Diagnosis of sub-clinical coccidiosis in fast growing broiler chickens by MicroRNA profiling.

Coccidiosis in broiler chickens, caused by infection with Eimeria spp. remains one of the most economically important production diseases. Development of a genetic biomarker panel of sub-clinical infection would be an important biological tool for the management of broiler flocks. We analysed expression of MicroRNAs (miRNAs) to determine the potential for these in diagnosing coccidiosis in broiler flocks. miRNA expression, in the ilea of Ross 308 broilers, was compared between chickens naturally clinically or sub-clinically infected with Eimeria maxima and Eimeria acervulina using NextSeq 500 sequencing. 50 miRNAs with greatest coefficient of variance were determined and principal component analysis showed that these miRNAs clustered within the clinical and sub-clinical groups much more closely than uninfected controls. Following false detection rate analysis and quantitative PCR we validated 3 miRNAs; Gallus gallus (gga)-miR-122-5p, gga-miR-205b and gga-miR-144-3p, which may be used to diagnose sub-clinical coccidiosis.

Differential immune response to Eimeria maxima infection in fast- and slow-growing broiler genotypes.

Very little has been reported comparing resistance to coccidiosis in fast or slow growing broilers, the latter of which are becoming more prevalent in the broiler industry. We examined mRNA expression in the intestines of fast and slow growing broilers following Eimeria infection. We show that by day 13 post-infection (d pi) with 2500 or 7000 oocysts of Eimeria maxima, slower-growing (Ranger Classic) broilers significantly (P < 0.01) upregulated expression of proinflammatory cyclooxygenase genes (LTB4DH, PTSG1 and PTSG2) above that detected in fast growing (Ross 308) broilers. Expression of CD8α mRNA was downregulated in Ross 308 at day 6d pi with either 2500 or 7000 oocysts of E maxima (P < 0.05), compared to uninfected controls, but was not differentially expressed in Ranger Classic. CD4 genes were not differentially expressed in either chicken line infected with either infectious oocyst dose at d6 pi, compared to uninfected controls. However, at d13 pi, CD4 expression was significantly upregulated in both chicken lines infected with either infectious oocyst dose, compared to uninfected controls (P < 0.05) but this was significantly greater in Ranger Classic broilers compared to Ross 308 (P < 0.05). At d13 pi, expression of CD3 chains (required for T lymphocyte activation) was significantly increased in Ranger Classic compared to Ross 308, infected with either oocyst dose (P < 0.05-0.01). Expression of IL-2 and IL-15 mRNA, required for T lymphocyte proliferation was also significantly upregulated, or maintained longer, in Ranger Classic broilers compared to Ross 308. These differences in immune response to E maxima corresponded with a reduction in E maxima genome detected in the intestines of Ranger Classic compared to Ross 308.

Differential gene response to coccidiosis in modern fast growing and slow growing broiler genotypes.

We analysed intestinal tissues from groups of fast growing (Ross 308) broilers with natural or experimental coccidiosis, by genomic microarray. We identified genes that were differentially expressed (DE) in all groups and analysed expression of a panel of these, by qPCR, in Ross 308 and slow growing (Ranger classic) broilers, infected with 2500 or 7000 oocysts of Eimeria maxima for 6 or 13 days post-infection (dpi). Four genes (ADD3, MLLT10, NAV2 and PLXNA2) were upregulated (P <0.05) in Ross 308 but were not DE in Ranger Classic at 6 dpi with 2500 oocysts. Six genes (PTPRF, NCOR1, CSF3, SGK1, CROR and CD1B) were upregulated (P <0.05) in both Ross 308 and Ranger Classic infected with 2500 oocysts at 6 dpi but were not DE at 6 dpi with 7000 oocysts. At 13 dpi with 7000 oocysts, NAV2 and NCOR1 were upregulated in Ross 308 (P <0.05) and PTPRF was upregulated in both genotypes (P <0.05). DE of immune genes within the biomarker panel also occurred, with CSF3 upregulated in both genotypes infected with 2500 oocysts at 6 dpi and in Ranger Classic infected with 7000 oocysts, at 6 and 13 dpi (P <0.05). IL-22 was down-regulated in Ranger Classic infected with 2500 or 7000 oocysts at 6 dpi (P <0.05) but upregulated in both genotypes at 13 dpi (P <0.05). CD72 was down-regulated in Ranger Classic infected with 2500 oocysts at 6 dpi and with 7000 oocysts at 6 and 13 dpi (P <0.05). CD72 was upregulated in Ross 308 infected with 2500 oocysts at 6 dpi but was down-regulated following infection with 7000 oocysts at 13 dpi (P <0.05). In conclusion, differential gene expression occurs in fast and slow growing broiler genotypes with coccidiosis. In addition, we highlight a potential genetic biomarker panel for early diagnosis of coccidiosis.

Risk factors for contact lens complications in US clinical practices.

PURPOSE: To determine significant risk factors for any inflammatory and infectious events with soft contact lenses (SCL) in a large retrospective clinical chart review. METHODS: Charts of patients who presented for SCL care from October 2005 through March 2006 were reviewed and observed for a potential of at least 2 years. Charts from those with office visits involving an event-requiring pharmacologic treatment and/or interruption of SCL wear were scanned and later adjudicated by a masked panel. Significant factors from a univariate analysis were included in a multivariate analysis for all events and subcategories of events separately. Overnight wear was not consistently recorded and was not analyzed. RESULTS: Charts from 1276 SCL wearers comprised 4120 visits and 1454 years of SCL wear (2908 eye/yr) and included 306 events of interest in 228 patients. In a multivariate analysis, age <25 years was significantly associated with presenting any event, inflammatory events, and infectious events that may or may not be CL-related [incidence rate ratio (IRR) = 1.3; 95% CI, 1.0 to 1.7; 2.6X, 1.5 to 4.6; and 2.0X, 1.2 to 3.3, respectively]. Ametropia >5.00 D increased risk of any event (IRR = 1.5; 1.2 to 1.9) and for other infectious events (IRR = 1.9; 1.2 to 3.2). Use of daily disposable lenses associated with lid irritation (IRR = 4.5; 2.1 to 9.8) but was not significantly associated with any other type of events. New and hydrogel lens wearers had a lower incidence of all event types (IRR = 0.07; 0.01 to 0.46 and 0.77; 0.59 to 0.99, respectively). CONCLUSIONS: Eighty-two percent of these SCL wearers did not present with any complications during the observation period >2 years. The risk factors for inflammatory and infectious events among SCL wearers in clinical practice are similar to those reported in prospective clinical trials. High ametropia and age <25 years are the risk factors that impact the most types of events.