Presence and coincidence of Listeria spp., motile aeromonads and Yersinia enterocolitica in a cold-smoked salmon processing and packing plant

Environmental samples and samples of partially processed fish from a cold-smoked salmon processing and packing plant, and product samples purchased from retail outlets, were examined for the presence of Listeria monocytogenes and other Listeria spp., motile aeromonads and Yersinia enterocolitica. Listeria spp. were not isolated from raw fish but were a contaminant of processed and partially processed fish. Listeria spp. were also detected in 18.8% of surfaces in contact with fish. Unlike Listeria spp., motile aeromonads were isolated from the raw fish but not from the finished product. They were more frequently isolated from environmental sources than Listeria spp. Yersinia enterocolitica was not isolated from any of the samples tested. Little evidence was found to show a coincidence of motile aeromonads and Listeria spp., since only one subset of samples showed such a link. It is concluded that contamination by Listeria spp. was from environmental sources at the processing plant at, or beyond, the slicing stage. Reducing the number of wet areas and special cleaning and sanitation considerations for a contaminated site (the freezer seal) are suggested as ways of reducing contamination of the product.     

The effect of cobalt topdressing in preventing cobalt deficiency disease of lambs in southland

Environmental samples and samples of partially processed fish from a cold-smoked salmon processing and packing plant, and product samples purchased from retail outlets, were examined for the presence of Listeria monocytogenes and other Listeria spp., motile aeromonads and Yersinia enterocolitica. Listeria spp. were not isolated from rati fish but were a contaminant of processed and partially processed fish. Listeria spp. were also detected in 18.8% of surfaces in contact with fish. Unlike Listeria spp., motile aeromonads were isolated from the raw fish but not from the finished product. They were more frequently isolated from environmental sources than Listeria spp. Yersinia enterocolitica was not isolated from any of the samples tested. Little evidence was found to show a coincidence of motile aeromonads and Listeria spp., since only one subset of samples showed such a link. It is concluded that contamination by Listeria spp. was from environmental sources at the processing plant at, or beyond, the slicing stage. Reducing the number of wet areas and special cleaning and sanitation considerations for a contaminated site (the freezer seal) are suggested as ways of reducing contamination of the product.     

Eosinophilic myositis and muscular sarcocystosis in the carcasses of slaughtered cattle and lambs

Fifty-three bovine and 7 ovine carcasses condemned for having eosinophilic myositis were evaluated. Four (7.3%) of the bovine carcasses had a few, large local lesions in skeletal muscles (category A), and 49 (92.7%) of the bovine carcasses and 7 (100%) of the ovine carcasses had multiple, small, disseminated lesions in tongue, esophagus, heart, diaphragm, or skeletal muscles (category B). Tissue from carcasses of category B were evaluated for bacteria, viruses, selenium, and pathologic changes. Pathogenic bacteria and viruses were not isolated and selenium concentrations were normal. In category B, all carcasses had granulomas; of the 49 bovine carcasses and 7 ovine carcasses, 38 (77.6%) and 7 (100%), respectively, had one or more granulomas with opened dead sarcocysts. The data indicated that opened sarcocysts killed the host myocyte and adjacent myocytes and stroma, thereby initiating granuloma formation.     

Experimental mixed infection of rabbits with Yersinia enterocolitica and Listeria monocytogenes

Experimental mixed infection was reproduced in rabbits after per os infection with Yersinia enterocolitica serotype 0:3 cells. Four days later some of animals were re-infected orally with Listeria monocytogenes serotype 4b cells. A third group of healthy rabbits was also infected per os with Listeria monocytogenes. The infectious process was followed dynamically from days 1-28. The experimental animals were examined for clinical, paraclinical and morphological findings. Augmentation of body temperature and alveolar macrophage number, a decreased number of peritoneal macrophages, leucopenia as well as purulent meningoencephalitis, catarrhal pneumonia, lienitis, lymphadenitis and enteritis were detected after experimental mixed infection. Both types of macrophages demonstrated a weak bactericidal activity against Yersinia enterocolitica and a highly expressed killing effect against Listeria monocytogenes. Yersinia and Listeria cells were isolated from the viscera and brain. Both species of bacteria were established intracellularly in the macrophages by electron-microscopic examination. The data received showed that mixed Yersinia enterocolitica 0:3 and Listeria monocytogenes 4b infection of rabbits runs with transitory hyperthermia as a generalized infection and is similar to the Listeria mono-infection. The immunosuppressive effect induced by oral Yersinia enterocolitica infection of rabbits promotes the expression of listerious agents.     

