Rodents play a major role in the transmission and maintenance of Salmonella contamination cycles in poultry facilities. However, very limited field data are available regarding the transmission routes, infection cycle, and shedding patterns of Salmonella by naturally infected wild rodents from commercial layer farms. In this study, a total of 128 resident wild roof rats (Rattus ratus) were captured from a Salmonella-contaminated layer facility. All roof rats were divided into 51 laboratory cages, and weekly monitoring of Salmonella fecal shedding patterns was conducted for 53 wk. Seven roof rats from cages that were observed to frequently shed Salmonella were isolated in individual cages, and daily Salmonella monitoring was performed for 35 days. At the end of monitoring, each roof rat was euthanatized, and isolation of Salmonella from different organs was performed. Results of weekly monitoring of Salmonella showed that 21 of 51 cages (41.2%) were positive for Salmonella Infantis, while two cages (3.92%) were positive for Salmonella Enteritidis. Moreover, 11 cages were positive for Salmonella for at least two sampling weeks. Isolation of Salmonella from fecal droppings was mainly observed during the first 12 wk of captivity. The longest interval between two Salmonella-positive fecal dropping was 24 wk. In the daily Salmonella monitoring, only Salmonella Infantis was isolated from fecal droppings, in which the highest number of Salmonella Infantis organisms per fecal dropping was at 1 x 10(8) colony-forming units (cfu), while the lowest measured quantity was 1 x 10(3) cfu. It was noted that the frequency of Salmonella shedding in fecal droppings appeared to have a linear correlation (r = 0.85) with the number of Salmonella organisms (cfu) per fecal pellet (P < 0.05). Moreover, pulsed-field gel electrophoresis analysis of Salmonella Infantis isolates revealed a single identical pulsed-field pattern. Salmonella Enteritidis isolates from fecal droppings and internal organs also generated a single identical pulsed-field pattern. Interestingly, Salmonella Infantis was not isolated from any of the organs examined, while Salmonella Enteritidis was isolated from the spleen and liver of one roof rat. These results may indicate that wild roof rats could persistently carry Salmonella and contaminate commercial poultry facilities through intermittent fecal shedding. Moreover, Salmonella Enteritidis in wild roof rats appears to be more of a systemic infection, in which isolation is most likely to occur in internal organs, whereas Salmonella Infantis is more likely an enteric type of infection, in which isolation is most likely to occur in the intestinal contents. It is very plausible that layer chickens could become infected with Salmonella through ingestion of Salmonella-positive fecal droppings or feeds contaminated with these fecal droppings from infected resident roof rats. This is likely one of the major reasons why layer houses can be persistently infected by Salmonella even if the facilities are thoroughly cleaned and ...

BackgroundNewcastle Disease (ND) is a highly contagious and economically devastating disease of poultry. At present, limited molecular epidemiological data are available regarding the causes of ND outbreaks in vaccinated commercial poultry farms. Knowing the genomic characteristics of Newcastle disease virus (NDV) infecting commercial poultry operations in spite of vaccination might give important insights on the infection dynamics of these viruses. In addition, molecular analyses at the subgenotype level and studies on the relationship of Japanese NDVs with other isolates from around the world are lacking. Therefore, in the present study, a molecular epidemiological investigation was conducted to characterize nine NDVs isolated from vaccinated commercial poultry flocks in five different Prefectures in non-epidemic areas of Japan between 1969 and 2002.MethodsNucleotide sequencing and phylogenetic studies were performed to characterize the complete fusion (F)-protein gene, 3-prime end of the nucleoprotein (NP)-gene and 5-prime end of the RNA dependent RNA polymerase (L)-gene. Sequence data were compared with 180 NDV strains from GenBank representing different NDV genotypes and subgenotypes from different regions of the world at different time periods. Deduced amino acids were analyzed for homologies, recombination and mutation. Recombination events were estimated using Recombination Detection Program (RDP) version 3.44. Phylogenetic trees were constructed to determine evolutionary relationships among strains.ResultsMean death time (MDT: 48-56 hr), Intracerebral Pathogenicity Index (ICPI: 1.7-1.9) and deduced amino acid sequences of the F0 proteolytic cleavage site (112RRQKR116) revealed that all nine field isolates were velogenic. Phylogenetic analysis showed that these isolates could be classified into two genetic lineages and three sublineages namely genotypes VIa (lineage 4a), VId (lineage 4d) and VIId (lineage 5d). No recombination events were observed but a point mutation in one of the neutralizing epitope of the F-protein was identified in the field isolates from Japan.ConclusionsAll field isolates from vaccinated commercial poultry in non-epidemic areas of Japan were part of much bigger outbreaks in provinces and regions and, in some cases, continents. In general, four ND panzootics occurred in Japan and that these outbreaks were mostly characterized by co-circulation of genetically distinct virus lineages due to involvements of infected wild birds. The point mutation identified in the field isolates from Japan may be due to escape from vaccine pressure. The identification of such mutation may be useful for future site-directed mutagenesis to understand the dynamics of NDV infection in vaccinated chickens.

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In total, 40 commercial layer farms and 32 replacement pullet farms with a combined population of 7.5 million adult layers and 6.6 million replacement pullets from six prefectures in eastern Japan were investigated for Salmonella Senftenberg contamination. We randomly collected 17,956 environmental samples, 5816 feed samples, and 218,470 egg samples from commercial layer farms; and 427 feed samples and 2896 environmental samples from replacement pullet farms. We monitored all samples for Salmonella. Samples were primarily enriched in Hajna tetrathinoate broth for 24 hr at 37 C followed by incubation in desoxycholate hydrogen sulfide lactose agar for 18 hr at 37 C. Salmonella colonies were confirmed and identified by biochemical tests and serotyped using Salmonella O and H antigens. We recorded 171 environmental samples (0.95%) and 10 feed samples (0.17%) that were positive for Salmonella spp. in which 36 environmental samples (0.20%) and six feed samples (0.10%) were identified as Salmonella Senftenberg. All Salmonella Senftenberg strains were isolated from nine replacement pullet farms. No Salmonella Senftenberg strains were isolated from adult layer farms and from eggs. Pulse field gel electrophoresis of BlnI-digested chromosomal DNA of 19 Salmonella Senftenberg isolates from feeds and environmental samples yielded a single identical DNA pattern. Traceback information showed that all positive feed samples were from a single feed source. Timeline studies showed that Salmonella Senftenberg contamination occurred first mostly in the feeds and then spread to the environment and other farms. This study demonstrated that the prevalence of Salmonella Senftenberg contamination in commercial layer facilities in eastern Japan is very low. Moreover, feed contamination played a major role in the epizootiology and spread of this pathogen in commercial poultry flocks. Given the resilient and persistent nature of this particular Salmonella serotype, routine monitoring and strict quality control measures at the feed level are recommended to prevent the colonization of poultry facilities with Salmonella Senftenberg that may lead to future outbreaks.