Mixed Giardia duodenalis assemblage A, B, C and E infections in pet chinchillas (Chinchilla lanigera) in Flanders (Belgium)

Fourteen isolates of Toxoplasma gondii were isolated from cats from 4 different geographic provinces (Anhui, Hubei, Shanxi and Guangdong) in China and their genetic diversity with 8 nuclear loci SAG2, SAG3, BTUB, GRA6, L358, PK1, c22-8, c29-2, and an apicoplast locus Apico, was analysed by restriction fragment length polymorphisms (RFLPs). Two genotypes from these 14 isolates were identified but none of them belongs to the typical genetic types (types I, II and III). It is unexpected that such high similarity was observed in these 14 isolates although their original regions are significantly distant. Our results strongly indicate that the three traditional clonal lineages of types I, II and III of this parasite may not be preponderant in China. In addition, our results show that the genotypes of T. gondii in China may be highly clonal with atypical genotypes and higher virulence.     

Laboratory diagnosis of Giardia duodenalis infection in dogs

Results of trichrome staining of fecal samples and intestinal contents preserved in polyvinyl alcohol fixative, fecal flotation utilizing unpreserved feces, and enzyme-linked immunosorbent assay of serum specimens were compared for the diagnosis of Giardia duodenalis infections in dogs. Trichrome staining of preserved fecal samples resulted in the identification of 44 (92%) of the 48 infected dogs from a group of 200 dogs. Trichrome staining of preserved intestinal contents resulted in the identification of 26 (54%) of the infected dogs, and fecal flotation resulted in the identification of 23 (48%) of the infected dogs. Giardia duodenalis antibodies were not detected consistently in the sera of infected dogs.     

Molecular identification of Cryptosporidium parvum and Giardia duodenalis in the Italian water buffalo (Bubalus bubalis)

Livestock are commonly infected with protozoan parasites of the genera Cryptosporidium and Giardia, and some of the species and genotypes found in these animals have zoonotic significance. We characterized isolates of both parasites recovered from the Italian water buffalo (Bubalus bubalis), an economically important species whose milk is used for the production of "buffalo mozzarella" fresh cheese. Molecular analysis of the Cryptosporidium small subunit ribosomal DNA gene and of the Giardia beta-giardin gene shows the presence of both zoonotic parasites (Cryptosporidium parvum and Giardia duodenalis assemblage A) and host-specific parasites (G. duodenalis assemblage E), suggesting that water buffaloes can contribute to environmental contamination with oocysts and cysts potentially infectious to humans if their faeces are improperly disposed of. On the other hand, mozzarella cheese is probably a safe product, given that its production involves the treatment of cheese curd at 85-95 degrees C, which is likely to kill or inactivate the parasites.     

Genotyping of Giardia duodenalis from Southern Brown Howler Monkeys (Alouatta clamitans) from Brazil

Giardia duodenalis is a widespread intestinal protozoan that can infect humans and animals, both domestic and wild. Independent of host, infections present with the same symptoms. However, based on host specificity, Giardia isolates have been grouped into genotypes A to G. Parasites of assemblage A and B are known to infect humans, in addition to primates and a wide variety of mammals. In Brazil, hitherto Giardia genotypes were defined only for humans and domestic animals. To evaluate the genotypes of different Giardia present among other animals, fecal samples from 28 Southern Brown Howler Monkeys (Alouatta clamitans) kept in captivity from South Brazil were screened for G. duodenalis using parasitological methods. All of them were asymptomatic, but positive for Giardia. The genotype of the G. duodenalis circulating among these animals was ascertained by molecular typing, performed using amplification and sequencing of the beta-giardin gene. Sixteen of 28 samples were successfully amplified by PCR and sequencing of this gene s revealed that all of them were of the genotype A1. These findings suggest that A. clamitans represent a potential risk of environmental contamination of a G. duodenalis genotype that also infect humans, and therefore can be considered a potential reservoir for G. duodenalis of a genotype that can also infects humans. Therefore, these results highlight a potential public health problem due to the epidemiological and molecular evidence for anthropozoonotic transmission.     