Demonstration of Listeria monocytogenes by immunohistochemistry in formalin-fixed brain tissues from natural cases of ovine and bovine encephalitis

In the present work, evidence of Listeria monocytogenes antigens based on the avidin-biotin complex (ABC) immunoperoxidase technique was performed on formalin-fixed central nervous system tissues (CNS) from a total of 23 natural cases of encephalitis (four ovine and 19 bovine). Listeria monocytogenes serotype 4 was isolated from 10 of 17 cultured specimens. Meningoencephalitis characterized by focal necrosis, microabscesses, perivascular cuffing, and gliosis with presence of macrophages and/or neutrophils was observed at histological examination. Positive L. monocytogenes antigens were successfully identified by immunohistochemistry (IHC) in the CNS of all 23 cases. Paraffin-embedded tissues assayed were stored up for 17 years. Morbidity of the outbreaks was between 0.3-3% and 0.1-1% for ovine and bovine cases, respectively. In all the ovine cases, flocks involved were under extensive grazing conditions. In nine of the 19 bovine cases (47.3%), supplementation with corn silage was used. The ABC test can help as a practical tool for the diagnosis of natural cases of L. monocytogenes encephalitis on formalin-fixed specimens from ovine and bovine.     

Prevalence of Campylobacter spp. and Yersinia spp. in the pig production

The aim of this study was to determine the prevalence of Campylobacter spp. and Yersinia spp. in a total of 1,040 faecal samples taken from animals at different ages from four farrowing and twelve fattening herds. In the farrowing unit, faeces were collected from 68 sows (faecal samples) and 256 suckling piglets (rectal swab samples). Further samples were collected from 362 growing and 354 finishing pigs (rectal swab samples). Additionally, 56 feed and environmental samples were collected. During the slaughtering process, 122 pigs and their carcasses respectively, were sampled three times. Finally, 86 meat and minced meat samples were taken from 34 retail stores. Campylobacter spp. were isolated in sows (33.8 %), piglets (80.9 %), growing (89.2 %) and finishing (64.7 %) pigs. Yersinia spp. were detected in growing (15.2 %) and finishing (13.3 %) pigs only. After twelve hours of chilling neither Campylobacter spp. nor Yersinia spp. were detected. In raw meat samples, Campylobacter spp. were isolated from one liver sample and Yersinia enterocolitica from two meat samples. Common slaughter techniques and hygiene procedures may be effective tools to reduce the risk of contamination and recontamination of meat products since Campylobacter spp. and Yersinia spp. were found only sporadically in raw meat samples.     

Demonstration of Listeria monocytogenes by Immunohistochemistry in Formalin‐Fixed Brain Tissues from Natural Cases of Ovine and Bovine Encephalitis

In the present work, evidence of Listeria monocytogenes antigens based on the avidin–biotin complex (ABC) immunoperoxidase technique was performed on formalin‐fixed central nervous system tissues (CNS) from a total of 23 natural cases of encephalitis (four ovine and 19 bovine). Listeria monocytogenes serotype 4 was isolated from 10 of 17 cultured specimens. Meningoencephalitis characterized by focal necrosis, microabscesses, perivascular cuffing, and gliosis with presence of macrophages and/or neutrophils was observed at histological examination. Positive L. monocytogenes antigens were successfully identified by immunohistochemistry (IHC) in the CNS of all 23 cases. Paraffin‐embedded tissues assayed were stored up for 17 years. Morbidity of the outbreaks was between 0.3–3% and 0.1–1% for ovine and bovine cases, respectively. In all the ovine cases, flocks involved were under extensive grazing conditions. In nine of the 19 bovine cases (47.3%), supplementation with corn silage was used. The ABC test can help as a practical tool for the diagnosis of natural cases of L. monocytogenes encephalitis on formalin‐fixed specimens from ovine and bovine.     

