Zinc Sulfate Centrifugal Flotation Research Articles

Detection of parasitic forms in feces of Chelonoidis carbonarius (Linnaeus, 1766) from Brazilian Cerrado.

Chelonoidis carbonarius is host to a great diversity of endoparasites. This research aimed to identify and taxonomically classify the parasitic forms of endoparasites found in the feces of C. carbonarius using coproparasitological methods. In total, 38 fecal samples were collected from animals captured in the municipalities of Cuiabá (n = 23) and Jataí (n = 15) in the Brazilian Ceraado. Fecal samples were submitted to at least one of the following techniques: Willis-Mollay flotation technique, zinc sulfate centrifugal flotation technique, and spontaneous sedimentation of Hoffman, and Pons, and Janer. Overall, 86.85% (33/38) of animals were positive in at least one of the diagnostic techniques. Of the 38 samples used, 50% of the animals were captive and 50% of the animals were free-living. Cysts, oocysts, and parasitic eggs were identified, as follows: Chapiniella spp. (65.78%; n = 24), Sauricola spp. (31.57%; n = 12), Ascaroidea (15.78%; n = 6), Oxyuridae (2.63%; n = 1), Trematoda (2.63%; n = 1), Acanthocephala (2.63%; n = 1), Balantidium spp. (39.47%; n = 15), Eimeria spp. (21.05%; n = 8), and Nyctotherus spp. (2.63%; n = 1). The study showed that both captive and free-living C. carbonarius are often parasitized by different endoparasite species, with nematode eggs being the most common.

Comparative study of a broad qPCR panel and centrifugal flotation for detection of gastrointestinal parasites in fecal samples from dogs and cats in the United States

BackgroundFor decades, zinc sulfate centrifugal fecal flotation microscopy (ZCF) has been the mainstay technique for gastrointestinal (GI) parasite screening at veterinary clinics and laboratories. Elsewhere, PCR has replaced microscopy because of generally increased sensitivity and detection capabilities; however, until recently it has been unavailable commercially. Therefore, the primary aim of this study was to compare the performance of real-time PCR (qPCR) and ZCF for fecal parasite screening. Secondary aims included further characterization of markers for hookworm treatment resistance and Giardia spp. assemblages with zoonotic potential and qPCR optimization.MethodsA convenience sampling of 931 canine/feline fecal samples submitted to a veterinary reference laboratory for routine ZCF from the Northeast US (11/2022) was subsequently evaluated by a broad qPCR panel following retention release. Detection frequency and agreement (kappa statistics) were evaluated between ZCF and qPCR for seven GI parasites [hookworm/(Ancylostoma spp.), roundworm/(Toxocara spp.), whipworm/(Trichuris spp.), Giardiaduodenalis, Cystoisospora spp., Toxoplasmagondii, and Tritrichomonasblagburni] and detections per sample. Total detection frequencies were compared using a paired t-test; positive sample and co-infection frequencies were compared using Pearson’s chi-squared test (p ≤ 0.05 significant) and qPCR frequency for hookworm benzimidazole (BZ) resistance (F167Y) and zoonotic Giardia spp. assemblage markers calculated. Confirmatory testing, characterization, and qPCR optimization were carried out with Sanger sequencing.ResultsqPCR detected a significantly higher overall parasite frequency (n = 679) compared to ZCF (n = 437) [p = < 0.0001, t = 14.38, degrees-of-freedom (df) = 930] and 2.6 × the co-infections [qPCR (n = 172) vs. ZCF (n = 66)], which was also significant (p = < 0.0001, X2 = 279.49; df = 1). While overall agreement of parasite detection was substantial [kappa = 0.74; (0.69–0.78], ZCF-undetected parasites reduced agreement for individual and co-infected samples. qPCR detected markers for Ancylostomacaninum BZ resistance (n = 5, 16.1%) and Giardia with zoonotic potential (n = 22, 9.1%) as well as two parasites undetected by ZCF (T.gondii/T.blagburni). Sanger sequencing detected novel roundworm species, and qPCR optimization provided detection beyond ZCF.ConclusionsThese results demonstrate the statistically significant detection frequency advantage offered by qPCR compared to routine ZCF for both single and co-infections. While overall agreement was excellent, this rapid, commercially available qPCR panel offers benefits beyond ZCF with detection of markers for Giardia assemblages with zoonotic potential and hookworm (A.caninum) BZ resistance.Graphical

