Shiga Toxin-Producing Escherichia coli Testing in New York 2011-2022 Reveals Increase in Non-O157 Identifications.

Shiga toxin-producing Escherichia coli (STEC) are an important cause of bacterial enteric infection. STEC strains cause serious human gastrointestinal disease, which may result in life-threatening complications such as hemolytic uremic syndrome. They have the potential to impact public health due to diagnostic challenges of identifying non-O157 strains in the clinical laboratory. The Wadsworth Center (WC), the public health laboratory of the New York State Department of Health, has isolated and identified non-O157 STEC for decades. A shift from initially available enzyme immunoassay testing to culture-independent diagnostic tests (CIDTs) has increased the uptake of testing at clinical microbiology laboratories. This testing change has resulted in an increased number of specimen submissions to WC. During a 12-year period between 2011 and 2022, WC received 5037 broths and/or stool specimens for STEC confirmation from clinical microbiology laboratories. Of these, 3992 were positive for Shiga toxin genes (stx1 and/or stx2) by real-time PCR. Furthermore, culture methods were utilized to isolate, identify, and characterize 2925 STEC from these primary specimens. Notably, WC observed a >200% increase in the number of STEC specimens received in 2021-2022 compared with 2011-2012 and an 18% increase in the number of non-O157 STEC identified using the same methodologies. During the past decade, the WC testing algorithm has been updated to manage the increase in specimens received, while also navigating the novel COVID-19 pandemic, which took priority over other testing for a period of time. This report summarizes updated methods for confirmation, surveillance, and outbreak detection of STEC and describes findings that may be related to our algorithm updates and the increased use of CIDTs, which is starting to elucidate the true incidence of non-O157 STEC.

Is the Medium Still the Message? Culture-Independent Diagnosis of Gastrointestinal Infections.

Infectious diarrhea is caused by a variety of pathogens, including viruses, bacteria, and parasitic organisms. Though the causative agent of diarrhea has historically been evaluated via stool cultures, recently, culture-independent diagnostic tests (CIDT) have been developed and utilized with increasing frequency. Current practice guidelines recommend their use as adjuncts to stool cultures for diagnosing acute and chronic diarrhea. The three principal CIDT are microscopy, enzyme-based immunoassays (EIAs), and molecular based polymerase chain reaction (PCR). This review explores the common causes of infectious diarrhea, the basics of stool culture, the diagnostic utility of these three culture-independent modalities, and the strengths and weaknesses of all currently available clinical techniques. It also outlines considerations for specific populations including returning travelers and those with inflammatory bowel disease.

Bovine tuberculosis in eastern Ethiopia: prevalence, risk factors and its public health importance.

BACKGROUND: Bovine tuberculosis is among the primary zoonotic disease caused by Mycobacterium bovis which has significant impact on the health of livestock and human. It has been significantly a cause for great economic loss in animal production. METHODOLOGY: A cross-sectional study was conducted from December 2014 to June 2016 on 315 cattle in selected areas of eastern Ethiopia, aiming to estimate the occurrence of bovine tuberculosis using comparative intradermal tuberculin skin test and assess cattle owners' awareness on its public health implication. Random sampling method was applied in order to select animals from farm/household and associated risk factors were recorded before purified protein derivative (PPD) injection. Forty three farm/household owners of tuberculin tested animals were interviewed using pre-tested structured questionnaires. RESULTS: The overall prevalence of bovine tuberculosis was 20.3% (n = 64) in dairy cattle at recommended cut off > 4 mm. From a total of 43 farms/households tested, 22 were positive; each farm exhibited at least one tuberculin positive reactor animal with a total herd level prevalence of 51.2%. The prevalence of bovine tuberculosis in individual animal level was significantly different (χ2 = 45.2; P-value = 0.000) in different sites with a higher prevalence (50%) in Dire Dawa. Farming system, herd size and other risk factors were significantly (p < 0.05) associated with bovine tuberculosis occurrence. Of the total interviewed farm owners, only 33% had the knowledge of or had heard about bovine tuberculosis and 23% respondents were aware of the zoonotic importance of the disease. More than 50% of the interviewees had shown their preference of raw milk consumption. Out of the total interviewed households, 3 (7%) farm workers had TB cases that had direct contact with the animals. CONCLUSION: The study showed bovine tuberculosis is highly prevalent. Associated risk factors contributed to the prevalence of the disease in cattle and its transmission. Moreover, the majority of cattle owners lack awareness about the disease and its public health significance. Awareness rising about the disease, its transmission and zoonotic implication is of great importance for reduction and control measures. Evidence of tuberculosis patient farm attendants calls also for further detail investigation.

