Determining the zoonotic significance of Giardia and Cryptosporidium in Australian dogs and cats

In a recent study of intestinal parasites in dogs and cats in Australia, Giardia was found to be the most prevalent parasite in dogs. The aim of the current study through the use of molecular tools was to determine the zoonotic significance of the Giardia and Cryptosporidium isolates recovered from dogs and cats during the Australian study. Of the isolates successfully amplified all but one of the Giardia from dogs was either Assemblage C and/or D, with one Assemblage A. Of the cat samples amplified all but one were Assemblage F, with one Assemblage D. We hypothesize that the lack of zoonotic Giardia Assemblages recovered is a result of their being a low prevalence of Giardia in the human population. The Cryptosporidium recovered from dogs and cats was determined to be C. canis and C. felis, respectively, a finding which supports growing evidence that Cryptosporidiumin companion animals is of limited public health significance to healthy people.     

Evaluation of immunofluorescence microscopy and enzyme-linked immunosorbent assay in detection of Cryptosporidium and Giardia infections in asymptomatic dogs

The performance of immunofluorescence microscopy (IF) and enzyme-linked immunosorbent assay (ELISA) in canine feces was evaluated. IF and Cryptosporidium ELISA detected 10(5)oocysts/g, while the detection limit for Giardia ELISA was 10(4)cysts/g. The Cryptosporidium ELISA showed 94% specificity but only 71% sensitivity. The Giardia ELISA correlated well with IF (sensitivity 100%, specificity 96%) and was capable of detecting animal specific Giardia duodenalis genotypes. Visual interpretation appeared appropriate for assessment of ELISA results. The proportion of positive samples and possible zoonotic character of Cryptosporidium and Giardia infections in 150 asymptomatic Finnish dogs from the Helsinki area were studied. The overall proportion of dogs positive for Cryptosporidium was 5% (7/150) and that for Giardia 5% (8/150). In dogs < or =12 months old, the corresponding proportions were 17% and 19% (n=36). Sequence analyses of the 18S rDNA gene identified the isolates as Cryptosporidium canis and animal specific genotypes of G. duodenalis (assemblages C-E), indicating restricted risk of zoonotic transmission.     

Evaluation of commercial ELISA kits for the detection of antibodies against bluetongue virus

The aim of this study was to estimate the diagnostic value of different commercially available ELISA kits for the detection of bluetongue virus (BTV) antibodies in infected and vaccinated animals. The relative specificity of ELISA kits was evaluated using a panel of sera originating from healthy cattle, never vaccinated nor exposed to BTV. All ELISA kits applied had a high relative specificity (99.3 - 100%). The relative sensitivity of ELISA kits assessed using a panel of sera collected from BTV infected cattle was also high and similar for all the kits (97.3 - 100%). However, the relative sensitivity evaluated on the basis of testing vaccinated animals was different: the highest sensitivity was found for Ingenasa, PrioCHECK and ID VET ELISAs (96.5 - 98.3%). Slightly lower sensitivity was calculated for Pourquier and LSI kits (82.8% and 85.4%, respectively) and much lower sensitivity was found for VMRD ELISA kit (69.5%). The repeatability of BTV ELISA kits was expressed as a coefficient of variation (CV) of results of sera tested 5 times in the same day and in different days by the period of 2 months, by the same person, in the same conditions, and by using the same equipment. The CVs of sera tested in all ELISA kits ranged from 6.1 to 9.8% and were below 10% threshold adopted as a maximum for the acceptable repeatability of the method. In conclusion, it can be stated that the applied ELISA kits can be a valuable diagnostic tool for the serological monitoring studies in the BTV contaminated premises. All the methods are very specific and sensitive when testing BTV infected animals. Nevertheless, the Ingenasa and PrioCHECK can be the most useful in sero-surveillance of livestock following vaccination.     

Analysis of commercial Entamoeba histolytica ELISA kits for the detection of Entamoeba invadens in reptiles

Entamoeba invadens is pathogenic in multiple reptile species and has caused severe outbreaks in zoos and other facilities worldwide. Infections can be difficult to diagnose and to differentiate from other reptilian Entamoeba species. The goal of this study was to determine if kits developed to identify the human pathogen Entamoeba histolytica could be used to detect E. invadens in reptile species at the Maryland Zoo. The E. histolytica II antigen enzyme-linked immunosorbent assay and the ProSpecT E. histolytica microplate assay did not react with cultured E. invadens controls or with fecal samples from multiple reptiles, demonstrating the need for a sensitive and specific test for E. invadens.     

