Parasitology Laboratory, Section for Microbiology, Immunology, and Parasitology, Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, P.O. Box 8146 Dep, 0033 Oslo, Norway. Lucy.Robertson@nvh.no
An outbreak of waterborne cryptosporidiosis in a town in northern Sweden during winter 2010 resulted in the potential exposure of cured meat products to Cryptosporidium oocysts during their manufacture. The purpose of this work was to develop a method for analyzing cured meat products for contamination with Cryptosporidium oocysts and use this method to analyze potentially contaminated product samples. A simple method of elution, concentration, separation, and detection was used, based on work with other food matrices but adapted for the relatively high fat content of cured meat surfaces. Using spiking experiments, the recovery efficiency of this method was found to be over 60%. In the analysis of the potentially contaminated products, only one putative Cryptosporidium oocyst was detected, and this was sufficiently deformed so that it could not be confirmed as an oocyst; if it was an oocyst, it was considered to have been probably deformed and inactivated prior to analysis. Based on the results of the analyses, together with data on the probable extent of contamination of the products and on our knowledge of factors, such as water activity, which affect oocyst survival, the products were safely released to the market.
Dirofilaria repens infection was diagnosed in a dog that had been imported to Norway from Hungary three years previously. The dog was a four-year-old castrated male mixed-breed dog and presented for examination of two masses on the right thoracic wall. Fine needle sampling from the subcutaneous nodules and subsequent cytological examination revealed a high number of microfilariae and a pyogranulomatous inflammation. At re-examination approximately 3 weeks later, both masses had apparently disappeared spontaneously, based on both inspection and palpation. However, examination of peripheral blood by a modified Knott's test revealed a high number of unsheathed microfilariae with mean length of 360 µm and mean width of 6-7 µm, often with the classic umbrella handle appearance of D. repens. Polymerase chain reaction and sequencing confirmed the D. repens diagnosis. Subcutaneous dirofilariosis caused by D. repens is probably the most common cause of human zoonotic dirofilariosis in Europe, but currently is rarely encountered in northern countries such as Norway. However, travelling, import and relocation of dogs have increased, and thus the geographical range of these parasites is likely to increase from traditionally endemic southern regions. Increasing numbers of autochthonous cases of D. repens infections in dogs have been reported in eastern and central Europe. Although infection with D. repens often induces only mild signs or subclinical infections in dogs, they nevertheless represent a reservoir for zoonotic transmission and thus a public health concern, and, in addition, due to the long prepatent period and the high frequency of subclinical infections or infections with unspecific clinical signs, could easily be missed. Lack of experience and expectation of these parasites may mean that infection is underdiagnosed in veterinary clinics in northern countries. Also, predicted climate changes suggest that conditions in some countries where this infection is currently not endemic are likely to become more suitable for development in the intermediate host, and thus the establishment of the infection in new areas.
Norwegian University of Life Sciences, Faculty of Veterinary Medicine, Department of Production Animal Clinical Sciences, Kyrkjevegen 332/334, 4325 Sandnes, Norway. Electronic address: email@example.com.
Int J Parasitol Drugs Drug Resist. 2018 08; 8(2):304-311
Ovine Eimeria spp. infections cause reduced welfare, increased mortality, and substantial economic losses, and anticoccidials are crucial for their control. Recent reports of toltrazuril resistance in pigs, and anecdotal reports of reduced anticoccidial efficacy in lambs, necessitate evaluation of anticoccidial efficacy. Due to the substantial lifecycle differences between nematodes and coccidia, current WAAVP methods for assessing anthelmintic efficacy are not suitable for such evaluations. Faecal samples were collected from 8 pairs of twin lambs from 36 Norwegian sheep farms 6-8 days after turnout. One twin of each pair was then treated with 20?mg/kg toltrazuril and a second faecal sample from all lambs was collected 7-11 days later. Oocyst excretion rate in all samples was determined using McMasters. Suitability of treatment timing was investigated by evaluating the increase in mean log oocyst excretion in untreated lambs. Based on comparisons between groups, a threshold of =0.75 (13 farms) was used to identify farms where drug efficacy could be assessed with confidence, drug efficacy on farms with increases of =0.5 but ?90%), and for 5 the results were inconclusive. This is the first evidence-based report of reduced anticoccidial efficacy in ovine Eimeria spp. Additionally, we highlight the problem of sub-optimal timing of treatment (16/36 farms), which could potentially result in incorrect conclusions being reached regarding lack of drug efficacy.
