An apple a day: an outbreak of cryptosporidiosis in Norway associated with self-pressed apple juice.

In the autumn of 2018, an outbreak of cryptosporidiosis affected adult employees from the same company in Western Norway. The organism was Cryptosporidium parvum, GP60 subtype IIaA14G1R1. All those infected had drunk from the same container of self-pressed apple juice. Incubation period (1 week) and clinical signs were similar among those infected, although some experienced a more prolonged duration of symptoms (up to 2-3 weeks) than others. The infections resulted after consumption from only one of 40 containers of juice and not from any of the other containers. It seems that although Cryptosporidium oocysts were detected in a sample from another container, the contamination did not affect the whole batch. This is perhaps indicative of a restricted contamination event, either from contaminated ground in the orchard, or during collection of the fruit, or during processing. Although outbreaks of food-borne cryptosporidiosis have previously been associated with consumption of contaminated apple juice, most of the more recent outbreaks of food-borne cryptosporidiosis have been associated with salad vegetables or herbs. This outbreak, the first outside USA reported to be associated with apple juice, is a timely reminder that such juice is a suitable transmission vehicle for Cryptosporidium oocysts, and that appropriate hygienic measures are essential in the production of such juice, including artisanal (non-commercial) production.

Apparent absence of Giardia infections among children under 5-years of age with acute watery diarrhoea in Abakaliki, Nigeria.

Although the impact of diarrhoeal disease on paediatric health in Nigeria has decreased in recent years, it remains an important cause of morbidity and mortality in children under 5 years. Rotavirus is recognised as an important aetiological agent, but information on the contribution of intestinal protozoa to watery diarrhoea in this age group in Nigeria is scarce. In this cross-sectional study, faecal samples from children admitted to healthcare centres in Abakaliki, Nigeria with acute watery diarrhoea (N = 199) and faecal samples from age-matched controls (N = 37) were examined for Cryptosporidium and Giardia using immunofluorescent antibody testing and molecular methods. Cryptosporidium was identified in 13 case samples (6.5%) and no control samples. For three samples, molecular characterisation indicated C. hominis, GP60 subtypes IaA30R3, IaA14R3 and IdA11. Giardia was not detected in any samples. This contrast in prevalence between the two intestinal protozoa may reflect their variable epidemiologies and probably differing routes of infection. Given that these two parasitic infections are often bracketed together, it is key to realise that they not only have differing clinical spectra but also that the importance of each parasite is not the same in different age groups and/or settings.

Changes in the seroprevalence of cysticercosis in suspected patients in Chandigarh, India between 1998 and 2014: analysis of 17 years of data.

Changes in seroprevalence of cysticercosis diagnosed in Chandigarh, India between 1998 and 2014 were investigated by extraction and analysis of data from records held at the Postgraduate Institute of Medical Education and Research in Chandigarh. Among the total number of samples for which cysticercosis had been suspected during this period (N = 9650), 1716 (17·8%) were seropositive. Adults were more likely to be seropositive than children, and women were more likely to be seropositive than men. In addition to there being fewer patients with suspicion of cysticercosis over the data analysis period, the proportion of patients seropositive also reduced significantly. Despite these reductions, which are probably associated with improved infrastructure and sanitation within Chandigarh, and despite meat consumption being relatively rare in this area, the extent of cysticercosis in this population remains problematic. Further efforts should be made to reduce transmission of this infection, with particular emphasis on women. Such efforts should follow the One Health concept, and involve medical efforts (including diagnosis and treatment of T. solium tapeworm carriers), veterinary efforts directed towards meat inspection and prevention of infection of pigs, and environmental health and sanitation engineers (to minimize environmental contamination with human waste).

Symptomatic and asymptomatic secondary transmission of Cryptosporidium parvum following two related outbreaks in schoolchildren.