3种ELISA试剂盒检测抗牛支原体抗体的比较

为比较3种抗牛支原体(M.bovis)血清抗体的ELISA试剂盒检测效果,本实验应用3种检测M.bovis血清抗体ELISA诊断试剂盒对38份阳性样品(自然感染19份,人工感染18份)和37份阴性样品进行检测.结果表明:本实验室制备的HVRI试剂盒与商品化试剂盒Kit 1的检测结果和综合检测结果符合率分别达到92%和96%;而商品化试剂盒Kit 2的检测结果与综合检测结果符合率仅为74.67%.一致性检验结果显示:HVRI试剂盒与Kit 1的一致性较高;Kit 2与HVRI试剂盒、Kit 2与Kit 1有中度的一致性.此外,3种试剂盒对牛传染性胸膜肺炎国际标准血清(PS2)的检测结果显示HVRI试剂盒和Kit 1均为为阴性,Kit 2为阳性.因此,HVRI试剂盒与Kit 1更适于M.bovis检测和开展流行病学调查.     

尿样中莱克多巴胺残留ELISA检测试剂盒的比对实验

酶联免疫吸附技术（ELIsA）由于其快速、简便、适合同时处理大量样品的特点，被国内外普遍应用于进行口一兴奋剂等各种兽药残留的筛选检测。国内目前销售的菜克多巴胺残留ELISA试剂盒主要有4种，据悉新的试剂盒还将推出。但国内外尚缺乏对不同莱克多巴胺ELISA试剂盒的检测性能的权威评估。由于不同试剂盒的检测效果可能存在差异，检测试剂不统一，可能导致检测结果差异较大，将不利于政府残留监控计划的实施和企业维护自身权益。本研究拟通过比对检测试验，对不同莱克多巴胺ELISA试剂盒进行评估和比对，为制定合理的莱克多巴胺残留检测技术方案和准确评判检测结果提供科学依据。     

Evaluation of two commercial test kits for detection of occult blood in feces of dogs

To evaluate the sensitivity and specificity of 2 commercial test kits for detection of occult blood in canine feces, various volumes of blood were administered to 6 dogs via orogastric tube. Blood volumes tested were chosen on the basis of hemoglobin quantities of 5, 10, 20, 200, 350, and 500 mg of hemoglobin/kg of body weight. Fecal specimens were collected twice daily and analyzed separately by 2 observers for the presence of occult blood by use of modified guaiac and orthotolodine tablet tests, and for melena by visual inspection. Five dogs given blood at the rate of 500 mg of hemoglobin/kg and 1 dog given blood at the rate of 350 mg of hemoglobin/kg developed melena. Results of both occult blood tests were positive in 2 of 6 dogs given blood at the rate of 5 mg of hemoglobin/kg. Five of 6, and 4 of 6 dogs given blood at the rate of 10 mg hemoglobin/kg had positive test results by modified guaiac and orthotolodine methods, respectively. Results of both methods were positive in all dogs given blood at the rate of 20 mg of hemoglobin/kg. There was 86% agreement between the 2 observers' results for the modified guaiac method, and 78% agreement for the orthotolodine method. There was 77% agreement of results between the 2 test methods. Gastrointestinal transit time decreased with increasing volumes of blood. Occult blood testing was found to be useful for detection of blood in feces at volumes 20 to 50 times less than that required to cause melena.     

Prevalence of Cryptosporidium, Giardia and Isospora species infections in pet cats with clinical signs of gastrointestinal disease

This study reports the prevalence of Cryptosporidium, Giardia and Isospora species in cats showing signs of gastrointestinal disease. Records from a United Kingdom commercial diagnostic laboratory between December 2003 and December 2005 were reviewed. Of 1355 cats, Cryptosporidium species oocysts were found in 13 cats (1%), Giardia species trophozoites in 74 (6%), and Isospora felis oocysts in 46 (3%). In a second group of 48 cats, prevalence of Giardia species was 15% using an immunoassay for detection of antigen compared to 4% detected with microscopy. Prevalence of Giardia (9%) and Isospora (9%) species was higher in cats less than 6 months old. Gender and breed did not affect prevalence. There was a trend for Cryptosporidium and Isospora species infections to be detected in late autumn and early winter. Regional differences in prevalence were not detected. None of these organisms show a characteristic pattern of clinical signs. This study demonstrates that enteric protozoal infection is common in domestic cats showing signs of alimentary disease.     

Giardia infection of cats and dogs in New Zealand

Giardia intestinalis is a pathogenic protozoan which infects humans and a wide range of animal hosts, including cats and dogs⁽¹⁾. However, the status of animals in New Zealand with respect to Giardia infection has not received much attention, so we undertook a preliminary study of cats and dogs in Palmerston North and Hamilton to determine the prevalence of infection, as indicated by the presence of cysts in faeces.     