Status of bovine tuberculosis and its zoonotic implications in Borana zone, Southern Ethiopia.

A cross-sectional study was carried out from April 2015 to June 2016 to estimate the status of bovine tuberculosis (BTB), assessment of community's current knowledge, and zoonotic importance on this disease in Borana zone, southern Ethiopia. In this study, comparative intradermal tuberculin (CIDT) test, structured questionnaires, and retrospective data were used, while the result indicated 3.8% prevalence at individual animal level with 5.6% (31/554) of doubtful reactors. Among related risk factors included, old animals were significantly infected by BTB than young one (χ 2 = 32.005, P = 0.001). Parity number again showed significant difference (χ 2 = 29.163, P = 0.001) where animals with many parity were more reactive to conducted test than few parity numbers. Animals born in the breeding center managed under semi-intensive production system were more infected (χ 2 = 10.795, P = 0.029) than those brought from outside of the center. Questionnaire survey in this study indicated that out of 130 interviewed respondents, only 30% pastoralists knew what BTB mean; whereas the level of individual knowledge from interviewed showed about 72.3% of respondents had poor understanding of BTB and only about 11.5% of them knew its zoonotic importance. Meat eating habit of communities in the area were culturally inhabited to eat cooked meat and only 12.3% (16/130) of respondents gave response on habit of eating both raw and cooked meat. Milk drinking habit of pastoralist in the area showed about 79.2% drunk raw milk and the rest 20.8% used both raw and boiled milk. A retrospective data from Yabello Hospital indicated that current prevalence of human TB as 38.79% and showing the disease was highly increasing from year to year in the study area. This implies a great importance of human tuberculosis and its future concern in Borana zone. From this, there should be detail awareness of communities on BTB, its zoonotic importance, and the need of further investigation to develop control and prevention strategies according to the pastoral settings.

Evidence of False Positivity for Vibrio Species Tested by Gastrointestinal Multiplex PCR Panels, Minnesota, 2016-2018.

BACKGROUND: Syndromic gastrointestinal multiplex polymerase chain reaction (PCR) panels (GMPPs) are used by an increasing number of clinical laboratories to identify enteric pathogens. Vibrio species are included on GMPPs, but because of the low prevalence of vibriosis, performance characteristics for these panels have been difficult to measure. METHODS: All Vibrio spp. cases identified by GMPPs in Minnesota during 2016-2018 (n = 100) were assessed to identify differences between culture-confirmed cases and those that were PCR-positive only. RESULTS: Overall, 47% of cases had Vibrio species recovered by culture. Two GMPPs were used in Minnesota, Verigene EPT and FilmArray GIP, and the recovery rate of Vibrio spp. was significantly different between these platforms (Verigene EPT 63%, compared with FilmArray GIP 28%). No distinct seasonality was identified among GMPP-positive, culture-negative cases, whereas culture-confirmed case incidence peaked during July and August. Among cases with no other pathogen detected by the GMPP, confirmed cases reported a lower rate of bloody diarrhea (odds ratio [OR], 0.7; P = .004) and were less likely to have a symptom duration >14 days (OR, 0.3; P = .04). Confirmed cases were also more likely to include reports of consuming food items typically associated with Vibrio spp. infection or to have another likely source of infection (eg, international travel or contact with an untreated body of fresh or salt water or marine life; OR, 9.6; P = .001). CONCLUSIONS: The combined findings indicate that cases identified by GMPP that did not have culture confirmation were less likely to include symptoms or exposures consistent with vibriosis. These findings emphasize the need for improvements to testing platform specificity and the importance of combining clinical and exposure information when diagnosing an infection. This study underscores the importance of maintaining the ability to culture Vibrio species to aid in accurate diagnoses.

Association of Ct Values from Real-Time PCR with Culture in Microbiological Clearance Samples for Shiga Toxin-Producing Escherichia coli (STEC).

Shiga toxin-producing Escherichia coli (STEC) are associated with acute gastroenteritis worldwide, which induces a high economic burden on both healthcare and individuals. Culture-independent diagnostic tests (CIDT) in frontline microbiology laboratories have been implemented in Alberta since 2019. The objectives of this study were to determine the association between gene detection and culture positivity over time using STEC microbiological clearance samples and also to establish the frequency of specimen submission. Both stx genes' amplification by real-time PCR was performed with DNA extracted from stool samples using the easyMAG system. Stools were inoculated onto chromogenic agar for culture. An association between gene detection and culture positivity was found to be independent of which stx gene was present. CIDT can provide rapid reporting with less hands-on time and technical expertise. However, culture is still important for surveillance and early cluster detection. In addition, stool submissions could be reduced from daily to every 3-5 days until a sample is negative by culture.