THE EFFECTIVENESS OF COMPOSTING USING STABILIZING URBAN SEWAGE SLUDGE COMPOUNDS

The purpose of this investigation was to verify the effectiveness of composting in the sanitization of sludge from urban sewage (SS). The treatments (T) used consisted of SS mixed with sugarcane bagasse (SB), tree pruning residues (TP), poultry litter (PL) or grass clipping (GC) at a carbon/nitrogen ratio of approximately 30/1 and ratios of T1 (SS+SB), T2 (SS+SB+TP), T3 (SS+SB+GC), T4 (SS+SB+PL) or T5 (SS+SB+TP+GC). Temperature was measured daily for over 280 days of composting. Every two weeks, the compounds were analyzed for thermotolerant coliforms and viable helminth eggs using the multiple-tube and Yanko techniques; and once each month to identify Cryptosporidium spp. and Giardia spp. (oo)cysts using the sucrose and zinc sulfate centrifugal flotation technique and nested PCR followed by sequencing. The maximum temperatures measured in the thermophilic phase of the compounds ranged from 55°C to 64.8°C, and at day 280, weight reductions of 27% (T1), 48% (T2), 63% (T3), 66% (T4) and 64% (T5) were observed. The absence of fecal coliforms was observed from day 98 (T4), day 126 (T1), day 196 (T3 and T5) and day 210 (T2). All treatments resulted in helminth egg inviability and the absence of protozoan (oo)cysts. Only T4 and T5 were positive for G. duodenalis according to the sequencing analysis. In conclusion, high temperatures during the composting thermophilic phase made the use of pathogens required by legislation unfeasible; therefore, it is important to investigate the viability of protozoa in compounds to ensure a safe final product for human and animal health.

The first study on the prevalence of gastrointestinal parasites in owned and sheltered cats in Yangon, Myanmar.

People who used to rear companion animals are healthier than others who do not. Gastrointestinal (GI) helminths are common in cats and serve as reservoirs for zoonotic diseases. However, the prevalence of GI parasites in cats in Myanmar has never been reported. This study aimed to estimate the prevalence of GI parasites in cats in Myanmar and to identify the potential risk factors associated with GI parasites. A total of 230 fecal samples were collected from seven veterinary clinics and two shelters within the Yangon region from January to May 2022. Sampled cats were classified according to age, gender, and the deworming and rearing practices. Fecal samples were analyzed by fecal wet mount, ethyl acetate centrifugal sedimentation, and zinc sulfate centrifugal flotation techniques. Descriptive data were described, and Pearson's χ2 test was used to identify associated risk factors, such as age, gender, and the deworming and rearing practices. The overall prevalence of GI parasites was 79.56%, and 57.82% of cats were infected with a diagnostic stage of more than one parasite species. Seven GI parasites were detected, including Ancylostoma spp. (55.65%), Toxocara spp. (46.08%), Trichuris spp. (20.86%), Platynosomum spp. (11.73%), Dipylidium caninum (7.39%), Taenia spp. (4.34%), and Cystoisospora spp. (32.17%). Based on statistical analysis, the deworming and rearing practices were significantly associated (p < 0.05) with GI parasitic infections. This study is the first to reveal the prevalence of GI parasites that could assist the need for effective control measures for zoonotic hookworm and roundworm infections in cats. Even with simple microscopic examination, the remarkably high prevalence of GI parasitic infections warrants regular deworming practice. Further molecular studies should also be performed to understand their genetic diversity.

Prevalence, risk factors and zoonotic potential of intestinal parasites in dogs from four locations in Morocco.

Dogs can harbor various intestinal parasites that have serious clinical, economic, and zoonotic impact. In Morocco, the epidemiological status of those parasites is largely unknown. This study aimed to obtain data on the prevalence of intestinal parasites in various Moroccan dog populations, to identify associated risk factors, to evaluate people's knowledge regarding zoonotic parasites and to estimate the risk of human infection. A total of 291 fecal samples were analyzed using a 33% Zinc Sulphate (ZnSO4) centrifugal flotation and a sheather's sugar simple flotation techniques. In addition, 100 dog owners were asked to fill out a questionnaire about their knowledge regarding canine zoonotic intestinal parasites. Overall, 58% of sampled dogs were positive for at least one parasite species. Ancylostoma/Uncinaria spp. (31.9%), Toxascaris leonina (27.4%), Toxocara canis (27.1%), Cystoisospora spp. (13.4%) and Giardia spp. (7.2%) were the most frequently isolated parasites. The overall prevalence was associated with the dogs' activity, and feces consistency. More specifically, the prevalence of Ancylostoma/Uncinaria spp., Toxascaris leonina and Cystoisospora spp. was associated with dogs' activity and feces consistency and, additionally with age for Cystoisosporaspp.Toxocara canis prevalence was associated with age and feces consistency, while Giardia spp. prevalence was associated with dogs' activity. A weak awareness among dog owners regarding the zoonotic potential of canine intestinal parasites was noticed (33%) especially when their knowledge was compared to another endemic zoonotic disease, i.e. rabies (85%). Furthermore, the rate of respecting the vaccination protocol (82%) was significantly higher than the rate of respecting the deworming protocol (47%). We conclude that intestinal parasites, including the zoonotic ones, were highly prevalent in the dogs sampled in this study whereas people's knowledge about those parasites was very limited. These findings suggest that the risk of human infection is very likely, which highlights the need for effective control programs and health education.