Most waterborne outbreaks of cryptosporidiosis are reported from the USA, and in Europe from the UK. However, since 2000 reports of foodborne cryptosporidiosis seem to be skewed towards Nordic countries, although consumers in these countries are apparently less concerned about microbiological contamination of food than consumers elsewhere. Possible reasons for this unexpected geographical distribution might include prolonged survival of oocysts in the Nordic climate, greater exposure due to elevated consumption of higher-risk products (possibly including imported foods), and better outbreak investigation and reporting. Although the risk of foodborne cryptosporidiosis is probably underestimated globally, we suggest that the next outbreak is no more likely to be in a Nordic country than anywhere else.
The effects of precipitation on the hygienic quality of water and blue mussels collected from five different localities in the urban areas in the Inner Oslofjord were investigated, with samples analysed for Escherichia coli, Salmonella spp., pathogenic Vibrio spp., Norovirus, Sapovirus, Cryptosporidium spp. and Giardia duodenalis. The sampling sites were located at varying distances from the outlet of combined sewer overflows (CSO)-impacted rivers/streams. In general, 1-3 log10 increases in fecal indicator bacteria and human pathogens were observed after heavy rainfalls. Blue mussels appeared to be a useful indicator of the impact of sewage at these sites, and generally a good correlation was identified between concentrations of E. coli and other human pathogens in the mussels. Provision of general advice to the public of avoiding areas near the outlets of CSO-impacted rivers after heavy rainfall may reduce the risk of gastroenteritis by bathers and others that may swallow water during recreational activities.
As part of larger survey of microbial contamination of fruits and vegetables in Norway, four different sprouted seed products were analysed for bacterial and parasitic contaminants (n = 300 for bacterial analyses and n = from 17 to 171 for parasite analyses, depending on parasite). Escherichia coli O157, Salmonella, Listeria monocytogenes, Cyclospora oocysts, Ascaris eggs and other helminth parasites were not detected in any of the sprout samples. Thermotolerant coliform bacteria (TCB) were isolated from approximately 25% of the sprout samples, with the highest percentage of TCB positive samples in alfalfa sprouts. Most TCB were Enterobacter spp. and Klebsiella. E. coli was isolated from 8 of 62 TCB positive mung bean sprout samples. Cryptosporidium oocysts were detected in 8% of the sprout samples and Giardia cysts were detected in 2% of the samples. All the Cryptosporidium positive samples, and most of the Giardia positive samples, were mung bean sprouts. Parasite concentrations in positive samples were low (between 1 and 3 oocysts/cysts per 50 g sprouts). Sprout irrigation water was also analysed for microbial contaminants. E. coli O157 and L. monocytogenes were not detected. TCB were isolated from approximately 40% of the water samples. Salmonella reading was isolated from three samples of spent irrigation water on 3 consecutive days. Cryptosporidium and Giardia were also isolated from spent irrigation water. Additionally, eight samples of unsprouted mung bean seed were analysed for Cryptosporidium oocysts and Giardia cysts. One or both of these parasites were detected in six of the unsprouted seed samples at concentrations of between 1 and 5 oocysts/cysts per 100 g unsprouted seed. Whilst our results support spent irrigation water as the most suitable matrix for testing for bacteria, unsprouted seed is considered the more useful matrix for analysing for parasite contaminants.