Two related outbreaks (in 2009 and 2012) of cryptosporidiosis in Norwegian schoolchildren during a stay at a remote holiday farm provided us with a natural experiment to investigate possible secondary transmission of Cryptosporidium parvum IIa A19G1R1. After the children had returned home, clinical data and stool samples were obtained from their household contacts. Samples were investigated for the presence of Cryptosporidium oocysts by immunofluorescence antibody test. We found both asymptomatic and symptomatic infections, which are likely to have been secondary transmission. Laboratory-confirmed transmission rate was 17% [4/23, 95% confidence interval (CI) 7·0-37·1] in the 2009 outbreak, and 0% (95% CI 0-16·8) in the 2012 outbreak. Using a clinical definition, the probable secondary transmission rate in the 2012 outbreak was 8% (7/83, 95% CI 4·1-16·4). These findings highlight the importance of hygienic and public health measures during outbreaks or individual cases of cryptosporidiosis. We discuss our findings in light of previous studies reporting varying secondary transmission rates of Cryptosporidium spp.

Second outbreak of infection with a rare Cryptosporidium parvum genotype in schoolchildren associated with contact with lambs/goat kids at a holiday farm in Norway.

In March 2012, a second outbreak of Cryptosporidium parvum affected children following a stay at a holiday farm in Norway; the first outbreak occurred in 2009. We studied a cohort of 145 schoolchildren who had visited the farm, of which 40 (28%) were cases. Cryptosporidium oocysts were detected in faecal samples from humans, goat kids and lambs. Molecular studies revealed C. parvum subtype IIa A19G1R1 in all samples including human samples from the 2009 outbreak. A dose-response relationship was found between the number of optional sessions with animals and illness, increasing from two sessions [risk ratio (RR) 2·7, 95% confidence interval (CI) 0·6-11·5] to six sessions (RR 8·0, 95% CI 1·7-37·7). The occurrence of two outbreaks 3 years apart, with the same subtype of C. parvum, suggests that the parasite is established in the farm's environment. We recommend greater emphasis on hand hygiene and routines related to animal contact.

Reproductive disorders in relation to Neospora caninum, Brucella spp. and bovine viral diarrhoea virus serostatus in breeding and dairy farms of central and southern Ethiopia.

Abortion and stillbirth are important reproductive disorders in the dairy industry and are often caused by infectious agents. This study investigated whether bovine viral diarrhoea virus (BVDV), Brucella spp., and Neospora caninum are associated with abortion and/or stillbirth in dairy cattle in Ethiopia. Dairy cattle from 99 farms were categorized as cases (n=134) or controls (n=268) according to reproductive data. Blood samples were screened for antibodies for these infectious agents. The overall proportion of cattle that were seropositive for BVDV, Brucella spp., and N. caninum was 11∙7%, 3∙2%, and 17∙2%, respectively. Seropositivity for BVDV and Brucella spp. was similar for cases and controls, but significantly more cases were seropositive for N. caninum (29∙8%) than controls (10∙8%). This is the first report demonstrating N. caninum is common in dairy cattle in Ethiopia, and is probably a greater impediment to reproductive success in Ethiopian dairy farms than either BVDV or Brucella spp.

Evaluation of the performance of a lateral flow device for quantitative detection of anti-SARS-CoV-2 IgG.

INTRODUCTION: The AbC-19™ lateral flow immunoassay (LFIA) performance was evaluated on plasma samples from a SARS-CoV-2 vaccination cohort, WHO international standards for anti-SARS-CoV-2 IgG (human), individuals ≥2 weeks from infection of RT-PCR confirmed SARS-CoV-2 genetic variants, as well as microorganism serology. METHODS: Pre-vaccination to three weeks post-booster samples were collected from a cohort of 111 patients (including clinically extremely vulnerable patients) from Northern Ireland. All patients received Oxford-AstraZeneca COVID-19 vaccination for the first and second dose, and Pfizer-BioNTech for the third (first booster). WHO international standards, 15 samples from 2 variants of concern (Delta and Omicron) and cross-reactivity with plasma samples from other microorganism infections were also assessed on AbC-19™. RESULTS: All 80 (100%) participants sampled post-booster had high positive IgG responses, compared to 38/95 (40%) participants at 6 months post-first vaccination. WHO standard results correlated with information from corresponding biological data sheets, and antibodies to all genetic variants were detected by LFIA. No cross-reactivity was found with exception of one (of five) Dengue virus samples. CONCLUSION: These findings suggest BNT162b2 booster vaccination enhanced humoral immunity to SARS-CoV-2 from pre-booster levels, and that this antibody response was detectable by the LFIA. In combination with cross-reactivity, standards and genetic variant results would suggest LFIA may be a cost-effective measure to assess SARS-CoV-2 antibody status.