Infection by intestinal parasites in disabled patients and their guardians

Introduction: intestinal parasitic infections are common major problem closely related to poverty, inadequate sanitation, insufficient health care and overcrowding. They cause significant morbidity among institutionalized patients, however, there are few studies that analyze the frequency of intestinal parasites in disabled patients that are not institutionalized. Objective: the aim of the present study was to determine the prevalence of intestinal parasitic infection in disabled patients and their guardians. Methodology: a total of 336 fecal samples were collected from 53 disabled patients and history of diarrhea during the study period and 31 guardians, parents and professional staff of Institution. Parasite research was carried out using zinc sulphate centrifugal-flotation technique, Lutz/Hoffman Pons and Janer method, Rugai method and Gram-Chromotrope, Leishman, Kinyoun, Kato-Katz and Trichrome stains were used. Results: we found 15.5% of positive sample for enteroparasites in all analyzed individuals (13/84), with 11.3% (6/53) of prevalence in disabled patients and 22.5% (7/31) for guardians, with significant difference. There was no difference between gender,but there was a higher number of positives in patients between 6 and 11 years of age. Monoparasitism and the presence of protozoa, especially Blastocystis hominis, were the most prevalent conditions. Conclusions: despite the aforementioned intrinsic susceptibility of patients with special needs, the prevalence of intestinal parasites was low. In guardians, the prevalence was higher, suggesting extreme attention to the care process, which may have prevented the transmission to their disabled patients contact.

Parasitological and bacteriological survey of liquid wastes from Naze industrial clusters in Owerri, Imo state

Liquid wastes are known to be highly contaminated and pose great public health risk, especially when indiscriminately discharged to the environment. The parasitological and bacteriological survey of liquid wastes from Naze industrial clusters in Owerri metropolis was carried out. Sewage wastewater and laundry wastewater were used as study samples. 100mls each of the samples were collected from the drainage sites with sterile screw-capped specimen bottles, and sent to the laboratory for analysis. The liquid wastes were filtered and the filtrates were used for the analysis. Parasitological analysis employed Zinc Sulphate Centrifugal Flotation Technique and Modified Ziel-Nielsen stain (Acid-Fast Staining). Parasitic ova, cysts, larva and oocysts isolated were identified using standard morphological and taxonomic keys. Bacterial isolates were identified using standard microbiological techniques and biochemical characteristics where necessary. Parasites such as Ascaris lumbricoides, Strongyloides stercoralis, Trichuris trichiura, Enterobius vermicularis, Schistosoma mansoni, Hymenolepsis nana, Entamoeba histolytica, Gardia lamblia, Balantidium coll and Isospora belli were isolated from the liquid wastes. Bacteria species isolated included Erwinia amylovora, Listeria monocytogens, Escherichia coli, Salmonella typhi, Bacillus cereus, Staphylococcus aureus and Proteus vulgaris. The results showed that the most prevalent parasite and bacteria isolated were Ascaris lumbricoides 49(37.6%) and Salmonella typhi (3.38x104 CFU/mL) respectively. These potentially pathogenic parasites and bacteria detected in these liquid wastes, pose serious threat to public health. Interventive measures such as public health education, adequate drainage and waste disposal system, regular and monitored environmental sanitation, and provision of safe and portable water supply to the study area, are recommended.