Yersiniosis in deer from the Otago-Southland region of New Zealand

Samples from 350 routine deer cases submitted to Invermay Animal Health Laboratory for diagnosis during 1979-1982 were examined specifically for the presence of Yersinia sp. An analysis of 57 cases of yersiniosis due to Yersinia pseudotuberculosis is made and two cases due to Yersinia enterocolitica are described. The occurrence of cases appeared strongly correlated with periods of stress, predominantly in winter when cool wet conditions and lack of grazing combined to precipitate the disease. Animals up to one year old were most commonly affected (64% of cases). Of the 61 strains of Yersinia pseudotuberculosis isolated 35 were of Serotype I, 17 of Serotype II and nine of Serotype III. All strains of Yersinia pseudotuberculosis and the strains of Yersinia enterocolitica isolated from the two cases described were Hela cell invasive and gave a positive autoagglutination test for virulence.     

Experimental Yersinia enterocolitica placentitis in sheep.

A strain of Yersinia enterocolitica of O serogroup 6,30 isolated from the liver of an aborted ovine fetus was inoculated intravenously into a group of pregnant ewes at about 90 days gestation and produced placentitis with abortion or delivery of infected lambs about 50 days later. Y. enterocolitica of the same serogroup was recovered from the necrotic placental cotyledons and most other fetal tissues and could be isolated from vaginal discharges of the ewes for a least 2 weeks after abortion. Histological changes were consistent with an acute bacterial necrotizing placentitis and systemic infection of the fetus. Subsequent pregnancies in the ewes proceeded to term without evidence of infection.     

The bacteriological and serological prevalence of Campylobacter spp. and Yersinia enterocolitica in fattening pig herds in Lower Saxony

This article presents the results of a study on the occurrence of two bacteria that cause zoonoses, Campylobacter spp. and Yersinia enterocolitica. The study was carried out in 30 fattening herds in Lower Saxony, Germany, in 2004 and compares the results of bacteriological and serological methods of detection. Bacteriological findings of Campylobacter spp. in the faeces indicated that 69.7% of the fattening pigs were positive, but 81.2% tested positive serologically. All herds tested here were both bacteriologically and serologically positive for Campylobacter spp. Furthermore, only 8.4% tested positive for Yersinia enterocolitica in the faecal samples, but 66.8% of the animals were serologically positive for that bacterium. While bacteriological examination did not detect Yersinia enterocolitica in 56.7% of the herds tested, serological testing showed that only 16.7% of the units were without reacting animals. The great difference between the results of bacteriological and serological testing, especially in the case of Yersinia enterocolitica, can be explained by the intermittent intestinal excretion and predominance of this bacterium in the animals' tonsils. Low faecal excretion is also the reason for the low detection rate of 3.4% of Yersinia enterocolitica in the environmental samples, while that of Campylobacter spp. was 33.3%. These results indicate that the environment plays only a secondary role in the distribution of Yersinia enterocolitica in pig herds.     

A simple technique to differentiate between animals infected with Yersinia enterocolitica IX and those infected with Brucella abortus.

While both Brucella abortus and Yersinia enterocolitica IX have O antigens in common, they differ significantly with respect to motility. Thus Br abortus is always non-motile while Y enterocolitica is motile when grown at room temperature. The presence of yersinia H agglutinins in serum can be shown to be evidence of previous exposure to Y enterocolitica. These agglutinins are not generated by brucella infection. A rapid H agglutination test will serve to provide this differentiation without interference from cross-reacting O antigens.     

