Rolf Nijsse

Reversible pulmonary hypertension associated with lungworm infection in a young cat

Two ten-week-old kittens presented with dyspnea. Two weeks later dyspnea had worsened and both kittens had developed a heart murmur. One kitten died and necropsy showed severe granulomatous pneumonia and moderate bronchi(oli)tis and peribronchi(oli)tis caused by Aelurostrongylus abstrusus. The results from echocardiography, thoracic radiography and the other kittens fecal examination were interpreted as severe parasitic pneumonia caused by A. abstrusus infection with pulmonary hypertension. Repeated administration of milbemycine-oxime and praziquantel resulted in cessation of larvae shedding and resolution of clinical, radiographic and echocardiographic signs of bronchopneumonia and pulmonary hypertension.

Coprophagy in dogs interferes in the diagnosis of parasitic infections by faecal examination

Many dogs display coprophagic behaviour. Helminth eggs can passively pass the dogs digestive tract and this may result in a false positive diagnosis of infection with gastrointestinal helminth parasites. For a period of one year, faecal samples of dogs were examined monthly using the Centrifugal Sedimentation Flotation (CSF) technique with a sugar flotation solution (s.g. 1.27-1.30 g/cm(3)). If a sample tested positive for canine helminth eggs, the owner was asked to submit another sample after preventing the dog from eating faeces for 3 days. If the second sample again tested positive for the same type of helminth egg, the dog was considered to have a patent infection. If the second sample tested negative, the first sample was considered a false positive due to coprophagy. The focus of this study was on dogs shedding Toxocara eggs. At the first examination, 246 samples (out of 308 samples testing positive for canine-specific helminth eggs) tested positive for Toxocara spp. Of these, 120 (49%) tested negative at the second examination. Coprophagic behaviour was recognized by 261 of the 564 owners that answered the accompanying questionnaire. This concerned 391 dogs. Coproscopical examination also provided proof of coprophagy (e.g. oocysts of Eimeria spp. or non-dog typical helminth eggs) in dogs belonging to owners that did not report coprophagic behaviour in their dogs. Results indicate that coprophagy in dogs may result in an overestimation of the prevalence of patent helminth infections and that dogs may serve as a transport host for helminth eggs.

Longitudinal Study of Extended-Spectrum-β-Lactamase- and AmpC-Producing Enterobacteriaceae in Household Dogs

ABSTRACT A longitudinal study was performed to (i) investigate the continuity of shedding of extended-spectrum-beta-lactamase (ESBL)-producing Enterobacteriaceae in dogs without clinical signs, (ii) identify dominant plasmid-mediated ESBL genes, and (iii) quantify ESBL-producing Enterobacteriaceae in feces. Fecal samples from 38 dogs were collected monthly for 6 months. Additional samples were collected from 7 included dogs on a weekly basis for 6 weeks. Numbers of CFU per gram of feces for non-wild-type Enterobacteriaceae were determined by using MacConkey agar supplemented with 1 mg/liter cefotaxime (MCC), and those for total Enterobacteriaceae were determined by using MacConkey agar. Cefotaxime-resistant isolates were screened by PCR and sequence analysis for the presence of blaCTX-M, blaCMY, blaSHV, blaOXA, and blaTEM gene families. Bacterial species were identified by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) analysis. PCR-negative isolates were tested by a double-disk synergy test for enhanced AmpC expression. A total of 259 samples were screened, and 126 samples were culture positive on MCC, resulting in 352 isolates, 327 of which were Escherichia coli. Nine dogs were continuously positive during this study, and 6 dogs were continuously negative. Monthly or weekly shifts in fecal shedding were observed for 23 dogs. Genotyping showed a large variety of ESBL genes and gene combinations at single and multiple consecutive sampling moments. The ESBL genes blaCTX-M-1, blaCTX-M-14, blaCTX-M-15, blaSHV-12, and blaCMY-2 were most frequently found. The mean number of CFU of non-wild-type Enterobacteriaceae was 6.11 × 108 CFU/g feces. This study showed an abundance of ESBL-producing Enterobacteriaceae in dogs in the Netherlands, mostly in high concentrations. Fecal shedding was shown to be highly dynamic over time, which is important to consider when studying ESBL epidemiology.