Rapid on-site diagnosis of canine giardiosis: time versus performance

BackgroundInfections by protozoans of the genus Giardia are a common cause of diarrhea in dogs. Canine giardiosis constitutes a disease with a zoonotic potential; however, it is often underestimated due to its challenging diagnosis. The objective of the study was to assess the diagnostic performance of an immunochromatographic strip test (SpeedTMGiardia, Virbac, France) comparing it with microscopy (zinc sulfate flotation) by utilizing the combination of an enzyme immunoassay (ProSpecTTMGiardia EZ Microplate Assay, Oxoid Ltd., UK) and the PCR as the gold standard. A positive result in both ELISA and PCR was set as the gold standard.MethodsInitially, fecal samples from dogs with clinical signs compatible with giardiosis were tested with the SpeedTMGiardia test and separated into two groups of 50 samples each: group A (positive) and group B (negative). Thereafter, all samples were examined by zinc sulfate centrifugal flotation technique and assayed by the ProSpecTTMGiardia Microplate Assay and PCR. The performance of the SpeedTMGiardia and zinc sulfate centrifugal flotation tests were calculated estimating sensitivity, specificity, and positive and negative likelihood ratio; the chi-square and McNemar tests were used for the comparison of the two methods.ResultsGiardia cysts were not detected by microscopy in 16 out of the 50 samples (32%) of group A and in none of group B samples. Eight out of 50 samples in group B (16%) were tested positive both with the ProSpecTTMGiardia Microplate Assay and PCR. Fecal examination with the SpeedTMGiardia test was more sensitive (86.2%) than the parasitological method (58.6%, P < 0.001) while the specificity of both methods was 100%.ConclusionsThe SpeedTMGiardia test is an easy-to-perform diagnostic method for the detection of Giardia spp., which can increase laboratory efficiency by reducing time and cost and decrease underdiagnosis of Giardia spp. infections. This immunochromatographic strip test may be routinely exploited when a rapid and reliable diagnosis is required, other diagnostic techniques are unavailable and microscopy expertise is inefficient. In negative dogs with compatible clinical signs of giardiosis, it is recommended either to repeat the exam or proceed with further ELISA and PCR testing.

Detection of gastrointestinal parasitism at recreational canine sites in the USA: the DOGPARCS study

BackgroundThe rapid growth in off-leash dog parks provides opportunity for canine socialization activities but carries risk of exposure to intestinal parasites. This study assessed the prevalence of these infections in dogs visiting off-leash dog parks.MethodsFresh defecations were collected from dogs visiting parks in 30 metropolitan areas across the USA. Samples were analyzed by coproantigen immunoassay (CAI) (Fecal Dx® and Giardia Test, IDEXX Laboratories, Inc.) and zinc sulfate centrifugal flotation (CF). Owners responded to a questionnaire on their dog’s signalment and use of heartworm/intestinal parasite control medications (HWCM).ResultsSamples were examined from 3006 dogs, 87.9% aged at least 12 months, visiting 288 parks. At least one intestinal parasite was detected in 622 (20.7%) samples, nematodes in 263 (8.8%), with hookworms, whipworms and ascarids in 7.1, 1.9 and 0.6% of samples, respectively. A sample positive for one or more intestinal parasites was found in 245 (85.1%) parks, with nematodes found in 143 (49.7%). Combined, CAI and CF detected 78.4% more intestinal nematode infections than CF alone. Hookworm and whipworm infections were detected in all age groups, but ascarids were only detected in dogs less than 4 years-old. Approximately 42% of dogs aged less than 1 year were positive for nematodes or Giardia. Based on owner reports, HWCM was current for 68.8% of dogs, dogs previously diagnosed with intestinal parasitism were more likely to be receiving a HWCM than those without such history, and a significantly lower (P = 0.0003) proportion of dogs receiving a HWCM were positive for intestinal nematodes compared with those not on such medication.ConclusionsIntestinal parasites, the most common of which were Giardia, Ancylostoma caninum and Trichuris vulpis, were found in 20% of dogs and 85% of dog parks across the USA. Enhanced detection of canine intestinal parasitism was achieved by combining CF and CAI. Canine intestinal parasites are common across the USA and dog health can be improved by regular testing of fecal samples and routine administration of medications effective against the most common infections.