Ready-to-eat salads and berry fruits purchased in Italy contaminated by Cryptosporidium spp., Giardia duodenalis, and Entamoeba histolytica.

Ready-to-eat (RTE) salads and berries are increasingly consumed in industrialized countries. These products can be contaminated by pathogenic parasites that have been responsible for foodborne outbreaks worldwide. In Italy, there are few data on contamination of RTE salads and berries with parasite transmission stages and this requires more-in-depth investigations. To estimate the prevalence of contamination with Cryptosporidium spp. and Giardia duodenalis in these fresh products, a total of 324 packages of local RTE mixed salads - belonging to three different industrial brands - and 324 packages of berries - blueberries from Peru, blackberries from Mexico, raspberries from Italy - were bought from supermarkets located in the Provinces of Bari and Foggia, Apulia, Italy. A pool size of nine packages was chosen and a total of 72 pools were processed in the whole year. After washing, the pellets were examined by microscopy (FLOTAC) and tested using conventional simplex PCR, targeting Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp., and sequencing. Several Cryptosporidium species and Giardia duodenalis assemblages, some of which are of potential zoonotic relevance, as well as Entamoeba spp., were identified in both matrices. By microscopy, Giardia-like cysts in local raspberries and Entamoeba-like cysts in imported blueberries were detected. Giardia duodenalis (Assemblages A, B and E) and Entamoeba histolytica were molecularly confirmed with overall prevalences of 4.6% (95% C.I. 3.0-6.8) and 1% (95% C.I. 0.3-2.1), respectively. Molecular methods identified Cryptosporidium ryanae, Cryptosporidium bovis, Cryptosporidium xiaoi, and Cryptosporidium ubiquitum in both matrices, with a prevalence of 5.1% (95% C.I. 3.3-7.3). A distinct seasonality in prevalence was observed for G. duodenalis, with most positives occurring in spring, whereas Cryptosporidium showed no significant seasonal variations. These results highlight that inadequate management of fresh produce, both locally produced and imported, along the food chain may have the potential for consequences on human health.

Unexpected results from large-scale cryptosporidiosis screening study in calves in Tanzania.

A study was undertaken to investigate Cryptosporidium infection in crossbreed dairy calves in two districts in Tanzania. A total of 943 fecal samples from 601 dairy calves were included in the study, with calves from both smallholder dairy farms and from large-scale and medium-scale dairy farms. The modified Ziehl-Neelsen (mZN) technique was used to examine 710 samples, and 13 of these were considered to be positive for Cryptosporidium. These 13 samples considered positive by mZN, along with the remaining 233 samples, were analysed by immunofluorescent antibody test (IFAT). Of these 246 samples examined by IFAT, 15 samples, 10 of which were considered positive by mZN, were also examined by the auramine phenol technique, and 5 samples, all of which were considered positive by mZN, were analysed by PCR. The results from the IFAT, auramine phenol and PCR analyses demonstrated that none of the samples contained Cryptosporidium oocysts, indicating that, cryptosporidiosis is currently not a problem in dairy calves in these regions of Tanzania. These unexpected results are discussed with respect to other reports on cryptosporidiosis in calves that suggest that this parasite is a serious calf disease globally, and particularly in relation to studies from Tanzania. We suggest that results from studies of cattle in Tanzania, in which mZN has been used as the sole analytical method, should be treated with caution.

Strengthening of district mental health services in Gauteng Province, South Africa.

In response to the Life Esidimeni tragedy, the Gauteng Department of Health established a task team to advise on the implementation of the Health Ombud's recommendations and to develop a mental health recovery plan. Consistent with international human rights and South African legislation and policy, the plan focused on making mental healthcare more accessible, incorporating a strategy to strengthen district mental health services to deliver community-based care for people with any type and severity of mental illness. The strategy included an organogram with three new human resource teams integrated into the district health system: a district specialist mental health team to develop a public mental health approach, a clinical community psychiatry team for service delivery, and a team to support non-governmental organisation governance. This article discusses the strategy in terms of guiding policies and legislation, the roles and responsibilities of the various teams in the proposed organogram, and its sustainability.

