-level identification of the vanA gene carrier strains was carried out by PCR using specific primers. In 2001, 25 out of the 289 isolated strains (8.6%) were vanA carriers (1 Enterococcus mundtii, 13 E. durans and 11 E.faecium). In 2002 (n = 87), 20 (23%) strains were vanA positive (11 E. durans and 9 E. faecium). In 2003 and 2004, none of the strains (n = 95 and 91, respectively) were positive for the most common van genes. In 2003, there was only one strain for which higher minimum inhibitory concentrations (MIC) of vancomycin (4 mg/L) and teicoplanin (8 mg/L) were found. In 2004 there were three strains for which the MIC of vancomycin was 8 mg/L, and 2 strains and 1 strain with teicoplanin MICs of 4 mg/L and 8 mg/L, respectively. The potential similarity of these strains was studied by pulsed-field gel electrophoresis (PFGE). The VRE strains were not closely related to one another. The annual data of vancomycin resistance indicate an association between the recovery of vancomycin-resistant enterococci and the use of avoparcin in animal feeds. This study indicates that with the reduced use of antibiotics in food animals, it is possible to decrease the rate of resistant bacteria. Although the use of avoparcin had been banned in 1998, the VRE strains disappeared only five years later.
[Show abstract][Hide abstract] ABSTRACT: The authors, on the basis of scientific literature and their own diagnostic results, summarise important knowledge about intestinal spirochaetosis in different poultry species (turkeys, geese, ducks, domestic fowls), compared to swine dysentery. Causative role of spirochaetas in swine dysentery was confirmed in 1970s, and they were also detected in domestic fowl and turkey flocks from diseases with diarrhoea in countries of developed poultry farming (USA, Great Britain, Australia). In 1990s severe necrotic colitis with deaths in connection with spirochaetas was described in American ostrich (Rhea americana). From water fowl species, spirochaetas were detected from domestic duck flocks and wild ducks free of clinical signs, abroad, while later Hungarian authors reported first time, also internationally, spirochaetosis in geese with fibrinous-necrotic colitis and severe nephropathy. The authors carried out examinations in 28 Hungarian goose, 3 duck and 3 fowl flocks. Chronic diarrhoea with clinical signs occurred in broiler parent flocks at egg-laying period, with reoccurrence in spite of treatment. Mortality has not increased significantly, diarrhoea affecting 60% of flocks caused severe contamination of the litter, environment and eggs, later weakening of the animals, decrease in egg production and hatchability and leg paresis and necrosis of the skin of sole. In duck and goose flocks diseased in intestinal spirochaetosis - often at the end of egg-laying period - mortality gradually increased without visible signs of diarrhoea. Post mortem examination revealed fibrinous or necrotic inflammation of the caecum, colon and rectum, and chronic nephropathy often with fibrosis, and in the ducks arthritis of the tarsus and toes. The diagnosis was based on epidemiological data, clinical signs and pathological and pathohistological changes, demonstration of brachyspiras in the changed intestinal mucosa by silver impregnation or immunohistochemical method, culture on selective medium, and identification by biochemical tests and sequence analysis. In goose flocks Brachyspira alvinipulli was the most frequently detected. From fowl flocks 8, and from duck flocks 21 Brachyspira strains were isolated, from which Brachyspira hyodysenteriae (and in duck flocks B. pilosicoli, too) was dominant.
Magyar Allatorvosok Lapja 01/2008; 130(11):663-670. · 0.19 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Ten one-day-old goslings were inoculated orally with a Brachyspira alvinipulli strain isolated from the large intestine of geese that had died of intestinal spirochaetosis (Group A), 10 day-old goslings were inoculated orally with a B. hyodysenteriae strain (Group B), and a third group of 10 goslings (Group C) served as uninfected control. The goslings were observed daily for clinical signs. They were sacrificed on days 7, 14, 21 and 35 days postinfection (PI), and necropsied. Segments of the large intestine were subjected to histopathological, immunohistochemical, electron microscopic (TEM, SEM) and microbiological examinations. Mortality did not occur during the experimental period. However, in both groups the caecum of the goslings killed by bleeding was slightly dilated, in its lumen there was a watery, yellowish and frothy content, and the mucous membrane was slightly swollen. By histopathological, immunohistochemical and electron microscopic examination, B. alvinipulli and B. hyodysenteriae could be detected in the caecum or colon, in the lumen of the glands and sometimes among the glandular epithelial cells in goslings of the respective groups, and could be reisolated from these organs by culturing. A mild inflammation of the intestinal mucosa was also noted. In transverse section of the brachyspirae, numerous (16-22) periplasmic flagella could be detected inside the outer sheath, also depending on the plane of section.
[Show abstract][Hide abstract] ABSTRACT: Isolates of Enterococcus spp. were collected from January 2001 to December 2004 from caecal samples of slaughtered poultry, swine and cattle in Hungary. The isolates were identified by their growth and biochemical properties and with PCR. The antibiotic susceptibility of a total number of 1272 isolates was tested with disk diffusion test to ampicillin, gentamicin, streptomycin, tetracycline, erythromycin and vancomycin.It was established that although ampicillin and amoxicillin are often used in veterinary practice its resistance rate was relatively low. In the case of tetracyclines and macrolides, a high incidence of resistance was found. Susceptibility of strains to tetracyclines and/or macrolides reduced in both 2003 and 2004 in all animal species, which may be due to the more frequent usage of these drugs in the veterinary practice following the ban of growth promoters.The annual data of vancomycin resistance point to an association between the recovery of vancomycin-resistant enterococci (VRE) isolates and the use of avoparcin.This study indicates that reducing antimicrobial resistance in food animals could be possible with lower usage of antibiotics, although variations can occur with different strains.
International Journal of Food Microbiology 05/2007; 115(1-115):119-123. DOI:10.1016/j.ijfoodmicro.2006.10.004 · 3.08 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The role of Brachyspira bacteria in the aetiology of increased mortality observed in two breeder goose flocks (Flock A consisting of 1,500 and Flock B comprising 4,500 laying geese) at the end of the first egg-laying season, in the period of moulting, was studied. In Flock A 415 geese (28%) died during an 8-week period while in Flock B 834 geese (18%) died during a 12-week period. On gross pathological examination, the geese were found to have haemorrhagic-to-necrotic inflammation of the large intestine (colon and rectum) and fibrinonecrotic typhlitis accompanied by severe degeneration. Often, fibrosis of the kidneys, and in five of the geese secondary visceral urate deposition ("visceral gout") was also observed. Histopathological examination consistently demonstrated spirochaetes in the mucous membrane of the affected large intestine. This was confirmed by the results of immunohistochemical and electron microscopic examination. In addition, Trichomonas stages were also detected from the large intestine of 11 geese. On the basis of their cultural and biochemical properties, and PCR sequencing analysis, eight out of the nine spirochaete strains isolated from the geese by culture on special media under anaerobic conditions were identified as Brachyspira alvinipulli. This is the first report on the isolation of B. alvinipulli from laying geese affected with fibrinonecrotic typhlocolitis.