Seasonal distributions and other risk factors for Giardia duodenalis and Cryptosporidium spp. infections in dogs and cats in Chiang Mai, Thailand

The objectives of this study were to explore risk factors associated with Giardia and Cryptosporidium infections in dogs and cats in Chiang Mai, Thailand, to describe the seasonal distributions of Giardia and Cryptosporidium prevalence, and to determine the potential for zoonotic transmission through genetic characterization of isolates. Fecal samples from 301 dogs and 66 cats were collected between August 2009 and February 2010. The presence of Giardia cysts and Cryptosporidium oocysts was determined using zinc sulfate centrifugal flotation and immunofluorescent assay (IFA). Genotype/species were determined by DNA sequence analyses of PCR products from Giardia glutamate dehydrogenase (gdh), beta-giardin (bg), and triosephosphateisomerase (tpi) and Cryptosporidium heat shock protein 70KDa (hsp70) and small subunit-rRNA (SSU-rRNA) genes. Information related to specific risk factors was collected from owners of each animal using a questionnaire. The risk factor data were analyzed for associations with Giardia and Cryptosporidium infections using logistic regression.The overall estimated prevalence of Giardia and Cryptosporidium in dogs was 25.2% and 7.6%, respectively and in cats, 27.3% and 12.1%, respectively. The estimated prevalence of Giardia infection in dogs in the rainy season (31.7%) was significantly higher than in the drier, winter season (17.2%) (p < 0.01). The estimated prevalence of Cryptosporidium infection in dogs and of Giardia and Cryptosporidium infections in cats was not associated with season (p > 0.05). Multivariable analysis indicated that Giardia cysts were more likely to be detected in fecal samples of dogs that resided in high-density environments, drank untreated water, were shedding Cryptosporidium oocysts, were having acute diarrhea or a history of chronic diarrhea, and were collected in the rainy season. All 19 Giardia PCR positive samples typed as G. duodenalis canine adapted genotypes (assemblages C or D). In cats, of six Giardia PCR positive samples, five typed as dog assemblages and one typed as assemblage AI. Of ten dogs with Cryptosporidium PCR positive samples, eight typed as C. canis, one as C. parvum (a zoonotic species) and one had both C. canis and C. parvum. Of three Cryptosporidium PCR positive samples in cats, one typed as C. felis and two typed as C. parvum.The presence of zoonotic G. duodenalis assemblage AI in a cat, and C. parvum in feces of dogs and cats suggests a potential role for a reservoir for zoonotic transmission. Whether or not these presences were from exposure to other animal or human hosts or environment are needed to be confirmed.

Retrospective and Comparative Study of Giardia sp. Prevalence in Dogs, Cats, and Small Ruminants in Endemic Areas in Different Brazilian States

Background: Giardia, an intestinal parasite of asexual reproduction, is an important etiologic agent of diarrhea in animals and humans, transmitted by orofecal route. The disease caused by this agent, giardiasis, is endemic in the world and representing an important public health problem. The aim of the present study was do a retrospective study to determine the prevalence of Giardia sp. in fecal samples from dogs, cats, sheep and goats (small ruminants) evaluated at FMVZ Unesp, Botucatu, SP. In addition, this study presents a review of the literature on the prevalence of this protozoan in dogs, cats and small ruminants in different states of Brazil, according to the environmental of these animals.Materials, Methods &amp; Results: During 2011 to 2017, 2,698 fecal samples of dogs, 359 fecal samples of cats and 320 fecal samples of goats and sheep were analyzed. A total of 18.9% of the dogs, 24.8% of the cats, and 6.6% of the goats and sheep were positive for Giardia sp. Only previous studies that used the zinc sulphate centrifugal flotation (Faust technique) were included for comparation of prevalence. The prevalence of Giardia sp. in samples of centrifugation-fluctuation in zinc sulfate was similar in dogs, higher in cats and lower for small ruminants when compared to previous investigations. 4.4% and 6.7% of the positive samples from dogs and cats, respectively, were associated with some co-infection. Co-infections by Cystoisospora spp., Ancylostoma spp. and/ or Toxocara spp. were the most common for dogs and cats.Discussion: This study reveals the presence of Giardia sp. in different animal species in an endemic área. The results are similar to the prevalence previous reported in dogs, and higher in cats using the same diagnostic technique (Faust technique). Co-infections by Cystoisospora spp., Ancylostoma spp. and/ or Toxocara spp. were the most common for dogs and cats, as well-known in previous studies. Investigations with household, shelter and stray dogs found a significantly lower occurrence of Giardia sp. in the group of household dogs. However, for cats these comparisons should made with caution. Despite having a owner, some of these cats have free access to the streets, so they might be more expose to the sources of infection. There are few previous reports of the presence of Giardia sp. in goats and sheep, which presented much higher prevalence when compared with this study. This variation in prevalance of Giardia sp. in small ruminants can occur due to differences inherent to each region in Brazil, as well as the presence of risck factors regarding animal age, type of raising of the animals, hygienic-sanitary and management conditions, and the presence of domestic animals inside sheep and goat installations, which can potentially favor the mechanical transmission of cysts our reservoirs of this parasite. Therefore, due to regional variations in this parasite prevalence, this information is more value in regional areas, reaffirming the importance of this kind of studies in brazilian states and cities. The intermittent elimination of cysts in feces, the low number of cysts in the samples and asymptomatic infections are relevant points to the diagnosis of giardiasis, making it a challenge. The technique of centrifugation-flotation technique in zinc sulfate gives the best results when compared to other techniques available for the diagnosis of Giardia infections, justifying the choice of this technique by this study. These evidences associated to molecular diagnoses are necessary to determine the real role of these animals in the epidemiology and zoonotic transmission, due to the close contact with humans. This may be considering as a start for further investigations of Giardia sp. at municipality of Botucatu.