Occurrence and genotype characterization of Giardia duodenalis in goat kids from the Canary Islands, Spain

Giardia duodenalis (syn. Giardia lamblia, Giardia intestinalis) is a wide-spread intestinal protozoa of both humans and animals. Although giardiosis in goat is commonly asymptomatic, young kids may bear an enteric disease associated with persistent diarrhoea and delayed weight gain. In the present study we have analysed the occurrence of Giardia in 315 young goat kids (2–6 months old) from Gran Canaria Island (Spain) through visualization of faecal cysts. The identification of genotypes of G. duodenalis among the farms was attained by nested PCR of the triophosphate isomerase (TPI) and single PCR of β-giardin genes and subsequent sequencing. Positive samples were found in 42.2% of the animals and 95.5% of the farms. Goat faecal specimens were positive for only livestock-associated G. duodenalis assemblage E genotype for both TPI and β-giardin genes. The genetic analysis of these two loci revealed the presence of different haplotypes among the farms included in the survey and high homology with homologous genes from cattle and sheep. Altogether, the data presented here provide additional information to the prevalence and genetic characterization of Giardia isolates. The absence of assemblages A and B in this study suggests that zoonotic transmission of Giardia from goats could be of low epidemiological significance, although these findings should be validated in studies including other geographical areas, age groups and larger number of samples.     

Giardia duodenalis in colony stray cats from Italy

Giardia duodenalis is the most common intestinal protozoan in humans and animals worldwide, including eight morphologically identical assemblages, infecting pets, livestock, wildlife and human beings. Assemblages A and B are those with the higher zoonotic potential, and they have been detected in several mammals other than humans; the others (C to H) show a higher host specificity. Cats can harbour both the specific Assemblage F and the zoonotic ones A and B. Several studies have been carried out on G. duodenalis genotypes in cats; however, the role of this species in the epidemiology of giardiasis is still poorly understood. In this scenario, the present study carried out the detection and genetic characterization at sub-assemblage level of G. duodenalis from colony stray cats in central Italy. In the period 2018–2019, 133 cat faecal samples were analysed for the presence of G. duodenalis cysts by a direct immunofluorescence assay. Positive samples were subsequently subjected to molecular analyses for assemblage/sub-assemblage identification. Forty-seven samples (35.3%) were positive for G. duodenalis cysts by immunofluorescence. G. duodenalis DNA was amplified at SSU-rDNA locus from 39 isolates: 37 were positive for zoonotic Assemblage A and 2 showed a mixed infection (A + B). Positive results for the β-giardin gene were achieved for 25 isolates. Sequence analysis revealed 16 isolates belonging to Sub-assemblage AII and 8 to Sub-assemblage AIII. One isolate resulted as ambiguous AI/AIII. Large sequence variability at the sub-assemblage level was detected, with several double peaks and mutations, making complex a proper isolate allocation. When compared with previous studies, the 35.3% prevalence of G. duodenalis in cats reported in the present article was surprisingly high. Moreover, all positive cats resulted to be infected with zoonotic assemblages/sub-assemblages, thus indicating stray cats as a possible source of human giardiasis and highlighting the sanitary relevance of cat colonies in the study area.     

Effects of oral cisapride administration in chinchillas (Chinchilla lanigera) with experimentally induced fecal output reduction