The tracheal mite, Acarapis woodi, may be one of many factors contributing to the decline in honey bee (Apis mellifera) populations. Databases on the widespread distribution of A. woodi exist, but the data seem patchy. Norway is not listed as being infested, although there have been at least two separate introductions of the parasite. Investigations in 2003, 2006, and 2009 using standard microscopy methods indicated persistence of A. woodi in honey bees in this region. In 2013, we conducted another survey. Samples were sent in from 335 beehives belonging to 39 apiaries, and all were asked to complete a questionnaire. Analysis for A. woodi in the submitted samples was by PCR, with sequencing of positive results. The results described in this article indicate that this parasite still persists in some apiaries in this region, but at a low, and possibly decreasing, level, with positive results obtained from just two (5.1%) of the apiaries. Of the 17 beekeepers that answered the questionnaire, none reported symptoms of infestation with A. woodi. Sequencing of PCR products indicated a difference between the two A. woodi isolates. Our results were generally encouraging regarding the apparent lack of spread of A. woodi, within the County. Furthermore, the sequencing results may indicate two separate introductions rather than spread. Nevertheless, the scarcity of data, the vulnerability of honey bee populations globally, and the potential contribution of this parasite to reduced survival, indicate that the situation should be continued to be monitored. In addition, Norwegian beekeepers should be made aware of, and follow, restrictions regarding import and transport of bees, both nationally and internationally.
The aim of this study was to investigate the occurrence of Giardia duodenalis and Cryptosporidium spp. in primates and determine their zoonotic or anthropozoonotic potential.
Direct immunofluorescence was used to identify Giardia and Cryptosporidium from faecal samples. PCR and DNA sequencing was performed on positive results.
Giardia cysts were identified from 5.5% (5/90) of captive chimpanzees and 0% (0/11) of captive mandrills in the Republic of Congo; 0% (0/10) of captive chimpanzees in Norway; and 0% of faecal samples (n?=?49) from wild Zanzibar red colobus monkeys. Two Giardia positive samples were also positive on PCR, and sequencing revealed identical isolates of Assemblage B.?Cryptosporidium oocysts were not detected in any of the samples.
In these primate groups, in which interactions with humans and human environments are quite substantial, Giardia and Cryptosporidium are rare pathogens. In chimpanzees, Giardia may have a zoonotic or anthropozoonotic potential.
Giardia duodenalis is an intestinal protozoan capable of causing gastrointestinal disease in a range of vertebrate hosts. It is transmitted via the fecal-oral route. Understanding the epidemiology of G. duodenalis in animals is important, both for public health and for the health of the animals it infects. We investigated the occurrence of G. duodenalis in wild Swedish red foxes ( Vulpes vulpes ), with the aim of providing preliminary information on how this abundant predator might be involved in the transmission and epidemiology of G. duodenalis . Fecal samples (n=104) were analysed for G. duodenalis using a commercially available direct immunofluorescent antibody test. Giardia duodenalis cysts were found in 44% (46/104) of samples, with foxes excreting 100 to 140,500 cysts per gram of feces (mean, 4,930; median, 600). Molecular analysis, using PCR with sequencing of PCR amplicons, was performed on 14 samples, all containing over 2,000 cysts per gram feces. Amplification only occurred in four samples at the tpi gene, sequencing of which revealed assemblage B in all four samples. This study provides baseline information on the role of red foxes in the transmission dynamics of G. duodenalis in Sweden.
Enteric viruses transmitted via the faecal-oral route occur in high concentrations in wastewater and may contaminate drinking water sources and cause disease. In order to quantify enteric adenovirus and norovirus genotypes I and II (GI and GII) impacting a drinking source in Norway, samples of surface water (52), wastewater inlet (64) and outlet (59) were collected between January 2011 and April 2012. Samples were concentrated in two steps, using an electropositive disc filter and polyethylene glycol precipitation, followed by nucleic acid extraction and analysis by quantitative polymerase chain reaction. Virus was detected in 47/52 (90.4%) of surface water, 59/64 (92%) of wastewater inlet and 55/59 (93%) of wastewater outlet samples. Norovirus GI occurred in the highest concentrations in surface water (2.51e + 04) and adenovirus in wastewater (2.15e + 07). While adenovirus was the most frequently detected in all matrices, norovirus GI was more frequently detected in surface water and norovirus GII in wastewater. This study is the first in Norway to monitor both sewage and a drinking water source in parallel, and confirms the year-round presence of norovirus and adenovirus in a Norwegian drinking water source.