Intestinal Parasites and the Occurrence of Zoonotic Giardia duodenalis Genotype in Captive Gibbons at Krabokkoo Wildlife Breeding Center, Thailand.

Intestinal parasitic infections can have an impact on health and growth of wildlife. The current study aims were to determine the prevalence of intestinal parasites and to molecular characterize Giardia duodenalis and Cryptosporidium spp. in captive gibbons at Krabokkoo Wildlife Breeding Center, Thailand. Fifty-five gibbons, 2 agile- (Hylobates agilis), 38 lar- (Hylobates lar) and 15 pileated gibbons (Hylobates pileatus) were included in this study. Fecal samples were collected individually at Krabokkoo Wildlife Breeding Center, Chachoengsao province, eastern Thailand, in November 2013. Intestinal parasitic infections were examined by zinc sulfate centrifugation flotation and by a commercially available immunofluorescent assay (IFA) for detection of G. duodenalis and Cryptosporidium spp.. Polymerase chain reaction targeting the Giardia glutamate dehydrogenase (gdh), beta- giardin (bg), triose phosphate isomerase (tpi) genes, and the Cryptosporidium small subunit-rRNA and heat-shock protein (hsp70) following by DNA sequencing were performed on the IFA positive samples. The overall prevalence of intestinal parasitic infection in gibbons at Krabokkoo Wildlife Breeding Center was 12.7% (95%CI: 5.3–24.5), Strongyloides spp. eggs or larvae were present in all positive samples. Co-infections with G. duodenalis were detected in 1.8% (95%CI: 0.1–9.7) of the samples. Based on the sequencing results of the three genes, the IFA Giardia positive isolate typed as the zoonotic genotype B. Since the data reveals the occurrence of zoonotic Giardia genotype, good hygiene management is suggested to prevent the transmission of this pathogen from gibbon to human, and vice versa.

Comparison of five diagnostic tests for Giardia duodenalis in fecal samples from young dogs

Five diagnostic tests were compared for the diagnosis of Giardia duodenalis in fecal samples of young dogs. Fecal samples were collected from 136 healthy dogs <1year old and examined using immunofluorescence antibody microscopy (IFA) after sucrose gradient centrifugation, zinc sulfate centrifugal flotation technique (ZSCT), SNAP®Giardia test, and ProSpecT®Giardia EZ Microplate assay. In addition, polymerase chain reaction (PCR) of the 16S rRNA gene was performed. Kappa (κ) statistic was calculated to assess diagnostic agreement between the IFA and each test. Using the IFA as the gold standard, the relative sensitivity and specificity of each test were determined. Subsequently, a Bayesian approach was used to estimate the sensitivity and specificity of each test in comparison to the IFA results. Giardia duodenalis was detected in 41% of the samples examined by IFA. The ZSCT resulted in 37% of positive samples, with a relative sensitivity and specificity of 86 and 98%, respectively. The SNAP®Giardia test was positive in 40% of the samples, with a relative sensitivity and specificity of 91 and 96%, respectively. The ProSpecT® test was positive in 51% of the samples, with a relative sensitivity and specificity of 100 and 83%, respectively. The relative sensitivity and specificity for PCR were 58 and 56%, respectively, with 55% of samples being PCR-positive. While the sensitivity and specificity estimates of each test in comparison to the IFA changed when using a Bayesian approach, the conclusions remained the same. While the ProSpecT® test was the most sensitive test in this study, it is not designed for dogs and more costly than the other tests. The SNAP®Giardia test performed similar to the ZSCT but may be more favorable because it is fast and easy to perform. Performance of the PCR was poor and the benefit of PCR may be in determining genotypes for evaluating zoonotic transfer between dogs and humans.

Prevalence and Multilocus Genotyping Analysis of Cryptosporidium and Giardia Isolates from Dogs in Chiang Mai, Thailand.