Mental health of healthcare workers during the COVID-19 outbreak: A rapid scoping review to inform provincial guidelines in South Africa.

COVID-19 is prevalent in sub-Saharan Africa. The healthcare response to the pandemic depends upon a mentally and physically healthy workforce. Infectious disease outbreaks cause high psychosocial stress among healthcare workers, which may impact negatively on workplace functioning. To understand which mental health conditions may occur and which interventions could be considered, we conducted a rapid scoping review. Using a 2018 systematic review as the starting point, PubMed, Cochrane, Web of Science and MEDLINE databases were searched for any type of evidence published in English between 2014 and 2020 on mental health of healthcare workers exposed to infectious disease outbreaks; 19 primary studies and 13 opinion pieces were included. Depression, anxiety, post-traumatic stress, and other mental health conditions were noted among healthcare workers exposed to COVID-19 and other outbreaks. Although no effectiveness studies were identified, certain proposed interventions may be implemented by healthcare leaders. Further research is recommended.

Giardia cysts in sewage influent in Bergen, Norway 15-23 months after an extensive waterborne outbreak of giardiasis.

AIMS: In autumn/winter 2004, a large outbreak of waterborne giardiasis occurred in Bergen, Norway. Over 1 year later, the concentrations and genotypes of Giardia cysts occurring in sewage influent were studied to investigate the impact of the outbreak event on Giardia infections in the community. METHODS AND RESULTS: Sewage influent samples from four sewage treatment works (STW) serving Bergen were analysed for Giardia cysts on four occasions between 15 and 23 months after the outbreak. Cysts genotypes were investigated at one to three genes. Data from influent analysis from one of the STW before the outbreak, and from patient faecal samples analysed during the outbreak, provided baseline comparative data. Relatively high concentrations of Giardia cysts of diverse genotypes, both from Assemblages A and B, were detected at all STW. CONCLUSIONS: Comparison of data suggests that although Giardia cyst concentrations in sewage influent returned to pre-outbreak levels within 18 months after the outbreak peak, the genetic composition of the isolates remained significantly influenced by the Assemblage B isolate associated with the outbreak. SIGNIFICANCE AND IMPACT OF THE STUDY: Genotypes associated with an extensive outbreak of giardiasis continued to occur in Giardia infections in Bergen's population many months after the outbreak was considered to be over.

Development and use of a pepsin digestion method for analysis of shellfish for Cryptosporidium oocysts and Giardia cysts.

Investigation of shellfish for Cryptosporidium oocysts and Giardia cysts is of public health interest because shellfish may concentrate these pathogens in their bodies, and because shellfish are frequently eaten raw or lightly cooked. To date, the methods used for the analysis of shellfish for these parasites are based on those originally designed for water concentrates or fecal samples; the reported recovery efficiencies are frequently relatively low and the amount of sample examined is small. Here, we describe the development and use of a pepsin digestion method for analyzing shellfish samples for these parasites. The conditions of the isolation method did not affect subsequent parasite detection by immunofluorescent antibody test, and allowed examination of 3-g samples of shellfish homogenate, with recovery efficiencies from blue mussel homogenates of between 70 and 80%, and similar recoveries from horse mussel and oyster homogenates. Although exposure of the parasites to the conditions used in the technique affected their viability, as assessed by vital dyes, the maximum reduction in viability after 1-h incubation in digestion solution was 20%. In a preliminary survey of shellfish collected from the Norwegian coast, Cryptosporidium oocysts were detected in blue mussel homogenates in 6 (43%) of 14 batches and Giardia cysts in 7 (50%) of these batches. However, this relatively high occurrence, compared with other surveys, may be due to the higher recovery efficiency of the new method, and the relatively large sample size analyzed. A more comprehensive study of the occurrence of these parasites in shellfish would be of pertinence to the Norwegian shellfish industry.

Mortality analysis of people with severe mental illness transferred from long-stay hospital to alternative care in the Life Esidimeni tragedy.

