The presence and characteristics of Clostridium difficile were investigated in 839 faecal samples from seven different animal species in the Netherlands. The number of positive samples ranged from 3.4% (cattle) to 25.0% (dogs). Twenty-two different PCR ribotypes were identified. Among 96 isolates, 53% harboured toxin genes. All C. difficile isolates from pigs, cattle and poultry were toxinogenic, whereas the majority of isolates from pet animals consisted of non-toxinogenic PCR ribotypes 010 and 039. Ribotype 012 was most prevalent in cattle and ribotype 078 in pigs. No predominant ribotypes were present in horse and poultry samples. Overall, PCR ribotypes 012, 014 and 078 were the most frequently recovered toxinogenic ribotypes from animal samples. Comparison with human isolates from the Dutch Reference Laboratory for C. difficile at Leiden University Medical Centre (LUMC) showed that these types were also recovered from human hospitalized patients in 2009/2010, encompassing 0.8%, 11.4% and 9.8% of all isolates, respectively. Application of multiple-locus variable-number tandem-repeat analysis indicated a genotypic relation of animal and human ribotype 078 strains, but a clear genotypic distinction for ribotypes 012 and 014. We conclude that toxinogenic C. difficile PCR ribotypes found in animals correspond to PCR ribotypes associated with human disease in hospitalized patients in the Netherlands. Contrary to PCR ribotype 078, significant genetic differences were observed between animal and human PCR ribotype 012 and 014 isolates.

Intestinal colonization and shedding of pathogenic bacteria in animal feces is an important factor in both human food safety and animal health. The effect of broiler feed additives flavophospholipol (FPL; Flavomycin, bambermycins) and salinomycin sodium (SAL; Sacox) given singly on the excretion of Salmonella enteritidis, Campylobacter jejuni, and Clostridium perfringens was studied following controlled infection. The incidence of shedding (number of birds with positive fecal cultures) and the degree of shedding (cfu per gram of feces in positive birds) were measured to determine the influence of these two common feed additive antibiotics on shedding rates of potential pathogens. A total of 216 Ross broiler chickens, housed in battery cages, were fed either an unmedicated feed (controls), feed containing FPL, or feed containing SAL. Feed treatment groups were subdivided into three bacterial challenge groups of 24 chicks, each receiving only one of the pathogens. Bacterial challenge was administered orally on Days 11 and 12 for Salmonella and Campylobacter and on Days 2 and 3 for Clostridium. Fecal samples were collected weekly up to 6 wk of age and cultured for presence of the target organism. The shedding rate was determined by decimal dilutions of the fecal samples. Feeding FPL resulted in a reduced (P < or = 0.05) degree and incidence of Salmonella and Clostridium shedding at 6 wk. Feeding SAL reduced (P < or = 0.05) the incidence of Salmonella shedding at 6 wk. Neither feed additive affected the incidence nor the degree of Campylobacter shedding. The results of this study indicate that these feed additives may reduce the incidence of these potential human and animal pathogens in preslaughter broilers.

The prevalence of granulomatous lesions in lymph nodes of pigs was studied. From January till August 2004 in two slaughterhouses in The Netherlands 2,116,536 pigs were examined for the presence of granulomatous lesions in the submaxillary lymph nodes. In 15,900 (0.75%) of these pigs, lesions could be detected. Nine farms with the highest incidence of lesions were selected for a more detailed pathological and bacteriological examination. On these farms, the prevalence of lesions in sub-maxillary lymph nodes ranged from 2.3 to 5.7% with a mean of 3.0%. From 1276 pigs that were sampled, 98 (7.7%) displayed granulomatous lesions in the sub-maxillary lymph nodes and one (0.1%) pig showed lesions in its mesenteric lymph node. Mycobacterium avium subsp. avium (MAA) could not be isolated from the lymph nodes of the 99 pigs with lesions and from a selection of lymph nodes (n = 61) of pigs without lesions. Rhodococcus equi was isolated from 44 out of 98 (44.9%) of the sub-maxillary lymph nodes with granulomatous lesions and from two mesenteric lymph nodes without lesions. A comparison of former studies and the current results indicate that the prevalence of MAA infections in slaughter pigs has strongly decreased over the last decade, whereas R. equi is highly prevalent. The high incidence of granulomatous lesions associated with the bacteriological presence of R. equi could be considered as a serious cause of misdiagnosis of MAA infections in cases where meat inspection is carried out by inspection for granulomatous changes of lymph nodes only. #

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Aims:  To study the effect of UV irradiation on the bacterial load of shell eggs and of a roller conveyor belt. Methods and Results:  The natural bacterial load on the eggshell of clean eggs was significantly reduced by a standard UV treatment of 4·7 s; from 4·47 to 3·57 log CFU per eggshell. For very dirty eggs no significant reduction was observed. Eggs inoculated with Escherichia coli and Staphylococcus aureus (4·74 and 4·64 log CFU per eggshell respectively) passed the conveyor belt and were exposed to UV for 4·7 and 18·8 s. The reduction of both inoculated bacteria on the eggshell was comparable and significant for both exposure times (3 and 4 log CFU per eggshell). Escherichia coli was reduced but still detectable on the conveyor rollers. The internal bacterial contamination of eggs filled up with diluent containing E. coli or S. aureus was not influenced by UV irradiation. Conclusions:  There is a significant lethal effect of UV irradiation on the bacterial contamination of clean eggshells and recent shell contamination, contamination of rollers can be controlled and the internal contamination of eggs is not reduced. Significance and Impact of the Study:  The penetration of UV into organic material appears to be poor and UV disinfection can be used as an alternative for egg washing.

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Salmonella infections originating from poultry are one of the major causes of food-borne disease. For the control of salmonella in poultry a multifactorial approach is more likely to be effective, and the genetic resistance of poultry breeds to salmonella infections may be a valuable contribution. Experimental Salmonella enteritidis infections were examined in three different broiler outbred lines: the FC line, which had been selected for feed conversion efficiency; the R line, which had been selected for growth rate; and the C line, a commercially available line. The FC line had the highest mortality rate after intramuscular inoculation with 5 3 10 6 colony forming units (CFU) of S. enteritidis at 2 weeks of age (40% versus 21 and 20% in the other lines). However, at slaughter age, the number of birds carrying salmonella in caecal contents, and the concentration of salmonella in the caecal contents, was lowest in the FC line. The FC and R lines were compared by inoculation with doses ranging from 10 2 to 10 7 CFU S. enteritidis. At sublethal doses (10 5 CFU or less), the FC line carried significantly less salmonella in caecal contents and the rate of systemic infection was lower. The start of shedding was also delayed compared with the R line. At doses of 10 6 CFU S. enteritidis or higher, there were no differences in salmonella carriage between the lines, and the FC line showed higher mortality. In conclusion, resistance to mortality and resistance to the carriage of S. enteritidis do not necessarily coincide within lines, as the FC line showed high mortality but low carriage, both in survivors of high infection doses and in all birds at lower infection doses.