Environmental contamination with Toxocara spp. eggs in public parks and playground sandpits of Greater Lisbon, Portugal

Toxocarosis is a zoonotic parasitic disease transmitted from companion animals to humans. Environmental contamination with Toxocara eggs is considered to be the main source of human infections. In Portugal, knowledge regarding the current situation, including density, distribution and environmental contamination by Toxocara spp., is largely unknown. The present study investigated environmental contamination with Toxocara spp. eggs, in soil and faecal samples collected from public parks and playground sandpits in Greater Lisbon, Portugal. A total of 151 soil samples and 135 canine faecal samples were collected from 7 public sandpits and 12 public parks, over a 4 month-period. Soil samples were tested by a modified centrifugation and sedimentation/flotation technique and faecal samples were tested by an adaptation of the Cornell-Wisconsin method. Molecular analysis and sequencing were performed to discriminate Toxocara species in the soil. Overall, 85.7% of the sandpits (6/7) and 50.0% of the parks (6/12) were contaminated with Toxocara spp. eggs. The molecular analysis of soil samples showed that, 85.5% of the sandpits and 34.4% of the parks were contaminated with Toxocara cati eggs. Faecal analysis showed that 12.5% of the sandpits and 3.9% of the parks contained Toxocara canis eggs. In total, 53.0% of soil and 5.9% of faecal samples were positive for Toxocara spp. Additionally, 56.0% of the eggs recovered from the samples were embryonated after 60 days of incubation, therefore considered viable and infective. The average density was 4.2 eggs per hundred grams of soil. Public parks and playground sandpits in the Lisbon area were found to be heavily contaminated with T. cati eggs, representing a serious menace to public health as the studied areas represent common places where people of all ages, particularly children, recreate. This study sounds an alarm bell regarding the necessity to undertake effective measures such as reduction of stray animals, active faecal collection by pet owners, awareness campaigns and control strategies to decrease the high risk to both animal and human health.

Urban Dog Parks as Sources of Canine Parasites: Contamination Rates and Pet Owner Behaviours in Lisbon, Portugal

Dog parks represent a recent trend in western countries, enabling owners to spend quality time with their pets in a controlled environment. Despite their growing popularity, few studies have been performed to date on these parks to investigate dog intestinal parasitic infections and soil contamination. The present study examined 369 faecal and 18 soil samples collected from 3 dog parks in Greater Lisbon, Portugal. Additionally, 102 interviews were performed with dog owners to assess dog-walking behaviours and parasite risk. In total, 33% of the faecal dog samples were infected with at least one parasitic agent: hookworms (16.5%), Cryptosporidium spp. (11.9%), Giardia spp. (11.4%), Toxascaris leonina (1.1%), Cystoisospora spp. (1.1%), Toxocara spp. (0.5%), and Sarcocystis sp. (0.3%). The soil of all the parks was contaminated with hookworm eggs. This is the first study performed in a European urban area to assess canine faecal contamination and parasitic agents in dog parks. Our results highlight the potential of these parks as a source of transmission for canine parasites, including some with zoonotic potential. Public awareness and effective preventive measures should be promoted to minimise the health-risk impact to both animals and humans, under the scope of environmental and public health.

Comparing four diagnostic tests for Giardia duodenalis in dogs using latent class analysis.

BackgroundTo accurately diagnose giardiosis in dogs, knowledge of diagnostic test characteristics and expected prevalence are required. The aim of this work was to estimate test characteristics (sensitivity and specificity) of four commonly used diagnostic tests for detection of Giardia duodenalis in dogs.MethodsFecal samples from 573 dogs originating from four populations (household dogs, shelter dogs, hunting dogs and clinical dogs) were examined with centrifugation sedimentation flotation (CSF) coproscopical analysis, direct immunofluorescence assay (DFA, Merifluor Cryptosporidium/Giardia®), a rapid enzyme immunochromatographic assay (IDEXX SNAP Giardia®) and qPCR (SSU rDNA) for presence of G. duodenalis. Bayesian latent class analysis was used to determine test performance characteristics and to estimate G. duodenalis prevalence of each of the four dog populations.ResultsAll tests were highly specific. IDEXX SNAP Giardia® showed the highest specificity (99.6%) and qPCR the lowest (85.6%). The sensitivities were much more variable, with qPCR showing the highest (97.0%) and CSF the lowest (48.2%) sensitivity. DFA was more sensitive than IDEXX SNAP Giardia®, but slightly less specific. Prevalences of G. duodenalis differed substantially between populations, with the hunting dogs showing the highest G. duodenalis prevalence (64.9%) and the household dogs the lowest (7.9%).ConclusionsThis study identifies qPCR as a valuable screening tool because of its high sensitivity, whereas methods using microscopy for cyst identification or cyst wall detection should be used in situations where high specificity is required. G. duodenalis is a prevalent gastro-intestinal parasite in Dutch dogs, especially in dogs living in groups (hunting and shelter dogs) and clinical dogs.

Additional file 4: of Comparing four diagnostic tests for Giardia duodenalis in dogs using latent class analysis

Text. Detection of Giardia duodenalis assemblages and other Giardia species with the qPCR. (DOCX 363 kb)