A mortality analysis of the Life Esidimeni tragedy was precluded during the investigation by the Health Ombud by the lack of data integrity. Information on the mental healthcare users (MHCUs) transferred out of Life Esidimeni hospitals between October 2015 and June 2016 was subsequently collected by the Gauteng Department of Health, permitting statistical analysis. Survival rates were calculated according to gender and transfer destination and adjusted for patient age. Mortality was compared with that of the general population for the calendar year of 2016. Of the 1 442 MHCUs, 15% were transferred to specialised psychiatric hospitals and 85% to a rehabilitation centre or non-governmental residential facility. By the end of August 2017, 9% (n=131) of the cohort had died. Significant predictors of survival were younger age (p<0.0001) and transfer to a psychiatric institution (p=0.004). The age-adjusted death rate was 63/1 000 and the overall standardised mortality ratio (SMR) was 4.9 (95% confidence interval (CI) 3.92 - 5.80), with an SMR of 3.9 (95% CI 2.95 - 4.86) for men and 6.3 (95% CI 4.22 - 8.38) for women. The excess deaths are therefore quantified, and the high-risk environment of the rehabilitation centre and residential facilities confirmed. High mortality among MHCUs is unlikely to be confined to the Life Esidimeni tragedy; monitoring of preventable deaths in this vulnerable population is recommended.

Giardia duodenalis in primates: Classification and host specificity based on phylogenetic analysis of sequence data.

Giardia duodenalis colonizes the gastrointestinal tract of a wide range of hosts, including humans and other primates. It is grouped into eight different Assemblages and, beyond that, into a number of sub-Assemblages, defined ad hoc on the basis of genetic differences; these various groups are often considered to be associated with a specific restricted host range. The aim of this study was to use publicly available genotyping data to investigate the relatedness of human and non-human primate (NHP) Giardia isolates in order to evaluate the usefulness of current taxonomic classification and to assess whether there is potential for zoonotic transmission between humans and NHP. Our final data set consisted of sequence data from 165 isolates, 111 from NHP and 54 from humans. Assemblages were well defined, but sub-Assemblages across Assemblage B were not resolved. Although sub-Assemblages AI and AII were resolved, the terms were not found to capture any useful molecular or host/deme properties. In the phylogenetic tree, NHP isolates were scattered among human isolates across Assemblages A and B, and were even found in Assemblage E. We conclude that there does not appear to be significant molecular distinction between human and NHP Giardia isolates across these four molecular markers. Thus, on the basis of these markers, we cannot exclude a risk for zoonotic and anthropozoonotic transmission of Assemblages A and B isolates, irrespective of sub-Assemblage classification. We further evaluated the relative merit of the four genes used in genotyping studies. The tpi, gdh and bg genes gave relatively congruent tree topologies, but the SSU gene did not resolve Assemblages according to the current classification. Future genotyping efforts should aim for multilocus or whole-genome approaches and, in particular, use of the SSU gene as the sole marker should be avoided when possible.

The potential for marine bivalve shellfish to act as transmission vehicles for outbreaks of protozoan infections in humans: a review.

Most marine molluscan bivalve shellfish feed on suspended phytoplankton which are trapped from water pumped across the gills by ciliary action. Pathogenic microorganisms in the water may be filtered by the gills during feeding, and become concentrated in the digestive glands/tract. If these pathogens are not excreted or inactivated by the shellfish, or in subsequent preparatory processes, they may be ingested by consumers, the shellfish thereby acting as vehicles of infection. The protozoan parasites Cryptosporidium spp., Giardia duodenalis and Toxoplasma gondii have the potential to be transmitted in this way, and here we review the accumulating knowledge on the occurrence and survival of the transmission stages of these parasites in shellfish, whilst also emphasising the considerable gaps in our knowledge. Relevant information is particularly lacking for T. gondii, which, in comparison with Cryptosporidium spp. and G. duodenalis, has been relatively under-researched in this context. Although it seems evident that these shellfish can accumulate and concentrate all three of these parasites from the surrounding water, whether Giardia cysts remain viable and infectious is unknown, and some evidence suggests that they may be inactivated by the shellfish. Although both Toxoplasma and Cryptosporidium apparently retain their infectivity for prolonged periods in shellfish, the actual public health threat posed by these parasites via these shellfish is unclear, largely because there is minimal evidence of infection transmission. Reasons for this apparent lack of infection transmission are discussed and it is recommended that the potential for transmission via shellfish consumption is recognised by those concerned with investigating transmission of these infections.