Differentiation of Brucella abortus and Yersinia enterocolitica serotype 09 infections in cattle: the use of specific lymphocyte transformation and brucellin skin tests

The lymphocyte transformation test (using an in vitro whole-blood lymphocyte stimulation procedure) and the Brucellin skin test were applied to five heifers infected with virulent Brucella abortus strain 544, five cows inoculated with Yersinia enterocolitica serotype 09, and four non-exposed cows. Lymphocytes from Brucella-inoculated animals persistently gave very high blastogenic reactions indicative of active Brucella infection. The test was persistently negative in Yersinia-infected and non-exposed cattle. Four of the five cows infected with Yersinia enterocolitica type 09 and all four control cattle were persistently negative to the delayed hypersensitivity skin reaction with brucellin. All cattle infected with Yersinia enterocolitica type 09 were strongly positive to the Rose Bengal, Serum agglutination, Complement fixation and Antibovine globulin tests using Brucella abortus antigens. One lactating cow infected with Yersinia enterocolitica type 09 was positive to Brucella milk ring test. These results indicate that standard Brucella serological tests are unreliable in differentiating the two infections in cattle and that both the Lymphocyte transformation and brucellin skin tests could be used to differentiate bovine brucellosis from yersiniosis.     

The prevalence of salmonella, yersinia and mycobacteria in slaughtered pigs in Switzerland

Clinically healthy food animals can be reservoirs for various foodborne pathogens. In general, such animals do not have lesions that are visible during meat inspection. Pigs are considered to be carriers of salmonella, yersinia and mycobacteria, but the risk of transmission to humans is difficult to assess. The aim of this study was to estimate the actual prevalence of the three above mentioned pathogens in the Swiss pig population and to comment on their significance. A total of 570 samples each of tonsils and mesenteric lymphnodes, were collected at two slaughterhouses from carcasses of apparently healthy pigs and analyzed for the presence of salmonella, yersinia and mycobacteria. The prevalence of salmonella (0.9%) was found to be lower than--while that of yersinia (8.1%) and mycobacteria (12.8%) about equal to--results reported from other European countries. Yersinia typing showed that serotype O:9 of Yersinia enterocolitica (2.5%) was 6 to 7 times more frequent than serotype O:3 (0.4%)--formerly the most frequent serotype. Mycobacterium avium was the most frequent isolate (90.7%) among the mycobacteria isolated. Although all three pathogens are present in the Swiss pig population, we consider the risk of transmission to humans via consumption of pork as low. Appropriate preventive measures and quality management should contribute to keep the risk under control.     

Studies of the efficacy of Enterocoliticin, a phage-tail like bacteriocin, as antimicrobial agent against Yersinia enterocolitica serotype O3 in a cell culture system and in mice

The efficacy of enterocoliticin, a phage tail-like bacteriocin, as antimicrobial compound against infections with pathogenic Yersinia enterocolitica serotype O3 strains was assessed. In cell cultures, which were infected with the Y. enterocolitica strains 13 169 or 6471/76, bactericidal activity of enterocoliticin was found for bacteria adhering to the surface of eukaryotic cells, whereas bacteria, which had invaded the eukaryotic cells, were not accessible to the bacteriocin. The interaction of enterocoliticin with Y. enterocolitica was further examined in animals. Female BALB/c mice were experimentally infected with the two Y. enterocolitica strains and enterocoliticin was applied as antimicrobial compound by the oral route. Experimental variations concerning the infectious doses of the Y. enterocolitica strains and the time points of application of the bacteriocin were investigated. The increase of the Yersinia CFU titre in animals was retarded at time points shortly after the application of enterocoliticin indicating that the particles were effective on recently introduced Yersinia. The repeated application of enterocoliticin, however, did not prevent the colonization of the gastrointestinal tract by Yersinia.     

