RESUMEN
Coccidiosis represents a major driver in the economic performance of poultry operations, as coccidia control is expensive, and infections can result in increased feed conversion ratios, uneven growth rates, increased co-morbidities with pathogens such as Salmonella, and mortality within flocks. Shifts in broiler production to antibiotic-free strategies, increased attention on pre-harvest food safety, and growing incidence of anti-coccidial drug resistance has created a need for increased understanding of interventional efficacy and methods of coccidia control. Conventional methods to quantify coccidia oocysts in fecal samples involve manual microscopy processes that are time and labor intensive and subject to operator error, limiting their use as a diagnostic and monitoring tool in animal parasite control. To address the need for a high-throughput, robust, and reliable method to enumerate coccidia oocysts from poultry fecal samples, a novel diagnostic tool was developed. Utilizing the PIPER instrument and MagDrive technology, the diagnostic eliminates the requirement for extensive training and manual counting which currently limits the application of conventional microscopic methods of oocysts per gram (OPG) measurement. Automated microscopy to identify and count oocysts and report OPG simplifies analysis and removes potential sources of operator error. Morphometric analysis on identified oocysts allows for the oocyst counts to be separated into 3 size categories, which were shown to discriminate the 3 most common Eimeria species in commercial broilers, E. acervulina, E. tenella, and E. maxima. For 75% of the samples tested, the counts obtained by the PIPER and hemocytometer methods were within 2-fold of each other. Additionally, the PIPER method showed less variability than the hemocytometer counting method when OPG levels were below 100,000. By automated identification and counting of oocysts from 12 individual fecal samples in less than one hour, this tool could enable routine, noninvasive diagnostic monitoring of coccidia in poultry operations. This approach can generate large, uniform, and accurate data sets that create new opportunities for understanding the epidemiology and economics of coccidia infections and interventional efficacy.
Asunto(s)
Coccidiosis , Eimeria , Parasitología , Enfermedades de las Aves de Corral , Animales , Pollos/parasitología , Coccidiosis/diagnóstico , Coccidiosis/veterinaria , Coccidiosis/epidemiología , Heces/parasitología , Oocistos/citología , Enfermedades de las Aves de Corral/diagnóstico , Enfermedades de las Aves de Corral/parasitología , Parasitología/instrumentación , Parasitología/métodos , Reproducibilidad de los ResultadosRESUMEN
One of the primary benefits associated with dietary resistant starch (RS) is the production of butyrate by the gut microbiome during fermentation of this fiber in the large intestine. The ability to degrade RS is a relatively rare trait among microbes in the gut, seemingly confined to only a few species, none of which are butyrate producing organisms. Thus, production of butyrate during RS fermentation requires a network of interactions between RS degraders and butyrate producers. This is further complicated by the fact that there are multiple types of RS that differ in their structural properties and impacts on the microbiome. Human dietary intervention trials with RS have shown increases in fecal butyrate levels at the population level but with individual to individual differences. This suggests that interindividual differences in microbiome composition dictate butyrate response, but the factors driving this are still unknown. Furthermore, it is unknown whether a lack of increase in butyrate production upon supplementation with one RS is indicative of a lack of butyrate production with any RS. To shed some light on these issues we have undertaken an in vitro fermentation approach in an attempt to mimic RS fermentation in the colon. Fecal samples from 10 individuals were used as the inoculum for fermentation with 10 different starch sources. Butyrate production was heterogeneous across both fecal inocula and starch source, suggesting that a given microbiome is best suited to produce butyrate only from a subset of RS sources that differs between individuals. Interestingly, neither the total amount of RS degraders nor butyrate producers seemed to be limiting for any individual, rather the membership of these sub-populations was more important. While none of the RS degrading organisms were correlated with butyrate levels, Ruminococcus bromii was strongly positively correlated with many of the most important butyrate producers in the gut, though total butyrate production was strongly influenced by factors such as pH and lactate levels. Together these results suggest that the membership of the RS degrader and butyrate producer communities rather than their abundances determine the RS sources that will increase butyrate levels for a given microbiome.
RESUMEN
ABSTRACT: Salmonella enterica subsp. enterica serovar Newport is a bacterial foodborne pathogen isolated from several environmental reservoirs on the Delmarva Peninsula and has been associated with several produce-related outbreaks. However, little is known about specific interactions between Salmonella Newport and soil amendments used as fertilizers. The purpose of this study was to determine Salmonella Newport persistence and resuscitation in raw poultry litter (PLR), a common biological soil amendment, and in soils containing poultry litter-based (heat-treated poultry pellets [HTPP]) or chemical fertilizer (urea [U]) amendments to provide equivalent levels of nitrogen to the soil. Inoculated samples were stored in a growth chamber and irrigated regularly over 4 weeks. Soil samples were collected every week for 4 weeks to determine moisture content and surviving Salmonella Newport populations (log CFU per gram dry weight). Data were analyzed by using a one-way analysis of variance and Student's t test. The PLR supported significantly higher (5.07 log CFU/g dry weight [gdw]) populations of Salmonella Newport than HTPP only (1.70 log CFU/gdw). However, PLR-amended (PRLA) soil (2.5 log CFU/gdw) samples had significantly (P < 0.05) lower Salmonella Newport populations compared with HTPP-amended (4.5 log CFU/gdw) and U-amended (4.0 log CFU/gdw) soil samples. The effect of irrigation on Salmonella Newport population levels in PRLA soils was significant, and in a comparative study, the overall increase in the pathogen levels in U-amended soil (mean = 1.12 log CFU/gdw) was significantly greater than that in PLRA soil (mean = 0.54 log CFU/gdw), whereas that in HTPP-amended soil (0.80 log CFU/gdw) was not significantly different from PLRA soil.