BackgroundCisapride, a prokinetic drug, has been anecdotally recommended in chinchillas at a dose of 0.5 mg/kg PO q8-12h to treat gastrointestinal hypomotility. However, studies in other rodent species suggest that higher doses are necessary to be effective.MethodsIn two randomized, placebo-controlled, blinded, crossover studies, the effects of cisapride (10 mg/kg PO q12h for four doses), administered with or without concurrent syringe feeding of a critical care formula (25 mL/kg PO q12h for two doses) were evaluated in chinchillas following recovery from sedation induced by alfaxalone-butorphanol. Food intake and fecal output were quantified to assess the effects of cisapride on these parameters.ResultsOver the first 24 hours after recovery from sedation, animals that received cisapride and syringe feeding had the least reduction in fecal output (-27 ± 23%) compared to the control treatment (-48 ± 22%, P = 0.008) or to animals which received syringe feedings alone (-40 ± 23%, P = 0.12). Cisapride administered without concurrent syringe feedings had no effect on fecal output. No adverse effects were recorded following the administration of cisapride.Conclusions and clinical relevanceThe oral administration of cisapride at 10 mg/kg q12h in conjunction with syringe feeding resulted in a slight, but not clinically relevant, attenuation of fecal output reduction by ~13% compared to syringe feedings alone and by ~20% compared to no treatment. Without the concurrent administration of syringe feedings, cisapride had no effect on fecal output in chinchillas.     

Use of radiographs and computed tomography for measurement of kidney size in healthy chinchillas (Chinchilla lanigera)

BackgroundChinchillas, being small mammals, are subject to many of the urinary disorders that affect other companion animals. In cats and dogs, relationships between renal length and the presence of renal pathology have been well documented through the use of diagnostic imaging. Radiography and computed tomography (CT) are two commonly utilized diagnostic imaging modalities for assessing renal pathology. Presently, there are no published data on chinchilla kidney size and appearance using radiography or CT. This study aimed to determine healthy adult chinchilla renal size and correlate renal size to L2 body length and aortic diameter by use of radiography and CT.MethodsTwenty-eight healthy adult chinchillas with no clinical evidence of renal disease were placed under sedation for acquisition of radiographs and CT imaging. Both kidneys were measured on the radiographic views (right lateral, left lateral and ventrodorsal), when identifiable. Renal length, L2 body length and abdominal aortic diameter were measured on CT multiplanar reformatted images in specified planes and windows.ResultsIn 25% (7/28) of chinchillas, the renal length of either kidney could not be obtained on any of the three radiographic views due to superimposition of fluid and ingesta within the alimentary tract. Both the right and left kidney were identified on CT images in 100% of chinchillas (28/28). The left renal and right renal 95% confidence intervals from the dorsal plane CT images were 2.26 – 2.33 cm and 2.31 – 2.39 cm, respectively. No correlation was found between kidney length, L2 body length and diameter of the abdominal aorta on CT images.ConclusionsBased on dorsal plane CT imaging, normal chinchilla renal length is proposed to be between 2.25–2.4 cm.Clinical RelevanceRadiographic evaluation of chinchilla renal length is greatly limited due to their large fluid- and ingesta-filled colon and cecum, as is typical of animals that are hindgut fermenters. Through establishment of a normal CT reference interval for renal length and the ever-increasing clinical utility of CT, clinicians can better evaluate chinchilla renal length and identify alterations that may indicate pathologic processes.     

牛十二指肠贾第虫的分子流行病学研究进展

贾第虫是一类在全球范围内广泛分布的寄生虫,包括7个虫种。其中的十二指肠贾第虫（Giardia duodenalis）是一种重要的肠道寄生虫,能感染人和大多数哺乳动物,可引起腹泻、营养不良和体重减轻等症状。分子分型工具的发展促进了贾第虫检测、基因分型和溯源的发展,大大改变了人们对贾第虫人兽共患潜力的理解。利用分子分型工具可将十二指肠贾第虫分为8种集聚体（A~H）,8种集聚体的宿主范围都存在差异,其中,集聚体A和B是人兽共患型。牛是十二指肠贾第虫的重要宿主,但目前对牛十二指肠贾第虫病的分子流行病学认识不够,其公共卫生意义也一直被忽视。本文汇总了国内牛十二指肠贾第虫的分子流行病学调查结果。结果发现,我国牛十二指肠贾第虫的感染是普遍存在的,其中,集聚体E为优势集聚体,集聚体A和B呈散发流行。近年来,一些国家出现了集聚体E感染人的报道,同时,集聚体A在牛中的感染率有上升的迹象,牛十二指肠贾第虫的人兽共患潜力正在逐步得到认识。     