The occurrence and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis isolated from dogs in Chiang Mai, Thailand were determined. Fecal samples were collected from 109 dogs between July and August 2008. Cryptosporidium spp. infection was determined by immunofluorescent assay (IFA), PCR assays that amplify Cryptosporidium heat-shock protein 70 kDa (hsp70), and two PCR assays that amplify a small subunit-ribosomal RNA (SSU-rRNA). Giardia duodenalis infection was identified using zinc sulfate centrifugal flotation, IFA, and four PCR assays that amplify the Giardia glutamate dehydrogenase (gdh), beta-giardin (bg), and generic and dog-specific assays of triosephosphate isomerase (tpi) genes. Overall prevalence of Cryptosporidium spp. and G. duodenalis was 31.2% and 45.9%, respectively. Sequence analysis of 22 Cryptosporidium-positive samples and 21 Giardia-positive samples revealed the presence of C. canis in 15, and C. parvum in 7, G. duodenalis Assemblage C in 8, D in 11, and mixed of C and D in 2 dogs. Dogs in Chiang Mai were commonly exposed to Cryptosporidium spp. and G. duodenalis. Cryptosporidium parvum can be isolated from the feces of dogs, and all G. duodenalis assemblages were dog-specific. Dogs could be a reservoir for a zoonotic Cryptosporidium infection in humans, but further studies will be required to determine the clinical and zoonotic importance.

Changes in biochemical analytes in female dogs with subclinical Ancylostoma spp. infection.

BackgroundAncylostoma spp. is one of the most prevalent canine intestinal nematode infections which usually causes subclinical disease in adult dogs and has zoonotic implications. Therefore, the aim of this study was to explore and evaluate the possible pathophysiological changes that Ancylostoma spp. could produce in female dogs naturally infected but without clinical signs of disease, by screening a wide variety of biochemical markers for potential changes. Samples of feces and blood of 45 dogs were collected and fecal flotation and zinc sulphate centrifugal flotation were performed. The biochemical analytes determined were: the acute-phase proteins C-reactive protein (CRP) and haptoglobin (Hp); the lipid profile (cholesterol, triglycerides, HDL, LDL); the serum iron profile: iron, unsaturated iron binding-capacity (UIBC), and ferritin; the enzyme butyrylcholinesterase (BChe); the pancreatic profile: amylase, lipase, and trypsin-like immunoreactivity (TLI); the oxidative stress markers: total antioxidant capacity (TAC) and paraoxonase −1 (PON-1), along with total protein, albumin, and insulin-like growth factor – 1 (IGF – 1). Ancylostoma spp. eggs were detected in 29/45 dogs (64.4 %). Dogs were divided into two groups according to the results of fecal flotation methods. Group 1: negative fecal floatation (n = 16), and Group 2: subclinical infection with the observation of Ancylostoma spp. type eggs/x 40 objective fields (n = 29).ResultsMann–Whitney U test was used to compare the biochemical analyte results between the two groups (P < 0.05). Significant increases in CRP (μg/mL) (median): non-infected dogs: 5.5; subclinically infected dogs 18.7; P = 0.03, Hp (g/L) (median): G1: 2.4; G2: 3.3; P = 0.03, and UIBC (μg/dL) (median): non-infected dogs: 139.4; subclinically infected dogs: 216; P = 0.0015, and significantly decreased iron (μg/dL) (median): non-infected dogs: 202.5; subclinically infected dogs: 125.7; P = 0.0041, IGF-1 (ng/mL) (median): non-infected dogs: 224; subclinically infected dogs: 123; P = 0.02, and albumin (g/dL) (median): non-infected dogs: 2.8; subclinically infected dogs: 2.5; P = 0.04 concentrations were observed in dogs with subclinical Ancylostoma spp. infection when compared to non-infected dogs.ConclusionThese findings provide an overview of the biochemical effects produced by patent Ancylostoma spp. in naturally infected dogs without any evident clinical signs of disease, which could be considered in differential diagnosis, especially in an endemic area for this parasite.