Giardia duodenalis cysts isolated from wild moose and reindeer in Norway: genetic characterization by PCR-rflp and sequence analysis at two genes.

There are few genotyping studies of Giardia duodenalis isolates from cervid hosts, although a previous study suggested that cervids may be a source of infection for humans and cattle. Giardia duodenalis isolates collected from wild moose (Alces alces) and reindeer (Rangifer tarandus) in Norway during 2002 and 2003 were characterized by polymerase chain reaction-restriction fraction length polymorphism (PCR-RFLP) at the beta-giardin gene, and sequence analysis at both the beta-giardin and glutamate dehydrogenase (gdh) genes. All results suggested that these isolates (n=25) belonged to assemblage A. Three different restriction patterns were obtained with PCR-RFLP, one of which has previously been associated with assemblage A. At the beta-giardin gene, sequences from six reindeer isolates and one moose isolate were identical to a previously published assemblage A sequence from G. duodenalis cysts isolated from dairy calves. The other 10 moose isolates could be divided into five groups, with between two and 14 single nucleotide polymorphisms (SNPs) from the published genotype A2. At the gdh gene, three different sequences were obtained, differing from each other by between one and 15 SNPs and which have all been previously published as genotype A1, but with different specific hosts. Grouping of the isolates based on the sequences from both genes gave complex results; whereas all the G. duodenalis isolates from reindeer grouped together, two moose isolates, which had identical sequences at the beta-giardin gene, had sequences that differed from each other by 15 SNPs at the gdh gene. The results of these studies, together with the large Norwegian populations of these cervids and the amount of fecal matter they produce, indicate that moose and reindeer may be significant reservoirs of G. duodenalis infection in Norway, which may be of importance to veterinary and public health.

The potential for acquiring cryptosporidiosis or giardiosis from consumption of mung bean sprouts in Norway: a preliminary step-wise risk assessment.

The current work evolved from a microbial survey of fruits and vegetables conducted in Norway between 1999 and 2001. This survey found that mung bean sprouts were more likely to be contaminated with Cryptosporidium and Giardia than the other produce included in the survey. To support this observation and to demonstrate to public health officials that this might be a risk warranting further attention, a simple risk assessment was initiated. Assuming that 60,000 people in Norway consume a single serving of bean sprouts per week, and contamination levels are similar to those found in the survey, it was calculated that there could be in the order of 20 cases of Giardia or Cryptosporidium infection per 100,000 population attributable to consumption of mung bean sprouts. A number of assumptions were made for the calculations, including parasite factors (e.g. viability, genotype), product factors (e.g. extent of product contamination) and host factors (e.g. composition and extent of consumer group). These assumptions and areas of uncertainty, where further data would improve the risk assessment, are highlighted throughout. Not only does the risk assessment identify new areas of research, but it also demonstrates how risk assessment can be used as a tool to try to influence public health surveillance.

Haemoglobin concentrations and concomitant infections of hookworm and Trichuris trichiura in Panamanian primary schoolchildren.

The blood haemoglobin concentrations and intestinal helminth infections were surveyed in 658 children aged between 6 and 12 years attending 4 primary schools in Coclé Province, Panama, in November 1987. 147 (22.3%) of the children were judged to have haemoglobin concentrations indicative of iron-deficiency anaemia. Blood haemoglobin concentrations were significantly lower in children with heavier Trichuris trichiura infections (> 5000 eggs/g) (P = 0.014), and in children with dual infections of both hookworm and T. trichiura (P = 0.005). Children with concomitant T. trichiura and hookworm infections were also significantly more likely to have blood haemoglobin levels indicative of anaemia than children who were uninfected or had single infections with either of these helminths (P < 0.005). In a longitudinal study involving 171 children selected from the original 658, blood haemoglobin concentrations were measured again in November 1988. Children who were considered to show an improvement in both hookworm and T. trichiura infections over this 12 month period showed a significantly greater increase in blood haemoglobin concentrations than children who had remained uninfected with either of these helminths throughout the study period (P < 0.05).