Effects of compounded metronidazole and metronidazole benzoate oral suspensions on food intake in healthy chinchillas (Chinchilla lanigera)

BackgroundThe administration of metronidazole in chinchillas remains controversial and some sources raise concerns regarding drug-induced anorexia or hyporexia. Therefore, the objective of this study was to evaluate if the oral administration of oral metronidazole has a significant effect on food intake in clinically healthy chinchillas, and if drug-induced reduction of food intake differs between metronidazole and metronidazole benzoate as well as between different concentrations of metronidazole benzoate suspensions.MethodsIn a series of randomized, blinded, crossover experiments, the effects of oral administration of metronidaole at 20 mg/kg was evaluated following q12h (for 3 days, n = 12) or q24h (for 5 days, n = 9) dosing using two different drug compounding protocols.ResultsFollowing administration of metronidazole, food intake was significantly reduced within 24 hours of drug administration, regardless of drug formulation type, compounding protocol, or drug concentration. Variation in metronidazole-induced food intake reduction differed widely between individual chinchillas, with a reduction of >50% of food intake reported in 50% of the animals following q12h administration, and 33% of animals after q24h administration of metronidazole. The reduction in food intake was self-limiting and return to normal food intake occurred after discontinuation of metronidazole administration. Neurological adverse effects were observed in one animal following q12h administration.Conclusions and clinical relevanceThe oral administration of metronidazole and metronidazole benzoate at published dosages can result in a marked reduction in food intake, and therefore, this drug should be used cautiously in chinchillas and food intake carefully monitored.     

Prevalence and intensity of infection of Cryptosporidium spp. and Giardia duodenalis in dairy cattle in Galicia (NW Spain)

Faecal samples were collected from 734 cattle selected at random from 60 dairy farms in Galicia (NW Spain). The animals studied were classified into 12 age groups: <1 month (53); 1-5 months (30); 6-11 months (31); 12-16 months (72); 17-20 months (64); 21-24 months (96); 3 years (94); 4 years (74); 5 years (67); 6 years (67); 7-8 years (63) and 9-13 years (23). Oocysts of Cryptosporidium spp. were identified in 104 animals (14.2%) distributed throughout all of the age groups and from 40 different farms (66.7%). The percentage of cattle infected ranged between 58.5% in calves <1 month and 7.9% in 7 to 8-year-old cows, i.e. the percentage of infection decreased significantly (P < 0.05) with increasing age. The intensity of infection in animals older than 1 month ranged between 10 and 5924 oocysts/g of faeces and there were no significant differences between the different groups. Cysts of Giardia duodenalis were identified in 221 animals (30.1%) from 56 farms (93.3%). The parasite was detected in all age groups, at rates of infection ranging between 21.8% (9-13 years) and 56.7% (1-5 months), although these differences were not statistically significant. The intensity of infection ranged between 7 and 15 412 cysts/g of faeces, with the number of cysts shed being significantly higher (P < 0.05) in calves <1 month than in calves aged 1-5 months. Significant associations between parasitisation by Cryptosporidium spp. or G. duodenalis and the consistency of the faeces were only found in calves aged <1 month and 1-5 months. Concurrent infections were more prevalent in the groups of calves of 1-5 months (23.3%) and 6-11 months (25.8%).     