Non-specific seroreactions against Brucella abortus in ruminants in New Zealand and the presence of Yersinia enterocolitica 0:9

The level of non-specific reactions found in the brucellosis serology of ruminants in New Zealand was very low until July 1992. This changed when, in an export consignment of 1071 deer, 35% reacted in the Brucella abortus tube agglutination test with titres varying from 50 to 200 IU. The reactors were also positive in the Rose-Bengal agglutination test and most of them reacted in the complement fixation test with titres varying from 10 to 80 IU. Yersinia enterocolitica 0:9 was later isolated from one deer of this consignment. It was the first isolate of this serotype recovered in New Zealand from an animal. Shortly after, false reactors occurred more frequently than before in sera from Brucella abortus accredited free cattle herds. As the involvement of Yersinia enterocolitica 0:9 was suspected in these cases, faecal samples from reactors and in-contact animals were cultured for this organism. Yersinia enterocolitica 0:9 was isolated from nine of 19 herds showing one or more false Brucella abortus seroreactions. Prior to 1990, Yersinia enterocolitica serotype 0:9 had not been isolated in New Zealand, despite the recovery of a number of other bio- or serotypes of the organism from humans and animals. From 1990 onward, serotype 0:9 began to be isolated from human faecal samples with increasing frequency. Since the first isolations from deer and cattle in 1992, it has now also been recovered from a cat and an alpaca and from cattle without any association with false positive Brucella abortus reactions. All serotype 0:9 isolates were of biotype 2.     

Microbiological identification and analysis of swine tonsils collected from carcasses at slaughter

The primary objective of this 7-month study was to determine the prevalence of porcine pathogens of the tonsil of the soft palate of swine at slaughter. Additional objectives were to determine if sampling the carcasses of normal or abnormal hogs provided different microbiological profiles and if the slaughter plant provides a feasible sampling frame and environment for detecting and monitoring important pathogens in tonsils that have health implications for both swine and humans. A total of 395 samples were collected from 264 farms. Of these, 180 tonsils were collected from normal carcasses and 215 tonsils were collected from carcasses that were diverted to the hold rail. Laboratory testing included bacteriological culture and identification as well as real time-polymerase chain reaction (PCR) testing for porcine reproductive and respiratory syndrome virus (PPRSV) and immunohistochemistry (IHC) for porcine circovirus-2 (PCV-2). The most commonly isolated bacteria included: Streptococcus suis (53.7%), Arcanobacterium pyogenes (29.9%), Pasteurella multocida (27.3%), and Streptococcus porcinus (19.5%). Virus screening revealed evidence of PRRSV and PCV-2 in 22.0% and 11.9% of the samples, respectively. Salmonella Typhimurium and Yersinia enterocolitica were isolated in 0.5% and 1.8% of the samples, respectively. Tonsils collected from the hold rail were more likely to be positive for Staphylococcus hyicus [odds ratio (OR) = 7.51, confidence interval (CI) = 2.89 to 19.54], Streptococcus porcinus (OR = 9.93, CI = 4.27 to 23.10), and Streptococcus suis (OR = 2.16, CI = 1.45 to 3.24). Tonsils collected from abnormal carcasses were less likely to be positive for Staphylococcus aureus (OR = 0.05, CI = 0.005 to 0.482).     

Prevalence of Yersinia enterocolitica in goat herds from Northern Germany

The prevalence of human pathogenic Yersinia enterocolitica isolates in livestock farming is of paramount interest. Raw goat milk has been proposed as a source of human yersiniosis; however, no data on the prevalence of human strains of Y. enterocolitica in goat herds are available. Therefore, fecal samples (n = 575) were collected from 24 goat herds from Lower Saxony, northern Germany. Pre-enrichment in peptone, sorbitol and bile salts broth was followed by plating on cefsuloidin irgasan novobiocin agar. Yersinia enterocolitica was isolated from 17 (3%) samples of five (21%) goat herds. All isolates were biovar 1A, but represented various serovars. PCR assays targeting Yersinia adhesin (yad) gene and the yopT gene, both associated with pathogenicity, produced no amplification products. Therefore, the isolates can be regarded as opportunistic apathogenic bacteria. Consequently, milk, cheese or meat from goats should not be considered as an important source for human yersiniosis.     