Prevalence and genetic characterization of Giardia and Cryptosporidium in cats from Italy

One hundred and eighty one cats living in central Italy were tested for the presence of Giardia and Cryptosporidium infection by IFAT test and specific PCRs. Overall eight (4.4%) samples were IFAT-positive for Giardia. All the IFAT-positive samples for Giardia scored positive for the PCRs, and three more samples IFAT-negative generated PCR products leading to a total 6.1% molecular positivity rate for Giardia. All the examined samples were negative for Cryptosporidium. Sequencing of samples molecularly positive to Giardia indicated that three cats harbored the zoonotic Giardia duodenalis Assemblage A, whereas all other positive animals were infected with the feline-specific G. duodenalis Assemblage F. Phylogenetic analysis carried out on the sequences obtained supported the clustering of the isolates within Assemblages A and F. The results here presented provide data on the occurrence of Giardia genotypes in cats living in close contact with humans highlighting the potential importance of this protozoan disease for the public health.     

Prevalence and genetic characterization of Giardia and Cryptosporidium in cats from Italy

One hundred and eighty one cats living in central Italy were tested for the presence of Giardia and Cryptosporidium infection by IFAT test and specific PCRs. Overall eight (4.4%) samples were IFAT-positive for Giardia. All the IFAT-positive samples for Giardia scored positive for the PCRs, and three more samples IFAT-negative generated PCR products leading to a total 6.1% molecular positivity rate for Giardia. All the examined samples were negative for Cryptosporidium. Sequencing of samples molecularly positive to Giardia indicated that three cats harbored the zoonotic Giardia duodenalis Assemblage A, whereas all other positive animals were infected with the feline-specific G. duodenalis Assemblage F. Phylogenetic analysis carried out on the sequences obtained supported the clustering of the isolates within Assemblages A and F. The results here presented provide data on the occurrence of Giardia genotypes in cats living in close contact with humans highlighting the potential importance of this protozoan disease for the public health.     

Comparison of commercial ELISA blood tests for early pregnancy detection in dairy cows

The objective of the present study was to compare two commercially available blood-based pregnancy tests, namely BioPRYN, an ELISA for pregnancy-specific protein B (PSPB), and an ELISA for pregnancy-associated glycoprotein (PAG), for early pregnancy diagnosis in dairy cattle using transrectal ultrasonography as a gold standard. Transrectal ultrasonography was conducted 26-58 days after artificial insemination (AI) in 197 cattle from 19 farms. Concurrently, a blood sample was collected for determination of serum PSPB and PAG. Transrectal palpation was performed approximately 120 days after AI to verify that pregnancy was maintained. For PSPB and PAG, there were no significant differences (P>0.05) in sensitivity (98.0 and 97.8%), specificity (97.1 and 91.2%), positive predictive values (99.3 and 97.8%), negative predictive values (91.9 and 91.2%) and accuracy (97.8 and 96.4%). In conclusion, the two blood pregnancy assays were equally efficacious and were highly accurate (based on transrectal ultrasonography as the gold standard).     

Survey and molecular characterization of Cryptosporidium and Giardia spp. in owned companion animal, dogs and cats, in Japan

Compared with other countries, surveys of these parasites have been rarely performed in companion animals of Japan in spite of their significance for public health. Here, we investigated pet dogs and cats in Japan for the first time, and genetically analyzed the isolates to evaluate the risk of zoonotic infections. Seventy-seven fecal samples were collected from privately owned dogs and 55 samples from owned cats in Osaka city, Japan. Cryptosporidium oocysts were identified in 3/77 dogs (3.9%) and 7/55 cats (12.7%), and Giardia infection in 2/77 dogs (2.6%) and 1/55 cats (1.8%). Amplification of the target regions for genotyping was successful, Cryptosporidium isolates in dogs and cats were identified as C. canis and C. felis, respectively, and those of Giardia in dogs and cats were G. intestinalis Assemblages D and F. The discharge period of the oocysts varied within 3-16 weeks and that of the cysts was 12 weeks. To date, zoonotic types of both parasites have been identified in other animals in Japan, and further large-scale studies are needed to determine the distribution of zoonotic genotypes in these animals, especially those closely associated with humans.     

3种口蹄疫O型抗体检测试剂盒对疫苗免疫牛血清抗体检测的比较试验

为了提供检测结果准确且操作便捷的试剂盒,本试验应用两种商品化试剂盒和本实验室研制的一种基于合成肽包被抗原的间接ELISA试剂盒同时对口蹄疫疫苗免疫后14、21和28 d的150份免疫牛血清进行口蹄疫O型抗体水平的检测,并对3种试剂盒测定结果的符合程度、重复性和便捷性进行了比较。试验结果表明3种试剂盒测得的抗体水平检测结果之间高度符合,且3种试剂盒各自的检测结果重复性良好,基于合成肽包被抗原的间接ELISA试剂盒在操作便捷性方面优势明显。