The Effect of chlorine on giardia cyst destruction isolated from different water sources in Maysan province

&lt;p&gt;G. lamblia was a binucleate flagellated protozoan parasite that infected the upper intestinal tract of human and many animal species. Giardiasis was the most frequently diagnosed water borne disease and the major public health concern of water utilities in the developed and developing nations. Water is an important vehicle for the transmission of Giardia to human and mammals. For identified the effect of chlorine on Giardia cyst. To detect viability of cyst in different chlorine concentration. To determine the threshold level of chlorine concentration that caused cyst destruction. Measure the pH, chlorine concentration, filtration processes and examined by zinc sulfate centrifugal flotation technique using. 50% of samples contain Giardia cysts which are untreated water sources. Cyst viability differs in different chlorine concentration in different period of time extend from few hours to more than twenty days. The extreme chlorine concentration which caused cyst destruction in hours is 1.5 mg/L.&lt;/p&gt;

Molecular typing of canine Giardia duodenalis isolates from Minas Gerais, Brazil

Giardia duodenalis (syn. Giardia intestinalis and Giardia lamblia) is a widespread intestinal parasite in mammals, including humans and pets worldwide. It should be considered a species complex and comprises eight assemblage (A–H). This works aimed to determine the genotypic variability among G. duodenalis isolates from dogs from Minas Gerais state, Brazil. Fecal samples of 97 dogs, from 1-to-10 months old from 15 commercial kennels, were collected and analyzed by the zinc sulfate centrifugal flotation technique, to determine their positivity for G. duodenalis cysts. Cysts pellets were stored and submitted to PCR and nested-PCR reactions with gdh and tpi primers, and then sequencing. Among positive samples (n = 19), fragment amplifications of gdh and tpi genes was observed in 16 (84,2%) and 14 (73,6%), respectively. In total, 30 sequences were obtained. Sequencing analysis showed that for gdh, all isolates were identified as host-specific genotype D, and for tpi, besides host-specific genotype C, were also observed zoonotic genotypes A and B. This study provides, for the first time, current information about genetic characterization of G. duodenalis isolates found in dogs in Minas Gerais state.

Prevalence and genotype of Giardia duodenalis in dairy cattle from Northern and Northeastern part of Thailand.

The aims of this study were to determine prevalence and genotype of Giardia duodenalis in feces of dairy cattle from the northern part and the northeastern part of Thailand. A total of 900 fecal samples were collected directly from rectum and examined by using zinc sulphate centrifugal flotation technique and polymerase chain reaction (PCR). The overall prevalence of G. duodenalis in dairy cows was 5.0 % (45/900) by zinc sulphate centrifugal flotation and 6.0 % (54/900) by PCR. Genotypes of G. duodenalis found in this study were Assemblage AI and E. The results indicated that dairy cattle may act as a potential risk of Giardia transmission among animals and humans (especially Assemblage AI).

Presença de Toxocara spp. e ancilostomídeos em áreas de peridomicílios de uma comunidade pesqueira no Centro-Oeste do Brasil

O objetivo do estudo foi verificar a contaminacao do solo de peridomicilios de uma comunidade pesqueira localizada no municipio de Bonito, Mato Grosso do Sul, Brasil, onde e frequente a ocorrencia de casos de Larva migrans cutânea. Utilizando-se das tecnicas de centrifugo-flutuacao, Baermann-Moraes e centrifugo- sedimentacao, amostras de solo e fezes de caes foram analisadas e os resultados evidenciaram altos indices de contaminacao por ovos de Toxocara spp., Ancilostomideos e larvas de nematoides. Estes achados podem estar associados aos casos de Larvas migrans cutânea e as elevadas taxas de eosinofilia em escolares daquela comunidade.

Performance of Glutamate Dehydrogenase and Triose Phosphate Isomerase Genes in the Analysis of Genotypic Variability of Isolates ofGiardia duodenalisfrom Livestocks

Giardia duodenalis is a small intestinal protozoan parasite of several terrestrial vertebrates. This work aims to assess the genotypic variability of Giardia duodenalis isolates from cattle, sheep and pigs in the Southeast of Brazil, by comparing the standard characterization between glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) primers. Fecal samples from the three groups of animals were analyzed using the zinc sulphate centrifugal flotation technique. Out of 59 positive samples, 30 were from cattle, 26 from sheep and 3 from pigs. Cyst pellets were stored and submitted to PCR and nested-PCR reactions with gdh and tpi primers. Fragment amplification of gdh and tpi genes was observed in 25 (42.4%) and 36 (61.0%) samples, respectively. Regarding the sequencing, 24 sequences were obtained with gdh and 20 with tpi. For both genes, there was a prevalence of E specific species assemblage, although some isolates have been identified as A and B, by the tpi sequencing. This has also shown a larger number of heterogeneous sequences, which have been attribute to mixed infections between assemblages B and E. The largest variability of inter-assemblage associated to the frequency of heterogeneity provided by tpi sequencing reinforces the polymorphic nature of this gene and makes it an excellent target for studies on molecular epidemiology.