Immunohistochemical identification of Campylobacter fetus in natural cases of bovine and ovine abortions

An immunohistochemistry (IHC) procedure for the detection of Campylobacter fetus antigens using an avidin-biotin complex technique was performed on formalin fixed bovine and ovine fetal tissues from 26 natural cases of Campylobacter spp. abortion (four ovine and 22 bovine). The species of Campylobacter isolated included C. fetus ssp. venerealis from 13 bovine fetuses, C. fetus ssp. fetus from two ovine and one bovine fetus, Campylobacter jejuni from seven bovine fetuses, Campylobacter lari from two ovine fetuses and an unspeciated Campylobacter species in one bovine fetus. Histologic lesions identified in the aborted fetuses included placentitis, serositis, pneumonia, gastroenteritis, hepatitis and encephalitis. Campylobacter fetus antigens were identified by IHC in 13 of 13 bovine fetuses from which C. fetus ssp. venerealis was isolated and in two of two ovine fetuses from which C. fetus ssp. fetus was isolated. The IHC stains were negative in tissues from seven bovine fetuses from which C. jejuni was isolated, one bovine fetus infected with C. fetus ssp. fetus, one bovine fetus infected with the unspeciated Campylobacter and two ovine fetuses infected with C. lari. In positive cases, the IHC stain most frequently identified bacteria in the lung and gastrointestinal tract. The C. fetus IHC procedure performed on formalin fixed tissues is a practical tool for the diagnosis of natural cases of ovine and bovine abortion caused by C. fetus.     

A cohort study of Yersinia infection in goats

OBJECTIVE: To determine the temporal pattern of Yersinia infections in three goat flocks and examine the influence of management and seasonal factors on the incidence of those infections over a 1-year period. METHODS: A longitudinal study involving monthly culture of faeces for Yersinia spp. from age groups of randomly selected goats on three farms in the Manawatu region of New Zealand. RESULTS: The incidence of excretion of potentially pathogenic Yersinia (Yersinia pseudotuberculosis and Y enterocolitica biotypes 2, 3 and 5) peaked in winter and fell in summer. In contrast, environmental Yersinia (Y enterocolitica biotype 1A, Y frederiksenii, Y intermedia and Y rohdei) showed no clear pattern of seasonal variation. Pathogenic Yersinia were more prevalent in young animals than in adults, while environmental Yersinia were more prevalent in adults. The same type was isolated from the same animal in two or more successive months in about 20 to 25% of cases, and in the remaining cases there was a gap of at least one month between successive isolations, with many animals yielding a particular type on only a single occasion. A notable difference was that with the potentially pathogenic types, no animal had more than one period of time when it was found to be excreting a particular type, suggesting that immunity develops following exposure. In contrast, it was common for environmental types to be isolated from the same animal throughout the study period. Two goats were suspected to have developed clinical yersiniosis but all remaining infected animals showed no clinical signs of infection. CONCLUSIONS: Asymptomatic Yersinia carriage was common in goats in New Zealand, with a clear seasonal and age group pattern of infection with potentially pathogenic types. There was evidence that immunity developed to potentially pathogenic types. This is the first time that Y rohdei has been isolated from goats.     

Isolation of Listeria monocytogenes from the skin of slaughtered beef cattle

We attempted to isolate Listeria monocytogenes from skin, contents of large intestines and carcasses of cattle introduced to a slaughterhouse in order to identify source of contamination for this pathogen. Sixty skin samples, 60 samples of the contents of large intestines and 30 carcass samples were colleted in June, August and November 2003 for use in this study. Listeria spp. and L. monocytogenes were isolated from 30 (50%) and 3 (5%) of the cattle skin samples, respectively. However, no Listeria spp., including L. monocytogenes, were isolated from intestinal contents or carcasses. Seven isolates were obtained, of which five and two strains were serotypes 1/2a and 1/2b, respectively. Genetic analysis suggested that there was persistent inhabitation of the pathogen around the area investigated in this study.