Prevalence and risk factors of Giardia duodenalis in dogs from Romania

The protozoan Giardia duodenalis is a mammalian-infecting parasite that produces diarrhoea and malabsorption in its hosts. A survey to investigate canine infections with G. duodenalis in Romania was undertaken between June 2008 and December 2009. The objectives of the study were to (i) estimate the prevalence of infection in different dog populations (kennels, shelters, shepherd, household) using microscopy and a commercially available enzyme-linked immunosorbent assay (ELISA) test kit; (ii) to establish the level of agreement and characteristics of the tests; and (iii) to identify risk factors for infection by multivariate logistic regression models. Faecal samples were collected from 614 dogs aged from 1 month to 16 years (mean ± SD=2.88 ± 2.86 years). Each sample was tested for the presence of cysts using a flotation method with saturated sodium chloride solution and 416 out of 614 stool samples were further examined for the presence of G. duodenalis specific antigens using Giardia Microwell ELISA (SafePath? Laboratories). Giardia cysts were identified in 8.5% of total dogs (52/614) and statistical significantly more frequently in dogs living in communities. The cysts prevalence according with dog populations was as follows: 7.2%(9/125) in kennel dogs; 16.5%(27/164 in shelter dogs; 4.3%(2/46) in shepherd dogs; 4.8%(4/84) in household dogs from urban areas; and 5.1%(10/195) in household dogs from rural areas. The overall prevalence of Giardia infection by ELISA was 34.6% (144/416). The prevalence was significantly higher in kennel dogs (50%; 13/26), shelter dogs (47.7%; 74/155) and shepherd dogs (40.5%; 17/42) than in household dogs from urban areas (34.1%; 15/44) and household dogs from rural areas (16.8%; 25/149). It was noticed poor agreement between microscopy and ELISA (k=0.19). The microscopy performed best, with an Youden Index of 0.74, a Se of 73.68% and a Sp of 100%. ELISA had 100% Sp, but only 19.44% Se. Young dogs (up to 12 months age) and living in communities were identified as risk factors for infection by multivariate logistic regression analysis. 71.2% (37/52) Giardia cysts positive dogs presented co-infections with other intestinal parasites: Toxocara canis (14/52; 26.9%), Isospora ohioensis (12/52; 23.1%), Ancylostoma caninum (9/52; 17.3%), Uncinaria stenocephala (7/52; 13.5%), Trichocephalus vulpis (6/52; 11.5%), Hammondia heydorni/Neospora caninum (5/52; 9.6%), Sarcocystis spp. (5/52; 9.6%), Isospora canis (4/52; 7.7%), Capillaria aerophila (3/52; 5.8%), Strongyloides stercoralis (2/52; 93.8%), Dipylidium caninum (1/52; 1.9%) and Toxascaris leonina (1/52; 1.9%).     

Genotypic analysis of Giardia duodenalis in domestic cats

BACKGROUND: Giardia duodenalis is an intestinal flagellated protozoan that affects many mammalian species often causing severe diarrheal disease. Several different genotypes have been identified (Assemblages A-G). Most isolates recovered from domestic cats have been assigned to either Assemblage A, the zoonotic form of the parasite, or Assemblage F, identified thus far only in cats. Genotypic variation within G. duodenalis may influence clinical presentation and course of disease. Therefore, host-adapted genotypes may not be responsible for diarrheal disease (eg, Assemblage F in cats). HYPOTHESIS: Multiple Giardia genotypes will be present in domestic cats, including Assemblage F, which will not be correlated with clinical signs. ANIMALS: 250 domestic cats from eastern Mississippi and northwestern Alabama. METHODS: Prevalence survey. Fecal samples evaluated for cysts using a centrifugation concentration technique and a commercially available direct immunoflourescent antibody kit. Giardia isolates were characterized by PCR amplification and sequencing of the glutamate dehydrogenase gene. RESULTS: Both Assemblage A-I (6/17) and Assemblage F (11/17) were identified. Although Assemblage was significantly associated with age and housing, no association was detected between Assemblage and a variety of other factors including the presence of gastrointestinal signs (acute vomiting, diarrhea, and constipation). CONCLUSIONS AND CLINICAL IMPORTANCE: The presence of diarrhea in domestic cats with Giardia cannot be used as a predictor of the presence of zoonotic genotypes in animals within the study area. Although Assemblage A was associated with age and housing, veterinarians should consider any isolation of Giardia from domestic cats as potentially zoonotic.     

Prevalence of Giardia duodenalis genotypes in adult dairy cows

The prevalence of Giardia duodenalis genotypes was determined in adult dairy cows. Fecal specimens were collected from two farms each in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Specimens, cleaned of fecal debris and concentrated using CsCl density gradient centrifugation, were subjected to PCR and DNA sequence analysis. The prevalence of G. duodenalis infection, ranged from 3% to 64%, with an average prevalence of 27% (144 positive cows out of 541 examined). DNA sequence analysis of the 16S rRNA gene revealed the presence of both Assemblage A and Assemblage E, G. duodenalis. Overall, Assemblage E was present in 25% of all animals tested and Assemblage A was present in 2% of the animals. As a percentage of G. duodenalis isolates, Assemblage E represented 94% and Assemblage A represented 6%. Although, most of the cows were infected with a genotype that is not known to be infectious for humans, adult cows on five farms did harbor varying levels of zoonotic Assemblage A, G. duodenalis. Therefore, although adult cows do not appear to be a significant source of human infectious cysts in the environment, the risk from this age group should not entirely be discounted.     

Multilocus genotyping of Giardia duodenalis in lambs from Spain reveals a high heterogeneity

Fecal specimens from 120 lambs in Valencia (Spain) were analyzed for Giardia duodenalis by IFA and nested-PCR using the beta giardin (bg), glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi) and small subunit ribosomal RNA (ssurRNA) genes. The highest prevalence was obtained using the ssurRNA gene (89.2%), whereas values from other techniques ranged from 64.1% to 69.2%. Sequences of the ssurRNA showed a high proportion of assemblage A or mixed assemblage A/E samples (55.1% and 25.2%, respectively). When the other 3 loci were analyzed, between 6.5% and 15.4% were found to be assemblage A or A/E, respectively. Nested PCR for the tpi gene was the most variable of the targets employed. Twelve new sequences of gdh and tpi for G. duodenalis from sheep were found. Multilocus genotyping resulted in 63 patterns from the 71 samples sequenced at the four loci. This high variability among isolates possibly reflects the high frequency of mixed infections.     

Prevalence of Giardia duodenalis genotypes in pre-weaned dairy calves

To determine the prevalence of Giardia genotypes in pre-weaned dairy calves, fecal samples were collected from a minimum of 18, 1-7-week-old dairy calves per farm on two farms each in the states of Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Samples cleaned of fecal debris and concentrated using CsCl density gradient centrifugation were stained and examined by immunofluorescence microscopy and also subjected to PCR and gene sequence analysis. Prevalence by PCR ranged from 9% on a farm in Pennsylvania to 93% on a farm in Vermont, with an average prevalence for 407 calves on 14 farms of 40%. Gene sequence analysis of the TPI, beta-giardin and 16S rRNA genes revealed 85% of the positive samples to be Assemblage E, while 15% were Assemblage A, although the percentages of these genotypes varied greatly from farm to farm. Some farms had no Assemblage A Giardia. Thus, while a majority of the calves were infected with a genotype that is not known to be infectious for humans, calves on 7 of 14 farms did harbor Assemblage A Giardia. Calves should be considered as a potential source of human infectious cysts in the environment.     

Prevalence and genotyping of Giardia duodenalis from beef calves in Alberta,Canada

Giardia infections in domestic cattle has come under increasing scrutiny owing to the potential contamination of surface and ground waters through manure distribution on fields and pasture runoff. The objective of the study was to determine the prevalence and genotypes of Giardia duodenalis in beef calves in major beef cow calf farms in Alberta, Canada. Fecal samples were collected from beef calves aged 2-10 weeks at nine farms in Alberta. Samples were examined for the presence of G. duodenalis cysts by immunofluorescent staining. Giardia cysts were found in 168 of the 495 fecal samples examined, with prevalence ranging from 7 to 60% among farms. Genotypic analysis of positive isolates utilizing PCR and sequencing of a 292 bp fragment of the 16S-rRNA locus, revealed the hoofed livestock genotype in 41 of the 42 isolates. One isolate was identical to the Assemblage A genotype. The results of this study demonstrate that beef calves in this area are primarily infected with the livestock genotype which is thought to be specific to artiodactyl hosts and non-infective to humans. This suggests that the Giardia carried by beef cattle may be a minimal zoonotic threat.